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Why beer bananas: The case for Rwanda
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The International
Journal on
Banana and
Plantain
The case for

growing beer
bananas
A new method
for producing
cell suspensions
Mycosphaerella
culture
Results for
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INIBAP
is 20 years old:
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Vol. 14 No.1
June 2005
InfoMusa
Vol. 14 No.1 Cover photo:
Fruit seller in the Philippines.
(A. Javellana)
INFOMUSA
Vol. 14, No. 1
Publisher:
International Network for the Improvement
of Banana and Plantain
Publishing director: Claudine Picq
Editor: Anne Vézina
Editorial Committee:
Jean-Vincent Escalant, Charles Staver, Jose Contents
Mauricio Rivera, Sebastiao de Oliveira e
Silva, Jorge Sandoval,Jean-Pierre Busogoro,
Catherine Abadie Why beer bananas? The case for Rwanda
S.V. Gaidashova, S.H.O. Okech, C.S. Gold and I. Nyagahungu 2
Layout:
Crayon & Cie
Printed in France Agronomic evaluation of production and quality of ‘Yangambi km 5’
ISSN 1023-0076 (AAA) and ‘Dátil’ (AA)
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A. Vargas and J.A. Sandoval 4
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[email protected] S. Rodríguez Morales and J.R. Gálvez Guerra 11
Subscriptions are free for developing
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INIBAP is a network of the International
MusaNews 44
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a Future Harvest centre. Forum 47
A new IPGRI home for INIBAP Editorial
A
s part of a major re-appraisal and re-structuring of its strategy, IPGRI launched the
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The editors
InfoMusa - Vol. 14 - No.1 1

Agricultural policy Why beer bananas? The case for Rwanda
S. V. Gaidashova, S. H.O. Okech, C. S. Gold and I. Nyagahungu
R
wanda is one of the major producers Agriculture (IITA) conducted a participatory
of bananas in the East African rural appraisal in the country’s key banana
Great Lakes region and has one of production zones. Its primary objective was
the highest consumption rates. Bananas to gain insight into the farmers’ perceptions
occupy 23% of the country’s arable land and of the relative profitability of the various crops
contribute more than 50% of annual crop available to them. The information generated
production in terms of fresh weight (Mpyisi et from this study is being used in shaping the
al. 2000). Banana is both a food and a cash national banana research strategy.
crop for most producers and, as such, is a
key component of Rwanda’s food security. Materials and methods
It is also the primary source of income for The participatory rural appraisal was
farmers in some of the most productive conducted in November and December
agricultural zones in Rwanda. Most 2000 in three villages in each of the
production is on small plots. A large number four major banana-growing regions of
of clones are grown, including the local East Rwanda — Cyangugu (in the southwest),
African highland cooking and beer cultivars Kibungo (in the southeast), Kigali Rural
(AAA-EAHB) and introduced beer (AB, ABB) (in central Rwanda) and Kivu Lake Border
and dessert (AAA, AB) types. For the last (in the northwest) — for a total of 12 sites
15 years, beer cultivars have dominated (Figure 1). The participatory rural appraisal
banana production (increasing from 67% to was conducted by a multidisciplinary team,
71% at the expense of dessert types) (Food including specialists in banana production,
Security Research Project 2000). socioeconomics and post-harvest. Activities
The predominance of beer bananas is included group and key informant interviews
controversial. High population pressure and on-farm visits.
makes land use and food production a key At each site, the group interview was the
concern. Some people at the Ministry of primary means of collecting information.
Agriculture have advocated replacing beer Regional agricultural extension officers
bananas with cooking bananas and/or more liaised with local community leaders to
nutritious annual food crops. Clearly, the assemble groups to meet with the survey
factors that have influenced the spread and team from ISAR to discuss agricultural
increased importance of beer bananas to the issues. As such, the participants represented
rural farming community need to be given a cross-section of villagers and were not
serious consideration in policy decisions. biased towards banana producers. Groups
In other words, do beer bananas hold a consisted of 30 to 154 farmers.
comparative advantage over other cash Group interviews were semi-structured
crops as a source of income to the rural and conducted in local language. The
poor? The economic and environmental interview teams solicited opinions from as
consequences of replacing beer bananas many farmers as possible to avoid certain
with other cash crops have not been properly individuals from dominating the discussion.
analysed at a time when the vision for a Where necessary, a special effort was made
regional agricultural policy is increasingly to draw out the opinions of women. Wherever
emphasising marketable crops and income possible a consensus was reached. Where a
generating opportunities for small farmers. consensus could not be reached, a show
There is a need to balance the government’s of hands was taken to reflect divergent
agricultural policies with food security needs, opinions.
rural household income, cultural values and Participants were asked to list all the
environmental concerns. banana cultivars by use and type. They
To address some of these issues, the discussed the importance, advantages
Banana Programme of the Institut des and disadvantages of each cultivar and the
Sciences Agronomiques du Rwanda (ISAR) criteria they use to decide which cultivar
and the International Institute of Tropical to grow. To obtain the opinions of farmers
2 June 2005
N
Ruhengeri Byumba
Umatara
Rubavu
Nyamuyumba
Gishenyi
Lake Kivu Kigali Rubungo
Kayonza
Bicumbi
Kanombe
Gitesi Gitarama Kibungo
Rukira
Kibuye
Kigali-Ngali Rusumo
Kagano
Cyangugu
Karengera
Gishoma Gikongoro Butare
Gishoma Participatory rural appraisal sites

Lakes
Feeder roads
Tarmac roads
0 20 40 60 80 Km Prefectutre boundaries
Figure 1. The banana growing sites surveyed
Country boundary during the 2000 participatory rural appraisal.
on the government’s policy of encouraging preference, food security and the relative
farmers to shift from beer bananas to performance of the cultivars across a range
cooking bananas, annual food crops or of criteria. Cooking bananas are widely
coffee, and how such a change might viewed as poorly adapted to withstand
affect them, we first asked which crops had stresses such as untimely rainfall, drought,
the best farm-gate price and which were declining soil fertility, intensive cropping
most profitable. We then asked why they systems (e.g. intercropping) brought on by
preferred beer bananas to cooking bananas, land pressure and reduced management
given that cooking banana bunches sold for levels. Cooking bananas also require a well-
more money. We also asked farmers how developed market infrastructure because of
replacing beer bananas with other crops their short shelf life and the lack of demand
might affect them. Detailed site descriptions for processed products. As a result, cooking
and methods are presented in Okech et al. bananas are predominant only where they
(2004). are the main staple food or in sites with
good access to the major market of Kigali. In
Results Kibungo, where soils tend to be more fertile
Beer bananas are the most common (Lassoudière et al. 1989) and farmers have
cultivars in three growing areas: Kivu Lake a good outlet for their cooking bananas in
(85 to 90% of cultivars), Cyangugu (60 Kigali, cooking bananas predominate.
to 80%) and, Kigali Rural (60 to 70%). In In contrast, beer bananas are considered
Kibungo, cooking bananas predominate more tolerant of adverse growing conditions
whereas beer types represent between 20 and low levels of management than cooking
to 30% of the cultivars (Figure 2). Dessert bananas. Moreover, they are considered
types never represent more than 10% of better suited to an underdeveloped market
the cultivars. infrastructure because the beer they
The proportions of the different banana produce has a longer shelf life and the
types within each region and at each site are demand for that product is higher than the
influenced by market opportunities, dietary one for cooking bananas. Farmers also

Figure 2. Proportion of beer, cooking and
dessert bananas grown in the regions surveyed
during the 2000 participatory rural appraisal.
Kivu Lake Border

5% Kigali-Rurale
90% 30%
5%
Kibungo
60% 10%
60%
10%
30%
20%
Cyangugu
10%
N
70%
Cooking
Brewing
Dessert
say that introduced beer bananas (ABB) Marketing

perform better on poor soils than AAA-EAHB Across the study regions, 80% of the beer
bananas (both cooking and beer type). ABB banana bunches are processed into beer at
and AB beer bananas do seem more efficient the farm level and sold to local consumers or
in producing on nutrient poor soils. Leaf intermediaries. The remaining 20% are sold
nutrient concentrations published by Lahav to local brewers. Banana beer is common
(1995) show that ABB bananas have much in Kigali, Cyangugu, Kibuye and Gisenyi.
lower plant nutrient concentrations (15% Gikongoro is the major market for the beer
less N and 50% less K) than AAA cultivars. bananas and beer produced in neighboring
Similarly, Bosch et al. (1996) found much Cyangugu. Some of the beer from the Kivu
lower leaf nutrient concentrations (up to Lake border region is also sold in Kigali.
23% less N and 50% less K) in the AB beer Kigali is the country’s major market for
banana ‘Kisubi’. cooking bananas. Much of the bananas
Farmers listed 14 criteria used to decide sold in Kigali are produced in the Kibungo
which cultivar to grow. They include bunch and Kigali Rural regions. Cooking bananas
produced in the Cyangugu region serve the
size (high yield) (12 sites), taste, as far as
smaller, but important, market in Cyangugu
cooking and dessert bananas are concerned
town, while farmers in Kivu Lake border
(12 sites), the quality of the juice produced
region sell in markets in Kibuye and Gisenyi.
by beer bananas (10 sites), marketability There is little movement of cooking bananas
(7 sites), resistance to pests and diseases from the Cyangugu and Kivu Lake border
(7 sites), multiple uses (6 sites), plantation regions to Kigali, which is also the primary
longevity (5 sites), finger size, as far as market for dessert bananas.
cooking bananas are concerned (3 sites), The farm-gate prices of a 20 kg bunch of
tolerance to poor soils (3 sites), maturation cooking banana are between 800 and 3000
rate (3 sites), availability of planting material Rwandese francs (1US$ = 450 francs),
(2 sites), resistance to toppling (2 sites), the while beer types sell for 100 to 800 francs.
quality of the wine produced by beer bananas Prices tend to be higher in sites with
(1 site) and tolerance to intercropping, as far developed market and road infrastructures
as cooking bananas are concerned (1 site). (e.g. around Kigali). Despite the higher
4 June 2005
prices paid for cooking bananas, farmers at profitable margins. This situation contrasts
nine sites reported that beer bananas are with the one in Kibungo, where soils are
more profitable and, of all the cash crops, more fertile and farmers are more open to
contribute the most to household income. change.
Farmers attributed the higher profitability to
the facts that: (1) bunch size requirements Discussion
are less stringent for beer than for cooking The participatory rural appraisal confirmed
and dessert bananas; (2) lower standards that beer bananas predominate in Cyangugu,
mean that beer bananas can do with less Kigali Rural and Kivu Lake Border regions,
management and inputs and are thus while cooking bananas are the most
cheaper to produce; (3) beer bananas important crop in Kibungo, in agreement
can withstand marginal soils, drought and with Bart (1993) and Kangasniemi (1998).
other adverse growing conditions and give In recent years, the Ministry of Agriculture of
appreciable yield throughout the year, Rwanda has expressed dissatisfaction with
thereby providing continuous income; the importance of beer bananas because
(4) beer bananas are readily available and these are seen as taking up land that could
have wider market than other crops; (5) the otherwise be turned over to food production.
processed products (juice, wine, and beer) Therefore, some members of the ministry
add value to the crop and hence higher profit advocated a drastic reduction in the area
margins (the 11 bottles of beer that can be cultivated with beer bananas, in favour of
obtained from a 20 kg bunch would sell for cooking bananas and annual crops.
about 1100 francs); and (6) the products can Our survey suggests that this will be
be stored for long periods, are more easily difficult to achieve. From the farmers’ point
transported over longer distances and have of view, beer bananas remain the most
lower transportation costs. appealing option. Nevertheless, we recom-
At the sites in Cyangugu, Kigali Rural mend studying the profitability of the different
and Kivu Lake border regions where beer banana types to verify this opinion. It is
bananas are important, farmers expressed possible farmers would find other types of
an unwillingness to switch from beer bananas banana more profitable if there were a
to cooking bananas or other crops. Firstly, more developed market for them and if the
cooking bananas require more fertile soils, requisite infrastructure and market access
higher levels of management and inputs, were developed. However, farmers are not
and a well-organised market and transport convinced about the government’s concerns
system. Producing cooking bananas entail over malnutrition, citing the availability of
greater risks and lower returns to farmers, beans (commonly intercropped with banana)
especially those further away from Kigali. and other starchy staples (cassava and
These disadvantages do not compensate sweet potato).
for the higher prices paid for cooking banana Moreover, there is no evidence that
bunches. Secondly, beer bananas are farmers are particularly poor or have had
available throughout the year and allow for problems with food security in regions
a better distribution of income than seasonal where beer bananas predominate. Many
crops. For example, although maize farmers use the income from the sale of beer
might bring higher returns, its seasonality, bananas to purchase beans, cassava, sweet
sensitivity to weather fluctuations and need potatoes and other sources of carbohydrate.
for greater levels of inputs make it less By contrast, problems with poverty and
desirable. Cassava and sweet potato can hunger are most pronounced in areas where
withstand adverse production conditions, but rainfall is low (e.g. Umutara and Bugesera in
the returns tend to be much lower than the eastern Rwanda). These areas are marginal
ones for beer bananas, whereas the market for many crops, including bananas.
for Irish potato, sorghum, and vegetables is There are also environmental aspects that
very small. Thirdly, beer bananas are seen need to be considered before recommending
as preferable to coffee because the latter the replacement of beer bananas by annual
requires higher levels of inputs and has a crops. Erosion is a major problem in
disorganized local market structure, which Rwandan and erosion from permanent
farmers see as having little potential to banana plots is less than three times that of
absorb increased production and generate annual cropped plots (Lufafa et al. 2003).

Possible approaches to achieving the Bosch C., A. Lorkeers, M.R. Ndile & E. Sentozi. 1996.
government’s wish to reduce the area on Diagnostic survey: constraints to banana productivity in
Bukoba and Muleba districts, Kagera region, Tanzania.
which beer bananas are grown would be:
Working paper No. 8. Royal Tropical Institute,
(1) to introduce high yielding disease Amsterdam.
resistant beer/juice cultivars and (2) to Food Security Research Project. 2000. Production par
promote market research and planning to préfecture, première saison, 1984-2000. Ministry of
facilitate the transport from remote areas, like Agriculture, Rwanda, unpublished data.
the Kivu Lake border region, to the lucrative Kangasniemi J. 1998. People and bananas on steep
market of Kigali and hence encourage the slopes: Agricultural intensification and food security
under demographic pressure and environmental
adoption of cooking bananas. In addition,
degradation in Rwanda. PhD dissertation. Michigan
farmers might take on other crops if they State University. East Lansing, Michigan.
could be assured reliable markets. Better Lahav E. 1996. Banana Nutrition. Pp. 258-316 in Bananas
infrastructures would provide farmers with and Plantains (S. Gowen, ed.). Natural Resource
greater options to improve their livelihoods. Institute and University of Reading Department of
Until these issues are addressed, however, Agriculture, UK.
we can expect many Rwandan farmers to Lassoudière A. 1989. Enquête diagnostique sur la culture
bananière en préfecture de Kibungo. Vol. 1, ISAR-
Svetlana V. Gaidashova and continue relying on beer bananas. IRFA-CIRAD.
Innocent Nyagahungu work Acknowledgments Lufafa A., A.M. Tenywa, M. Isabirye, M.J.G. Majaliwa &
at the Institut des Sciences P.L. Woomer. 2003. Prediction of soil erosion in Lake
Agronomiques du Rwanda, We wish to thank the Rockefeller Victoria basin catchment using a GIS-based Universal
P. O. Box 138, Butare, Foundation for providing funding for this Soil Loss model. Agricultural Systems 76: 883-894.
Rwanda, and Suleman H.O. study through a grant to IITA. We also Mpyisi E., J.B. Nyarwaya & E. Shyringiro. 2000.
Okech and Clifford S. Gold thank Piet van Asten and Rupert Best Statistiques agricoles: production agricole, élevage,
for their critical comments on an earlier superficies et utilisation des terres. Année agricole
(author for correspondence,
2000. MINAGRI-FSRP-USAID. 43pp.
[email protected]) at the version of this paper.
Okech S.H., S.V. Gaidashova, C.S. Gold, I. Nyagahungu
International Institute of
Tropical Agriculture, Eastern
References & J.T. Musumbu. 2004. The influence of socioeconomic
Bart F. 1993. Montagnes d’Afrique. Terres paysannes. and marketing factors on banana production in
and Southern Africa Regional Le cas du Rwanda. Centre d’études de géographie Rwanda: Results from a Participatory Rural Appraisal.
Center, P.O. Box 7878, tropicale. Presses Universitaires de Bordeaux. International Journal of Sustainable Development and
Kampala, Uganda. Espaces tropicaux No. 7. World Ecology (in press).
Agronomic evaluation Agronomic evaluation of production and quality of

‘Yangambi km 5’ (AAA) and ‘Dátil’ (AA)
A. Vargas and J. A. Sandoval
T
he dessert banana ‘Yangambi km 5’ countries for study and evaluation (Daniells
(AAA) is known in the Democratic and Bryde 1995).
Republic of Congo, its country of The dessert banana ‘Dátil’ (AA) originates
origin (Daniells et al. 2001a), as ‘Ibota’ from Malaysia (Daniells et al. 2001b) and
meaning “many small fruits” (Daniells and is commercially important in southeast
Bryde 1995). The fruits have a slightly acid, Asia. The cultivar is called ‘Baby banana’
or ‘Lady finger’ in Costa Rica, ‘Pisang mas’
pleasant flavour (Daniells and Bryde 1995,
in Indonesia and Malaysia (Valmayor et al.
Menon 2000). Its resistance to black leaf
1990), ‘Sucrier’ in Australia (Daniells 1986),
streak (caused by Mycosphaerella fijiensis), ‘Bocadillo’ in Colombia (Buitrago et al.
Sigatoka disease (caused by Mycosphaerella 1994) and ‘Titiaro’ in Venezuela (Haddad
musicola),the borer nematode (Radopholus and Borges 1974). The flesh of the fruit is
similis), and possibly to banana borer weevil whitish-yellow, smooth, soft and sweet, and
(Cosmopolites sordidus), has meant that has a distinct aroma. The skin is very thin
this cultivar has been distributed to many and the cuticle prone to bruising, making
6 June 2005
transport and preservation difficult. Fruits production cycles of ‘Yangambi km 5’ and
achieve twice the price of better known types ‘Datil’ respectively according to the method
of banana. Despite the growing interest in described by Villardebo (1973).
Costa Rica of cultivating them for export, The bunch was bagged 15 days after
there is a lack of data on the agronomy and emergence of the inflorescence. Only the
production of ‘Yangambi km 5’ and ‘Dátil’. false hands were removed. The bunch was
The objective of this work was to evaluate harvested 10 and 8 weeks after flowering for
the growth, production and fruit quality of this ‘Yangambi km 5’ and ‘Dátil’ respectively. The
type of musaceae. plants were not propped up.
The following agronomic parameters
Materials and methods were measured: days to flowering, height
The study took place at the Centro de (measured from the base to the point where
Investigación Agrícola 28 Millas, Corporación the last two emerged leaves overlapped),
Bananera Nacional (CORBANA S.A.). The girth (measured on the first third of the
experimental field was located in Limón pseudostem, using a vernier caliper), number
province, Cantón de Matina, at an altitude of leaves of the motherplant at flowering and
of 25 metres. The investigation was carried at harvest, number of days from planting to
out between September 2000 and October first flowering, number of days from the first
2002, a period covering two production to second flowering, height of the sucker,
cycles. Up to 1990, the experimental site had number of foliage leaves on the sucker,
been planted with pejibaye (Bactris gasipaes weight of the bunch and rachis, number of
K.), also known as peach palm, to produce hands and fruits per bunch; diameter of the
palm hearts and left unused thereafter. The middle fruit of the second, fourth and sixth
soil texture is a clay loam (sand 35.8%, clay hands, and length of the fruit of the middle
34.9%, silt 29.2 %), with pH 6.2, extractable fruit of the second, fourth and sixth hands.
acid 0.23, organic matter content 2.2%, Ca The parameters pertaining to the
27.7 cmol/L, Mg 11.7 cmol/L, K 0.96 cmol/L postharvest quality of the middle fruit of the
and a cation exchange capacity of 40.4 cluster’s outer row were Brix, flesh firmness
cmol/L. and skin colour. This was carried out by
The planting materials were corms of harvesting five plants per cultivar, each one
the cultivars ‘Dátil’ and ‘Yangambi km 5’ filling a box from which were removed two
weighing between 1 kg to 3 kg. The materials clusters for evaluation. Brix determination
were arranged in a randomized complete was by means of an Atago refractometer
block design with six replicates or plots. The model Palette–PR 100; firmness was
usable part of the experimental plot contained measured with a Chatillon penetrometer with
12 plants arranged in double furrows spaced a tooth-shaped point and measurements
2.75 m apart, with 1 m between the rows of were recorded in Newton (N), equivalent
the double furrow and 2.15 m between the to (m ∙ kg ∙ s-2). Skin colour was measured
plants of the same furrow, for a density of with a Minolta CR-200 colorimeter and
2480 plants/ha. Fertilizer, 0-46-0 (N-P2O5- calibrated according to the L, a, b scale of
K2O), was applied at 22 g per plant one Hunter where L is a measure of the level
month after planting and then monthly at 47 of light or dark (from L=0 for an absence of
g per plant of 15-3-31 (N-P2O5-K2O) until the reflection, or black, to L=100 for a perfect
end of the study. reflecting diffuser). For the a and b scales,
Chemical treatments to control black a negative a is green and a positive one is
leaf streak were not applied. Sampling for red. A positive b is yellow and a negative one
nematodes was performed on the suckers is blue. Postharvest quality was measured in
of recently flowered plants in the second the postharvest laboratory of the university
production cycle. Sampling was repeated of Costa Rica on fruits produced during the
on two occasions on different groups of second production cycle.
plants. The first sampling was on 9 plants for The mean values were calculated for each
each cultivar and the second on six plants plot and production cycle. The data were then
for each cultivar. The damage by larvae of subjected to an ANOVA by means of PROC
the banana borer weevil was estimated on MIXED of SAS (1999-2001), assuming that
recently harvested plants of the first (68 and the interactions cultivar*block and production
41 plants) and second (68 and 31 plants) cycle*block are random.

Results the production cycles in the length of the
In both ‘Yangambi km 5’ and ‘Dátil’, same fruit from the same hands (Table 2).
pseudostem height and girth increased In ‘Dátil’, the weight of the bunch and rachis
during the second production cycle. There and the number of hands were similar over
were no differences between cultivars in the the two production cycles, but there were
number of leaves at flowering and harvest, more fruits in the second production cycle.
or in the height and number of foliage leaves With the exception of the outer length of the
of the sucker between the first and second middle fruit of the sixth hand, there was a
production cycles (Table 1). reduction in the thickness and length in the
In ‘Yangambi km 5’ pseudostem height fruits of the evaluated hands (Table 2).
and girth were lower than in ‘Dátil’ during In the first production cycle, neither cultivar
the first production cycle only. There were no differed in rachis weight and number of
differences between cultivars in the numbers hands, but ‘Yangambi km 5’ presented higher
of leaves at flowering and at harvest, or values of these parameters than ‘Dátil’ in the
in the height of the sucker and its number second production cycle. ‘Yangambi km 5’
of foliage leaves during the first or second had thinner and longer fruits than ‘Dátil’ over
production cycles. There were no differences both production cycles (Table 2).
in the number of days from planting to first The flesh of ‘Yangambi km 5’ was less
flowering or from the first to the second sweet and firmer than the one of ‘Dátil’ and
flowering (Table 1). the yellow colour of the skin was not as
During the second production cycle, intense as in ‘Dátil’ (Table 3).
‘Yangambi km 5’ showed an increase in the ‘Yangambi km 5’ had 2.1 and 2.6 times
weight of the bunch and rachis, and in the more total and functional roots respectively
number of hands and fruits. There was a than ‘Dátil’, and 1.2 times functional roots
reduction in thickness of the central fruit of in percentage terms (Table 4). Nematodes
the outer row of the second, fourth and sixth (Radopholus spp., Helicotylenchus spp.,
hands. There were no differences between Meloydogine spp., Pratylenchus spp.) were
Table 1. Mean values of various agronomic parameters in ‘Yangambi km 5’ (AAA) and ‘Dátil’ (AA) over two production cycles (n=6 plots).
Production Pseudostem Numbers of leaves Number of days Sucker
cycle height girth at at from from first height number
(m) (cm) flowering harvest planting to second (m) of foliage
to first flowering leaves
flowering
Yangambi km 5 1 2.2 11.8 12.0 8.3 280 2.1 8.0
Yangambi km 5 2 3.7 15.3 12.4 8.3 261 2.4 9.1
Dátil 1 3.2 12.8 11.9 8.0 307 2.0 8.0
Dátil 2 3.9 15.5 11.8 7.8 255 2.3 8.6
Standard error* 0.1 0.2 0.2 0.2 14.9 12.8 0.2 0.5
Pr> F Yangambi km 5 1 vs 2 0.0002 0.0005 0.2273 0.9169 0.1952 0.0717
Pr> F Dátil 1 vs 2 0.0051 0.0012 0.7518 0.6590 0.3110 0.2656
Pr> F Yangambi km 5 vs Dátil 1 0.0012 0.0457 0.8285 0.3041 0.2404 0.7919 0.9174
Pr> F Yangambi km 5 vs Dátil 2 0.2984 0.5218 0.1168 0.1978 0.7706 0.5062 0.4139
*Standard error of mean.
Table 2. Mean values of various production parameters in ‘Yangambi km 5’ (AAA) and ‘Dátil’ (AA) over two production cycles (n=6 plots).
Production Bunch Rachis Number Number Diameter of middle fruit Length of middle fruit
cycle weight weight of hands of fruits of outer row of nth hand of outer row of nth hand
(kg) (kg) per bunch (mm) (cm)
Second Fourth Sixth Second Fourth Sixth
Yangambi km 5 1 8.1 0.9 5.9 95 30.1 29.6 28.9 14.2 14.0 12.6
Yangambi km 5 2 11.7 1.2 8.3 165 28.0 28.0 27.9 14.4 14.0 12.8
Dátil 1 11.0 1.0 6.5 122 33.6 31.9 30.4 13.3 11.9 10.4
Dátil 2 9.7 0.9 7.2 150 30.4 29.1 28.7 12.1 10.9 10.0
Standard error* 0.49 0.06 0.26 7.6 0.2 0.3 0.3 0.2 0.2 0.20
Pr> F Yangambi km 5 1 vs 2 0.0062 0.0188 0.0014 0.0030 0.0033 0.0134 0.0563 0.3727 0.8970 0.4178
Pr> F Dátil 1 vs 2 0.1518 0.5393 0.0717 0.0620 0.0008 0.0016 0.0110 0.0044 0.0172 0.2183
Pr> F Yangambi km 5 vs Dátil 1 0.0139 0.3888 0.2064 0.0678 0.0006 0.0049 0.0150 0.0231 0.0014 0.0016
Pr> F Yangambi km 5 vs Dátil 2 0.0459 0.0243 0.0438 0.2455 0.0021 0.0638 0.0649 0.0007 0.0003 0.0006
*Standard error of mean.
8 June 2005
not found in the roots of either cultivar Table 3. Mean values for Brix, firmness of flesh and skin colour using the Hunter color
(Table 4). Weevils (Cosmopolites sordidus) in ‘Yangambi km 5’ (AAA) and ‘Dátil’ (AA) (n=10 clusters).
Brix Firmness Hunter colour scale
were not found in the corms of ‘Yangambi
(%) (N) L a b
km 5’, whereas a small quantity of weevils Yangambi km 5 18.2 ± 0.5 4.4 ± 0.4 79.6 ± 0.5 -5.1 ± 0.5 49.8 ± 1.3
was found in the ones of ‘Dátil’ (Table 5). Dátil 25.4 ± 0.5 3.6 ± 0.2 82.4 ± 0.7 -0.1 ± 0.6 55.8 ± 1.1
Pr> F 0.0001 0.0637 0.0087 0.0001 0.0075
Discussion Standard error * 0.5 0.3 0.4 0.4 0.9
According to Shepherd et al. (1986), triploid *Standard error of mean.
cultivars, particularly those in commercial
plantations, are generally superior to Table 4. Root content (mean ± standard deviation) in ‘Yangambi km 5’ (AAA) and ‘Dátil’
diploids in terms of vigour, productivity and (AA) (n=15 plants).
acceptability. In the present work, whilst Total roots Functional roots Functional roots
(g) (g) (%)
‘Dátil’ was more vigorous and productive Yangambi km 5 119 ± 27 112 ± 28 94 ± 2
than ‘Yangambi km 5‘ during the first Dátil 57 ± 8 43 ± 4 76 ± 3
production cycle, the data from the second
production cycle confirm this observation,
with the exception of the variables related to Table 5. Estimated damage (mean ± standard deviation) by weevil larvae in ‘Yangambi
fruit quality which, in ‘Yangambi km 5’, were km 5’ (AAA) and ‘Dátil’ (AA) (n=68 plants in each production cycle for ‘Yangambi km 5’,
and n= 41 and 31 plants in first and second production cycles respectively for ‘Datil’).
inferior to the ones observed in ‘Dátil’. Production cycle Infestation Degree of rot
The increases in plant height, pseudostem (%)
girth and numbers of hands per bunch Yangambi km 5 1 0.0 0
observed in both cultivars during the second Dátil 1 2.8 ± 3.2 0
Yangambi km 5 2 0.0 0
production cycle, translated into a higher Dátil 2 2.7 ± 4.2 0
bunch weight only in ‘Yangambi km 5’. This
was mainly due to an increase in the number
of fruits per bunch, which were thinner and of specific to the genotype, probably as a result
similar length to those in the first production of a reduced root system.
cycle. This was different from ‘Dátil’, whose The root system, together with the
smaller fruits in the second production increase in height observed in the second
cycle were not more numerous. Given that production cycle, increased the susceptibility
both cultivars had a similar and adequate of the plants to toppling. Given that the land
number of leaves at flowering and harvest, had not been cultivated for some time, it
the differences in productivity were not due is likely the populations of phytoparasitic
to black leaf streak. nematodes affecting Musa were reduced, a
Paradoxically, the increased vigour situation that could have been responsible
of ‘Dátil’ did not translate into a higher for their absence in the roots of ‘Dátil’,
increased productivity, suggesting that in a cultivar considered susceptible to R.
this cultivar there is a similar decline in similis (Stoffelen et al. 1999), Meloidogyne
productivity to that in plantain type False incognita, Meloidogyne javanica (Stoffelen
horn (AAB). In these materials (Perea 2003, et al. 1999, De Waele and Davide 1998) and
Pantoja et al. 1995, Swennen et al. 1984), Pratylenchus coffeae (Stoffelen et al. 1999).
the decline in productivity increases with However, this latter would not be important in
the number of production cycles. Perea ‘Yangambi km 5’, a cultivar considered to be
(2003) and Pantoja et al. (1995) report not resistant to R. similis and P. coffeae (Viaene
knowing the cause of such a decline and et al. 2000, Sarah et al. 1996) nor, in the
suggest that the plant could be weakened case of weevil, to which both are considered
by diseases, weevils and nematodes and to be resistant (Gold et al. 2002, Hasyim and
by environmental factors or agronomic Gold 1998).
management. Nevertheless, given the The better fruit quality of ‘Dátil’ observed in
favourable conditions, whether agronomic or this study, as well as the greater interest in
climatic and related to crop sanitation under exporting it as fresh fruit (Buitrago et al. 1994,
which the present work was conducted, and Jamaluddin 1990, Daniells 1986, Contreras
as reflected in increased plant vigour over 1982), suggests that there is a need to
the production cycles, it is very unlikely that further evaluate the intensive production
the decline in ‘Dátil’ was associated with the strategies such as the one developed for
previously mentioned factors. It could be plantains (Belalcázar 1991, Vargas 1994).

This should include consideration of high sordidus Germar, with natural enemies from Asia
(with emphasis on Indonesia). Pp. 59-71 in Mobilizing
plantation densities, replanting the field IPM for sustainable banana production in Africa
after each production cycle and spreading (E. Frison, C.S. Gold, E. Karamura and R. Sikora, eds).
out planting over time, practices which, for International Network for the Improvement of Banana
plantain, have been shown to be a more and Plantain. Montpellier, France.
Jamaluddin S.H. 1990. Bananas and plantains in
profitable and reliable management option Malaysia. Pp.71-86 in Banana and Plantain R&D in
that traditional system. Asia and the Pacific (R. Valmayor, ed.). International
Network for the Improvement of Banana and Plantain.
Acknowledgements Montpellier, France.
The authors thank Ing. Marco Vinicio Menon R. 2000. Preliminary evaluation of some banana
introductions in Kerala (India). INFOMUSA 9(2):27-28.
Sáenz MSc., Laboratorio Poscosecha de la
Pantoja A, L. Chyuan & L. Jang. 1995. Factores que
Universidad de Costa Rica and Ing. Mauricio causan la decadencia del platanal. Journal of
Serrano for their collaboration. Agriculture of the University of Puerto Rico 79(3-4):
187-193.
References Perea M. 2003. Biotecnología Bananos y Plátanos.
Belalcázar S. 1991. El cultivo del plátano en el trópico. Editora Guadalupe Ltda. Bogotá, D.C., Colombia.
Manual de Asistencia NO 50. Instituto Colombiano 228pp.
Agropecuario, Centro Internacional para el Desarrollo, Sarah J.L., J. Pinochet & J. Stanton. 1996. The
Red Internacional para el Mejoramiento del Banano y burrowing nematode of bananas: Radopholus similis.
el Plátano. Cali, Colombia. 376pp. Musa Pest Fact Sheet No. 1. INIBAP. Montpellier,
Buitrago L., G. Garcia & M. Arcilla. 1994. Mejoramiento France.
de la producción de plátano. Segundo Informe Técnico Shepherd K., J.L. Loyola Dantas & E.J. Alves. 1986.
1984-1994. Instituto Colombiano Agropecuario, Mejoramiento genético del banano. Pp. 1-19 in
ICA, Corporación Colombiana de Investigación Mejoramiento genético de banano y plátano en Brasil
Agropecuaria, CORPOICA. Armenia, Colombia. Y Honduras (S. Pons, R. Jaramillo & J. Pinché, eds).
256pp. Unión de Países Exportadores de Banano, Panamá.
Contreras M. 1982. Identificación y caracterización de 16 Stoffelen R., V.T. Tan, R. Swennen & D. De Waele. 1999.
clones de plátano en Tabasco. Universidad Autónoma Host plant response of banana (Musa spp.) cultivars
de Chapingo. Chapingo, México. 78pp. from Southeast Asia to nematodes. International
Daniells J. 1986. Banana cultivars in Australia. The Journal of Nematology 9(2):130-136.
Banana Bulletin (May/June):24-26.
Swennen R., G. Wilson & E. De Langhe. 1984. Preliminary
Daniells J.& N. Bryde. 1995. Semi-dwarf mutant of investigation of the effects of gibberellic acid (GA3) on
Yangambi km 5. INFOMUSA 4(2):16-17. sucker development in plantain (Musa cv. AAB) under
Daniells J., C. Jenny, D. Karamura & K. Tomekpe. 2001a. field conditions. Tropical Agriculture 61(4):253-256.
Diversity of the genus Musa. Cultivated varieties AAA.
Valmayor R.V., S. Silayoi, S. Jamaluddin, R. Kusumo &
P. 72 in Musalogue. A catalogue of Musa germplasm
O. Pascua. 1990. Commercial banana cultivars in
(E. Arnaud and S. Sharrock, eds). IPGRI/INIBAP, CTA,
Cirad-Flhor, Montpellier, France. ASEAN. Pp. 23-32 in Banana. Fruit development,
postharvest physiology, handling and marketing in
Daniells J., C. Jenny, D. Karamura & K. Tomekpe. 2001b.
Diversity of the genus Musa. Cultivated Varieties AA. ASEAN (H. Abdullah, E.B. Pantastico, eds). ASEAN
P. 49 in Musalogue. A catalogue of Musa germplasm Food Handling Bureau, Kuala Lumpur, Malasia.
(E. Arnaud and S. Sharrock, eds). IPGRI/INIBAP, CTA, Vargas A. 1994. Validación de tecnología de producción
Cirad-Flhor, Montpellier, France. para alto rendimiento en el cultivo del plátano Curraré
De Waele D. & R Davide. 1998. The root-knot nematodes o Falso Cuerno (Musa AAB) en el Atlántico de Costa
of banana. Musa Pest Factsheet No. 3. INIBAP. Rica (primera cosecha). CORBANA 19(42):17-24.
Montpellier, France. Viaene N., F. Duran, J. Dueñas, M. Rivera, D. De Waele
Gold C.S., B. Pinese & E. Peña. 2002. Pest of banana. & P. Rowe. 2000. Reacción de híbridos y genotipos
The authors work at naturales de Musa al ataque de los nematodos
Pp. 13-56 in Tropical Fruits Pests and Pollinators (E.
CORBANA S.A., Peña, L. Sharp and M. Wysoki, eds). CAB International. Radopholus similis y Pratylenchus coffeae en
Apdo 390-7210, Guápiles, Wallingford, UK. ambiente de casa de sombra in Memorias XIV Reunión
Limón, Costa Rica. Haddad O. & O. Borges. 1974. Los bananos en ACORBAT. San Juan, Puerto Rico.
Venezuela. Estudio y descripción de clones de plátano Villardebo A. 1973. Le coefficient d’infestation, critère
E-mail: y cambur. CONYCIT. Caracas, Venezuela. 105pp. d’évaluation du degré d’attaques des bananeraies par
[email protected]; Hasyim A. & C.S. Gold. 1998. Potential of classical Cosmopolites sordidus Germar, le charançon noir du
[email protected] biological control for banana weevil, Cosmopolites bananier. Fruits 28(6):417-426.
10 June 2005
Production of banana and plantain hybrids in Cuba Breeding
T. Ramirez Pedraza, L. González Díaz, J.de la C. Ventura Martín, S. Rodríguez Morales
and J.R. Gálvez Guerra
G
enetic improvement of banana and carbonates (Hernández 1995). Management
plantain in the various programmes was according to technical instructions for
operating around the world is the cultivation of plantain (MINAGRI 1994).
based on crosses between commercial Ten months after planting, at the peak of
triploids and improved diploids, with the flowering, male flowers with abundant pollen
objective of developing higher yielding sacs were selected and spread vigorously
cultivars that are more resistant to the main over the stigma of female flowers that
diseases (Sigatoka disease caused by had opened in the morning. The process
Mycosphaerella musicola, black leaf streak was repeated with flowers that were in the
caused by Mycosphaerella fijiensis, fusarium process of opening (Silva et al. 1997).
wilt caused by Fusarium oxysporum f.sp. The pollinated bunches thus obtained
cubense), and pests (nematodes and were harvested when they reached maturity.
weevils). They were transferred to a maturation
In the last years, various research groups room, grouped according to their crossing
throughout the world have made concerted scheme, where they remained until
efforts to increase genetic variability, which complete maturation. The fingers were cut
is of major importance for the selection of longitudinally to remove the seeds. The
clones with higher productivity and resistance seeds were sieved (3 mm mesh) under
to the main diseases (López 1989). a steady flow of running water in order to
The current study was to determine remove the flesh, and transferred to flasks
the combinations of parents that provide of distilled water to start disinfection and
the best opportunity for producing new
dissection.
hybrids possessing the desirable agronomic
The regenerated plants were established in
characters.
polythene bags, with a substrate composed
Materials and methods of 50% soil and 50% organic matter from a
The present work was carried out at the partially decomposed by-product of sugar
Instituto de Investigaciones en Viandas cane, and transferred to the field when
Tropicales (INIVIT), Santo Domingo, Cuba ready.
from January 1995 to December 2003 using The best combinations were determined on
conventional hybridization methods. Table 1 the basis of the numbers of seeds obtained
describes the genomic group of the parents and the value of the progenies obtained.
used. The Musa balbisiana used is a wild
diploid from the INIVIT collection, originally
Results and discussion
from Vietnam, which is very resistant to In the first crosses, Musa balbisiana (BB)
toppling and the main banana diseases. or Musa acuminata ssp. malaccensis (AA),
A total of 3500 crosses representing 17 was crossed with ‘Highgate’ or ‘Hembra 3⁄4’
combinations were obtained. as the female parent. The progenies showed
Selected parents were planted in the resistance to Fusarium wilt and black leaf
field in alternate furrows at a distance of streak but inherited the defects of the male
3.6 m x 2 m. The soil had brown mottles and parent, such as asymmetrical bunches,
short fingers and high seed content. These
progenies were discarded as having no
Table 1. Name and genomic group of parents
used. commercial value. Table 2 presents data on
Male parents Female parents the numbers of good and bad seeds.
Paka (AA) Highgate (AAA)
Pisang jari buaya (AA) Hembra 3⁄4 (AAB)
Crosses between SH-3436-L9 and Musa
Calcutta 4 (AA) Pelipita (ABB) balbisiana (BB)
SH-3142 (AA) Saba (ABB) No satisfactory results were obtained in
SH-3362 (AA) Saba somaclone (ABB) the process of improving AAA and AAAA
Musa balbisiana (BB) SH-3436-L9 (AAAA)
cultivars.

Table 2. Mean number of seeds per bunch (n = 20 pollinated bunches) obtained from (1 to 2 seeds per pollinated bunch).
various crosses. Progenies from these crosses died at the
Female parent Male parent Mean number of seeds
per pollinated bunch greenhouse stage.
Good Bad Crosses between ‘Highgate’ and
SH-3436-L9 (AAAA) Musa balbisiana (BB) 0.0 1.5
SH-3436-L9 (AAAA) Pisang jari buaya (AA) 0.0 3.0
‘Calcutta 4’ or ‘Paka’
SH-3436-L9 (AAAA) Paka (AA) 0.0 4.6 The use of ‘Calcutta 4’ produced progenies
Highgate (AAA) SH-3142 (AA) 2.5 3.0 with seedless but small fruits and dwarf
Highgate (AAA) SH-3362 (AA) 1.0 4.5
plants that were therefore discarded. No
Highgate (AAA) Calcutta 4 (AA) 5.0 11.0
Highgate (AAA) Paka (AA) 0.0 0.0 seeds were produced when ‘Highgate’ was
Hembra 3⁄4 (AAB) SH-3142 (AA) 0.0 18.0 crossed with ’Paka’.
Hembra 3⁄4 (AAB) SH-3362 (AA) 0.0 15.0
Hembra 3⁄4 (AAB) Calcutta 4 (AA) 0.0 11.0
Crosses between ‘Hembra 3⁄4’ and
Pelipita (ABB) Pisang jari buaya (AA) 14.5 2.0 SH-3142, SH-3362 or ‘Calcutta 4’
Pelipita (ABB) Calcutta 4 (AA) 6.4 3.0 When ’Hembra 3⁄4’ was crossed with
Saba (ABB) Pisang jari buaya (AA) 3.7 7.5 SH-3142 or SH-3362, one to two seeds
Saba somaclone (ABB) Pisang jari buaya (AA) 2.9 5.3
per bunch were obtained, as when it was
pollinated with ‘Calcutta 4’. The seeds from
The tetraploid SH-3436 (AAAA) is a product of these crosses were not viable.
the improvement programme of the Fundación Crosses between ‘Pelipita’ and ‘Pisang
Hondureña de Investigación Agrícola (FHIA). jari buaya’
Starting with this hybrid, INIVIT obtained through The genetic improvement of cooking
tissue culture the somaclonal variant SH-3436- bananas (ABB) involved crossing the cultivar
L9 . This clone originates from ‘Highgate’, as ‘Pelipita’, which is resistant to black leaf
do most of the tetraploids produced by FHIA. streak, Fusarium wilt and nematodes, with
When used as a female parent and pollinated ‘Pisang jari buaya’, which is highly resistant
by a diploid, this semi-dwarf mutant of ‘Gros to nematodes. The result was a tetraploid
Michel’ (AAA) produces non-reduced gametes progeny of considerable height (more than
to generate tetraploids.
3.5 m) with small bunches and asymmetrical
When SH-3436-L9 was crossed with
fingers of unacceptable quality. Flowering
M. balbisiana, not only was the number of seeds
took place 15 to 18 months after planting.
small (1 seed/bunch), but the seeds also had
The progeny from this cross was discarded
abnormal endosperm and embryos and did not
for these reasons. On the other hand, the
germinate. The use of SH-3436-L9 as a male
descendents had inherited the resistance to
parent was not promising. None of the progenies
survived the selection processes or showed any nematodes.
agronomic value. The primary tetraploids obtained from
Using wild Musa acuminata, Larter (1947) these crosses could be used as female
obtained tetraploids that were reasonably parents in crosses with improved diploids to
satisfactory from a commercial point of view, obtain secondary triploids and to increase
but that had inherited the non-parthenocarpic genetic diversity.
characteristics of the male parent. Since seeds Crosses between ‘Pelipita’ and
are an undesirable character, the tetraploids ‘Calcutta 4’
should be female sterile. When ‘Pelipita’ was crossed with ‘Calcutta 4’,
Crosses between SH-3436-L9 and ‘Pisang the progeny had thick leaves and a dwarf
jari buaya’ or ‘Paka’ habit, and did not survive in the greenhouse.
Crossing SH-3436-L9 with ‘Pisang jari buaya’ The plants appeared to be heptaploid
and ‘Paka’ produced only a few seeds that although this was not verified.
did not germinate and had abnormal embryos Crosses between ‘Saba’ or a
and endosperms, for reasons that are not somaclone of ‘Saba’ and ‘Pisang jari
understood. buaya’
Crosses between ‘Highgate’ and SH-3142 Crossing a ‘Saba’ somaclone with ‘Pisang
or SH-3362 jari buaya’ yielded a small number of seeds
Crosses between ‘Highgate’ and SH-3142 that had reduced endosperms and abnormal
and SH-3362 produced very few seeds embryos, and did not germinate.
12 June 2005
Crosses between a somaclone of ‘Saba’ production cycle undergoing evaluation for
and SH-3362 or SH-3142 their commercial potential and their response
When a ‘Saba’ somaclone was crossed to different pests and diseases.
with SH-3362, which is highly resistant to When the ‘Saba’ somaclone was crossed
black leaf streak, 14.6 seeds per pollinated with SH-3142, 350 seeds were obtained, of
bunch were obtained. The seeds had which only 100 proved viable; the remainder
normal endosperms and embryos. The were eliminated as having dry and abnormal
seeds were opened in the laboratory and embryos. Mean seed numbers per bunch
produced 70 plants, of which 12 survived varied between 14 and 20 seeds.
in the greenhouse, when they were put
on culture medium. The remaining plants References
were discarded as having undesirable Hernández A. 1995. Nueva Versión de Clasificación de
los Suelos de Cuba. Instituto de Suelos de Cuba, La
characteristics such as thick leaves and a Habana.
dwarf habit. The plants that were planted in Larter L.N.H. 1947. Report on banana breeding. Dep.
the field were tetraploids that had acceptable Agric. Jamaica Bull.34:24. The authors work at the
bunch characters inherited from the male Silva S. de O., A.P. de Matos, E.J. Alves & K. Shepherd. Instituto de Investigaciones en
1997. Mejoramiento de bananos diploides (AA) en
parent (pendulous with 8 or 9 hands, 90 to EMBRAPA/CNPMF. INFOMUSA 6(2):4-6. Viandas Tropicales (INIVIT),
100 fingers per bunch). At the present time, MINAGRI. 1994. Instructivo Técnico para el Cultivo del Sto. Domingo, Apdo 6, Villa
the plantings in the field are in their second Plátano. Clara, Cuba.
New methodology for the establishment of cell Embryogenic cell suspensions

suspensions of ‘Grande naine’ (AAA)
B. Chong Pérez, R. Gómez Kosky, M. Reyes Vega, I. Bermúdez Carballoso, J. Gallardo
Colina, M. Freire Seijo, L. Posada Pérez, I. Herrera O’Farril and R. Swennen
M
ost research groups initiate cell Effect of the immature male flower’s
suspensions of the cultivar ‘Grande position on cell suspension initiation
naine’ (AAA) from immature Only male flowers in the 5th to 15th position
male flowers. However, the formation of were taken. Ten immature male flowers from
calli with embryogenic structures takes the same position were taken from 10 male
five to six months and the success rate buds and transferred to 50 ml Erlenmeyer
is between 3% and 10%. Furthermore, flasks containing 5.0 ml of a MA1 liquid
after obtaining these structures, the cultivation medium prepared with salts and
proportion of established embryogenic cell MS vitamins (Murashige and Skoog 1962)
suspensions do not exceed 30% in the two supplemented with 4.09 µM biotin, 5.7 µM
indole-3-acetic acid (IAA), 18.1 µM 2,4-
months needed to become homogenous.
dichlorphenoxyacetic acid (2,4-D), 5.37 µM
The objectives of this work were to
naphleneacetic acid (NAA) and 87.6 mM
establish cell suspensions by cultivating
saccharose. The pH was adjusted to 5.7
immature male flowers of the cultivar before sterilization with 1 N NaOH and 1 N
‘Grande naine’ directly on liquid culture HCl (Escalant et al. 1994). Erlenmeyer flasks
medium, and to achieve the formation of were placed on an orbital shaker model
somatic embryos from these suspensions INFORS (HT), in constant dark, at a speed of
and regenerate the plants. 90 rpm and a temperature of 27±0.2°C. Half
of the culture medium was replaced every 15
Materials and methods days until the appearance of cells; from then
Immature male flowers of the banana cultivar on the culture medium was changed weekly.
‘Grande naine’ were used as vegetative After 45 days, the presence or absence of
material. Immature male flowers were yellow globular structures on the surface of
extracted according to the methodology the immature male flowers was evaluated.
described in Escalant et al. (1994). After 70 days of culture, the quality of the cell

suspensions was evaluated by determining The somatic embryos were transferred to
cell viability with fluorescein diacetate (FDA) the culture medium described in Gómez et
and cell counts were made in a Neubauer al. (2000) for maturing. The culture medium
chamber. Similar evaluations were made in was prepared with salts and MS vitamins
all experiments as required. supplemented with 4.09 µM biotin, 2.22 µM
Effect of culture medium on cell 6-benzylaminopurine, 1.1 µM IAA, 130 mM
suspension initiation saccharose and 2.0 g/L Gelrite, with the pH
adjusted to 5.8 before sterilization. Culture
Immature male flowers from the 7th to the conditions were total darkness at 27±2°C
11th position, ten per position, were collected and the embryos remained in the culture
and transferred to 50 ml Erlenmeyer flasks medium for 30 days.
containing 5 ml of culture medium. The MA1 Mature somatic embryos were then
liquid culture medium was compared to the placed on the germination culture medium
MA2 medium for the establishment and (M5) described in Gómez et al. (2000)
multiplication of cell suspensions described and containing salts and MS vitamins
in Côte et al. (1996). The latter contained supplemented with 4.1 µM biotin, 1.1 µM
salts and MS vitamins supplemented with IAA, 0.2 µM 6-benzylaminopurine, 0.01 mg/L
4.09 µM biotin, 4.5 µM 2,4-D, 680 µM Biobras-6, 87 mM saccharose, solidified
L-glutamine, 100 mg/L malt extract and with 2 g/L Gelrite, and adjusted to pH 5.8
130 mM saccharose. The pH was adjusted before autoclaving. The culture flasks had
to 5.3 before sterilization by autoclaving. a capacity of 250 ml and contained 30 ml
The culture conditions and variables in of culture medium. Flasks were placed in
evaluation were the same as in the previous growth chambers with sunlight and exposed
experiment. to a flux of photosynthetic photons of
Multiplication of embryogenic cell 50 - 62.5 µmol m-2 s-1 and a temperature of
suspensions and formation of somatic 27±2°C.
embryos Data were analysed by means of the
After 100 days from the start of the statistical packages SPSS ver. 9.0 and
StatGraphics Plus ver. 4.1.
experiment, cell suspensions were sieved
on a metal mesh with a pore size of 500 µm. Results
The filtrates were the cell suspensions
which were transferred to a MA2 culture Effect of the immature male flower’s
position
medium for maintenance or multiplication.
Two hundred µl of settled cells from three Immature male flowers cultivated directly
on liquid culture medium responded after
different cell lines, all obtained by using our
two weeks in culture. Phenol formation was
methodology, were plated on Petri dishes
observed in the cortex zone followed by
9 cm in diameter and containing the MA3
enlargement of the explants. The biggest
culture medium described in Côte et al.
response occurred with flowers from the
(1996): salts (Schenk and Hildebrandt
8th to 15th position, with no significant
1972), MS vitamins, 680 µM L-glutamine, differences between them (Table 1).
2 mM L-proline, 1.1 µM NAA, 0.7 µM Flowers from positions 5 and 6 did not
adenine N6-(2-isopentenol), 0.5 µM kinetin, respond satisfactorily, with the majority
29 mM lactose, 0.2 µM zeatin, 100 mg/L observed to have total tissue necrosis.
malt extract, 130 mM saccharose and After five weeks of culture, yellow globular
solidified with 3.0 g/L Gelrite. The pH was structures appeared on the surface of the
adjusted to 5.3 before sterilization. explants, which rapidly separated from the
Petri dishes were sealed with latter (Figure 1A). There were significant
Parafilm® and placed in total darkness statistical differences between the different
at a temperature of 27±2°C. Visual positions with regards to the quantity of
observations were made daily after the explants that formed such types of structure
fifteenth day of culture to evaluate the (Table 1). Immature male flowers from
appearance of the first somatic embryos. positions 7 to 11 had the largest quantity
Counts of somatic embryos were made of explants with such structures (70-90%),
after 45 days of culture and no subculturing with no significant differences between
was done. positions. Explants from positions 12 to 15
14 June 2005
had a disorganised growth but formed very Influence of culture medium
few of these structures. There were significant differences, for all
After 10 weeks of culture, turbidity was variables evaluated, between treatments
observed at the bottom of all Erlenmeyer looking at the interaction between the
flasks. Observation under the optical position of the immature male flowers and
microscope at 200x magnification revealed the type of culture medium (Table 2). The
the presence of small spherical embryogenic best responses with regards to the formation
cells with dense cytoplasm, starch granules of yellow globular structures were obtained
and various aggregates of embryos. Cell with immature male flowers from positions
vacuoles were also observed. 8, 9, 10 and 11 in MA1 culture medium, and
Counts of cell aggregates, single cells position 10 in MA2 culture medium, with
and vacuolate cells, showed that flowers 85% to 88% of the explants producing yellow
from the 8th position yielded the largest globular structures.
quantity of embryogenic cell aggregates After 10 weeks of culture, cells were
(3.0x105) and the least quantity of vacuolate observed in all the treatments. The
cells (1.4x105) (Table 1). Positions 5 and 6 treatment with the best results was position
generally produced very few cells, whereas 8 in culture medium MA1, with the biggest
positions 11 to 15 produced large quantities quantity of embryogenic cells (3.1x105)
of single cells and vacuolate cells, which is and the lowest quantity of vacuolate cells
not desirable. (1.3x105) per millilitre of culture medium
Table 1. Effect of the position of ‘Grande naine’ (AAA) immature male flowers after 5 weeks in a liquid
culture medium, and composition of cell suspensions after 10 weeks of culture (n=10).
Position % of explants % of explants Number (x105) of Number (x105) of Number (x105) of
of male enlarged with globular single cells per ml cell aggregates vacuolate cells
flower structures of culture medium per ml of culture per ml of culture
medium medium
5 10 c* 0.0 b* 1.6 i** 0.1 g** 5.3 f**
6 10 c 7.5 b 2.2 h 0.1 g 4.5 g
7 80 b 72.5 a 8.6 e 2.4 b 2.5 i
8 90 ab 90.0 a 8.9 e 3.0 a 1.4 j
9 90 ab 90.0 a 1.2 c 1.7 c 2.1 h
10 100 a 70.0 a 15.2 b 1.8 c 5.4 f
11 100 a 87.8 a 18.3 a 1.3 d 7.4 e
12 100 a 17.5 b 14.9 b 1.2 d 11.3 d
13 100 a 2.5 b 11.0 d 0.5 e 17.2 c
14 100 a 2.5 b 6.6 f 0.5 e 24.5 b
15 100 a 2.5 b 5.6 g 0.3 f 26.4 a
SE ±5.2 ±4.2 ± 5.4 ± 1.0 ± 8.0
*Values followed by different letters in the same column are significantly different at p< 0.05 according to Tukey’s test.
** Values followed by different letters in the same column are significantly different at p< 0.05 according to Dunett´s C test.
Table 2. Effect of culture medium and ranking of immature male flowers of ‘Grande naine’ (AAA) after 5 weeks culture and composition of cell
suspensions after 10 weeks culture (n=10).
Position Culture % of explants % of explants Number (x105) of Number (x105) of Number (x105) of
of male medium enlarged with globular single cells per ml vacuolate cells cell aggregates
flower structures of culture medium per ml of culture per ml of culture
medium medium
7 MA1 92.5 a* 77.5 cd* 8.6 ± 1.2 d** 2.6 ± 1.6 d** 2.5 ± 0.6 b**
8 MA1 92.5 a 85.0 ab 8.9 ± 1.5 d 1.3 ± 0.7 e 3.1 ± 1.2 a
9 MA1 95.0 a 87.5 a 12.1 ± 2.1 c 2.0 ± 0.5 c 2.0 ± 1.2 c
10 MA1 95.0 a 85.0 a 15.2 ± 2.1 b 5.7 ± 2.9 b 1.9 ± 0.5 c
11 MA1 97.5 a 87.50a 18.2 ± 1.7 a 7.7 ± 1.1 a 1.3 ± 0.5 e
7 MA2 25.0 b 40.0 e 1.9 ± 0.7 h 3.9 ± 1.3 c 0.8 ± 0.4 fg
8 MA2 47.5 b 65.0 d 5.8 ± 1.4 f 2.3 ± 0.9 d 1.6 ± 0.4 d
9 MA2 85.0 a 80.0 bc 6.2 ± 1.7 f 2.3 ± 0.5 d 0.9 ± 0.4 f
10 MA2 97.5 a 85.0 ab 5.9 ± 1.3 f 6.0 ± 2.2 b 0.6 ± 0.3 g
11 MA2 100 a 37.5 e 2.8 ± 1.3 g 7.8 ± 1.9 a 0.4 ± 0.2 h
SE ±5.8 ±9.2 ±5.2 ±2.32 ±0.9
*Values followed by different letters in the same column are significantly different at p< 0.05 according to Tukey’s test.
** Values followed by different letters in the same column are significantly different at p< 0.05 according to Dunett´s C test.

(Table 2). These results support those The observation of cell suspensions
obtained in the previous experiment where verified that there had been changes
position 8 gave the best results in terms in their composition with aggregates of
of the establishment of embryogenic cell embryogenic cells predominating and
suspensions. No references were found on the quantity of single and vacuolate
the use of immature male flowers directly cells eventually decreasing to zero with
on a liquid culture medium. each subculturing. These aggregates of
These results may be due to the stage embryogenic cells occupied 90% to 95%
of differentiation of immature male flowers of the suspension and their size varied
and to the composition of the MA1 culture between 80 µm to 300 µm, resulting in a
medium. This culture medium has a higher homogeneous cell suspension (Figure 1B).
concentration of auxins than the MA2 The first embryos were observed 20
medium, in addition to having a mixture of days after plating the three cell lines
them (2,4-D, IAA and NAA), whereas the on MA3 culture medium. The number
culture medium MA2 only has one auxin, of embryos was counted after 45 days
2,4-D, at a lower concentration (1 mg/L). culture (Figure 1C). All cell lines had a
good embryogenic response as determined
According to Parrot (1993), the type,
by the quantity of somatic embryos
physiological state and degree of differenti-
formed. There were significant statistical
ation and polarization of the tissues used
differences (p<0.05 according to Duncan’s
as initial explants are some of the factors
test, SE=102.3) between cell line 2, with
that favour or impede the formation of
the formation of a mean of 517 somatic
calli with embryogenic structures. In an embryos per 200 µl of plated settled cells,
experiment using ‘Grande naine’, immature and cell lines 1 and 3, with the formation of
male flowers from positions 7 to 13, 74% 662 and 761 somatic embryos respectively.
of the calli formed embryogenic structures These differences could have resulted from
in a semisolid culture medium (Escalant et differences in the embryogenic potential of
al. 1994). Noceda et al. (1999) studied the the cell lines
effect of various factors on callus production After 17 days of culture, the first somatic
with embryogenic structures in ‘Grande embryos started to germinate with the
naine’ and FHIA-18 (AAAB), and the factor emergence of leaves and roots. Of the
that most affected the process was the time somatic embryos, 80.5% had completely
when male buds were harvested and the germinated (Figure 1D) after 45 days of
next most important factor was the position culture. It should be pointed out that no
of the flower, with positions 5 to 8 giving the morphological changes were observed in
best results. the regenerated plants (Figure 1E).
Multiplication of embryogenic cell Discussion
suspensions and formation of somatic
Our methodology made it possible to obtain
embryos
homogeneous suspensions of embryogenic
Given the results of the previous cells in 20 weeks (about five months) after
experiments, cell suspensions established the start of culture, in 70% of the explants
from immature male flowers from the 8th inoculated. The methods proposed by Côte
position were inoculated directly onto MA1 et al. (1996) and Grapin et al. (1996) take
culture medium in 50 ml Erlenmeyer flasks. between four and seven months just for
The heterogeneous suspensions obtained the formation of calli with embryogenic
were filtered to remove the globular structures, and a further two months for
structures and proembryos. After 20 weeks the establishment of cell suspensions.
from the start of culture, and subculturing Furthermore, it is necessary to take into
every 12 days, cell suspensions account that the embryogenic response of
reached a high degree of homogeneity, male flowers cultured on semisolid medium
mainly with aggregates of embryogenic was low. Escalant et al. (1994) reported
cells characterized by small spherical that 0 to 7% of the explants had calli with
embryogenic cells with dense cytoplasm, embryogenic structures in five banana
small vacuoles, starch granules and a well cultivars, including ‘Grande naine’. Daniels
defined nucleus/cytoplasm. et al. (2002) reported a success rate of
16 June 2005
fa
g
A B C
D E
Figure 1. Different stages in the process of obtaining plants from immature male flowers of ‘Grande naine’ (AAA). A) Formation of yellow
globular structures after 5 weeks of culture (g: globules, fa: flower aggregates) (4x). B) Cell aggregates present in cell suspensions
20 weeks after start of culture (20x). C) Somatic embryos at the globule stage formed on germination semisolid culture medium after 45
days of culture (4x). D) Germination of somatic embryos and formation of small plantlets in M5 culture medium. E) Developing plantlets.
6.5% with the hybrid FHIA-21 (AAAB). With (82.5%) of somatic embryos for the hybrid
‘Grande naine’, the mean was 8% (Strosse et FHIA-21 using a maturation medium before
al. 2003). Khalil et al. (2002) obtained better germination.
results (58.8%) but with the cultivar ‘Brazilian
dwarf’ (AAB). Moreover, the proportion of
Acknowledgements
embryogenic cell suspensions established This work was carried out with the
from ideal embryogenic calli was between 10% economic support of the VLIR for project
and 30% in ‘Grande naine’ (R. Domergue and AEIN2000PR230 on the development
colleagues, Cirad, unpublished results). of transgenic lines with resistance to
The number of somatic embryos formed Mycosphaerella fijiensis.
in each ml cells varied between 7835 and References
11 530, which demonstrates the quality Barranco L.A. 2001. Embriogénesis somática en banano
of the cell suspensions obtained with our (Musa AAAB, cv. FHIA-18) empleando medios de
cultivo líquidos. Tesis en opción del Grado Científico de Borys Chong Pérez*, Rafael
methodology. Values of 100 and 300 000 Doctor en Ciencias Agrícolas. Instituto de Biotecnología Gómez Kosky, Maritza Reyes
somatic embryos per ml have been reported de las Plantas, Cuba. 97pp.
Vega, Idalmis Bermúdez
in the literature (Côte et al. 1996, Grapin Cabrera M. 2001. Embriogénesis somática en Musa
Carballoso, Jorge Gallardo
(AAB) cv. Navolean empleando medios de cultivo
et al. 1996, Schoofs 1997, Daniels et al. líquidos. Tesis en opción del Grado Científico Maestro Colina, Marisol Freire Seijo,
2002). The levels of germination of somatic en Ciencias. Instituto de Biotecnología de las Plantas. Laisyn Posada Pérez y Idalia
embryos obtained after being in a maturation Santa Clara, Cuba.
Herrera O’Farril work at the
Côte F., R. Domergue, S. Monmarson, J. Schwendiman,
medium were on average 80.5%, a value C. Teisson & J.V. Escalant. 1996. Embryogenic cell Instituto de Biotecnología
higher than the 40.6% germination rate for suspensions from male flower of Musa AAA cv. ‘Grand de las Plantas, Universidad
the hybrid FHIA-18 on semisolid culture Naine’. Physiol Plant 97:285-290.
Central “Marta Abreu” de Las
medium obtained by Barranco (2001) and Daniels D., R.G. Kosky & M.R. Vega. 2002. Plant
regeneration system via somatic embryogenesis in Villas. Carretera a Camajuaní
the 49.3% for the cultivar ‘Navolean’ (AAB) the hybrid cultivar FHIA-21 (Musa sp. AAAB Group). In Km 5.5, Santa Clara,
obtained by Cabrera (2001). Escalant Vitro Cell. Dev. Biol. 38:330-333. Villa Clara, Cuba CP 54830;
et al. (1994) obtained between 60% and Escalant J.V., C. Teisson & F.X. Côte. 1994. Amplified
somatic embryogenesis from male flowers of triploid
Rony Swennen works at the
70% germination in different cultivars banana and plantain cultivars (Musa sp.). In Vitro Cell Laboratory of Tropical Crop
with a system of temporary immersion of Dev. Biol. 30:181-186. Improvement, KULeuven,
type RITA and Navarro et al. (1997) 25% Gómez R., T. Gilliard, L.A. Barranco & M. Reyes. 2000. Kasteelpark Arenberg 13,
Somatic embryogenesis in liquid media. Maturation
germination in ‘Grande naine’. Daniels et and enhancement of germination of the hybrid cultivar B-3001 Leuven, Belgium.
al. (2002) obtained high germination rates FHIA-18 (AAAB). INFOMUSA 9(1):12-16. *Email: [email protected]

Grapin A., J. Schwendiman & C. Teisson. 1996. Somatic and a triploid, Cavendish banana. Plant Cell, Tissue
embryogenesis in banana plant. In Vitro Cell Dev. Biol. and Organ Culture 51:17-25.
32:66-71. Schenk R.U. & A.C. Hildebrandt. 1972. Medium
Khalil S.M., K.T. Cheah, E.A. Perez, D.A. Gaskill & J.S. and techniques for induction and growth of
Hu. 2002. Regeneration of banana (Musa spp. ABB cv. monocotyledonous and dicotyledonous plant cell
Dwarf Brazilian) via secondary somatic embryogenesis. culture. Canadian Journal of Botany 50:199-204.
Plant Cell Rep. 20:1128-1134. Schoofs H. 1997. The origin of embryogenic cells in Musa.
Murashige T. & F. Skoog. 1962. A revised medium for Thesis PhD. KULeuven, Belgium. 257pp.
rapid growth and bioassays with tobacco tissue Strosse H., R. Domergue, B. Panis, J.V. Escalant & F.X.
cultures. Physiol Plant 15:473-497. Côte. 2003.Banana and plantain embryogenic cell
Navarro C., R. Escobedo & A. Mayo. 1997. In vitro plant suspensions (A. Vézina and C. Picq, eds). INIBAP
regeneration from embryogenic cultures of a diploid Technical guidelines 8. INIBAP, Montpellier, France.
Tissue culture Effect of an analogue of brassinosteroid on FHIA-18

plantlets exposed to thermal stress
J. L. González-Olmedo, A. Córdova, C. E. Aragón, D. Pina, M. Rivas and
R. Rodríguez
E
nvironmental stress factors, such as (Murashige and Skoog 1962) supplemented
drought, cold and salinity, impose with 6-benzylaminopurine (BAP) according
a cost on plants trying to survive to the protocol of Barrera et al. (2001).
these severe conditions. Many genes have At the end of the in vitro culture phase,
been implicated in the various physiological plantlets were transferred to ex vitro
and biochemical responses induced in conditions according to the protocol
plants following a stress (Shinozaki and described in Anon. (2003). After four weeks
Yamaguchi-Shinozaki 1997, Shinozaki and in acclimatization houses, the plantlets were
Yamaguchi-Shinozaki 2000, Nuotio et al. sprayed with a trihydroxilated spirotane
2001). analogue of brassinosteroid (C27O6H42),
In bananas, growth ceases at about 10°C, (molecular mass = 462.606) at a rate of
irreversible damage happens at 0°C, and 2.2 ml per plant and a concentration of
severe foliar damage is also observed at 0.1 mg/L. The control group was sprayed
37°C. However little is known about the with water only. Two hours later, each group
molecular, biochemical and physiological was subdivided into three and transferred for
changes associated with disorders resulting 72 h to different chambers with temperatures
from temperature extremes. In other of 7°C, 27°C and 34°C. The variables
species, damage has been mitigated by measured were: rate of survival after 72 h in
the exogenous application of plant growth the chambers, after 10 days and at the end
regulators (González-Olmedo and Borroto of acclimatization period; free proline content
1987). The use of brassinosteroids to that after 72 h in the chambers, as described in
end in horticulture is recent but so far it has Bates et al. (1973); and at the end of the
given satisfactory results in reducing the acclimatization period, number of necrotic
effects of environmental stress (Núñez 1999, leaves, plant fresh weight, number of roots,
González-Olmedo et al. 2003). number of leaves and plant height.
The present work was carried out with the Only fully unfolded leaves were used
objective of controlling the effects of thermal to cover the container (PLC6, 2.5 cm2).
stress on banana plantlets by the application Measurement was by means of a CIRAS_2
of a brassinosteroid analogue during the (Europa, PP Systems, UK) portable
acclimatization phase photosynthesis system. Under controlled
light, the concentration of carbon dioxide
Materials and methods was 375 µmol per mol of air and the relative
Plantlets of FHIA-18 were produced by humidity 80%. Measurements were made
micropropagation in temporary immersion on 5 plants with 10 replicates, 24 hours and
bioreactors by using a liquid MS medium 7 days after the application of the thermal
18 June 2005
stress. Statistical analysis was by parametric Table 1. Effect of a trihydroxilated spirotane analogue of brassinosteroid (BRAS) and
analysis (ANOVA, Tukey test, p< 0.05) after temperature on FHIA-18 plantlets during acclimatization (n= 40).
Temperature BRAS Survival (%) % of plants with Number of leaves
checking the data for normal distribution spots with spots
(°C) (mg/L) 72 hours 10 days
(Kolmogorov-Smirnov) and homogeneity of per plant
variance (Bartlett). 7 0 97 97 97 a 4
7 0.1 100 100 92 b 4
Results and discussion 27 0 100 100 0e 0
Table 1 presents the data on plantlet survival. 27 0.1 100 100 0e 0
34 0 100 97 38 c 2
Since the banana plantlets were already 34 0.1 100 100 18 d 1
established and adapted to the temperatures Values followed by different letters in the same column are significantly different at p< 0.05 according to Tukey’s test.
outside the acclimatization chambers,
changing the temperature did not noticeably
affect their survival. Nevertheless, extreme Table 2. Effect of a trihydroxilated spirotane
temperatures resulted in some deaths, analogue of brassinosteroid (BRAS) and
which occurred earlier in the treatments with temperature on the levels of free proline in
FHIA-18 plantlets 72h after treatment (n=15).
cooler temperatures. No plantlets died in the
Temperature BRAS Concentration
treatments with the brassinosteroid. (°C) (mg/L) (µg/ml)
Extreme temperatures increased the 7 0.0 0.54 a
numbers of plants with stress symptoms, as 7 0.1 0.23 bc
27 0.0 0.22 bcd
measured by necrotic areas on the leaves,
27 0.1 0.11 e
which were 38% and 97% at 34°C and 7˚C 34 0.0 0.30 b
respectively. In the plants treated with a 34 0.1 0.15 cd
brassinosteroid analogue the effects of the Values followed by different letters in the same column are
thermal stress were significantly reduced, significantly different at p< 0.05 according to Tukey’s test.
with more than 53% and 5% of plants free of

symptoms at 34°C and 7°C respectively.
According to the free proline content, (Table 3). The brassinosteroid analogue only
the most stressed plants were those resulted in an increase in this variable in
submitted to temperatures of 7°C (Table plants placed in the chamber at 34°C.
2). The significant reduction in the levels None of the treatments resulted in
of this indicator following spraying of a significant changes in root numbers.
brassinosteroid is in keeping with the Similarly, morphogenesis appears to be
previously observed properties of this less affected than growth. Plant height was
group of plant growth regulators (Sasse significantly reduced by both temperature
1997, Núñez 1999, González-Olmedo et al. extremes, whereas the application of
2003). Similar results were obtained with the brassinosteroids was effective only in plants
plants placed in chambers with the highest exposed to the warmer temperature.
temperature and with the plants in the control As regards fresh weight, temperature had
group (27°C). no direct effect, with low values in all groups,
According to Sasse (1997), brassinosteroids except when sprayed with brassinosteroids
play an independent role in the first stages of in chambers at 27°C and 34°C, when there
vegetative growth, in particular as growth were significant increases. Brassinosteroids
promoters. Their characteristic is stimulating appear to exercise a major role on water
cellular elongation and division, vegetative regulation in treated plants, with no loss
growth, reproduction, interacting with in the assimilation of photosynthates in
other hormones, increasing yield and the control plants as demonstrated by the data
production of biomass in different cultivars in table 3.
and acceleratiing maturation. Furthermore, It is important to note the marked
they increase plant resistance to pests and photosynthetic activity evident in the
different stress factors such as high salinity, banana plantlets after only four weeks of
drought, high temperature and powerful acclimatization, as indicated by results in
chemical agents such as pesticides and the chamber at 27°C. Extreme temperatures
herbicides (Sasse 1997). significantly reduced photosynthesis after
Cool temperatures affected leaf emer- 24 hours of exposure. The brassinosteroid
gence, with a significant reduction in their had marked positive effects only at
number in comparison with the control the highest temperature. In the week

Table 3. Effect of a trihydroxilated spirotane analogue of brassinosteroid (BRAS) and temperature
on the number of leaves and roots, plant height and fresh weight of FHIA-18 (n=30) and on net
photosynthesis (n=50).
Temperature BRAS Number Number Height Fresh Net photosynthesis
C° (mg/L) of leaves of roots (cm) weight (µmol CO2 m-2 s-1)
(g) 24 hours 7 days
7 0.0 4.2 c 5.3 a 3.6 b 4.5 b 2.1 c 4.7 cd
7 0.1 4.4 bc 6.1 a 3.8 b 4.6 b 0.5 e 4.6 d
27 0.0 4.5 ab 5.5 a 4.6 a 5.0 b 8.8 a 8.9 b
27 0.1 4.8 a 5.2 a 4.5 a 7.0 a 8.8 a 10.3 a
34 0.0 4.3 bc 5.2 a 3.5 b 4.8 b 1.5 d 2.5 e
34 0.1 5.1 a 5.5 a 4.8 a 6.5 a 3.3 b 5.0 c
SE 0.2 0.5 0.2 0.34 0.1 0.2
Values followed by different letters in the same column are significantly different at p< 0.05 according to Tukey’s test.
in which the acclimatization conditions Barrera L., M. Daquinta, Y. Lezcano, O. Mosqueda & M.
Escalona. 2001. Manejo de bananos tetraploides en
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Physiol. Plant. 15:473-479.
known as choking (Daniells 1993), except for
Núñez M. 1999. Aplicaciones prácticas de los
differences in age and development. Results brasinoesteroides y sus análogos en la agricultura.
with the brassinosteroid analogue suggest Cult. Tropicales 20(3):63-72.
a potential for its use, pending further Nuotio S., P. Heino & E.P. Palva. 2001. Signal transduction
laboratory and field experiments. under low-temperature stress. Pp 151-175 in Crop
Responses and Adaptations to Temperature Stress.
Acknowledgements (A.S. Basra, ed.). Food Products Press, New York.
To the memory of Dr Rodolfo Maribona Sasse J.M. 1997. Recent progress in brassinosteroid
who promoted this work. research. Physiol. Plant. 100:696-701.
Shinozaki K. & K. Yamaguchi-Shinozaki. 1997. Gene
The authors work at the References expression and signal transduction in water-stress
Centro de Bioplantas, Anon. 2003. Fase IV. Adaptación o aclimatación a response. Plant Physiol. 115:327-334.
condiciones ambientales. Pp. 18-20 in Instructivo Shinozaki K. & K. Yamaguchi-Shinozaki. 2000. Molecular
Universidad de Ciego de Avila, técnico para la micropropagación de plátanos y responses to dehydration and low temperature:
PO Box 69450, Cuba. bananas. Capitulo 8. Empresa de semillas, La Habana, difference and cross-talk between two stress signaling
E-mail: [email protected] Cuba. pathways. Cur. Opin. Plant Biol. 3:217-223.
20 June 2005
Optimizing the culture conditions of Mycosphaerella Mycosphaerella
fijiensis Morelet
M. Puch-Ceh, K. García-Sosa and L. M. Peña-Rodríguez
O
ne of the main problems that affects fungus might produce other metabolites
the production of Musa sp. in the that could be primary determinants of the
tropics is black leaf streak disease, disease. It is well known that some micro-
caused by Mycosphaerella fijiensis Morelet, organisms producing metabolites under
a fungus recognized as the most aggressive in vitro conditions do not produce them in
form of the Mycosphaerella leaf spot disease vivo, or vice versa (Shaw 1991). Culturing
complex. The disease has been reported to conditions, e.g. medium composition,
cause a 30% reduction in crop yield when the aeration and light, under which a fungus
disease is not controlled and it is estimated is grown, are the most important factors
that the cost of using fungicides to control affecting the production of phytotoxins
the disease in Central America, Colombia (Stierle et al. 1992). This study is part of a
and Mexico has reached 350 million dollars project aimed at isolating and identifying host
in eight years (Stierle et al. 1991). specific phytotoxins produced by M. fijiensis.
Fungal pathogens are known to produce In order to determine the best culturing
phytotoxins that are harmful to the host plant conditions for a high phytotoxicity of the
in low concentrations. These metabolites fungal filtrates and a high yield of organic
can diffuse from the site of infection to crude extracts, we evaluated the growth of
adjacent tissues causing necrosis, chlorosis,
M. fijiensis in eleven liquid media, under two
withering, or a combination of these
lighting and two aeration conditions.
symptoms, in a susceptible host (Wheeler
1981). Phytotoxins are classified as host- Materials and methods
specific, or primary determinants of the The strain of M. fijiensis (W6) was provided
disease, and non-host specific, or secondary by Dr. Andrew James of the Centro de
determinants of the disease (Scheffer and Investigación Científica de Yucatán in
Briggs 1981). The potential applications Mexico. It was originally isolated from
of phytotoxins include their use as probes infected Musa acuminata plants.
to study the molecular basis of disease
A portion of the mycelium culture was
resistance and susceptibility in plants, as
placed in slant tubes containing 10 ml of
tools for the in vitro selection of disease
potato dextrose agar (PDA) culture medium
resistant plant lines, and as possible agents
and the tubes were incubated for 30 days
for weed control (Buiatti and Ingram 1991).
at room temperature and under a 12:12
Most of our knowledge on phytotoxins
comes from those produced by fungi of photoperiod. After growth was completed,
the Alternaria and Cochliobolus genus, but tubes were kept at 4°C until utilized.
studies carried out on the fungal metabolites An aqueous suspension of spores/
produced by M. fijiensis resulted in the mycelium was prepared by adding 2 ml
identification of a number of phytotoxins, of distilled sterile water to a slant tube
including the non-host specific fijiensin containing a parent culture of M. fijiensis
(Stierle et al. 1992, Upadhyay et al. 1991) and removing the mycelium by using a wire.
and the host-specific tetralones (Stierle et One ml of the aqueous suspension was
al. 1991). However, attempts to generate used to inoculate a PDA plate, which was
disease resistant banana plants, using both incubated for 20 days at room temperature
semipurified fractions and pure phytotoxins under natural light conditions. Five ml of
from M. fijiensis as selection tools, have not sterile water was added to each plate and
been successful (Harelimana et al. 1997, a spore suspension was prepared by gently
Okole and Schulz 1997). scratching the surface with a paintbrush.
These results suggest that neither One ml of the spore suspension was used
fijiensin nor tetralones are essential for the to inoculate a Roux flask containing liquid
pathogenicity of M. fijiensis and that the culture media.

Five liquid media, prepared as reported also done using four Erlenmayer flasks that
in the literature, were initially selected for were maintained under a 12:12 photoperiod
evaluating the culturing conditions of M. for 30 days at 26°C.
fijiensis: Once the culture period was over, the
• V-8 juice medium (Peña-Rodriguez et al. culture broth was separated from the
1988) mycelial mat by filtration through two layers
• Potato-dextrose broth (Natural 1989) of cheesecloth. A 350 ml portion of the culture
• Nutritious synthetic medium (Natural 1989) filtrate was lyophilized and resuspended in
• Czapek-Dox medium (Natural 1989) distilled water to a concentration of 100 mg/ml.
• Synthetic M1D medium (Pinkerton and The remaining filtrate was kept in a freezer.
Strobel 1976).
Leaf spot assay
However, since it has been reported that
the addition of an infusion of the host plant Leaves of susceptible banana plants
to the culture medium promotes sporulation (‘Grande naine’) were used to evaluate
and phytotoxin production (Durbin 1981), the the phytotoxic activity. The leaves were
growth of M. fijiensis was also evaluated in a obtained from plants growing in pots under
banana infusion medium prepared by adding greenhouse conditions.
200 g of chopped banana leaves to 800 ml The first or second youngest leaf of four-
of boiling water and boiling for 30 min. After month-old banana plants was exscinded and
cooling, the mixture was filtered through disinfected for 60 seconds with a 5% sodium
cheesecloth and the volume adjusted to hypochlorite solution. The leaves were rinsed
one liter. The infusion was then sterilized. with distilled water, dried between two paper
Each of the liquid media was combined with towels and placed in a plastic container,
a banana infusion in a 2:1 ratio. The cultures previously disinfected with 70% ethanol,
were evaluated under still and shaked lined with two layers of moist filter paper.
conditions, and under total darkness and The activity of the culture filtrates was tested
light/dark conditions. by placing 20 µl of either the culture filtrate
For the still cultures of each culture medium, (3% and 1.5%), the uninoculated medium
250 ml of liquid medium were added to four (3% and 1.5 %) or sterile water, on two
250 ml Roux bottles. Three of the flasks were incisions made with a scalpel on either
inoculated with 1 ml of a M. fijiensis spore/ side of the adaxial face of the leaf. Two
mycelium suspension and the remaining leaves were utilized for each treatment and
flask was kept as control. The four flasks the containers were maintained at room
were maintained in the dark for 45 days, at temperature under natural light conditions.
26±2°C. Similar preparations were also done The effect (the area of the necrotic lesion)
using four Roux bottles that were kept under a was registered at 24, 48 and 72 hours.
12:12 photoperiod for 45 days at 26±2°C.
For the shaked cultures of each culture Results and discussion
medium, 300 ml of liquid medium were added Although both the still and shaked V8
to four 500 ml Erlenmeyer flasks. Three cultures showed abundant mycelial growth
of them were inoculated with 1 ml of a M. under the two lighting conditions (data not
fijiensis spore/mycelium suspension and the shown), their yield of organic crude extract
remaining flask was kept as control. Cultures was higher under still and dark conditions
were maintained in the dark at 100 rpm for (Table 1). Conversely, while the M1D cultures
30 days, at 26°C. Similar preparations were did not show discernible mycelial growth
Table 1. Quantity (mg/L) of organic crude extract obtained by culturing Mycosphaerella

fijiensis under various conditions.
Culture medium* Still Shaked
12:12 Darkness 12:12 Darkness
photoperiod photoperiod
V8 21.3 83.0 8.6 11.0
Potato-dextrose broth 18.0 9.6 23.6 54.6
Synthetic M1D 122.6 46.3 69.6 86.3
Nutritious synthetic –** – 11.3 13.0
Czapek-Dox 11.0 11.6 12.6 35.3
* Only the media for which fungal growth was observed are listed.
** No significant growth observed.
22 June 2005
Table 2. Necrotic area (cm2) as a measure of the phytotoxic activity caused by 3% and
1.5% culture filtrates of Mycosphaerella fijiensis grown under different conditions and in
various culture media.
Culture medium Light conditions Culture filtrate
3% 1.5%
Still cultures
V8 12:12 photoperiod 0.138 –*
V8 darkness 0.207 –
Czapek-Dox 12:12 photoperiod 0.898 0.249
Czapek-Dox darkness 0.243 –
Shaked cultures
V8 12:12 photoperiod 1.182 0.376
V8 darkness) 0.237 –
Czapek-Dox 2:12 photoperiod 0.146 –
Czapek-Dox darkness 0.267 –
* No activity.
(data not shown), their yield of organic crude Grayer R.J. & T. Kokubun. 2001. Plant-fungal interactions:
extract was consistently high (Table 1), in the search for phytoalexins and other antifungal
compounds from higher plants. Phytochemistry 56:
agreement with the high yields reported in 253-263.
the literature (Upadhyay et al. 1991). The Harelimana G., P. Lepoivre, H. Jijakli & X. Mourichon.
results suggest that there is no relation 1997. Use of Mycosphaerella fijiensis toxins for the
between the amount of mycelium generated selection of banana cultivars resistant to black leaf
in a given culture medium and the yield of streak. Euphytica 89:125-128.
Natural M.P. 1989. The development of an in vitro
organic crude extract obtained.
screening procedure to Sigatoka leaf disease of
It is interesting to note that neither the banana. Pp. 208-230 in Sigatoka leaf spot diseases of
banana infusion nor any of the media banana. Proceedings of the 1st International workshop
containing banana infusion showed fungal on Sigatoka leaf spot diseases held in San José, Costa
growth. This suggests the presence of Rica (R.A. Fullerton & R.H. Strover, eds). INIBAP,
Montpellier, France.
phytoalexins with antifungal activity in the
Okole B.N. & F.A. Schulz. 1997. Selection of
culture medium, particularly in the banana Mycosphaerella fijiensis-resistant cell lines from
infusion (Grayer and Kokubun 2001). The micro-cross sections of banana and plantain. Plant Cell
isolation and identification of the metabolites Reports 16:339.
responsible for the antifungal activity is being Peña-Rodríguez L.M., N.A. Armingeon & W.S. Chilton.
done. 1988. Toxins from weed pathogens. I. Phytotoxins
from a Bipolaris pathogen of Johnson grass. Journal of
The strongest phytotoxic activities were Natural Products 51:821-828.
observed in the filtrates of the V8 shaked Pinkerton F. & G. Srobel. 1976. Proceedings of the
cultures and the Czapek-Dox still cultures National Academy of Sciences USA 74:4007-4011.
kept under light/dark conditions (Table 2). Scheffer R.P. & S.P. Briggs. 1981. Introduction and
Given that the V8 shaked cultures kept perspective of toxin studies in plant pathology. Pp. 1-17
in Toxins in plant disease (R.D. Durbin, ed.). Academy
under light/dark conditions had a higher yield Press, New York.
of organic crude extract, and that many of Shaw P.D. 1981. Production and isolation. Pp. 21-44 in
the phytotoxic metabolites reported to date Toxins in plant disease (R.D. Durbin, ed.). Academic
are readily extracted into organic solvents, Press, New York.
these conditions were chosen as optimum Stierle A., G. Strobel, D. Stierle & F. Sugawara. 1992.
for culturing M. fijiensis. Accordingly, we are Analytical methods for phytotoxins. Pp. 1-32 in Methods
of plant analysis (H.F. Linskens & J.F. Jackson, eds).
presently working towards the isolation and Springer Verlag, Berlin.
identification of the phytotoxic metabolites Stierle A., R. Upadhyay, J. Hershenhorn, G. Strobel &
produced by the fungus; the results of which G. Molina. 1991. The phytotoxins of Mycosphaerella The authors work at the
will be published in due course. fijiensis, the causative agent of black Sigatoka disease Unidad de Biotecnología,
of bananas and plantains. Experientia 47:853-858. Centro de Investigación
References Upadhyay R., G. Strobel, S. Coval & J. Clardy. 1991. Científica de Yucatán
Buiatti M. & D.S. Ingram.1991. Phytotoxins as tools in Fijiensin, the first phytotoxin from Mycosphaerella (CICY), Calle 43 # 130,
breeding and selection of disease-resistant plants. fijiensis, the causative agent of black sigatoka disease.
Experientia 47:982-984.
Col. Chuburná de Hidalgo,
Experientia 47:811-819.
Durbin R.D. 1981. Applications. Pp 495-505 in Toxins in Wheeler H. 1981. Role in Pathogenesis. Pp. 477-494 in 97200, Mérida, Mexico.
Plant Disease. (R.D. Durbin, ed.). Academic Press, Toxins in plant disease. (R.D. Durbin, ed.). Academic Author for correspondence:
New York. Press, New York. [email protected]

Mycosphaerella culture Effect of light and sealing pattern on sporulation and
growth of Mycosphaerella fijiensis
E. Etebu, C. Pasberg-Gauhl, F. Gauhl and L. Ayibo Daniel-Kalio
B
lack leaf streak disease, caused with 8 fluorescent tubes (OSRAM, L8W/20).
by Mycosphaerella fijiensis, is the One of the three Petri dishes was sealed with
most destructive leaf spot disease parafilm throughout the incubation period, one
of plantains and bananas. Yield losses was left unsealed throughout the incubation
between 20 to 90% have been reported period, and one was sealed with parafilm for
(Stover 1983, Fouré 1985, Pasberg-Gauhl the first 14 days and unsealed for the next
1989, Mobambo et al. 1993). 7 days of incubation. The same procedure
Disease severity varies between cultivars, was repeated with the same isolates
in part because of differences in virulence incubated for 21 days under blacklight, 2
between the pathotypes (Fullerton and Olsen Sylvania fluorescent lamp tubes (F40/350
1991). Consequently, the need to test on a BL) positioned 40.5 cm upward away from the
given cultivar the aggressiveness of different culture dishes. Temperature was maintained
M. fijiensis isolates has often been proposed. between 26 to 29°C for each type of light.
This, however, has been difficult to achieve The experiment was replicated four times.
because of the slow-growing nature of the Linear colony size (the mean of the largest
fungus which precludes the production of and smallest diameters out of a sample of
enough inoculum for artificial inoculation 10) was measured at the end of the 21-day
(Mobambo 1993, Pasberg-Gauhl 1994). incubation period. Each colony was viewed
Moreover, most M. fijiensis isolates produce under a Leitz larbolux S light microscope
very few or no conidia at all in culture. (20x) fitted with a Leica Wild MPS 52 camera
Allowing some air circulation is one of to determine the number of sporulating
the most important factors for successful isolates. Sporulation was expressed as a
sporulation of various fungi (Henry and percentage that was arcsine-transformed
Andersen 1948, Lilly and Barnett 1950, 1951), (Gomez and Gomez 1984) and analysed by
a factor not investigated with M. fijiensis using Statgraphics version 2.1.
(Mobambo 1993, Pasberg-Gauhl 1994). This
study was carried out to study the effect of light
Results and discussion
and of the sealing pattern on the sporulation Sporulation of M. fijiensis was observed to be
and linear growth of M. fijiensis. dependent on both light and sealing pattern.
Sporulation was significantly higher under
Materials and methods blacklight than whitelight (Table 1). Exposure
Unexposed parts of stage 2 leaves (Ganry to UV light has been reported to induce
and Laville 1983, Gauhl et al. 1993) from sporulation in fungi (Ramsey and Bailey 1930,
an ‘Agbagba’ False horn plantain were cut McCallan and Chan 1944), but recent studies
into pieces of about 2 cm2, placed on a have also shown that wavelengths between
solid potato dextrose agar (PDA) medium 300 to 380 nm, near the UV region, induce
and autoclaved to form a PDA autoclaved more sporulation than those in the UV range
plantain leaf (PAL) medium. (200-300 nm). This latter region could be lethal
Three-month-old ascospore cultures derived or mutagenic (Leach 1971). Consequently,
from 30 M. fijiensis isolates were multiplied blacklight is preferred to whitelight in inducing
and subcultured on fresh PDA medium and sporulation of M. fijiensis isolates. Colony size
incubated in a culture room at 25 to 29°C was also significantly greater under blacklight
for 10 to 20 days. Three pieces of about than whitelight.
0.5 cm to 1.0 cm by 0.5 cm to 1.0 cm were Sealing patterns were also observed to
cut from the actively growing mycelium of significantly (P<0.01) affect sporulation of
each isolate, placed in separate Petri dishes M. fijiensis (Table 1). Sporulation was better
containing PAL medium and incubated for in the Petri dishes that had been unsealed for
21 days under continuous whitelight in a the duration of the experiment or for the last
Gallenkamp incubator (INF-781-T) equipped 7 days. The accumulation of carbon dioxide
24 June 2005
and the production of inhibitory metabolic Table 1. Effect of light (under all sealing patterns) and sealing pattern (under both
products such as ammonia could explain the types of light) on the percentage of sporulation (original data and arcsine-transformed
data that have been transformed back) and mean linear colony size (diameter) of
lower level of sporulation in the airtight Petri Mycosphaerella fijiensis.
dishes (Henry and Andersen 1948, Barnett Treatments N Mean sporulation Transformed Mean linear
and Lily 1950, Cochraine 1958). Although (%) back arcsine colony size
air circulation was an important factor, the data (%) (mm)
Whitelight 12 14.4 21.9 a 17.0 a
percentage of sporulation was higher in the
Blacklight 12 25.7 29.7 b 18.6 b
Petri dishes that had been sealed for the first Sealed for 21 days 8 10.9 19.1 a 15.0 a
14 days of incubation and unsealed for the Unsealed for 21 days 8 19.9 25.9 b 21.4 c
remaining 7 days than in those that had not Sealed for 14 days and
unsealed for 7 8 29.5 32.3 c 17.0 b
been sealed for 21 days. This suggests that
Means followed by the same letter in a column are not significantly different at P<0.01 according to Duncan’s Multiple
the metabolic products thought to accumulate Range Test.
in the sealed Petri dishes favour sporulation,
but that their continuous presence has an
inhibitory effect. Mean colony size, however, Lilly V.G. & H.L. Barnett. 1951. Physiology of the fungi.
McGraw-Hill Book Company Inc. USA.
was significantly higher in Petri dishes left McCallan S. E. A. & S. Y. Chan. 1944. Inducing sporulation
unsealed for the duration of the experiment. of Alternaria solani in culture. Boyce Thompson Inst. 13:
Table 2 shows that blacklight significantly 323-336.
affected the growth of M. fijiensis colonies Mobambo K.N. 1993. Factors influencing the development of
black Sigatoka disease on plantain and plantain hybrids.
but only when the Petri dishes were sealed
PhD Thesis, Rivers State University of Science and
for 21 or 14 days. In the Petri dishes that Technology, Nkpolu, Port Harcourt, Nigeria.
were not sealed, linear colony size was not Mobambo K.N., F. Gauhl, D. Vuylsteke, R. Ortiz, C. Pasberg-
significantly different between the two types Gauhl & R. Swennen. 1993. Yield loss in plantain from
of light. The effect of aeration in enhancing black Sigatoka leaf spot and field performance of resistant
hybrids. Field Crops Research 35:35-42. Ebimieowei Etebu (author for
the growth of M. fijiensis was more Müller R., C. Pasberg-Gauhl, F. Gauhl, D. Kaemmer & G.
pronounced under whitelight than blacklight. correspondence,
Kahl. 1995. Tracing microsatellite polymorphism within
Further studies are needed to evaluate the the Nigerian population of Mycosphaerella fijiensis. e-mail: [email protected])
effect of light and of the sealing pattern on INFOMUSA 4(1):9-11. works at the Department of
the in vitro production of conidia. This may Pasberg-Gauhl C. 1989. Untersuchungen zur Symptom- Biological Sciences, Niger
entwicklung and Bekaempfung der Schwarzen Sigatoka Delta University, Wilberforce
facilitate the testing of the aggressiveness of Krankheit (Mycosphaerella fijiensis Morelet) an Bananen
different pathotypes on different cultivars. (Musa spp.) in vitro und im Freiland. Goettinger Beitraege
Island, Bayelsa State, Nigeria
zur Land- und Forstwirtschaft in den Tropen und and Lloyd Ayibo Daniel-
References Subtropen, Heft 40. 142pp Kalio at the Department of
Barnett H.L. & V.G. Lilly. 1950. Nutritional and environmental Pasberg-Gauhl C. 1994. Symptom development of black Biological Sciences, Rivers
factors influencing asexual sporulation of Choanephora Sigatoka leaf spot on young or adult banana and plantain
cucurbitarum in culture. Phytopathology 40:80-89. plants after natural inoculation. Pp 261-274 in Biological
State University of Science
Cochraine V.W. 1958. Physiology of fungi. John Wiley and and integrated control of highland banana and plantain and Technology, P. M. B.
sons. New York and London. pests and diseases (Gemmill B. & C. Gold, eds). IITA 5080, Nkpolu, Port Harcourt,
Fouré E. 1985. ‘Les cercosporioses du bananier et leur Proceedings. Cotonou, Benin. Rivers State, Nigeria. Cornelia
traitement. Etude de la sensibilité variétale des bananiers Ramsey G.B. & A. Bailey. 1930. Effects of ultra-violet
et plantain à Mycosphaerella fijiensis Morelet au Gabon. Pasberg-Gauhl and Friedhelm
radiation upon sporulation in Macrosporium and Fusarium.
(suite III). Fruits 40(6):393-399. Botan. Gaz. 89:113-136. Gauhl work at Chiquita
Fullerton R.A. & T. L. Olsen. 1991. Pathogenic variability Stover R.H. 1983. Effet de la cercosporiose noire sur les International Brands,
in Mycosphearella fijiensis Morelet. Pp. 105-115 in
Banana diseases in Asia and the Pacific: Proceedings
plantains en Amérique centrale. Fruits 38:326-329. Costa Rica.
of a technical meeting on diseases affecting banana and
plantain in Asia and the Pacific (R.V. Valmayor, B.E. Umali
& C.P. Bejosano, eds). INIBAP, Montpellier, France. Table 2. Effect of light and sealing pattern on the percentage of sporulation (original
Ganry J. & E. Laville. 1983. Les cercosporioses du bananier data and arcsine-ransformed data that have been transformed back) and the mean linear
et leur traitement. Evolution des méthodes de traitement. colony size (diameter) of Mycosphaerella fijiensis (n=4).
Fruits 38:3-20 Treatments Mean Transformed back Mean linear
Gauhl F., C. Pasberg-Gauhl, D. Vuylsteke & R. Ortiz. 1993. sporulation arcsine data colony size
IITA research guide 47. International Institute of Tropical (%) (%) (mm)
Agriculture, Ibadan, Nigeria. 49 pp. Whitelight
Gomez K.A. & A.A. Gomez. 1984. Statistical procedures Sealed for 21 days 8.9 17.3 13.3
for Agricultural research (2nd ed.). John Wiley and sons, Unsealed for 21 days 13.1 21.2 21.3
Singapore. Sealed for 14 days and unsealed for 7 21.3 27.4 16.3
Henry W.B. & A.L. Andersen. 1948. Sporulation by Piricularia Blacklight
oryzae. Phytopathology 38:265-278. Sealed for 21 days 12.9 21.0 16.6
Leach C. M. 1971. A practical guide to the effects of visible Unsealed for 21 days 26.7 31.0 21.5
and ultraviolet light on fungi. Pp. 609-664 in Methods Sealed for 14 days and unsealed for 7 37.6 37.3 17.6
in microbiology (C. Booth, ed.). London and New York
Least significant difference at P<0.05 6.0 1.1
Academic Press.

Farmer training Designing farmer training programmes using a
participatory approach
C. Staver
F
arm households grow Musa under improve production systems based on local
widely varying conditions: under high variability.
to moderate rainfall or in irrigated Precision agriculture. Computerized
desert environments, in poor to very fertile spatial information on soils, drainage, yields
soils, and anywhere between sea level to and other characteristics is being used to
altitudes of 1500 meters. Bananas are also fine tune the application of inputs for reduced
grown under different cropping systems such environmental pollution and a more efficient
as a monocrop, in association with annual or yield response (Stoorvogel et al. 2004).
perennial crops, or in rotation with other However, predicting the yield response is
crops. In each zone farmers face specific made difficult by weather unpredictability
problems, including how to manage pests while the costs of mapping and computer
and diseases, and plant nutrition, which technology are justified primarily on large
are all influenced by weather variability. farms with high yield potential.
In recent decades, new pests have been Using local resources. In response to the
introduced and certain existing pests have cost of purchased inputs, rural communities
been favoured by changes in cropping have begun to recover traditional technology
practices. Over the same time period, prices and search for substitutes based on local
for agricultural products have fluctuated resources, such as green manures and
wildly, but new markets have also opened. botanical brews, and soil conservation
As a result of all these factors, farmers make practices. Viewed at first with scepticism,
crop and pest management decisions under these techniques are now being validated in
extreme uncertainty. scientifc trials and incorporated into training
programmes.
What experts have offered Harnessing ecological processes in
farmers locally-designed cropping systems.
During much of human history, farmers In this approach, farmers learn about
have done their own research. Rural ecological processes such as food webs,
communities domesticated the major crops nutrient cycles, and habitat management
and designed a host of cropping systems. and use these concepts to redesign and
Only since the mid-1800s have governments, fine tune local cropping systems for pest
universities and scientists become involved suppression, more efficient nutrient cycling,
in agricultural research, especially with the and a healthier crop. The farmers are not
advent of fertilizers and pesticides (Staver simply following best average practice, but
2001, 2003). combine diverse strategies of crop and soil
The approaches which scientists and agro-ecosystem management (Figure 1).
development projects have offered farmers Based on their increasing knowledge of
have evolved. During the early Green ecological processes and their ability to
Revolution, plant breeders developed reason ecologically, they can innovate to
varieties responsive to fertilizers which they manage variability and uncertainty and
tested under various input levels to identify to use inputs that enhance ecological
the best average performance. Farmers processes rather than substitute for them.
were expected to apply the same practices, The increased recognition of the role
including pesticides, year in and year out. played by variability has begun to broaden
In later years, scientists targeted their the focus of development projects from
recommendations according to the soil and inputs and technologies to include the skills
weather conditions and farmer resources. of the persons using the technologies, their
The continuing search for greater information base for decision-making and
productivity and lower environmental their strategies for incorporating alternative
impacts has led to other approaches to practices. In a technological focus, scientific
26 June 2005
trials are used to generate and validate the
practices which on average are the best. Fostering
These are then promoted through field days Fostering an Direct a favorable
Crop unfavorable control
(Figure 2A). However, from the perspective environment
strengthening environment measures for beneficial
of the technology users, such field days are for pests against pests organisms
just one of many inputs into their decision
making processes (Figure 2B). Farmers Primary producers Herbivores Consumers
are experimenters and networkers. They Virus
Crops Virus
filter information and experience from a wide Bacteria Bacteria
Weeds Fungi Fungi
range of sources and adapt them to their Nematodes Nematodes
farming problems and opportunities. This Insects Insects
Birds Birds
switch from technology promotion to the Mammals Mammals
strengthening of the farm community’s ability
to adopt new technologies and integrate Decomposers
them into their livelihood strategy calls for
alternative training approaches. Improving nutrient cycling,
water infiltration, root penetration
Participatory group learning and
experimentation
Adult training differs from the formal
education most of us experienced as farmer’s experience is a case of variability Figure 1: A simplified food web (boxes
and arrows) and possible agroecological
children. Firstly, farmers participating in and therefore a resource for the trainer. management strategies.
training are immersed in their subject. They Secondly, farmers are highly motivated to
have abundant experience producing the learn about a crop which is often part of their
crop on their own farms with the resources income and food security strategy, although
available in their region under variable they each have their own set of concerns.
weather and fluctuating markets. Each An adult training process should use daily
A Figure 2. (A) The scientific trials used to

Demonstration plots generate and validate the practices developed
Field days by scientists and the field days organized to
Multi-year, promote them are one of many information
Research Multi-location
station trials Demonstration plots sources (B) famers have access to.
Validation Field days
On-farm
Demonstration plots
Field days
B
Scientists
Farmer D Farmer C
Popular
media
Farmer A:
Farmer G experience, experiments
Traders information search Farmer F
Farmer E Farmer B:
experience, experiments
Technical information search
media
NGOs
Commercial Extension Farmer
agricultural staff organizations
inputs

life situations as learning laboratories, create their experiments. At each meeting they also
a collaborative environment that emphasises conduct field visits and discovery exercises
the different experiences and approaches to strengthen their ecological knowledge of
the participants bring and incorporate them the crop, other primary producers, pests and
in a learning cycle (analysis, planning, beneficial organisms and soil factors. At the
action, observation). end of the cycle, the group reviews what they
These principles were applied to an have discussed, seen, done and learned.
integrated pest management project in They identify major lessons learned to be
Nicaragua financed by Norway and run used in the next crop cycle and problems for
by the Centro Agronómico Tropical de further experimentation or study.
Investigacíon y Enseñanza (CATIE) to The approach uses participatory methods
come up with an approach for training rather than conventional lectures with
farmers in agro-ecological crop and pest audiovisual materials for practical and
management known as participatory farmer philosophical reasons. In rural areas,
group learning and experimentation by crop electricity is often unreliable and equipment
stage (Staver 2004). In the case of Musa, is expensive and difficult to transport. On the
the project worked with MusaNic, a working other hand, Musa fields of different ages with
group of research, teaching and extension different pest problems are readily available
organizations and grower associations for field laboratory exercises. Farmers are
dedicated to more effective research and more comfortable in informal settings than
training (CATIE 2003a). This approach in a classroom. In addition, retention and
draws on methods from FAO farmer field learning are more effective when active
schools (www.communityipm.org) and rather than passive methods are used.
farmer participatory research (Okali et al. The group approach multiples the learning
1994; Haverkort et al. 1991). opportunities for each participant. A group
The objectives for the participants are to: of 15-20 farmers meeting 4-6 times during
• learn about the ecology of the main pests a crop cycle may discuss up to 60-80
and how to identify them; crop and pest scouting reports and 30-40
• strengthen their observation skills linked to experiments.
pest and crop management decisions; The approach emphasises both learning
• strengthen their agro-ecological reasoning and experimentation. Experimentation is a
capacity (food webs, life cycles, nutrient more formal process, while learning is much
cycles, the role of abiotic and biotic broader and can be achieved with many
factors in population dynamics, and the different approaches – comparisons of in-
factors influencing the variability observed field variability, group analysis of monitoring
between farms, seasons and fields); data, farm visits, review of decision making
• increase their awareness of non-pesticide at the end of the crop cycle, extrapolation of
management alternatives; lessons learned from the small scale to the
• use experiments to test and adapt whole field and from putting into practice
management alternatives; principles.
• promote informal farmer networks; Finally, a training process by crop stage
• seek and evaluate information from many builds on numerous learning cycles that
sources. progress from observation and analysis to
In participatory farmer group learning planning and action. The crop cycle itself is
and experimentation by crop stage, an a learning cycle from which farmers learn for
extensionist meets with a group of farmers the next crop cycle. During the crop cycle,
before crop planting to discuss pest and each farmer meeting is also a learning cycle
crop problems and their experience with based on the issues of the particular crop
different practices, to prioritize problems phase.
and possible alternatives for testing, and to
identify indicators for evaluating the group’s Applying the approach to Musa
progress. Volunteers offer to establish The Pacific plains of Nicaragua have six
experiments and to scout for pests in their months of rain and six months with minimal
fields. At key moments in the crop cycle, or no rain. This is not the ideal climate for
they meet to compare plant growth and pest Musa production, but plantains and cooking
levels. They also discuss the progress of bananas are an important cash crop in
28 June 2005
certain zones. The participatory farmer reached the field, what causes it to increase
group learning and experimentation by crop and how it affects production. In the same
stage in this region has seven meetings field, farmers estimate the availability of
(Table 1) covering about 18 months (CATIE poor, average and good planting material.
2003a). Three events are planned prior to They end the event by planting a small
planting, one as a final evaluation and three experiment with planting materials that have
during the crop cycle. Each event has a been pared or not. Volunteers offer to do the
main theme corresponding to the decision experiment on their own farm or to estimate
making needs of the moment. sucker quality.
In the first meeting, farmers review their In the third meeting, farmers first report
current production practices, the major on the quality and number of suckers they
problems they face and the alternatives they found in the field from which they plan to
are testing. They also answer ten questions obtain their planting material. In a nearby
on their agro-ecological knowledge using live field that is at least in its third production
samples and photographs. As the meeting cycle, small groups quantify disease and
closes, they plan what they want to test in pest problems, plant vigour and bunch
their fields and learn about the Musa agro- quality. Based on these data, they discuss
ecosystem. possible measures to apply during planting
The second meeting focuses on planting to avoid the major problems found in old
material, frequently a source of disease and Musa fields. Next, they join the host farmer
pest infestations. Farmers walk a nearby whose field is scheduled to be planted in
field and gather plant samples that show the upcoming season. They map the field
problems. The group gathers to discuss and its surroundings, identifying the factors
what is the cause of each problem, how it conducive to certain problems, and then
Table 1. Example of a farmer training process in a wet-dry zone of Nicaragua.

Crop Theme and timing of meetings Topics to be discussed
stage
Problems and priorities 1. How do we produce Musa?
(March - dry season) 2. What are our main problems?
Preparation for
3. What do we know about Musa and its pests?

planting
4. What will we test/what do we want to learn?

(April - dry season) 1. Diagnosing pests and diseases in new and old fields – origin of
Source of planting material pests, their movement, the conditions favouring their build-up
2. Estimating how many suckers are available in each field
3. Estimating sucker quality and preparation of planting material
Design of new field 1. Number and quality of suckers present in fields
(Early May – before the first rains) 2. Analysis of old fields (problems related to planting practices)
Planting
3. Analysis of the favourable and unfavourable factors in the new

fields
4. Evaluating the suckers planted at the previous meeting
5. Possible experiments, including new cultivars
Status of new plants 1. Which experiments were set up
Vegetative growth
(August – rains) 2. Analysis of new plantings – effect of sucker and field

characteristics on crop vigor and future pest problems
3. Weeds – growth, life cycles, problems and benefits, management
options
4. Ideas for experiments – weeds, plant health, pest propagation
Plan for next rains 1. Analysis of what is happening in experiments
(February - dry season) 2. Dry season soil cover and nutrient input
3. Ideas and plans for rainy season management
Managing production 1. Analysis of what is happening in fields and experiments
(July – rains) 2. Evaluating plant and root health
Harvest
3. Projected harvest and costs

Final evaluation 1. Analysis of plant growth, yield and profits
(October – rains) 2. Analysis of whether the experiments addressed the identified
problems
3. What was learned
4. What can be scaled up and what should be tested during next
cycle

discuss management alternatives. They management, market orientation with limited
close the day by observing and then digging inputs and no irrigation, market production
up the suckers planted in the previous with moderate inputs and irrigation).
meeting. They identify desirable suckers 2. Identify problems in key zones
and discuss the effect of paring on sucker Based on the experience of the assembled
root growth and vigour. The extensionist and group of scientists and extensionists, each
the farmers suggest alternative practices key zone is characterized according to the
which might be tested. average yield, agronomic practices, main
In meetings four, five and six, the group pest problems, extent of pesticide use, yield
monitors the progress of the plants, the reducing problems, main production costs
increase in pest problems and the response and particular problems which farmers face
of the crop to varying rainfall. They to achieve a greater value for their crop.
complete short learning exercises and set up
experiments with alternative management. 3. Identify key alternatives to move
At each meeting, farmers also discuss the from current to improved situation
conditions in their own fields and report on The group then describes an improved
the progress of their experiments. situation which farmers should be able to
At the evaluation meeting, the extensionist achieve without major changes in invested
and the farmers review weather, pest levels resources – perhaps yield increase of
and crop growth and yield during the cycle. 30%, reduction in production cost of
They calculate costs and yields and discuss 25%, increased value of the crop of 40%,
how to improve their decision-making. They elimination of a toxic pesticide. The group
repeat the test on their agro-ecological identifies what needs to be done to achieve
knowledge and reflect on what they have the improved situation, such as applying
learned. They also analyse their experiments different cultural practices, acquiring new
to determine if alternatives can be scaled up skills and doing additional research.
and what themes need attention in the next 4. Organize available information by
crop cycle. crop stage
With the information from the previous steps,
Designing a training for other the design group then lays out a matrix by
zones crop stage. The group begins by identifying
Because farmers face a different array of basic information by crop stage such as
problems depending on where they live, water and nutrient needs and the crop’s
the content of the training process used in response to deficit and excess. Pest-crop
Nicaragua may not apply. In this section, interactions for each key pest can also be
we describe six broad steps which a group laid out by crop stage. How does the pest
of trainers and scientists with practical reach the field, what conditions favour and
experience in Musa can use to develop the hinder its increase? Are there natural control
curriculum for farmer participatory group organisms, what conditions favour and hinder
learning and experimentation for their own their activity? Any relevant management
conditions (CATIE 2003b). practices leading to the elimination of toxic
1. Define the main production zones pesticides, a reduction in production costs or
by type of technology and agro-climate an increase in productivity are also identified.
For the area of interest, the group identifies With this array of background information,
the range of production technologies used the group then identifies the key moments
and the salient weather, climate and soil when the farmers can meet for maximum
conditions under which the crop is grown. learning. Many of the key decisions occur
There may be two or three major production before planting.
zones with a similar range of technologies. 5. Identify what farmers will test out
MusaNic in Nicaragua defined three zones in their field
(mid-altitude with a 3-month dry season, At this stage in the design of the training
Atlantic lowlands with short dry season curriculum, the group should identify
and Pacific lowlands with a 6-month dry which learning activities farmers might do
season) and three production technologies when they return to their farms after each
(home consumption with minimal inputs and meeting. Trainers should bear in mind that
30 June 2005
farmers will report on this activity at the fields between each meeting to verify the
next meeting. The activity might include experiments, check their understanding of
monitoring pest levels, plant vigour or the training process and learn more about
sucker quality, a learning exercise on the resources and constraints of particular
sucker paring, debudding, estimating households.
earthworm populations, weevil trapping, By the end of the participatory group
experimenting with alternative management learning and experimentation process, the
practices such as new variety, green trainers will have numerous inputs to monitor
manure, planting density or selective weed the effectiveness of their work – how regular
management. The set of activities for the was farmer attendance, how much farmers
entire crop cycle should be closely linked learned, how many farmers did experiments,
to the key changes previously identified for structured observation or learning exercises
moving the crop production from the current in their own fields and which practices
to an improved situation in terms of farmer farmers expect to scale up in the next
skills and practices.
planting cycle. This experience can then be
6. Design learning activities to used to modify the design of the new training
motivate and prepare farmers to act cycle to more effectively strengthen farmer
Once the group has identified the activities decision making in agro-ecological crop
farmers could do, the meeting is designed management (Staver and Guharay 2004).
to prepare farmers to return to their farms
with the necessary skills, materials and References
enthusiasm to do the activity. A meeting CATIE. 2003a. Guia para el Manejo Agroecologico de
Musáceas. CD. Managua, Nicaragua
of 4-5 hours should schedule time at the
CATIE. 2003b. ¿Cómo Analizar el Estado de Tecnologías
beginning to review, discuss and analyse y Conocimientos para un Manejo Agro-ecológico?.
what farmers have done since the last CD. Managua, Nicaragua.
meeting and leave time at the end to identify Haverkort B., J. van der Kamp, & A.Waters-Bayer.
who will do what before the next meeting. 1991. Joining Farmers’ Experiments: Experiences in
Participatory Technology Development. Intermediate
The middle section of 2-3 hours can be Technology Publications, London.
dedicated to new learning activities. In a Okali C., J. Sumberg & J. Farrington. 1994. Farmer
training process with 6 farmer meetings, Participatory Research. Intermediate Technology
only about 8-12 hours will be available Publications, London.
Staver C. 2001. Knowledge, science and practice in
for these activities. The exercises must ecological weed management: farmer-extensionist-
be carefully chosen and well designed for scientist interactions. Pp. 99-138 in Ecological
maximum learning value. Training events Management of Agricultural Weeds (M. Liebman, C.
that are too long and complicated may be Mohler and C. Staver, eds). Cambridge University
Press,
well intentioned, but are less effective than a Staver C. 2003. Farmer learning linked to ecological
well designed process with a moderate level processes for better pest management: Challenges
of content. to CATIE and its partners (in Spanish). Revista MIP y
Agroecología 65:21-33.
Monitoring outcomes and Staver C. 2004. El MIP eficaz: aprendizajes
impacts metodológicos sobre la relación familias rurales
– extensionistas en los proyectos CATIE/NORAD
The procedure described above prepares en Nicaragua durante los 90’s. In Procesos de
trainers and extensionists to work with Innovacion Rural en America Central - Reflexiones
farmer groups in a particular region. The y Aprendizajes (C. Prins, ed.). CATIE, Turrialba,
Costa Rica.
design is a first approximation that can be Staver C. & F. Guharay. 2004. Beyond integrated pest
modified during the process itself. After management: From farm households to learning
the diagnostic farmer meeting, activities capacity and innovations systems. In Participatory
can be adjusted based on the results of Research and Development for Sustainable
Agriculture and Natural Resource Management: A
the test on agro-ecological knowledge Sourcebook. (J. Gonsalves, T. Becker, A. Braun,
and the prioritized problems. After each D. Campilan, H. De Chavez, E. Fajber, M. Kapiriri, Charles Staver works at
subsequent meeting, activities can continue J. Rivaca-Caminade and R. Vernooy, eds). CIP- the international Network for
to be adjusted based on the pest problems, UPWARD, Laguna, Philippines; IDRC, Ottawa, the Improvement of Banana
Canada; IFAD, Rome, Italy.
or any other problem, that appeared Stoorvogel J., J. Bouma & Ortiz, R. 2004. Participatory
and Plantain (INIBAP), Parc
during the crop cycle. The trainers and research for systems analysis: prototyping for Costa Scientifique Agropolis II,
extensionists can also visit farmers in their Rica. Agronomy Journal 96(2): 323-336. Montpellier, France

Focus on the creation of INIBAP How INIBAP was born
Although bananas and plantains (Musa fungus Mycosphaerella fijiensis, that was
spp.) are reputedly the fourth most devastating banana fields.
important food staples in the world, until In Latin America and the Caribbean,
fairly recently these crops were largely which were free of black leaf streak
ignored by national agricultural research disease at the time, the call for international
institutes in developing countries. In the assistance originated with ACORBAT,
early 1980s, the long-established banana a regional association of scientists
breeding programme in the Caribbean involved in research or extension work
was dormant. Only Brazil, India, Nigeria, on Musa and their production systems.
the Philippines and the East Caribbean The Government of Jamaica also raised
had research programmes on Musa, the need for international action at a
albeit inadequately staffed and funded meeting of the United Nations Conference
ones. Elsewhere, research on Musa was on Trade and Development in November
mainly the work of a handful of scientists 1982. In West Africa, support came from
working on individual research projects. the International Institute of Tropical
The one exception was Honduras, where Agriculture, which had established the
West African Regional Cooperation for
a private company had just handed over
Research on Plantain (WARCORP) to
its breeding and pathology research
conduct a series of collaborative projects
programme on export bananas to the
in several countries with the aim of
national government.
increasing the productivity of Musa in
In developed countries, the Centre de traditional farming systems. In Asia, the
coopération internationale en recherche pressure for international action came
agronomique pour le développement from scientists at the University of the
(Cirad) had the most important research Philippines at Los Baños (UPLB). These
programme on Musa with antennas in Musa scientists had seen the impact on
Montpellier, the Guadeloupe and French- national programmes of the International
speaking Africa. Australia had a smaller, Rice Research Institute (IRRI), also based
but nevertheless important research at UPLB.
programme. These regional interests directed their
A major reason for this relative neglect requests for a coordinated international
was the widespread misconception in the approach and funding of Musa research to
developed world that the banana was the International Development Research
an export crop whose marketing and Centre (IDRC). In the previous decade,
production is mainly the responsibility IDRC had shown considerable interest in
of large multinational companies, even initiating research on smallholder crops
though in the 1980s the proportion of that had no home in the international
the global Musa production that was agricultural research system.
exported as dessert bananas was, at 7%,
even lower than today’s. As is still the
Gathering support
case, most of the bananas and plantains IDRC responded positively and contracted
produced in the tropics are consumed a consultant to prepare a short briefing
locally. paper on the need for some form of action
Around the same time, however, on Musa by the donor community. In
scientists were cumulating achievements November 1983, the paper was presented
at the International Centres Week held in
with the other major crops, and interest
Washington. The meeting was attended by
was gathering to develop, fund and
representatives from 15 donor agencies,
coordinate actions on Musa at the
5 producing countries and 3 International
international level. The main concerns
Agricultural Research Centres. The
were the narrow genetic base of the participants agreed that the time had come
crop and the rapidly spreading black for some form of international initiative
leaf streak disease, caused by the to support the genetic improvement of
32 June 2005
Musa. A clear preference was expressed the localization of the central secretariat.
for a research network that would link This had also been an issue during the
donors with national programmes rather regional consultations, at which a number
than an international agricultural centre of participants had suggested their
similar to the ones that are members of country as a suitable place to set up the
the Consultative Group on International headquarters of the network.
Agricultural Research (CGIAR). IDRC
was asked to consult representatives of Working out the last details
national Musa research programmes, The briefing paper proposing the
Musa specialists, and donors, and to establishment of a network was presented
present a formal proposal at the meeting to the donor group meeting in Rome in
of the CGIAR donor group to be held in May 1984. The donor group approved
Rome in May 1984. the paper in principle and asked IDRC
At the regional consultations for Africa, to undertake more consultations to
the participants expressed strong support obtain funding commitments from
for the network approach and a strong donors, determine the localization of the
breeding component. After visiting, in early headquarters and the membership of the
1984, the Philippines, Thailand, Malaysia, Board of Trustees and provide a list of
Indonesia, Papua New Guinea and the potential Directors Generals in time for
South Pacific, where black leaf streak Centres Week in November 1984. At the
disease originated, scientists funded by meeting, the recommendation to create
the Australian Centre for International a network backed by a donor support
Agricultural Research reached the group, rather than a CGIAR centre,
same conclusion about the need for an was accepted, in light of the constraints
international Musa programme. At the on CGIAR funding and the number of
regional consultations for Latin America institutions vying for CGIAR membership.
and the Caribbean, which took place At the first meeting of the donor support
in Miami in April and May 1984, the group, an agreement was reached on the
participants made detailed suggestions in composition of the Board of Trustees,
support of the network approach. the appointment of Edmond de Langhe
In addition to these regional as Director and the name of the new
consultations, three leading Musa institute, the International Network for
experts were asked to advise on the the Improvement of Banana and Plantain
broad shape of the research programme. (INIBAP), which was to be located in
Jean Champion, Norman Simmonds Montpellier. Some donors pledged funds
and Edmond de Langhe met at Gatwick, for the first year of operations and IDRC
in the UK, in December 1983. They agreed to be the executing agency until
suggested that the main priorities were the institute was up and running.
to identify and evaluate banana clones The next step was the signing with the
in producing countries; to gain a socio- Government of France of an international
agricultural understanding of Musa treaty granting INIBAP an international
production systems and their utilization; status. Signed in December 1988, the
and to develop an international breeding treaty had to be ratified by at least four
programme aimed primarily at the other countries. By 1990, it had been
smallholder crop. Structurally the group ratified by Belgium, Canada, Colombia,
foresaw the need for an international the Philippines and Senegal.
breeding programme, three regional INIBAP operated as an autonomous and
networks and a sound governance nonprofit intergovernmental organization
ensured by a small secretariat supported until 1990, when it was invited to join
by a scientific advisory body. the CGIAR system. The next turning
A number of meetings bringing point was 1995, when INIBAP joined the
together representatives of interested International Plant Genetic Resources
donor countries and agencies were also Institute (IPGRI), another member of the
organized by the IDRC consultant. Most CGIAR.
of these focused on structural matters. Of Barry Nestel
particular concern to some donors was Consultant

Focus on the Musa collection Ploidy levels revealed
The results are in. After years of work, the demonstrated that the ploidy level of banana
ploidy levels of all the accessions in the plants could be accurately determined by
world’s largest collection of Musa germplasm using flow cytometry (Dolezel et al. 1994).
are known. Flow cytometry measures the content of
The International Musa Germplasm nuclear DNA, which is directly proportional to
Collection is held at the INIBAP Transit Centre the number of chromosomes. More recently,
(ITC), which is hosted by the Katholieke this technique has also been successfully
Universiteit Leuven (KULeuven) in Belgium. used on Musa embryogenic cell suspensions
The accessions are maintained under slow (Roux et al. 2004). Since preparing the
growth conditions in the form of proliferating sample is easy and the analysis is performed
shoot tips (Van den houwe et al. 1995). The in just a few minutes, the method is suitable
ITC currently stores 1175 accessions, each to screen a large number of plants. Unlike
of which is represented by 20 tissue culture chromosome counting, cytometry can be
plantlets. A list of the accessions available in performed with any type of tissue, a feature
the collection can be found on the INIBAP that facilitates the detection of mixoploidy,
website (http://www.inibap.org). i.e. plants that contain cells of different ploidy
As part of its mandate to facilitate access levels.
to genetic diversity, the ITC also distributes
samples free of charge to bona fide users,
The tally
together with the data available on the At the end of the project in 2004, the 1150
accessions. Over the past 20 years, it has accessions held in the collection had been
provided more than 60 000 samples of virus- analysed (25 accessions have since been
free accessions to researchers worldwide. added to the collection). Since the IEB is
In order stimulate a still wider use of the located some 1000 km east of Leuven,
accessions, in the spring of 1999, the ITC in Olomouc, Czech Republic, in vitro
undertook the task of determining the ploidy rooted plants were shipped in Cultu saks®
level of all the accessions in the collection (Figure 1) in batches of 50 accessions, with
using the best available technique. five plants per accession. Upon arrival, the
Ploidy, or the number of chromosome sets plants were transplanted and maintained
in a cell, is one of the defining characters in a greenhouse for up to eight weeks until
of a species or cultivar. Wild species and their analysis. A small piece (about 50 mg)
subspecies are all diploid, whereas cultivars of young leaf tissue was used to prepare the
can be diploid, triploid or tetraploid. Knowing nuclei suspension to be analysed by flow
the ploidy level of an accession is not only
useful to confirm its classification, but it
can also reveal whether the plant has been
changed by in vitro conservation (in vitro
storage has been implicated in mutations,
epigenetic changes and changes in
chromosome structure and number).
Ploidy has conventionally been determined
by counting chromosomes in dividing root tip
cells, a labour intensive procedure that is
made difficult in Musa by the fact that the
chromosomes are small and numerous. A
further disadvantage of this approach is that
the number of chromosomes in the root cells
is not necessarily representative of the one
in the rest of the plant.
About ten years ago, the Laboratory of
Molecular Cytogenetics and Cytometry at
the Institute of Experimental Botany (IEB) Figure 1. Cultu saks® containing in vitro plantlets.
34 June 2005
cytometry (Dolezel et al. 1997). At least four Table 1. Examples of accessions in which the expected ploidy was not confirmed.
plants per accession were analysed. The ITC Code Accession name Group Expected ploidy Ploidy estimated
by flow cytometry
accessions for which the results were not 0127 Kamaramasenge AB 2x 3x
consistent were re-analysed and the sample 0051 Foulah 4 ABB 3x 4x
size increased to at least 10 plants. 1261 PA03-22 AAAB 4x 3x
1065 Pisang slendang AABB 4x 3x
The data are now available in the
1027 Asupina Fe’i 2x 3x
INIBAP Musa Germplasm Information
System (MGIS), a database holding the
characterization and evaluation data of the Table 2. Examples of accessions in which mixoploid plants and plants that turned out to
Musa accessions maintained in 16 banana have a ploidy level different from the expected one were found.
ITC Code Accession name Group Expected ploidy Ploidy estimated
collections around the world. The data are by flow cytometry*
accessible online at http://mgis.grinfo.net 0983 Auko AB 2x 2x+3x (15)
under cytological characters. 0987 Auko AB 2x 2x (10); 2x+3x (13)
0796 Kirkirnan AA 2x 2x (10); 4x (2); 2x+4x (2)
The analysis confirmed the ploidy of
0043 Champa nasik AAAA 4x 2x (7); 4x (10)
958 accessions and revealed the level of 1465 Ibwi AAAh 3x 2x (8); 3x (8)
81 accessions for which it was unknown *The number of plants is given in parentheses.
(Figure 2). Not only did this work support the
ploidy classification, it also confirmed that mixoploid plants whose ploidy level was
maintaining plants under in vitro conditions different from the expected one.
does not lead to large-scale changes in the Finally, 14 accessions had no mixoploid
genome. In nearly 10% of the accessions, plants but were represented by plants
however, ploidy turned out to be different with different ploidy levels, such as ‘Ibwi’
from the previously accepted level. For (ITC1465), which had 8 diploid plants and
example, the cultivar ‘Kamaramasenge’ 8 triploid plants (Table 2). Such accessions,
(ITC0127) is now classified as a triploid whose difference in ploidy level cannot be
rather than a diploid (Table 1). attributed to polyploidy (a doubling of the
Mixoploidy, a clear sign of genetic genome), represent 85% of the accessions
instability, was observed in nine accessions that ended up with a ploidy level different
such as ‘Auko’ (ITC0987), which was found to from the expected one. One possible
consist entirely of mixoploid plants containing explanation is that ploidy was incorrectly
both diploid and triploid cells (Table 2). The determined the first time around. Another
other accessions in which mixoploidy was is an accidental interchange of plants from
detected, were also represented by non- different accessions during the subculturing
process.
To our knowledge, this work is the largest
Confirmed exercise ever undertaken to determine the
(83.3%)
ploidy levels of a collection of vegetatively
propagated crop. The results also provide Marie Dolezelovà and
quantitative data on the extent of genetic Jaroslav Dolezel work at
variation in plant materials maintained the Laboratory of Molecular
under in vitro conditions. As to the nature Cytogenetics and Cytometry,
and origin of the variation observed, these Institute of Experimental
Determined
for the first questions will be answered by the ongoing Botany, CZ-77200 Olomouc,
time efforts to characterize the accessions by Czech Republic, Ines Van den
(7.04%)
using molecular markers and by planting the Houwe at the INIBAP Transit
Mixoploidy accessions in the field to evaluate them in Centre, Kasteelpark Arenberg
(0.79%)
situ. It is also planned to assess the ploidy of 13 - 3001 Leuven, Belgium,
each incoming accession. Nicolas Roux at INIBAP,
Parc Scientifique Agropolis II,
Mixed ploidy
(1.22%)
Other ploidy
(7.65%) Acknowledgements 34397 Montpellier Cedex 5,
We thank our colleagues Jan Bartoš, France, and Rony Swennen
Figure 2. Distribution of 1150 Musa accessions in relation to their Nikol Gasmanová, Pavlína Kovárová, Martin at the Laboratory of Tropical
ploidy level before and after flow cytometry analysis. Mixoploidy
refers to a plant containing cells of different ploidy (e.g. 2x+3x).
Lysák, Pavla Suchánková, Katerina Vlácilová Crop Improvement, KULeuven,
Mixed ploidy refers to accessions represented by plants of different and Jan Vrána for the flow cytometric Kasteelpark Arenberg 13 -
ploidy. analyses, and Els Kempenaers, Ronald 3001 Leuven, Belgium.

Bogaerts and Jeroen Mertens for the in vitro bananas (Musa acuminata and Musa balbisiana).
Biologia Plantarum 36:351-357.
manipulations. This work was supported Dolzezel J., M.A. Lysák, I. Van den houwe, M. Dolezelová
by INIBAP (research contracts INIB 98/21 & N. Roux. 1997. Use of flow cytometry for rapid ploidy
and INIB 2001/06) and by the International determination in Musa species. InfoMusa 6: 6-9.
Roux N., H. Strosse, A. Toloza, B. Panis & J. Dolezel.
Atomic Energy Agency (research contract 2004. Detecting ploidy level instability of banana
No. 12230) and was conducted within embryogenic cell suspension cultures by flow
cytometry. Pp. 251-261 in Banana Improvement:
the framework of PROMUSA, a Global Cellular, Molecular Biology, and Induced Mutations (S.
Programme for Banana and Plantain Mohan Jain and R. Swennen, eds). Science Publishers
Improvement. Inc., Enfield, USA.
Van den houwe I., K. De Smet, H. Tézenas du Montcel
References & R. Swennen. 1995. Variability in storage potential
of banana shoot cultures under medium term storage
Dolezel J., M. Dolezelová & F.J. Novák. 1994. Flow conditions. Plant Cell, Tissue and Organ Culture 42:
cytometric estimation of nuclear DNA amount in diploid 269-274.
Focus on IPGRI Results of survey on IPGRI’s commodity work
In December 2004, a stakeholder survey networks, PROMUSA and the Global Musa
was conducted as part of IPGRI’s Genomics Consortium, made up 64% of
commissioned external review of its new the respondents, of which around one-
Commodities for Livelihoods Programme. third belonged to more than one network.
The survey comprised 31 questions in However, only 7 people responded from the
three languages aimed at drawing comment two African regional networks.
on various aspects of IPGRI’s work on The following brief summary provides
cacao, coconut, and banana and plantain: those findings from the survey that relate to
the coordination of networks, information INIBAP:
products, quality of partnerships and future
priorities. Approximately 300 participants Networks
were asked to complete the survey and an When asked how adequately networks have
open invitation to take part in the survey was served a function, respondents replied that
posted on the INIBAP web site. In total, 128 they had served or partly served a large
people responded by completing the survey range of uses, most significantly providing
either on the Internet or by sending it through a forum for meetings. Responses in some
e-mail. areas gave a clear indication of where
Most of the respondents said being in network members consider improvements
regular contact with IPGRI (twice or more should be made (Table 1).
a year) and being employed in public- Members of all networks indicated that
funded research institutes (62%). Three- “finding funds from external sources” was
quarters were biological or agricultural a high priority to make networks function
scientists. Each banana-producing region more effectively (Table 2). By contrast,
had roughly equal numbers of respondents. there were mixed responses to the idea of
Members of networks, including regional restricting or expanding membership, and
a large number of respondents (40%) were
Table 1. Percentage of respondents who considered that the network served the various not in favour of “asking members to pay
functions adequately (as opposed to needing improvement or development). Figures in fees or in-kind contributions”.
bold indicate the largest proportion of respondents. When asked whether IPGRI was a good
Network functions PROMUSA Musa Genomics Regional banana partner to work with, 80% agreed and
(31 respondents) (9 respondents) networks
(34 respondents) 17% partly agreed. The large majority of
Providing a forum for meetings 68% 67% 68% partners expressed a positive impression
Developing priorities at a of their working relationship with IPGRI
regional/global levels 29% 44% 50%
Capacity building/training 27% 11% 26% (Table 3).
Establishing complementarities The majority of respondents indicated
among organizations 10% 22% 42% that IPGRI had performed adequately its
36 June 2005
obligations in coordinating projects (e.g. Table 2. Percentage of respondents considering the various mechanisms to be high
administration, communication, planning, priority (as opposed to medium, low or not a priority). Only the mechanisms considered
as high priority by more than 70% of respondents are presented. Figures in bold
monitoring and evaluation, developing indicate the largest proportion of respondents.
shared aims and visions, establishing Mechanisms for PROMUSA Musa Genomics Regional banana
decision-making that is fair etc.). Three rendering networks (32 respondents) (9 respondents) networks
more effective (32 respondents)
areas were highlighted by a majority of
Finding funds from external sources 88% 89% 90%
respondents considering improvement or Collecting baseline information for the
development to be necessary in INIBAP’s development priorities and decisions 81% 22% 68%
project coordination: “establishing effective Providing incentives for information
exchange 72% 78% 58%
mechanisms for managing data or results” Developing data management tools 61% 33% 71%
(54%), “planning” (51%), and “establishing Involving members in functioning and
an effective framework for project monitoring coordination of the network 52% 22% 72%
and evaluation” (50%).
All INIBAP information products, including
databases and web sites, were considered
“very useful” or “quite useful” by 75% or more Table 3. Percentage of respondents who strongly or partly agreed or disagreed with
various statements concerning partnerships with IPGRI.
of the respondents. INFOMUSA attracted the
Characteristics of the partnership with IPGRI Strongly agree Partly agree
strongest response with 75% of respondents IPGRI is a good partner 80% 17%
considering it to be “very useful”. IPGRI develops trust and shared interests with its partners 66% 31%
IPGRI is a communicative partner 59% 35%
Future priorities IPGRI works under equal partnership with my organization 49% 36%
The large majority of respondents (84-
96%) considered that IPGRI’s existing
and proposed activities in conservation
were medium or high priority: “providing a Table 4. Percentage of respondents considering the various germplasm conservation,
network for germplasm conservation and exchange and evaluation activities to be high priority (as opposed to medium, low or not
management” and “building capacity to a priority). Figures in bold indicate the largest proportion of respondents.
conserve and distribute germplasm in NARS Germplasm conservation, exchange and evaluation Proportion of 72
respondents who
and other partners” were the most popular considered activities
responses (Table 4). to be high priority
There was more divergence in opinion Network for germplasm conservation and management 74%
Building capacity in NARS and partners to conserve and distribute germplasm 74%
concerning genetic research and Collecting wild or cultivated germplasm 67%
improvement activities. Although a majority Ensuring long term security of ex situ collections 67%
of respondents considered research into Conserving germplasm in situ or on farm 64%
genetic manipulation and embryogenic cell Research on diseases and therapy for ex situ collections 62%
Providing incentives for germplasm exchange 61%
suspensions to be medium or high priority,
Acting on genetic erosion 57%
a notable percentage considered it to be Molecular characterization 54%
“not a priority” (26% and 21% respectively). Research on ex situ conservation methodologies 52%
Research in genomics and bioinformatics
received slightly more support (Table 5).
As with genetic improvement, there is
strong support for promoting the use of
Table 5. Percentage of respondents considering the various genetic research and
biodiversity in production systems (Table 6). improvement activities to be high priority (as opposed to medium, low or not a priority).
There is more ambivalent support for less Figures in bold indicate the largest proportion of respondents.
conventional areas of development work Genetic research and improvement activities Proportion of 71 respondents
(e.g. strengthening community based who considered activities
to be high priority
organizations, developing processing Research on pest and disease resistance 86%
technologies and promoting enterprise Use of biodiversity in genetic improvement 76%
development). Supporting breeding programmes 76%
Building capacity to carry out genetic research & improvement in NARS 71%
Conclusions Pooling research resources 68%
Strengthening networking 61%
The survey has provided valuable Producing guidelines 55%
feedback to assist INIBAP in confirming Research in genomics 53%
where its strengths lie, in suggesting where Bioinformatics 48%
improvements in networking are demanded Research in genetic manipulation 35%
and in attaining stakeholders’ perspectives Research in embryogenic cell suspensions 36%

Table 6. Percentage of respondents considering the various activities related to of where our priorities lie. We would like to
production systems and postharvest to be high priority (as opposed to medium, low or thank wholeheartedly those who took part
not a priority). Figures in bold indicate the largest proportion of respondents.
in the exercise and assure them that their
Production systems and post-harvest activities Proportion of 70 respondents
who considered activities input has been registered and valued, by
to be high priority the review panel of the CCER and IPGRI’s
Use of biodiversity in production systems 66% Board of Trustees as well as by the staff at
Research in integrated pest management 68% INIBAP. We hope that we can continue to
Improving access to other resources (e.g.infrastructure) 59%
depend on their support.
Monitoring incidence of pests and diseases 64%
Promoting farmer participation 61%
Investigating uptake pathways for new technologies 58%
Research on soil properties and root systems 50% For more information:
Strengthening community based organizations 40% Charlotte Lusty INIBAP
Trials on processing technologies 40% Montpellier, France
Promoting enterprise development 27%
[email protected]
Focus on INFOMUSA Highlights from the reader survey
Who are the respondents? What kind of INFOMUSA?

A total of 326 respondents, or 12% of Only a few respondents would not mind if
our individual subscribers, returned the INFOMUSA was published only in English
questionnaire. Of these, 54% filled in the and available only electronically: being
English version, 35% the Spanish one available in print was rated as very important
and 11% the French one, a distribution by 74% of the respondents, and important by
that more or less reflects the proportion of 25%, and being available in three languages
INFOMUSA printed in each language. The was rated as very important by 65% of the
greatest contingent of respondents is based respondents, and important by 31%.
in the Latin America and Caribbean region
(41%) followed by Africa (27%), Asia (19%),
Should INFOMUSA be a
Europe (7%), the Pacific Islands (3%), North recognized peer-reviewed
America (1.5%) and the Middle East (1.5%). journal?
A little over 60% of the respondents are A majority of respondents (69%) would like
scientists. Most of the people who returned to see INFOMUSA become a peer-reviewed
the questionnaire (76%) are subscribers. journal. When broken down by region, the
The others access it from the Internet results show that the greatest proportion of
(11%) or from a library (8%), borrow it from respondents in favour of such a move (47%)
a colleague (2%) or get it through some are based in Africa (82% of the respondents
another unspecified way (3%). from this region said yes) followed by Latin
America (68% of the respondents from this
How do you rate INFOMUSA? region said yes). Asians were split 50:50
Most of the respondents think that on the question, whereas only 17% of the
INFOMUSA is a useful source of information European respondents and none of the
and appreciate its format and contents: 80% respondents from North America and the
of the respondents rate its usefulness in Pacific Islands think INFOMUSA should
keeping up with research on Musa as strong become a peer-reviewed journal.
and the remaining 20% as average, while the
quality of the writing and of the presentation Which sections do you read and
were respectively rated as strong by 72% how often?
and 76% of the respondents and as average Eighty-seven percent of the respondents
by 28% and 22% of the respondents. The always read the scientific articles, 80%
quality of the scientific content was rated Musanews, 65% the Focus on section,
as strong by 65% of the respondents and as 60% the thesis abstracts and 47% the
average by 32%. editorial. The vast majority of the remaining
38 June 2005
respondents said they sometimes read these The greatest demand was for articles
sections. on integrated pest management (62%),
pests and diseases (60%) and uses and
Are you satisfied with the products (54%). Many of the respondents
spectrum of articles? who answered the question about the type
Only a minority of respondents would like to of information they would like to see added
see more articles on genetic transformation were also mainly interested in seeing more
and germplasm screening (31%), molecular practical information and articles on applied
biology (34%) and tissue culture (40%). research.
Variability in reproductive fitness and pathogenicity Thesis

of Radopholus similis in Musa: effect of biotic and
abiotic factors
Thomas Moens
PhD thesis submitted in October 2004 To screen the resistance of Musa
to the Katholieke Universiteit Leuven, varieties to R. similis, parameters like
Belgium inoculum number, substrate type, nematode
Nematodes are one of the most important population, exposure time and pot volume
constraints in Musa production after leaf are important. However, published
spot diseases. These tubular, vermiform literature show that these parameters are
animals, especially Radopholus similis, not standardized. Therefore, the effect
parasitize the root system and can reduce of each of these factors on reproductive
yield by up to 80%. Nematodes induce root fitness and pathogenicity were studied to
damage directly, and indirectly by facilitating optimize screening protocols. Cavendish
the entry of fungi and bacteria. This results in plants grown in river sand supported the
lower nutrient uptake, extends the harvest- highest R. similis numbers and had the
to-harvest interval and increases plant lowest root weight, compared to different
toppling, when the plant is not propped banana soils that varied in sand, silt or clay
up. This research was conducted in Costa content. This was probably due to a higher
Rica at the Corporación Bananera Nacional macroporosity and a better aeration. When
(CORBANA). comparing exposure times between 2 to
The population growth of 11 R. similis 16 weeks, the highest increase in R. similis
populations extracted from roots of plants in population was observed between 6 and
a commercial plantation, and of 7 R. similis 12 weeks after inoculation. Root weight of
isolates extracted from the roots of one plant Cavendish plants grown in pots of 4 different
in a field of 4.5 ha, was evaluated on carrot volumes, and inoculated with identical
disks and ‘Grande naine’ potted plants. densities, was significantly higher in 1.8
Differences in reproductive fitness among the and 3.6 L pots, compared to 0.4 and 0.8 L
populations and isolates were observed on pots. Application of these factors in a time
carrot disks and in potted plants. Variations experiment confirmed the susceptibility of
were expected among the populations but ‘Grande naine’ and resistance of ‘Yangambi
not among the isolates. These may be due km 5’ and FHIA-23 to R. similis. Based on
to the presence, among the populations or these results, Musa plants grown in local
isolates, of subpopulations better adapted soil, initially inoculated with 500 R. similis
to growth on carrot disks, to the number of and exposed during 8 to 12 weeks in 1.8 L
subcultures on carrot disk, or to differences pots, gave consistent screening results.
in female reproductive fitness. Under our Thereafter, the protocol was applied for
experimental conditions, no relation was resistance testing against R. similis. Twenty-
found between reproductive fitness and six Musa varieties, belonging to different
pathogenicity, as reported in the scientific genome groups (AA, AB, BB, AAB, ABB,
literature. AAAA, AAAB and AABB) and 10 F1 lines

of a cross between Musa acuminata ssp. In a microplot experiment using 200 L
burmannicoides ‘Calcutta 4’ (AA) and ‘Pisang drums filled with sterilized soil, inoculated
Berlin’ (AA) were tested for resistance to ‘Grande naine’ plants were followed until
R. similis under in vitro conditions and in harvest and compared with uninoculated
pot plants. In vitro, no difference in final plants. Only R. similis significantly reduced
nematode numbers was found among the root weight by 66%, while root damage
tested varieties, including ‘Grande naine’, was higher and bunch weight was lower
‘Pisang jari buaya’ and ‘Yangambi km in R. similis and P. coffeae inoculated
5’, 30 days after inoculation. In contrast, plants, compared with the control. Also
‘Yangambi km 5’, FHIA-23, ‘Kunnan’, ‘Paka’ M. incognita reduced bunch weight. Probably
and ‘Pisang lemak manis’ in pots supported the detrimental effect of M. incognita is more
lower nematode numbers, compared to associated with physiological alterations,
‘Grand naine’, 8 weeks after inoculation. The changing the cell structure and metabolism,
longer exposure time probably allowed the than physical root deterioration. Due to their
expression of the resistance mechanisms negative effect on roots or yield, not only
in these varieties grown in pots. Also ‘Tjau R. similis, but also P. coffeae, M. incognita
lagada’ proved, for the first time, to be and H. multicinctus have to be considered in
resistant to R. similis. These results need to future breeding programmes for nematode
be confirmed under different soil and climatic resistance.
conditions in the field. Finally, the biodegradation of 6 nema-
Helicotylenchus multicinctus, Meloidogyne ticides was studied alone or in rotation,
incognita and Pratylenchus coffeae were under field conditions and in a R. similis-
studied in pot and microplot conditions. maize laboratory test. In the field, only
In pots, H. multicinctus suppressed root terbufos (Counter®) consistently reduced
weight, while M. incognita stimulated it and the number of nematodes per 100 g
P. coffeae did not influence it over the tested of roots after 5 consecutive applications.
time intervals. In a pot experiment, the mean Carbofuran (Furadan ® ), ethoprophos
number of R. similis in plants that had been (Mocap®), fenamiphos (Nemacur®) and
inoculated with M. incognita were always the untreated plants supported the highest
lower than in noninoculated plants. This numbers of nematodes per 100 g of roots.
illustrates the suppressive impact of this The mean percentage of functional roots
nematode on R. similis, probably due to a for all nematicide treatments was 7%
reduction in feeding sites. H. multicinctus higher than in the untreated plants. This is
and M. incognita numbers were not affec- reflected in a 38% higher bunch weight of
ted by increased R. similis inoculum the treated plants. Of all nematicides, only
numbers, while this was the case for carbofuran resulted in a significantly lower
P. coffeae. The suppressive effect of R. similis bunch weight, which was still higher than the
on P. coffeae can be due to competition for one of the untreated plants. Nevertheless,
the same feeding site, i.e. the root cortex. carbofuran, ethoprophos, fenamiphos and
In the screening experiments, ‘Yangambi oxamyl showed enhanced biodegradation
km 5’, ‘Niyarma yik’ and ‘Pisang Berlin’ in the biotest after 5 consecutive field
supported the lowest numbers of applications. Contrasting results are probably
H. multicinctus per 100 g of roots, together due to the variation in the start of enhanced
with ‘Pisang bungai’ and ‘Tjau lagada’. All biodegradation for the tested nematicides
tested varieties proved to be susceptible to and the slow root health deterioration.
M. incognita, while FHIA-01 and FHIA-18 Nematicide rotation gave the highest bunch
were equally resistant to P. coffeae, weight and a high proportion of functional
compared to ‘Yangambi km 5’, a variety root, because of the low probability of
with known resistance to this nematode. developing enhanced biodegradation.
Also ‘Tjau lagada’, ‘Pisang mas’ and ‘Pisang Further research is needed to study the
bungai’ were less susceptible to P. coffeae development of enhanced biodegradation
than ‘Grande naine’. These characteristics after increasing nematicide applications in
of ‘Tjau lagada’, together with its resistance different soil conditions. Nevertheless, due to
to R. similis and its partial resistance to the future ban on nematicide use in banana,
black leaf streak disease, make this variety the development of alternative nematode
interesting for further research. management techniques is urgent.
40 June 2005
Breeding for resistance to Radopholus similis in East Thesis
African highland bananas (Musa spp.)
Carine Dochez
PhD thesis submitted in November male and female fertility. Selected tetraploids
2004 to the Katholieke Universiteit are then crossed with improved diploids to
Leuven, Belgium produce sterile secondary triploids.
The East African highland bananas (Musa The objective of this study was to identify
spp. AAA) are the most important staple food sources of resistance to R. similis in existing
crop in the East African Great Lakes Region. Musa germplasm and newly bred hybrids.
East Africans cook the fruit as ‘matooke’ or This study also looked at the variability
brew beer from it. In Uganda, East African in reproductive fitness and virulence of
highland bananas are divided in five clone different R. similis populations from Uganda
sets: four clone sets cover the cooking and aimed to better understand host plant
types (Nfuuka, Nakitembe, Nakabululu and resistance to R. similis through genetic
Musakala) and one clone set covers the beer analysis of a segregating banana population
brewing types (Mbidde). and preliminary studies on the mechanisms
Uganda is the leading regional producer of resistance.
and consumer. Until the 1970s, East African In the first part of this research, a new
highland bananas were traditionally grown method was developed for screening Musa
in central Uganda. However, since that germplasm for resistance to R. similis.
time, banana production has declined by This method is based on the inoculation
more than 25%. This decline has led to the of individual roots with a small number of
replacement of cooking bananas by exotic R. similis females. The individual root
banana cultivars and annual food crops. screening method has several advantages
At the same time, cultivation of the crop compared to the standard greenhouse
has shifted to the southwest of the country. screening method. Fewer plants and a
Nematodes are considered one of the lower nematode inoculum are needed. By
major constraints causing this decline. The using individual roots, the evaluation of the
burrowing nematode, Radopholus similis, host response to nematode infection is not
has been identified as the most destructive influenced by differences in root growth rates
species in Uganda. among Musa genotypes. Moreover, primary
Nematodes can be reliably controlled roots of the same age can be selected
by nematicides. However, the use of for inoculation, avoiding bias caused by
nematicides has adverse environmental differences in host response to R. similis
effects and is too costly for resource-poor related to root age. In addition, this method
farmers. A promising alternative is the use seems to be able to pick up both constitutive
of nematode resistant cultivars. Improving and induced resistance.
the East African highland bananas through In the second part of this research,
breeding was identified by the Ugandan this new screening method was used to
National Agricultural Research Organisation evaluate available Musa germplasm and
(NARO) as the most appropriate strategy for newly developed hybrids for resistance to
addressing pest and disease problems. The R. similis. East African highland bananas are
Musa breeding program of the International susceptible to R. similis. Tetraploid hybrids
Institute of Tropical Agriculture (IITA), in which are resistant to R. similis have been
collaboration with NARO, aims at developing developed by crossing susceptible East
improved Musa genotypes, preferably African highland bananas with the resistant
triploids, with resistance to multiple pests and wild diploid Calcutta 4. This diploid has been
diseases, high and stable yield, improved widely used in Musa breeding programs as a
agronomic traits and acceptable fruit quality. male parent. Resistance was also identified
This usually involves crossing triploid in several diploid hybrids, which were used
cultivars with fertile diploids to produce to further improve the tetraploid hybrids.
tetraploids that generally display greater TMB2x 9128-3 is the most resistant diploid

identified so far and is often used as a parent may have serious implications for farmers
in the breeding program. Tetraploid hybrids as Mbarara is the main banana growing
have been further crossed with improved area in Uganda. Recent observations
diploids, resulting in secondary triploids. To in farmers’ fields in Mbarara show that
date, five secondary triploids that are resistant R. similis infestation is localized for the
to R. similis and seven that are partially moment but these farms are heavily infested
resistant have been identified. Three of the and the incidence of toppling is high.
secondary triploids with partial resistance to In the fourth part of this research, we
R. similis have good bunch characteristics studied the genetic analysis of segre-
and a matooke-like taste, while one resistant gation for resistance to R. similis in a
hybrid is recommended for juice production. diploid banana hybrid population. This
In addition, new sources of resistance to diploid banana hybrid population was
R. similis have been identified, mainly among derived by crossing the diploid hybrids
germplasm from Papua New Guinea. TMB2x 6142-1 and TMB2x 8075-7. The
In the third part of this research, different female parent TMB2x 6142-1 is susceptible
populations of R. similis from Uganda to R. similis and derived from the cross
were compared for their variability in between the East African highland banana
reproductive fitness and virulence. Four ‘Nyamwihogora’ (AAA) and the wild
R. similis populations of different locations banana ‘Long tavoy’ (AA), which are both
within Uganda (Namulonge, Mbarara, susceptible to R. similis. The male parent
Ikulwe and Mukono) were collected and TMB2x 8075-7 is resistant to R. similis
cultured monoxenically on carrot discs. and derived from the cross between the
The reproductive fitness of these four bred hybrid SH-3362 (AA) and the wild
R. similis populations was compared on banana Calcutta 4 (AA), which are both
carrot discs as a function of time and resistant to R. similis. The diploid banana
inoculum level. These in vitro experiments hybrid population was evaluated with the
showed that the R. similis population from individual root inoculation method using
Mbarara had the highest reproduction the R. similis population from Namulonge.
ratio. This was shown by comparing the Of the 81 hybrids evaluated, 37 hybrids
final nematode population densities and were resistant, 13 were partially resistant
by calculating growth curves using the and 31 were susceptible to R. similis. A chi-
Gompertz equation. The population from square analysis indicated that resistance to
Mukono had the lowest reproduction ratio. R. similis is controlled by two dominant genes,
Pathogenicity experiments on host A and B, both with additive and interactive
plants were carried out in pot trials. Both effects, whereby recessive bb suppresses
the final nematode population densities dominant A (either A- or B- required for
and percentages root necrosis on different partial resistance, both A- and B- confer full
host plants were higher for the R. similis resistance, but bb suppresses A-).
population from Mbarara than for the In the last part of this research, potential
populations from Namulonge, Ikulwe and mechanisms of resistance to R. similis were
Mukono. The R. similis population from studied. Knowledge on the mechanisms of
Mbarara managed to break the resistance of nematode resistance may help the breeder
‘Pisang jari buaya’, known to be resistant to to select for a desired characteristic for
R. similis. The diploid hybrid TMB2x 9128- the breeding program, and it may also
3 and ‘Yangambi km 5’ showed resistance assist in the identification of resistance
against the four R. similis populations. markers to facilitate screening of Musa
These results indicate that differences in germplasm. In a first set of experiments,
pathogenicity among different R. similis the attraction and penetration ability of R.
populations exist and should be taken similis was compared among resistant and
into consideration in a breeding program. susceptible Musa cultivars. No significant
It is recommended to use the R. similis differences in attraction and penetration
population from Mbarara in routine of R. similis were observed between
screening for identification of resistance resistant and susceptible cultivars. Similar
in Musa germplasm. The fact that the R. invasion rates of R. similis on resistant
similis population from Mbarara is more and susceptible Musa cultivars, suggests
pathogenic than the other populations that the resistance is not due to physical
42 June 2005
or mechanical barriers. Subsequently, phenolic compounds in healthy roots do
histochemical experiments were carried out not contribute to the constitutive resistance
to detect whether there are differences in of banana to R. similis. Lignification of the
lignin and phenolic compounds between endodermis was observed in the susceptible
susceptible and resistant Musa cultivars. cultivars early in time. Lignification in the
After R. similis infection, a higher number of resistant cultivars was only observed after
phenolic cells were observed in the resistant 12 weeks. No lignified cells were observed
cultivars compared to the susceptible in the cortex of any cultivar. More detailed
cultivars. Phenolic cells were also observed studies are needed to understand the role of
in healthy plants, though the number was phenolic compounds and lignin formation in
lower compared to nematode infected relation to host plant response to nematode
plants. It is assumed that preformed infection.
Random amplified polymorphic DNA analysis of Thesis

28 clones
C. Rajamanickam
PhD thesis submitted in 2004 to Kerala ‘Red banana’ and ‘Vellakappa’ (AAA)
Agricultural University, Kerala, India as well as ‘Pisang Ceylon’ and ‘PKNNR’
The genetic diversity of 28 banana clones (AAB) formed clusters at a distance of
belonging to different genomic groups 0.12. ‘Kunnan’ and ‘Njalipoovan’ (AB)
(AAA, AAB, ABB, AA, AB and BB) and formed a cluster at a distance of 0.22.
ploidy groups (triploids and diploids) ‘Kadali’ and ‘Pisang lilin’ (AA) as well
was evaluated based on morphological as ‘Quintal’ and ‘Padalamurian’ (AAB)
traits and random amplified polymorphic formed clusters at a distance of 0.24. The
DNA (RAPD) profiles was evaluated. The triploids ‘Attu nendran’ (AAB), ‘Monthan’,
bananas were collected from the Banana
‘Robusta’ (AAA), ‘Koonoor ethan’ (AAB),
Research Station in Kannara, the Banana
and ‘Vellapalayankodan’ (AAB) and the
Nursery Farm in Thiruvananthapuram, and
the College of Agriculture in Vellayani. diploid ‘Ilavazha’ (BB) formed individual
Out of the 41 decamer primers screened for clusters and diverged the most.
RAPD analysis, 34 produced amplification. Tocher’s method was used to analyse
Of the 123 bands generated, 116 were morphological characters. The cultivars
polymorphic and 7 were monomorphic. formed 6 clusters. ‘PKNNR’, ‘Pisang
Finally, 6 primers (OPA-01, OPA-03, OPA- Ceylon’, ‘Motta poovan’, ‘Chandra bale’
13, OPB-04, OPB-10 and OPB-12) were and ‘Palode palayankodan’ (Palayankodan,
used to analyse the 28 clones. These AAB), ‘Kadali’ and ‘Pisang lilin’ (AA),
primers yielded 46 scorable bands with an ‘Kunnan’ and ‘Njalipoovan’ (AB)
average of 7.66 bands per primer. In the formed the first cluster. ‘Red banana’
dendrogram, the 28 clones clustered into ‘Vellakappa’ and ‘Robusta’ (AAA),
14 groups at a distance of 0.20. ‘Myndoli’, ‘Mysore ethan’, ‘Changanasseri
Eight out of 12 Nendran clones (AAB),
nendran’, ‘Kaliethan’, ‘Chengazhikodan’,
‘Chengazhikodan’, ‘Myndoli’, ‘Kalie-
‘Manjeri nendran’, ‘Padalamurian’ and
than’, ‘Vellayani nendran’, ‘Zanzibar’,
‘Mysore ethan’, ‘Manjeri nendran’ and ‘Attu nendran’ (AAB), ‘Monthan’ and
‘Changanasseri nendran’ formed a ‘Peyan’(ABB), and Ilavazha (BB) formed
single cluster at a distance of 0.25 the second cluster. ‘Vellayani nendran’
and Palayankodan clones (AAB) like, and ‘Zanzibar’ (AAB) formed the third
‘PKNNR’, ‘Pisang Ceylon’, ‘Motta cluster. ‘Quintal’ (AAB) formed the fourth
poovan’, ‘Chandra bale’ and ‘Palode cluster, ‘Vellapalayankodan’ (AAB) the
palayankodan’ formed another cluster. fifth and ‘Koonoor ethan’ (AAB) the sixth.

Musanews Rediscovery of Musa splendida A. Chevalier and
description of two new species (Musa viridis and
Musa lutea)
An article by Ramon Valmayor, Le Dinh home of M. splendida and discovered large

Danh and Markku Hakkinen published in populations still thriving in the Red River
the March 2004 issue of the Philippine Valley, near Lao Cai. The suspicion that
Agricultural Scientist (Vol. 87(1):110-118) M. splendida was a mere synonym of either
illustrates the distinguishing characteristics M. sanguinea or M. laterita was dismissed
of the newly rediscovered Musa splendida with the demonstration of their distinguishing
and two recently described Musa species characters.
from Vietnam. Recent characterization studies of Musa
Musa splendida, known in Vietnam as accessions at the Phu Ho Fruit Research
‘Chuoi gai’, is a very rare species of wild Center in Vietnam describe two new species,
banana that was drifting toward oblivion. Musa viridis and Musa lutea. The former is
Cheesman, who revised the classification known locally as ‘Chuoi rung hoa sen’ and
of bananas, did not include its description the latter as ‘Chuoi rung hoa do’. The word
in his monumental series “Critical Notes on Chuoi, meaning banana, is part of the actual
Species”. Simmonds doubted its status as name which is often descriptive. ‘Chuoi rung’
a valid species and Champion associated means jungle banana. ‘Chuoi rung hoa sen’
it with Musa sanguinea and Musa laterita. means jungle banana with lotus colored
The leading banana taxonomists of the flower and ‘Chuoi rung hoa do’ means jungle
world seemed were prepared to relegate banana with red flower.
M. splendida as species ignota. The article also presents a diagnostic char-
A comprehensive report on the Musa acterization of M. viridis and M. lutea to
germplasm resources of Vietnam listed differentiate them from Musa balbisiana,
banana cultivars, ornamental species Musa acuminata and Musa itinerans
and wild relatives but never mentioned and local species such as M. sanguinea,
M. splendida. An illustrated flora of Vietnam M. splendida and M. laterita. Finally,
showed drawings of indigenous Musaceae, M. viridis and M. lutea are differentiated from
but M. splendida was not included. However, each other by the color of their fruits and
some elderly people in Vietnam maintained male buds. The Latin terms viridis and lutea
that ‘Chuoi gai’ existed between Lao Cai were selected to highlight the difference
and Sa Pa. Inge Van den Bergh, currently in colour of the immature fruits, which are
an expert associate at the INIBAP regional silvery green in M. viridis and yellow in
office in the Philippines, surveyed the original M. lutea.
Musanews Presence of banana bunchy top virus in Angola

Banana bunchy top disease, caused by Recently, symptoms of bunchy top
the Banana bunchy top virus (BBTV), were observed in the fields of small-scale
is one of the most serious diseases farmers in Mabuia (48 masl: S09º01’,
affecting banana worldwide (Dale 1987). E013º41’) and Boa Esperanca (52 masl;
Symptoms include dark green spots S08º57’, E013º40’) in Bengo Province,
along the leaf veins, especially the midrib South of Luanda, in Angola. The disease
and petiole, upright leaves with wavy was found on plantains (False horn) and
margins, stunted growth and leaves the Cavendish cultivar ‘Poyo’. It is known
more erect than normal, giving the plant that cultivars in the Cavendish subgroup
a rosetted, or ‘bunchy top’, appearance are highly susceptible to bunchy top
(Robinson 1996, Jones 2000). (Thomas and Iskra-Caruana 2000). This
44 June 2005
is the first report of the disease in Angola. Jones D. 2000. Diseases of banana, abacá, and enset.
CAB International, Wallingford, Oxon, UK.
Countries in Africa where BBTV has been
Robinson J.C. 1996. Bananas and plantains. CAB
reported include Burundi, Central African International, Wallingford, Oxon, UK.
Republic, Congo, Democratic Republic of Thomas J.E. & M. L. Iskra-Caruana. 2000. Diseases
Congo (DRC), Egypt, Gabon, Malawi and caused by viruses. Pp. 241-253 in Diseases of banana,
abaca and enset (D. R. Jones, ed). CAB International,
Rwanda (Thomas and Iskra-Caruana 2000). Wallingford, Oxon, UK.
Angola and the DRC share common borders
and it is likely that infected planting materials For more information:
were exchanged. BBTV is transmitted M. Pillay, International Institute of Tropical
through conventional planting material such Agriculture (IITA), Uganda. P.O. Box 7878,
as corms, corm pieces that have a growing Kampala, Uganda, G. Blomme, International
point and suckers. The vector of BBTV is the Network for the Improvement of Banana and
banana aphid. Plantain (INIBAP), P.O. Box 24384, Kampala,
Destroying the infected material is the Uganda, E. Rodrigues, Co-operative League
main way of controlling the disease and the of the USA (CLUSA), Rua Custodio Bento
method advocated in Angola. Azevedo, Bairro, Valodia, Sambizanga,
Luanda, Angola, and L. Ferreira, Instituto
References
de Investigacao Agronomica (IIA),Avenida
Dale J.L. 1987. Banana bunchy top: an economically
important tropical virus disease. Advances in virus Deolinda Rodrigues, km 5 Caixa Postal
research 33:301-325. n° 2104, Angola.
Containing banana xanthomonas wilt Musanews
A workshop on “Developing a coherent • Throughout the region: to coordinate and

regional response to the banana monitor efforts, exchange information and
xanthomonas wilt epidemic in East and facilitate policy dialogue.
Central Africa”, was held in Kampala, • The technical core of the disease
Uganda, on 14-18 February 2005. Organized management campaign, which needs to be
with the support of the Food and Agriculture developed and implemented using farmer
Organization of the United Nations (FAO), the participatory approaches, will involve:
International Development Research Centre • Debudding (removal of male bud) and field
of Canada (IDRC) and the International sanitation, reinforced by:
Network for the Improvement of Banana - statutory measures at national and
and Plantain (INIBAP), it brought together local levels to slow the spread of
regional and national stakeholders from disease (quarantine regulations) and
containment and control measures
Uganda, Ethiopia, the Democratic Republic
(by-laws);
of Congo, Kenya, Tanzania and Rwanda, as
- awareness raising efforts at the
well as international specialists.
international, regional, national and
The participants considered priority needs local levels, directed towards decision-
for research, outreach and policy activities makers and the general public, in order
to address this regional threat and identified to mobilize the necessary resources
and agreed on the following key issues for and ensure support for the campaign;
short- to medium-term action. - improved ‘seed’ systems for supplying
• In advance of the epidemic: to track its clean, high quality planting material;
spread and prepare for its arrival. and
• At the advancing disease front: to slow its - improved agronomic practices to
advance and mitigate its impact. increase productivity and sustainability,
• In areas where the disease had become combined with the dissemination
established: to rebuild production systems of utilization options to improve
and improve livelihoods. livelihoods.

• Evaluation and introduction of new Given the scale of the problem and the
varieties which are acceptable to farmers seriousness of the threat to livelihoods, the
and consumers and are more tolerant participants invited national governments
to BXW. and donors to invest in the proposed
Other actions needed include: framework “as offering the best strategy
• research to ensure the soundness of the for containing the spread of the disease,
current control measures and to generate mitigating its immediate effects on livelihoods
new options for the future; and eventually restoring the productivity and
• mechanisms for planning, information sustainability of banana-based production
exchange and coordination at the national systems”.
and regional level to ensure the most cost-
effective use of resources; For more information:
• and evaluating the impact of the activities Eldad Karamura, Moses Osiru or
to inform policy and strengthen strategies Guy Blomme, INIBAP-Eastern and
for managing trans-boundary pests and Southern Africa regional office, Kampala,
diseases. Uganda.
Musanews Controlling banana xanthomonas wilt through

debudding
Banana xanthomonas wilt, which is caused by first, he used a knife attached to a stick but he
the bacterium Xanthomonas campestris pv. was advised to use a forked stick (Figure 1)
musacearum ,was first observed in Uganda to avoid spreading the disease through
in 2001 and has since spread to 31 out of contaminated knives. He also says that he
56 districts. Technologies used to combat sees fewer insects, probably because the
similar bacterial wilt diseases in Latin America male buds have been removed.
and Asia have been introduced to Ugandan Some Ugandan farmers participating
farmers and campaigns have been conducted in a banana germplasm evaluation and
to raise awareness of the disease and ways to multiplication project, funded by the Common
control it. Since the bacteria are believed to be Fund for Commodities and managed by the
transmitted by insects and contaminated farm Uganda National Agricultural Programme
tools, the removal of diseased plants, early and INIBAP, started debudding their plants
debudding and the use of clean farm tools when the disease closed in on their fields.
could thus significantly reduce new infections. Their experience shows that prevention
Small-scale Ugandan farmers have started – debudding combined with controlling
to apply these techniques and have been sources of infection – can protect the plants
able to prevent new infections or have even even when the surrounding fields are heavily
eradicated the disease from their farms. infected. This is especially important in
Fredrick Kisegerwa has a field of ‘Pisang sizeable plantations where the eradication
awak’ (ABB) in Central Uganda. He started of a large number of sick plants is costly and
debudding his banana plants in January 2004 labour-intensive.
after observing a strange disease on several
of his plants. He got the information about the For more information:
disease and its control measures from the G. Blomme and H. Mukasa,
sub-county agricultural extension officer and INIBAP-Eastern and Southern Africa
had heard about it on the radio. At that time regional office, P.O.Box 24384, Kampala,
there were about 20 sick plants on his farm. Uganda - [email protected],
He rouged these plants by digging them up, [email protected]
chopping them into pieces and burying them. S. Mpiira and R. Ssemakadde,
In the year after he started debudding, which National Agricultural Research Organisation,
he does twice a week, only four of his plants National Banana Research Programme,
contracted the disease, three of them in the P.O.Box 7065, Kampala, Uganda -
Figure 1. Debudding using a wooden stick first two months after he started debudding. At [email protected]
46 June 2005
Survey on banana bract mosaic disease in Kerala Musanews
A survey on the incidence of banana bract 3 = Reddish brown streaks over 51-75 % of
mosaic disease (BBrMD), locally known the pseudostem
as Kokkan disease, was carried out in 4 = Reddish brown streaks over more than
50 randomly selected farms in Kalliyoor 75 % of the pseudostem
Panchayat, southern Kerala, India. Ten
The plants were observed at weekly
plants from each site were sampled. The
cultivars sampled were ‘Nendran’ (AAB), intervals and symptoms appeared three to
‘Red banana’ (AAA), ‘Robusta’ (AAA), four months after planting. Over 130 plants
‘Palayamkodan’ (AAB) and ‘Rasakadali’ scored 1, more than 160 scored 2, nearly
(AB). 100 scored 3 and less than 70 scored 4. The
incidence of disease appeared greater on
BBrMD was first reported on ‘Nendran’
‘Red banana’.
in Kerala by Samraj et al. (1966). The first
symptoms appear on the leaf sheath as References
longitudinal, irregular, reddish or pinkish Estelitta S., A. Suma, G. Zachariah & K. P. Pradeep. 1996.
streaks of varying sizes (Estelitta et al. Light and fluorescent microscopic studies on Kokkan
1996). Later on, the pseudostem becomes disease of banana (Musa AAB Nendran). J. Trop. Agric.
34:155-156.
abnormally red in colour and spongy in
Samraj J., M.R. Menon, S.P. Christudas & P.K. Satyarajan.
texture (Estelitta et al. 1996). The symptoms 1966. Kokkan a new disease of banana (Musa
were scored using the following scale: paradisiaca Linn). Agric. Res. J. Kerala 4(1):116.
0 = No symptom For more information:
1 = Reddish brown streaks over 1-25% of Smitha Rose Gasper or M. Suharban,
the pseudostem Plant pathology department, College of
2 = Reddish brown streaks over 26-50% of Agriculture, Thiruvananthapuram 695 522,
the pseudostem India
The story behind the name ‘Yangambi km 5’ Forum
‘Yangambi km 5’ (AAA) has numerous, the northern bank of the Congo River,
small fruits that have a very pleasant taste while the second axis was perpendicular
when ripe. It is a vigorous plant that remains and stretched northwards for about 25 km.
productive on poor soils and which has The main crop divisions (oil palm, rubber,
become well-known for being resistant coffee, etc.) were clustered around the fifth
to black leaf streak disease, caused by kilometre of this axis, with the consequence
Mycosphaerella fijiensis. that the banana field, with its ‘Gros Michel’,
Its somewhat peculiar name has recently Silk, Prata, Cavendish and this mysterious
caused people to speculate about its cultivar, were located at ‘km 5’.
meaning and origin. It just happens that in So why ‘Yangambi km 5’ and not a more
the 1950s I was in charge of the banana convenient cultivar name? Simply because
programme at Yangambi and may be able to we were not able to identify the plant. It did
shed some light on the matter. not fit any of the descriptions and illustrations
The name comes from the location of the available at the time. For a while we thought
fruit gardens at the “Institut National pour it was a diploid, but chromosome counting
les Etudes Agronomiques au Congo Belge” at the division of plant genetics showed that
(INEAC), which is now called INERA. The ‘Yangambi km 5’ was a triploid.
research divisions, with their laboratories After a visit to Yangambi, in 1957 if I
and experimental fields, were built along two remember well, the regretted Jean Champion
main road axes. The original axis followed introduced ‘Yangambi km 5’ in Guinée’s

banana collection. He too was unable to local scientists have classified numerous
identify it. French scientists distributed cultivars from Southeast Asia. In Thailand,
the cultivar in their research stations as Dr. Silayoi identified a ‘Kluai thong ruang’,
‘Yangambi km 5’, and the name stuck. the description of which strongly reminds me
I have since learned that the plant had of ‘Yangambi km 5’ except for its deciduous
been introduced in Yangambi before World fruits, but the most serious candidate for
War II from Kilo-Moto in the far northeastern synonymy with ‘Yangambi km 5’ is ‘Kluai
corner of Congo. Since I could not find any hom bao’ from southern Thailand. Other
trace of the introduction in the archives, it possible synonyms in Thailand are ‘Kluai
looks as if it was brought by the spouses of nam nom’, ‘Kluai chak nuan’ and ‘Kluai hom
the laborers recruited by INEAC. I was also hak’. It is possible that ‘Yangambi km 5’ was
told that local people call the cultivar ‘Ibota brought from Thailand directly to Congo by
bota’, which means ‘very fertile’ in reference a miner working in one of the many mines in
to the large number of fruits on a bunch. the Kilo-Moto area.
The question then is was the plant grown We may have to live with the name
in East Africa or was it directly introduced ‘Yangambi km 5’ for a while still, unless
from Asia? I found no trace of the cultivar in molecular tools establish that it is a Thai
none of my explorations in Congo, Rwanda cultivar, in which case the Thai appellation
and Burundi and on visits to collections in would be preferable.
Uganda and Tanzania. Edmond De Langhe
Meanwhile, under the impulse of Ramon Professor emeritus
Valmayor, an authority in banana taxonomy, Leuven, Belgium
Announcements Conference on black leaf streak Symposium on plant

disease biotechnology
An international congress on the The VII International Symposium of Plant
management of black leaf streak disease Biotechnology will be held 17-21 April 2006
in Latin America and the Caribbean will be at the Institute of Plant Biotechnology, Villa
held in San Jose, Costa Rica, 1-2 November Clara, Cuba. Among the themes covered will
be genetic transformation, bioinformatics,
2005. The conference is organized by
tissue culture, genetic improvement,
CORBANA, INIBAP and MUSALAC. The
biosafety and intellectual property rights.
four sessions will be devoted to the impact A workshop will be devoted to bananas
of the disease on production and quality; and plantains. For more information,
the epidemiology, biology and ecology of contact Orlando Gregorio Chaviano at
the fungus; chemical control; and biological [email protected].
control and genetic improvement. For more
information consult the INIBAP website at
www.inibap.org.
48 June 2005
Instructions to authors
I
NFOMUSA is an international journal published twice Discussion: The discussion should not contain extensive
a year in English, French and Spanish. Our focus is to repetition of the results section nor should it reiterate the
provide an outlet for research results and reports of interest introduction. It can be combined with the results section.
to the Musa community. As INFOMUSA publishes articles on References: All references to the literature made in the
any Musa-related issue, authors should aim for simple and text should be referred to by author(s) and year of publication
clear phrases that avoid unnecessary jargon in order to make (e.g.: Sarah et al. 1992, Rowe 1995). References to not
their paper accessible to readers in other disciplines. widely circulated documents, such as annual reports, and
Manuscripts should be prepared in English, French or citations of personal communications and of unpublished
Spanish and should not exceed 2500 words, including data should be avoided. A list of references, in alphabetical
references. They should be double-spaced throughout. All order, should be provided at the end of the text.
pages (including tables figures, legends and references) Please follow the style shown below:
should be numbered consecutively. Periodicals: Sarah J.L., C. Blavignac & M. Boisseau. 1992.
Include the full name of all the authors of the paper, Une méthode de laboratoire pour le criblage variétal des
together with the addresses of the authors at the time of bananiers vis-à-vis de la résistance aux nématodes. Fruits
the work reported in the paper. Indicate also the author 47(5):559-564.
nominated to receive correspondence regarding the paper. Books: Stover R.H. & N.W. Simmonds. 1987. Bananas (3rd
Manuscripts can be sent as e-mail attachments or put on edition). Longman, London, United Kingdom.
a 3.5-inch disk for PC-compatible machines. Please indicate Articles (or chapters) in books: Bakry F. & J.P. Horry.
the name and version of the word processing software used 1994. Musa breeding at CIRAD-FLHOR. Pp. 169-175 in
and the author’s e-mail address. In either case, we will need The Improvement and Testing of Musa: a Global Partnership
to receive by mail two printed copies of the manuscript. (D.R. Jones, ed.). INIBAP, Montpellier, France.
Title: The title should be as short as possible and should Illustrations and tables: These should be numbered
not have numbers, acronyms, abbreviations or punctuation. consecutively and referred to by these numbers in the text.
Abstract: An abstract, not exceeding 200-250 words, Each illustration and table should include a clear and simple
should be provided. It should concisely summarise the basic caption. Figures and tables should be inserted after the
contents and should be sent in the same language as the references or in separate files.
manuscript. Translations (including the title) into the two Graphs: provide the corresponding raw data with the
graphs, if possible in Excel format.
other languages should also be sent if this is possible.
Drawings: provide originals if this is possible.
Key words: Provide a maximum of six key words, in
Photographs: We prefer hard-copy printouts of
alphabetical order, below the native-language abstract.
photographs (bright paper with good contrast for black and
Introduction: The introduction should provide the rationale
white photographs; good quality proofs and films or original
for the research and any relevant background information. slides for colour photographs), but please remember that
Since it is not meant to be an exhaustive review of the topic, we will not return them. We will publish pictures that have
the number of references should be kept to a minimum. been scanned or taken with a digital camera as long as the
Introductions on the importance of bananas as a staple food resolution is high enough (1 million pixels or a minimum of
or a traded commodity should be avoided, unless they are 300 dpi when the photograph is in real size). Acceptable
absolutely necessary for the comprehension of the article. file types are JPEG, TIFF and EPS. Avoid sending photos
Materials and methods: The authors should provide inserted in a Word or Power Point document, unless they are
enough details of their experimental design to allow the reader accompanied by a better quality alternative.
to gauge the validity of the research. For commonly used Acronyms: These should be written in full the first time they
materials and methods, a simple reference is sufficient. appear in the text, followed by the acronym in parenthesis.
Results: The unit should be separated from the number Cultivar names: The name of the cultivar should be placed
by a single space and follow SI nomenclature, or the between single quotation marks. If the name is a compound
nomenclature common to a particular field. Unusual units or noun, only the first word starts with a capital letter, unless the
abbreviations should be defined. other refers to a place or person. Use the most commonly
Present data in the text, or as a figure, or a table, but agreed upon name, such as ‘Grande naine’ and avoid local
never in more than one of these ways. Avoid extensive variations or translations, such as ‘Gran Enano’.
use of graphs to present data that could be more concisely Note: When plant material used for the experiments
presented in the text or in a table. Limit photographs to those reported originates or is registered in the INIBAP genebank,
that are absolutely necessary to show the experimental its accession number (ITC code) should be indicated within
findings. the text or in a tabular form.
Thank you in advance for following these instructions. This will facilitate and accelerate the editing work.
INIBAP Publications In press
D.W. Turner and F.E. Rosales (eds). 2005. Banana Root System: towards a better
understanding for its productive management. Proceedings of an International Symposium
held in San José, Costa Rica, 3-5 November 2003.
Recent publications
INIBAP 2004. INIBAP Annual Report 2003. International network for the Improvement of Banana
and Plantain, Montpellier, France.
S. Mohan Jain and R. Swennen (eds). 2004. Banana improvement, cellular, molecular biology,
and induced mutations. This 392-page book, co-published by FAO/IAEA and INIBAP, presents
the results from the FAO/IAEA
Coordinated Research Project
entitled “Cellular biology and
biotechnology including mutation
techniques for creation of new
useful banana genotypes”. The
book also contains several review
papers providing up-to-date
information on biotechnological
tools that can be used to produce
new Musa varieties with desirable
characters in a more rapid and
efficient way.
To obtain a complete list of
our publications, consult our
website or contact Leila Er-
rachiq at INIBAP headquarters
in Montpellier.
E-mail : [email protected]
www.inibap.org
INIBAP addresses
• Headquarters: Associate Scientist, Musa technology transfer:

Parc Scientifique Agropolis II Dr Inge Van Den Bergh
34397 Montpellier Cedex 5 - FRANCE c/o IRRI, Rm 31, GS Khush Hall
e-mail: [email protected] Los Baños, Laguna 4031
Fax : (33) 467 61 03 34 Philippines
Director: Dr Richard Markham Fax: (63-49) 536 05 32
e-mail: [email protected] e-mail: [email protected]
Coordinador, Musa Genetic Improvement:
Dr Jean-Vincent Escalant
• Regional Office for West and Central Africa
e-mail: [email protected] Regional Coordinator: Dr Ekow Akyeampong
Coordinator, Musa Genomics and Genetic Resources Regional information officer for Africa:
Conservation: Dr Nicolas Roux Mr Josué Tetang Tchinda
e-mail: [email protected] Associate Scientist, Musa technology transfer:
Coordinador, Musa Agroecosystems and Channels for Added Ms Kim Jacobsen
Value: Dr Charles Staver c/o CARBAP - BP 12438
e-mail: [email protected] Douala, Cameroon
Coordinator, Information/Communications: Tel./Fax: (237) 342 91 56
Ms Claudine Picq E-mail: [email protected]
e-mail: [email protected]
Officer in charge MGIS: Ms Elizabeth Arnaud • Regional Office for Eastern and Southern
e-mail: [email protected] Africa
Accountant: Mr Emmanuel Gonnord Regional Coordinator: Dr Eldad Karamura
e-mail: [email protected] Associate Scientist: Musa technology transfer:
Impact assessment specialist: Ms Charlotte Lusty Dr Guy Blomme
e-mail: [email protected] PO Box 24384
Kampala, Uganda
• Regional Office for Latin America and the Fax: (256-41) 28 69 49
Caribbean e-mail: [email protected]
Regional Coordinator: Dr Franklin E. Rosales
Associate Scientist, Musa technology transfer: Dr Luis • INIBAP Transit Center (ITC)
Pocasangre Officer in charge: Ms Ines Van Den Houwe
c/o CATIE Katholieke Universiteit Leuven
Apdo 60-7170 Turrialba, Costa Rica Laboratory of Tropical Crop Improvement
Tel./Fax: (506) 556 2431 Kasteelpark Arenberg 13,
e-mail: [email protected] B-3001 Leuven
Belgium
• Regional Office for Asia and the Pacific Fax: (32-16) 32 19 93
Regional Coordinator: Dr Agustín Molina e-mail: [email protected]
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Why beer bananas: The case for Rwanda

Article · January 2005

Svetlana V. Gaidashova Suleman H. O. Okech

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The case for

Publishing director: Claudine Picq

Editor: Anne Vézina

InfoMusa - Vol. 14 - No.1 1

Gishoma Participatory rural appraisal sites

InfoMusa - Vol. 14 - No.1 3

Kivu Lake Border

say that introduced beer bananas (ABB) Marketing

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Agronomic evaluation Agronomic evaluation of production and quality of

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New methodology for the establishment of cell Embryogenic cell suspensions

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Tissue culture Effect of an analogue of brassinosteroid on FHIA-18

with more than 53% and 5% of plants free of

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Table 1. Quantity (mg/L) of organic crude extract obtained by culturing Mycosphaerella

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A Figure 2. (A) The scientific trials used to

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Table 1. Example of a farmer training process in a wet-dry zone of Nicaragua.

3. What do we know about Musa and its pests?

4. What will we test/what do we want to learn?

3. Analysis of the favourable and unfavourable factors in the new

(August – rains) 2. Analysis of new plantings – effect of sucker and field

3. Projected harvest and costs

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InfoMusa - Vol. 14 - No.1 35

Focus on IPGRI Results of survey on IPGRI’s commodity work

InfoMusa - Vol. 14 - No.1 37

Focus on INFOMUSA Highlights from the reader survey

Who are the respondents? What kind of INFOMUSA?

Variability in reproductive fitness and pathogenicity Thesis

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Random amplified polymorphic DNA analysis of Thesis

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An article by Ramon Valmayor, Le Dinh home of M. splendida and discovered large

Musanews Presence of banana bunchy top virus in Angola

Containing banana xanthomonas wilt Musanews

A workshop on “Developing a coherent • Throughout the region: to coordinate and

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Musanews Controlling banana xanthomonas wilt through

The story behind the name ‘Yangambi km 5’ Forum

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