Protección de Cultivos / Crop Protection

Acta Agron. (2018) 67 (2) p 368-374 ISSN 0120-2812 | e-ISSN 2323-0118 https://doi.org10.15446/acag.v67n2.61334

Identification of new sources of resistance to RHBV- rice hoja

blanca virus
Identificación de nuevas fuentes de resistencia al virus hoja blanca en arroz-RHBV

Maribel Cruz-Gallego1,2*, Camila Rebolledo2, Juan Cuasquer2, Daniel Cruz3, Alexandra Lorena Peña-Fernández2,
Constanza Quintero2, Alexander Silva-Córdoba2, María Fernanda Álvarez2, Sebastián Jojoa-Cruz4, Mathias
Lorieux5,2, Jeffrey J. Stuart6 and Fernando Correa2

FLAR-Fondo Latinoamericano para Arroz de Riego. 2CIAT- Centro Internacional de Agricultura Tropical. Cali, Valle-Colombia.3
1

Ghent University, Flanders, Belgium.4The Scripps Research Institute. Florida, EE. UU. 5IRD- Institut de Recherche pour le
Développement. Marseille, France.6 Department of Entomology, Purdue University. West Lafayette, EE. UU.

Rec.: 30.11.2016 Accep.: 27.06.2017

Abstract
With the aim to find new sources of resistance to rice hoja blanca (white leaf) disease, transmitted by the insect
Tagosodes orizicolus, 660 genotypes were evaluated under greenhouse and field conditions. Seven resistant
genotypes were identified, and genomic studies were performed to demonstrate that the resistance in these sources
is genetically different from that of Fedearroz 2000, which is currently the variety with the most resistance to hoja
blanca. These new resistance sources constitute a resource that can be used to sustainably extend hoja blanca
disease management throughout all of the rice-growing regions of tropical America. This is the first report of hoja
blanca resistance in indica rice and different from that of Fedearroz 2000.
Key Words: Oryza sativa L.; phenotyping; plant-virus-vector pathosystem; Sogata; Tagosodes; Tenuivirus.

Resumen
Con el objetivo de encontrar nuevas fuentes de resistencia a la enfermedad de la hoja blanca del arroz, transmitida
por el insecto Tagosodes orizicolus, se evaluaron 660 genotipos en condiciones de invernadero y campo. Se
identificaron siete genotipos con resistencia a la enfermedad y se realizaron estudios del genoma para evidenciar
que eran genéticamente diferentes a Fedearroz 2000, la variedad de mejor comportamiento ante el virus, en el
momento. Estas nuevas fuentes de resistencia constituyen un recurso que puede utilizarse para extender un
manejo sostenible de la enfermedad, en todas las regiones productoras de arroz en América tropical. Este es el
primer reporte de fuentes de resistencia, tipo indica, diferentes a Fedearroz 2000.
Palabras clave: Fenotipado; Oryza sativa L.; patosistema planta-virus-vector; Sogata; Tagosodes; Tenuivirus.

Introduction segment of ssRNA. Currently, this disease affects

rice fields in Colombia, Costa Rica, Ecuador,
Rice hoja blanca disease (RHB) is a highly Guyana, Panama, Peru, Dominican Republic,
important economic constraint to rice (Oryza Nicaragua, and Venezuela. Disease epidemics
sativa L.) production. Yield losses can reach are intermittent or cyclical, probably because
100% when susceptible varieties are sown and of the population dynamics of its insect vector,
infected vectors are present in the field (Morales and the interaction between the virus and the
& Jennings, 2010). The causal agent of RHB is a vector insects (Jennings & Pineda, 1971; Morales
Tenuivirus named Rice hoja blanca virus (RHBV). & Jennings, 2010). RHBV is closely related to
It is composed of four different filamentous Echinochloa hoja blanca virus (EHBV), which
particles that are 3 nm in diameter. Each particle is transmitted by a different Tagosodes species
has a different length and contains a different (Madriz, De Miranda, Cabezas, Oliva, Hernandez

368
 Identification of new sources of resistance
to RHBV- rice hoja blanca virus

& Espinoza, 1998). RHBV is detectable in both resistant lines, most plants remain symptom-
plants and insects through ELISA (enzyme- free, and ELISA fails to detect virus in the plants.
linked immunosorbent assay). Virus incubation “However, RHBV-resistance in the cultivated rice
requires 20 to 22 days in the insect and 7 to species O. sativa, especially among cultivars that
9 days in 10-day-old plants (Pantoja, Fisher, produce the indica grain type preferred in Latin
Correa, Sanint & Ramírez, 1997). The virus America, is rare and a major obstacle in the
cannot be transmitted mechanically or through implementation of integrated RHB management
seeds (Malaguti, Diaz & Angeles, 1956). The programs” (Morales & Jennings, 2010, p.13).
virus also causes disease in the vector insect, Thus far, a single cultivar, Fedearroz 2000, is
shortening its life and decreasing its fecundity the best source of resistance to RHB disease.
(Jennings & Pineda, 1971). Symptoms of RHB Other sources of resistance to RHBV have been
depend on the variety and age of the plant. Young identified in O. sativa japonica germplasm and
leaves have short chlorotic stripes, parallel to the transferred to indica backgrounds suitable
midrib. Older plant leaves lose their green color for use in Latin American breeding programs
as the chlorotic stripes coalesce into wide white (Zeigler, Pantoja, Duque & Weber, 1994). The
(or pale yellow) bands. These symptoms give the International Center for Tropical Agriculture
disease its name (hoja blanca means white leaf in (CIAT) and the Latin American Fund for Irrigated
Spanish). Eventually, the virus causes necrosis, Rice (FLAR) have been evaluating thousands of
which begins from the apical part of the leaves lines for years. However, no sources have yet
and extends toward the base of young plants. provided RHB resistance that is as effective as
Plant tillering decreases, growth is poor, and, that of Fedearroz 2000. Thus, only a few sources
sometimes, the disease causes the death of the of resistance have been used to produce all of the
plant (Pantoja et al., 1997). Normally, during commercially available RHB-resistant cultivars.
late infection, some tillers are also affected, Without the discovery of new efficacious sources
and the panicles are sterile and deformed. Poor of resistance, rice production will be at risk when
root development has also been observed under the virus breaks the resistance of Fedearroz 2000.
controlled greenhouse conditions (unpublished
Here, we report the discovery of new RHB-
data). Unfortunately, once infection occurs, there
resistance sources in a diverse indica panel of O.
is no chemical treatment to control or cure the
sativa. These new resistance sources constitute a
disease (Pantoja et al., 1997).
resource that can be used to sustainably extend
Sogata is the common name of the RHBV hoja blanca disease management throughout all
insect vector, Tagosodes orizicolus (Müir) of the rice-growing regions of tropical America.
(Hemiptera: Delphacidae). This species is
distributed in almost all of the tropical and
subtropical rice-growing areas of the Americas. Material and methods
Sogata sucks nutrients from phloem and excretes Evaluations were performed with methodologies
honeydew on the leaves, which both encourages commonly used at CIAT (Triana, Cruz & Meneses,
fungal growth and reduces the photosynthetic 2003). Indica genotypes were evaluated in
capacity of the plant. In the absence of RHBV, four sequential experiments: (1) initial RHBV
high insect populations cause severe crop damage screening, (2) individual RHBV evaluation, (3)
or hopper burn. Sogata adults are relatively field trials, and (4) insect resistance screening.
sedentary, but disperse easily with strong winds In each evaluation, Sogata insects were obtained
or water currents (Heinrichs, 1994). Sogata from colonies maintained at CIAT. The RHBV-
transmits RHBV persistently and the virus can harboring colony contained insects that were
be acquired either by feeding on infected plants fed on RHBV-infected plants and allowed to
or transovarially. Interestingly, not all Sogata are reproduce on a rice variety (Bluebonnet 50) that
capable of virus transmission (Galvez, Thurston is susceptible to both the insect and RHBV. To
& Jennings, 1961). However, even when the determine the percentage of virulent insects
proportion of Sogata capable of transmission (insects capable of transmitting the disease) in
is as low as 1%, serious damage results when this colony, 200 individual nymphs were tested
susceptible varieties are sown (Calvert & Reyes, for virulence on separate, caged, RHB-susceptible
1999). Moreover, both chemical and biological 8-day-old seedlings. The nymphs were permitted
control are often ineffective in managing RHBV to feed and transmit virus to the plants for 3 days.
transmission in Sogata populations. Therefore, After 11 days, the number of plants displaying
RHBV-resistant lines and cultivars are the most disease symptoms was determined and the
effective and economical means of managing hoja percentage of virulent nymphs in the colony
blanca disease in rice. (62.3%) was extrapolated. This percentage is very
Rice hoja blanca disease resistance has proven high compared to the natural conditions where
to be an effective crop management tool. In virulence ranges from 1% up to 5% when the risk
of epidemic is severe (Pantoja et al., 1997).

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Acta Agronómica. 67 (2) 2018, p 368-374

The RHBV-free colony was begun using non- the CIAT screening (Zeigler, Rubiano & Pineda,
vector insects, insects that did not transmit 1988). This method releases RHBV-vectoring
disease to susceptible rice seedlings for at least Sogata that are maintained in greenhouse
three generations. This colony was maintained colonies to ensure that RHBV incidence is high.
separately on RHBV- and insect-susceptible rice Genotypes that exhibited a low percentage of
(Bluebonnet 50) and closely monitored for any infection in initial screening experiments were
signs of RHB disease symptoms. examined in randomized complete blocks with
ten replications. Replications consisted of rows
Initial RHBV screening 0.5 m in length, with 0.15 m between rows and
100 plants per row. Rows were planted using
To identify new sources of resistance against dry seed. One row was the experimental unit.
RHBV, 660 genotypes were evaluated in cages, Fedearroz 2000, Colombia 1, and Bluebonnet 50
under greenhouse conditions (27 °C, 80% relative were included as checks in each block. Eighteen
humidity, and 12 daylight/12 night hours). We days after planting, an average of two insects per
evaluated 295 genotypes belonging to a diverse plant were released. Three days later, the insects
indica rice panel from the International Rice were killed with imidacloprid insecticide. The
Research Institute (Group 1) and a second set insects came from an RHBV-harboring colony in
of lines of diverse origin that were sequenced which 48.6% of the insects were virulent. Forty
in “The 3000 Rice Genomes Project” (Li et al., days later, disease incidence was evaluated using
2014) (Group 2). The genotypes were screened in a visual scale of damage with five levels (1, 3, 5,
plastic trays containing sterilized soil arranged 7, and 9): Level 1, rows exhibiting less than 10%
in two randomized complete blocks, with three of plants with symptoms; Level 3, >10% and
replications and 20 plants per replication. Each <30%; Level 5, >30% and <50%; Level 7, >50%
block included check varieties Fedearroz 2000 and <70%; and Level 9, >70%. Levels 1 and 3 were
(resistant), Colombia 1 (intermediate), and classified as resistant, level 5 was classified as
Bluebonnet 50 (susceptible). Nymphs collected intermediate, and levels 7 and 9 were classified as
from the RHBV-harboring colony were used to susceptible. The data were recorded and analyzed
transmit RHBV to the plants. An average of four as a percentage of incidences in which the rows
nymphs per plant were released into each tray 18 of each genotype exhibited these reactions.
days after planting. The nymphs were permitted
to feed on the plants for 3 days and were then Some phenotypical traits were determined
killed with imidacloprid insecticide. The number under CIAT-Palmira field conditions in 2013 and
of plants showing disease symptoms per genotype 2014 as part of a complementary study. In 2013,
was determined visually 40 days after infestation. rows were established through transplantation,
using five plants per linear meter. In 2014, the
Individual RHBV evaluation rows were established through direct seeding with
20 plants per linear meter. Data are registered in
Individual evaluation was performed on genotypes the results section in the interest of rice breeding.
that exhibited a percentage of infected plants
lower than that of Fedearroz 2000 in the initial Reaction to insect damage
RHBV screening. The main difference in this
evaluation was that the insects were not free to Ideally, commercial varieties targeted for RHB-
choose the plants on which they fed. Instead, four prone areas must be RHBV resistant as well as
insects from the RHBV-harboring colony were tolerant of the mechanical damage caused by
confined together on individual caged plants. T. orizicolus. As a consequence, RHB-resistance
The cages were constructed of acetate tubes, sources were tested for tolerance of the mechanical
covered with tulle fabric to prevent the insects damage caused by the insects. The experimental
from escaping and to provide aeration. Genotypes design was completely randomized with three
and the checks were planted in a completely replications and Makalioka (resistant), Cica 8
randomized design with ten replications in one (intermediate), and Bluebonnet 50 (susceptible)
experiment and with 20 replicates in a second were included as checks. The experimental unit
experiment. The insects were allowed to feed on was a set of ten plants that were planted in trays
the plants for 3 days and were then killed using with sterilized soil. Ten nymphs from the virus-
imidacloprid insecticide. The number of plants free colony were released per plant 15 days after
displaying RHB symptoms was determined 40 planting. The insects were allowed to feed on
days after infestation. the plants until the time of evaluation: the day
when 100% of the plants of the susceptible check
Trials under field conditions died (about 8 days after infestation). Evaluation
was performed using the Standard Evaluation
The sources of RHBV resistance were evaluated System for Rice (www.knowledgebank.irri.org/
under semi-controlled field conditions based on images/docs/rice-standard-evaluation-system.

370
 Identification of new sources of resistance
to RHBV- rice hoja blanca virus

pdf), in which ratings of 1 and 3 are a resistant parameters recommended in Duitama, Quintero,
reaction, a rating of 5 is an intermediate reaction, Cruz, Quintero, Hubmann, Foulquié-Moreno,... &
and ratings of 7 and 9 are susceptible reactions. Tohme. (2014). Parameters used for the selection
The maximum value exhibited observed in a of SNPs were as follows: (1) minimum genotype
replication was reported as the final result. quality 40, (2) SNPs genotyped in all accessions,
and (3) no heterozygous SNPs. All the SNPs located
Statistical analysis in the selected region were used for the calculation
of a p-distance matrix and the construction of an
Statistical analysis, including linear mixed model unrooted tree following the neighbor joining (NJ)
and least significant difference (LSD), was carried method with the software SplitsTree4 (Huson
out to test the differences between the current & Bryant, 2006). Based on the SNPs obtained
source of resistance (Fedearroz 2000) and each by WGS data, eight polymorphisms within the
one of the evaluated genotypes. The statistical analyzed region of chromosome 4 were developed
analysis included a test of the hypothesis to as Fluidigm SNPtype assays (Fluidigm®, San
measure the genotype effect through a linear Francisco, USA) to build haplotypes and confirm
mixed model. Adjusted means were also estimated the differences between Fedearroz 2000 and the
for each genotype. In addition, a comparison test seven new sources of resistance.
of adjusted means against Fedearroz 2000 was
done through LSD method to identify genotypes
better than or equal to Fedearroz 2000. Results

Molecular analysis and genetic variability Initial RHBV screening

Population structure for the diverse indica rice Through mass evaluation, one genotype belonging
panel of 295 genotypes (Group 1) was assessed. to Group 2 was potentially RHBV resistant, and
The analysis was carried out with the package nine from Group 1 were statistically as resistant
Structure V2.3 using 83,374 SNPs (Rebolledo, as the check, Fedearroz 2000 (data not shown).
Peña, Duitama, Cruz, Dingkuhn, Grenier &
Tohme, 2016). Parameters were K=1 to 10 Individual RHBV evaluation
subpopulations, tested with 16 replications,
a burning period of 20,000 iterations, and After individual confirmation of RHBV reaction,
sampling period of 40,000 iterations. The final seven genotypes from Group 1 remained resistant
result of the population structure was obtained under greenhouse conditions (Table 1). The
using the online tool Structure Plot (Ramasamy, remainder of the genotypes in both groups were
Ramasamy, Bindroo & Naik, 2014). Based on susceptible (data not shown).
population structure analysis, five rice genotypes
showing resistance to RHBV and belonging to
Table 1. Reaction of indica genotypes to RHBV and insect damage under
different subpopulations were chosen for WGS greenhouse conditions.
(whole-genome sequencing) at BGI Americas.
RHBV-infected Mechanical damage*
RHBV-resistance sources
To determine whether the new resistance plants (%) (visual scale)
sources were different from Fedearroz 2000, Badkalamkati::IRGC 45011-1 9.8 a 3
single nucleotide polymorphisms were analyzed IR77384-12-35-3-12-1-B::IRGC 117299-1 5.5 a 3
within and surrounding the QTL associated with
RHBV resistance on chromosome 4, previously Pokkali::IRGC 108921-C1 8.6 a 3

reported by Romero, Lozano, Garavito, Carabali, PTB 25::IRGC 6386-1 9.4 a 3

PTB 9::IRGC 6274-1 11.0 a 5
Triana, Villareal… & Lorieux (2014). The genomic
Sahel 108::C1 7.3 a 3
region spanning from 2.81 Mbp to 3.8 Mbp was
WAS 208-B-B-5-1-1-3::C1 5.5 a 3
compared between Fedearroz 2000, the five
Fedearroz 2000 (virus-resistant check) 5.7 a NE
new sources, and Colombia 1 (intermediate),
Colombia 1 (virus-intermediate check) 43.2 b NE
Fedearroz 50 (highly antibiotic effect against the
Makalioka (mechanical damage-resistant
vector), and Bluebonnet 50 (RHBV-susceptible check)
NE 3
check). The whole genome sequences (WGS) of Cica 8 (mechanical damage-intermediate
NE 5
the former three genotypes were obtained from check)
previous work (Duitama, Silva, Sanabria, Cruz, %OXHERQQHWbVXVFHSWLEOHFKHFN 97.7 c 9
Quintero, Ballen… & Tohme, 2015); the WGS of 9DOXHVIROORZHGE\GL΍HUHQWOHWWHUVDUHVWDWLVWLFDOO\GL΍HUHQWIURP)HGHDUUR]
Colombia 1 (IRGC 116970-1-1) and Bluebonnet ȭ/6'PHWKRG
50 (IRGC 1799-1-1) were obtained from the NE: Non-evaluated.
3K RGP (Li, Wang & Zeigler, 2014). The reads *Maximum value registered in at least one of the replicates.
were analyzed using NGSEP (Next Generation
Sequencing Eclipse Plugin) software with the

371
Acta Agronómica. 67 (2) 2018, p 368-374

Field evaluation Table 2. Frequency (%) of responses of indica genotypes exposed to RHBV
XQGHUVHPLFRQWUROOHGȴHOGFRQGLWLRQV

Under semi-controlled conditions in the field, four RHBV-resistance sources Resistant Intermediate Susceptible
genotypes exhibited a high frequency of resistant Badkalamkati::IRGC 45011-1 80 20 0
rows compared with resistant check Fedearroz IR77384-12-35-3-12-1-B::IRGC
2000: Badkalamkati, IR77384-12-35-3-12-1-B, 80 20 0
117299-1
Pokkali, and PTB 25. The genotypes PTB 9 and Pokkali::IRGC 108921-C1 90 10 0
WAS 208-B-B-5-1-1-3 exhibited 50% and 20% PTB 25::IRGC 6386-1 100 0 0
of susceptible rows, respectively. And, the total PTB 9::IRGC 6274-1 30 20 50
number of rows of Sahel 108 was intermediate.
Sahel 108::C1 0 100 0
None of the genotypes tested was as susceptible
WAS 208-B-B-5-1-1-3::C1 40 40 20
as Bluebonnet 50 (Table 2). Important traits for
Bluebonnet 50 (susceptible
breeding purposes (Number of tillers, Number check)
0 0 100
of panicles, 1000-grain weight, Number of total Colombia 1 (intermediate
grains, Fertility, Plant height, Flowering time, 50 50 0
check)
and Cycle length) are shown in Tables 3 and 4. Fedearroz 2000 (resistant
100 0 0
check)

Table 33KHQRW\SLFDOWUDLWVRI5+%9UHVLVWDQWJHQRW\SHVWUDQVSODQWHGWRȴHOG\HDU

Number of Number of 1000-grain Number of Plant height Flowering Cycle length

Designation Fertility (%)
tillers panicles weight (g) total grains (cm) time (days) (days)
Badkalamkati::IRGC
69.0 61.5 27.0 111.9 70.7 112.0 98.7 146.7
45011-1
IR77384-12-35-3-12-1-
80.5 70.5 23.1 88.8 64.9 89.8 91.6 139.0
B::IRGC 117299-1

Pokkali::IRGC 108921-C1 54.0 50.6 26.5 84.1 61.9 146.6 85.0 127.3

PTB 25::IRGC 6386-1 70.6 64.6 23.1 102.5 66.4 143.4 82.6 129.0
PTB 9::IRGC 6274-1 66.9 62.6 19.5 93.8 56.9 148.0 76.3 135.3
Sahel 108::C1 109.2 96.3 22.0 101.8 79.8 87.9 86.3 136.0

WAS 208-B-B-5-1-1-3::C1 91.9 74.3 20.3 147.9 77.0 84.1 86.2 140.7

Table 4. Phenotypical traits of indicaJHQRW\SHVVRZQLQȴHOGWULDO\HDU

Number of Number of 1000-grain Number of Plant height Flowering Cycle length

Designation Fertility (%)
tillers panicles weight (g) total grains (cm) time (days) (days)
Badkalamkati::IRGC 45011-1 57.7 53.3 22.3 99.6 79.8 126.3 106.0 152.0
IR77384-12-35-3-12-1-B::IRGC
60.1 54.6 23.9 113.5 84.9 93.4 90.3 138.3
117299-1
Pokkali::IRGC 108921-C1 48.2 44.3 26.2 83.1 67.1 128.5 86.6 132.5

PTB 25::IRGC 6386-1 67.7 63.6 23.3 78.3 83.3 121.6 86.0 136.3

PTB 9::IRGC 6274-1 92.3 86.6 19.6 86.3 81.9 104.3 77.7 117.0

Sahel 108::C1 79.8 75.8 21.1 70.9 91.4 66.9 84.7 132.6

WAS 208-B-B-5-1-1-3::C1 69.1 65.9 18.1 94.0 84.1 66.2 88.0 138.3

Reaction to insect damage

Six genotypes were as resistant to mechanical
damage as Makalioka (Table 1). PTB 9 displayed
an intermediate reaction, like that of Cica 8.

Molecular Analysis and Genetic Variability

The structure analysis revealed that the seven
new sources of resistance belonged to three Figure 1.3RSXODWLRQVWUXFWXUHN RI*URXSJHQRW\SHV1HZVRXUFHV
of resistance to RHBV are indicated within subpopulations.
different subpopulations (Figure 1).

372
 Identification of new sources of resistance
to RHBV- rice hoja blanca virus

Five genotypes representing the three subpopulations Discussion

were submitted for WGS (Badkalamkati, IR77384-
12-35-3-12-1-B, Pokkali, PTB 9, and WAS 208-B- After Colombia 1, an unacceptable commercial
B-5-1-1-3). variety, and Fedearroz 2000, the most RHBV-
resistant variety widely sown in Colombian fields,
A total of 1514 SNPs was identified in the region there has been a limited quantity of sources with
between 2.81 Mbp and 3.80 Mbp of chromosome high RHBV resistance (Morales & Jennings, 2010)
4, which includes the RHBV-resistance QTL region and no RHBV-resistant indica type has been
of Fedearroz 2000 and Fedearroz 50 (Romero identified. In crop management, this constitutes
et al., 2014). According to the distance-based a latent risk, especially since the disease is
dendrogram (Figure 2), nine accessions analyzed currently spreading in tropical America. Moreover,
were classified into four groups. Fedearroz 50 and past experience demonstrates that monocultures
Colombia 1 were close to Fedearroz 2000. can suffer a breakdown of resistance because
of genetic changes in the pathogen. The risk is
even greater when plant health relies on a single
genetic source of resistance (Kiyosawa, 1982).
Fedearroz 2000, Fedearroz 50, and Colombia
1 clustered together and were distant from the
new indica accessions that show high levels of
resistance to RHBV. The analysis of the genomic
region surrounding and harboring the QTL
suggested that the new sources of resistance
are probably unique and different from the one
in Fedearroz 2000 and Fedearroz 50. The origin
of the resistance in these new sources needs to
be clarified. It is still unclear whether it could
be an allelic variation of the Fedearroz 2000
QTL or new genomic regions that are involved in
resistance responses. Several biparental mapping
populations are under development to distinguish
between these possibilities.
Figure 2. Neighbor joining dendrogram for the WGS accessions comparing
the RHBV-resistance QTL region of Fedearroz 2000 (FA2000) and Fedearroz The new RHBV-resistance sources identified
50 (FA50).
in this research are from India, the Philippines,
and Senegal, and belong to the indica type.
On the other hand, the new indica RHBV- Thus, it is feasible to use these potential sources
resistance genotypes were classified into groups in crosses targeting the tropics. In addition to
II and III, distant from Fedearroz 2000 in group RHBV resistance and tolerance of mechanical
IV. Furthermore, the eight SNPs developed as damage, the new sources are long-grained with
Fluidigm SNPtype assays allowed us to identify intermediate to high amylose content as preferred
four haplotypes in the region on chromosome by most Latin American consumers (Calingacion,
4 (Table 5). Fedearroz 2000, Fedearroz 50, and Laborte, Nelson, Resurreccion, Concepcion,
Colombia 1 share the same haplotype, while the Daygon,… & Fitzgerald, 2014). Those are
seven indica accessions were different. important advantages for rice breeding programs
in the tropical zone. Future investigations will
examine the putative antibiotic effects of the
resistance to the insects observed in this study,
the inheritance of the resistance and antibiotic
traits, resistance gene discovery, and the design
of molecular markers to accelerate plant breeding
with these new resistance sources.

Conclusion
The resistance sources identified in this research
could play a key role in defeating rice hoja blanca
Table 5. Haplotype from the seven indica sources of RHBV resistance disease. This is the first report of rice hoja blanca
compared with the current sources used by rice breeders, Fedearroz 2000 resistance in indica rice types with different
and Fedearroz 50 (na, missing data). genetic resistance from that of Fedearroz 2000.

373
Acta Agronómica. 67 (2) 2018, p 368-374

Acknowledgments Malaguti, G., Diaz, C. & Angeles, N. (1956). Ensayos

preliminares sobre el uso de insecticidas en el
We are grateful to Efren Córdoba and Rodrigo control de los vectores de la “Hoja Blanca” del
Moran for their valuable technical support, and arroz. Agronomía Tropical, 7(3), 157௅163. http://
to CIAT, FLAR, and Purdue University for support www.sian.inia.gob.ve/revistas_ci/Agronomia%20
Tropical/at0703/pdf/at0703_nota2.pdf.
for this study.
Morales, F. & Jennings, P. (2010). Rice Hoja Blanca:
A complex plant-virus-vector pathosystem. CAB
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