Plant Pathol. J.

22(3) : 278-282 (2006)

The Plant Pathology Journal

© The Korean Society of Plant Pathology

Mini-Review

Bacillus vallismortis EXTN-1-Mediated Growth Promotion and Disease

Suppression in Rice
*
Kyungseok Park , Diby Paul and Wan-Hae Yeh

Plant Pathology Division, National Institute of Agricultural Science and Technology, RDA, Suwon 441-707, Korea
(Received on August 4, 2006; Accepted on August 12, 2006)

Bacillus vallismortis EXTN-1, a biocontrol agent in effects are brought about by different mechanisms as
cucumber, tomato and potato was tested in rice patho- reported by many researchers (Vessey et al., 2003; Defago
system against rice fungal pathogens viz. Magnaporthe et al., 1990). Biocontrol organisms act as antagonists of the
grisea, Rhizoctonia solani and Cochliobolus miyabeanus. targeted pathogen or induce systemic resistance to the host
Apart from increasing the yield in the bacterized plants plant (Cook and Baker, 1983), with systemic biochemical
(11.6-12.6% over control), the study showed that EXTN- and ultra structural changes in the plant system that lead to a
1 is effective in bringing about disease suppression greater ability of the host plant to defend itself against
against all the tested fungal pathogens. EXTN-1 treat- pathogens.
ment resulted in 52.11% reduction in rice blast, 83.02%
reduction in sheath blight and 11.54% decrease in The present study dealt with rhizobacteria-mediated plant
brown spot symptoms. As the strain is proven as an growth promotion and disease suppression in rice. The
inducer for systemic resistance based on PR gene rhizobacteria used in the study was Bacillus vallismortis
expression in Arabidopsis and tobacco models, it is strain, EXTN-1. The strain had been demonstrated to
supposed that a similar mechanism works in rice, induce systemic resistance in many cropping systems lead-
bringing about disease suppression. The strain could be ing to significant disease suppression and enhanced plant
used as a potent biocontrol and growth-promoting growth (Park et al., 2001).
agent in rice cropping system.
Keywords : Bacillus vallismortis, biological control, induced Materials and Methods

systemic resistance, rice diseases

The microorganisms and the plant source. The rhizo-
bacteria, EXTN-1 that previously showed significant induced
Rice is highly vulnerable at all stages of growth to systemic resistance (ISR) activity on other crops (Park et
pathogens that affect the quality and quantity of its yield. al., 1998) was used in the study. For long-term storage, the
Rice blast caused by Magnaporthe grisea is a problem strain was maintained at −80 C in tryptic soy broth (TSB)
o

almost everywhere that rice is grown. This fungal disease is with glycerol (20%). The fungal pathogens used in the
estimated to cause production losses of US $ 55 million experiments, Magnaporthe grisea, Rhizoctonia solani and
each year in South and Southeast Asia. The losses are even Cochliobolus miyabeanus were maintained on potato
higher in East Asia and other more temperate rice growing dextrose agar (PDA) slants at 15 C. Rice variety cv.
o

regions around the world (Herdt et al., 1991). Sheath blight Choochung, which is susceptible to the fungal pathogens
(caused by Rhizoctonia solani) is distributed in 1.2 to 1.4 tested, was used in the experiment.
million ha which is about 32-50% of the world’s total rice
cropping area. Rice brown spot, caused by Cochliobolus Treatment of the plants and challenge with the patho-
miyabeanus has been severely increased recently in Korea. gen. The treatment included bacterization of rice seeds with
Sustainable and environmental friendly control measures the bacterial strain B. vallismortis EXTN-1 (5 × 10 cells/
10

are a preferred strategy for plant disease control. Beneficial ml). The inducer of systemic disease resistance, benzo
microorganisms form an important component of Integ- (1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (BTH)
rated Pest Management (IPM). The use of rhizobacteria for (100 mM) (Lawton et al., 1996) was included in the
controlling soil borne plant diseases has been well treatments as a positive control. Bacterial inoculum was
documented (Weller and Cook, 1986). The beneficial prepared by harvesting cells from Tryptic Soy Agar (TSA)
plates (incubated at 28 C for 48 h) and re-suspended in 10
o

*Corresponding author. mM MgSO . Seed-bacterization was carried out by soaking

4

Phone) +82-31-290-0424, FAX) +82-31-290-0406 the seeds in the bacterial suspension for 4 h. Seeds soaked
E-mail) [email protected] in 10 mM MgSO served as control. The seeds were sown
4
 Bacillus vallismortis EXTN-1-Mediated Growth Promotion and Disease Suppression in Rice 279

on non-sterile seedbed nursery. Five weeks after seeding, Results

the rice seedlings were transplanted to the field. One set of
plants were maintained for measurement of growth para- Plant growth promotion by EXTN-1. Treatment of rice
meters and the experiment was carried out in field. The plants with EXTN-1 resulted in growth promotion in the
field trial was carried out with a completely randomized green house and field condition. In the green house, the root
block design (CRBD) and repeated in two consecutive proliferation was multifold in the EXTN-1-treated plants
years and the yield data obtained. Three other sets of the when compared with the control (Fig. 1). There was signifi-
same treatments were maintained in the green house for cantly increased growth in the EXTN-1-treated plants in the
challenge inoculation with the three different fungal field in terms of height (10.5% over control) and number of
pathogens. tillers (15.56% over control) (Fig. 2 & 3). Treatment with
Plant growth promotion by the bacterial strains. To
evaluate the effect of bacterial treatment, the growth of
plants was measured in terms of height of the plant and the
number of tillers upon a month after transplanting. Also the
yield parameters were recorded upon harvest.
Challenge inoculation with the fungal pathogens and
scoring disease development. M. grisea was plated on
rice-bran-agar for spore production. The plates were incu-
bated in diurnal condition at 25oC. The inoculum (2×105
conidia/ml) with Tween-20 (0.05% v/v) and gelatin (0.25%
w/v) was sprayed until leaves were covered with fine
droplets. From the first 20 h in darkness followed by Fig. 1.The root proliferation in EXTN-1-treated rice seedlings in
continuous light, inoculated plants were kept at 100% the nursery.
relative humidity (RH) until the end of the experiment.
Blast disease was quantified by counting the number of
acute lesions with a gray, sporulating center 5 days post
inoculation.
For challenge inoculation with R. solani, a rice grain:rice
hull (1:3) substrate was prepared and sterilized. Mycelial
plugs of R. solani were introduced into this mixture and
allowed to grow for 10 days at 28oC. Before inoculation
onto the rice plants, the culm region of the rice tillers of
each plant was covered with aluminium foil to form a cone.
For artificial inoculation, 40 g of the above inoculum was
dropped into the cone at the base of each rice plant. The Fig. 2. EXTN-1 mediated growth promotion in paddy field.
Different letters indicate significant differences among treatments
results on severity of sheath blight development were according to a least significant difference.
scored 15 days after inoculation. C. miyabeanus was raised
in potato dextrose agar (PDA) plates. Conidia were collect-
ed in sterile water. Inoculum was adjusted to 105 conidia/ml
and Tween-20 was added to a final concentration of 0.05%
(v/v). The plants were sprayed with the conidial suspension.
Plants were kept for 24 h in a dew chamber at 100% RH
under fluorescent light, with a 12 h photoperiod, and then
maintained in a greenhouse for 7 days. Percent disease (brown
spot) suppression obtained in treatments was compared.
Statistical analysis. All data were analyzed with JMP (a
PC-version of SAS) (SAS Institute. 1995) software. EXTN-1 treatment increase proliferation of rice-tillers in
Fig. 3.
Significant differences in treatments were determined using paddy field. Different letters indicate significant differences
Student’s T test as LSD at P = 0.05. among treatments according to a least significant difference.
280 Kyungseok Park et al.

Table 1. EXTN-1 mediated yield-increase in paddy

Yield (kg/10 a)
Treatments
Year 2004 Year 2005
Control 588.4 (0.0) a
681.6 (0.0) a

BTH 589.1 (0.1) a

747.6 (8.0) ab

EXTN-1 665.8 (11.6) b

780.0 (12.6) b

The same letters do not differ significantly at P = 0.05. The numbers

in parenthesis are percent values over control

BTH also resulted in enhanced height (8.23%) and number

of tillers (4.70%) in comparison with the control plants. The Fig. 4. EXTN-1 mediated sheath blight suppression in paddy field
increased yield in the EXTN-1 treated plants over the against R. solani.Bars corresponding to different letters differ
significantly at P = 0.05.
untreated control plants were 11.6 and 12.6% in the year
2004 and 2005 respectively (Table 1).
control had 83.02% diseased plants, the BTH treatment had
Disease suppression by EXTN-1.The B. vallismortis 63% disease plants while the EXTN-1 treatment had only
strain, EXTN-1 showed consistent disease protection 28.04% diseased plants.
against tested fungal pathogens. Compared with the control There was 11.54% reduction in the number of lesions
plants, the average reduction in rice blast was 52.11% after (brown spot) in the EXTN-1 treated plants compared to the
treatment with EXTN-1 (Table 2). ISR by treatment with untreated control (Fig. 4). Here treatment with BTH also
BTH also significantly reduced rice blast (by 78.81%). effected significant levels of disease suppression.
The sheath blight symptoms were significantly less in the
EXTN-1 treated plants (Fig. 4 and 5). When the untreated Discussion
Rice disease management strategies are mainly through the
Table 2. Disease (Rice blast and Brown spot) suppression effect- use of chemical pesticides. The persistent, injudicious use
ed in rice plants by treatments with EXTN-1 of chemicals has toxic effects on non-target organisms and
Treatment Lesion area Brown spot can cause undesirable changes in the environment. Bio-
Rice blast Lesion number/Leaf logical control agents have been used successfully in
Control 7.1 (0.00)
a
7.8 (0.00)
a
several pathogen/host systems to enhance plant growth and
BTH (1 mM) 1.5 (78.81) b
7.5 (3.85)
a
bring about disease control. The present study revealed the
EXTN-1 3.4 (52.11) b
6.9 (11.50) ab
potential of the rhizobacterial strain, EXTN-1 to bring
The same letters do not differ significantly at P = 0.05. The numbers about significant protection against three different fungal
in parenthesis are percent values over control pathogens, M. grisea, R. solani and C. miyabeanus and also

Fig. 5. EXTN-1 mediated sheath blight suppression by seed soaking with the bacteria.
 Bacillus vallismortis EXTN-1-Mediated Growth Promotion and Disease Suppression in Rice 281

to enhance plant growth promotion. Plant growth pro- mechanisms of disease suppression by rhizobacteria (Zehnder
motion by rhizobacteria could be due to the production of et al., 2001; Vessey, 2003), antibiosis being one of the
growth hormones, Indole-3-Acetic Acid (IAA) & Gibberellic important mechanism (Duffy and Defago, 1999). Homma
Acid (GA), suppression of deleterious organisms and (1984) observed certain soil bacteria degrade the conidia of
promotion of the availability of uptake of mineral nutrients C. miyabeanus. Also, chitinase-mediated cell wall lysis of
(Kloepper et al., 1980). The exact mechanisms are unclear R. solani has been reported by Lorito et al. (1998). Our
yet. recent studies proved that the different fractions of the
EXTN-1 treatment resulted in 52.11% reduction in rice metabolites produced by EXTN-1 considerably brought
blast, 83.02% reduction in sheath blight and 11.54% down the mycelial development of several fungal plant
decrease in brown spot symptoms in the crop. Reports are pathogens (data unpublished).
few on potential of a strain of biocontrol agent suppressing As EXTN-1 effectively brought about multiple disease
several disease of single crop, which is an important suppression in rice via different mechanisms of action, the
strategy to reduce the cost of the technology. The ability of strain could be effectively explored in large scale for the
EXTN-1 to suppress multiple diseases proves its multi- benefit of rice farming community.
farious modes of action. It could also be supposed that the
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