In Potential Rhizobacteria Different isolates of the rhizobacteria

In a separate plated
medium, Xoo suspensions (approx. 106cfu/mL) were seeded uniformly using pour plate method. Five equidistant
wells were made in the previously plated medium seeded with Xoo by using a
sterilized cork borer and using a sterile syringe of a 0.1 mL of rhizobacteria
(108cfu/mL) was placed on
the wells. The assayed plates were incubated for 24 to 48 hours and the zones
of growth inhibition of Xoo around the rhizobacterial antagonists were measured
(Beric et al., 2012).

 

In vivo Screening of the Potential Rhizobacteria

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Different isolates of the rhizobacteria that inhibit the growth of Xoo under in vitro
test were separately applied to rice seeds of NSIC Rc224. The seeds were soaked
in 0.01% polyoxyethylene–sorbitan monolaurate (Tween20, Sigma, Aldrich) containing 108cfu/ mL of
the rhizobacterial antagonist, and then
kept at 28°C for 48 hours. The control
seeds (without rhizobacteria) were soaked in sterile water. Subsequently, both
control and treated seeds were sown in seedbeds
using natural field soil. Seedlings were transplanted into plastic pots after
21 days. At the time of transplanting, the seedlings were root dipped into
rhizobacterial antagonist suspension containing 108cfu/mL and
sterile distilled water for control. Foliar sprays of each rhizobacterial antagonist
were applied 28 and 35 days after transplant ng (DAT). Inoculation of bacterial
leaf blight (BLB) pathogen was done at 40 and 45 DAT using leaf cutting method
(Gnanamanickam, Velusamy, & Immanuel, 2013, pp.356362). Percent of disease
suppression was calculated by measuring the mean lesion length of the
inoculated control plants minus the mean lesion length of the rhizobacterial
antagonist treated plants over the mean lesion length of the inoculated control
plants multiply by 100. There were 20 random plants with three replications in
these experiments. Table 1 shows that the disease severity was evaluated 18
days after inoculation (DAI) up to 56 DAI with seven days interval following
the modified rating scale of International Rice Research Institute –Standard Evaluation
System for BLB percent infection following the formula: 

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