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A Simplified Protocol for Reversing Phenotypic Conversion of Ralstonia solanacearum during Experimentation.

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Title A Simplified Protocol for Reversing Phenotypic Conversion of Ralstonia solanacearum during Experimentation.
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Creator Sahu, P.K.; Singh, S.; Gupta, A.; Singh, U.B.; Paul, S.; Paul, D.; Kuppusamy, P.; Singh, H.V.; Saxena, A.K.
 
Subject Ralstonia solanacearum; virulence; virulence induction protocol;Phyllanthus emblica;nutrient deprivation
 
Description Not Available
Ralstonia solanacearum has the problem of losing the virulence in laboratory conditions, during prolonged experimentation. Since pure colonies of R. solanacearum contain cell fractions differing in virulence, it was considered worthwhile to find a way of selecting the cells with lower attenuation. Therefore, a methodology for inducing virulent-type colonies occurrence in Ralstonia solanacearum was developed. Methods: Nutrient gradient was created by swabbingR. solanacearumculture in a slanted KMTTC medium, and Phyllanthus emblica extract was givenby well diffusion. Live–dead cell imaging using BacLight, effects of ascorbic acid on cell viability, and production of virulence factors (exopolysaccharides, cellulase, and pectinase) supported this hypothesis. The tagging of R. solanacearum with green fluorescent protein and further confocal scanning laser microscopic visualization confirmed the colonization in vascular bundles of tomato. Results:P. emblica extract suppressed R. solanacearum initially in well diffusion, but further developed virulent-type colonies around the wells. Nutrient deprivation was found to have synergistic effects with P. emblica extract. The converted fluidal (virulent type) colonies could be able to colonize vascular bundles and cause wilting symptoms. Conclusion: This method will be useful in the laboratories working on biocontrol of R. solanacearum for maintaining virulent-type colonies. Moreover, it could form the basis for studies on the stability of phenotypic conversion and cell fractions in R. solanacearum.
The experimental facilities and funding received from ICAR-NBAIM, Mau, UP, India grantnumber IXX11672.
 
Date 2021-07-28T06:27:01Z
2021-07-28T06:27:01Z
2020-06-15
 
Type Article
 
Identifier Not Available
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http://krishi.icar.gov.in/jspui/handle/123456789/50558
 
Language English
 
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Publisher International Journal of Environmental Research and Public Health