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Development of a scar marker for the detection of Ralstonia solanacearum (Smith) Yabuuchi

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Title Development of a scar marker for the detection of Ralstonia solanacearum (Smith) Yabuuchi
 
Creator Sarita.V.Gund
 
Contributor Narayan.Moger
 
Subject Plant Biotechnology
 
Description Bacterial wilt disease caused by Ralstonia solanacearum is one of the most important
constraint in the production of solanaceaous vegetable crops. It has been divided into five
races based on their host range, geographical distribution and ability to survive under
different environmental conditions.
One hundred fourteen strains of R. solanacearum were isolated from wilt affected
plants of brinjal, chilli and tomato were collected from Karnataka and Goa. The identification
and confirmation of collected isolates were done by cultural characteristics, pathogenicity test
and PCR amplification of 16S rDNA region. Leaf infiltration test on tobacco leaves was
performed for race identification of bacteria. It was found that among 114 isolates, forty two
were belonged to Race 2 and seventy two were belonged to Race 3. Ten isolates from each
group were screened with seventy five RAPD primers as an initial step to develop SCAR
markers.
It was found that OPAB-08 RAPD primer gave amplicon of size 600 bp only in Race
2 isolates. The species specific amplicon of size 950 bp and 900 bp was observed with OPA-
09 and OPG-07 primers respectively. These specific amplicon with respective RAPD primers
were sequenced to develop SCAR marker. Race 2 specific SCAR marker, RS-R2 and two
species specific SCAR markers RS-1 and RS-2 were developed. Further confirmation of
designed primer was done with all collected isolates (114 isolates). It was found that RS-R2
showed an amplicon of size 600 bp in Race 2 isolates. This indicated that the RS-R2 can be
used for unbiased identification of Race-2. Similarly, RS-1 and RS-2 primers gave an
amplicon of size 920 bp and 900 bp respectively in all collected isolates. So, these two SCAR
markers will help in the diagnosis of R. solanacearum.
 
Date 2016-11-09T18:17:04Z
2016-11-09T18:17:04Z
2010
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/84768
 
Format application/pdf
 
Publisher UAS, Dharwad