Anastomosis grouping, molecula r characterization and development of markers for detection of Rhizoctonia solani causing wet root rot in chickpea
KrishiKosh
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Title |
Anastomosis grouping, molecula r characterization and development of markers for detection of Rhizoctonia solani causing wet root rot in chickpea
Ph.D. |
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Creator |
P. GANESHAMOORTHI
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Contributor |
S. C. DUBEY
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Subject |
packaging, flavouring, animal husbandry, productivity, application methods, fats, proteins, economic systems, storage, milk products
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Description |
T-8828
Chickpea ( Cicer arietinum L.) is one of the most important pulse crops grown in I ndia. Among the diseases affecting chickpea, wet root rot caused by Rhizoctonia solani Kühn is one of the major yield limiting factors and it is prevalent in almost all chickpea growing states of India. Fifty chickpea isolates of R. solani collected from d ifferent states of India were included in the present study. Most of the isolates produced light brown mycelium with fluffy growth and were grouped into six categories on the basis of their growth rate. Sclerotia formed in different isolates were highly va riable innumber as well as size. The isolates were grouped into five and six categories based on the number and size of sclerotia, respectively. The sclerotial were scattered in most of the isolates followed by in circle at middle and periphery in Petri pl ates. Irrespective of host of origin and anastomosis groupings (AGs), the isolates were proved to be pathogenic on chickpea, urdbean ( Vigna mungo ) and mungbean ( Vigna radiata ). The isolates were also variable in respect of their virulence and they were gr ouped into three categories as high, moderate and less virulent isolates. Based on AGs the isolates were variable in their hyphal anastomosis reactions and they were grouped into seven AGs as AG1, AG2 - 2, AG2 - 2LP, AG2 - 3, AG3, AG4 and AG5. Neighbor - joining tree analysis categorized the isolates into 8 major groups using three sets of markers, namely, RAPD, ISSR and SSR evaluated. These clusters of the isolates were partially corresponding to the geographical origin and AGs. Inter transcribed spacer (ITS) reg ions sequences of 12 representative isolates of R. solani , namely, RKNG9 AG 1, RAPG11 AG 3, RKNG10 AG 2 - 2L, RAPG14 AG 2 - 3, RAPG9 68 AG 2 - 2, RMHG23 AG 3, RKNG11 AG 4, RTNG4 AG 5, RUPG107 AG 2 - 2, RAPG13 AG 2 - 3, RAPG15 AG 4 and RTNG7 AG 1 were analyzed. The sequ ence homology between 12 isolates R. solani (7 different AGs) range of 93 to 100% in conserved regions of ITS. The phylogenetic analysis showed the Indian population of R. solani had variable ITS sequences, therefore, grouped separately in one major cluste r. The RAPD primer OPA 11 consistently amplified ≈1700bp product during PCR only from the DNA of R. solani isolated from chickpea. The common DNA fragment was sequenced and used to design a pair of oligonucleotide primers amplifying a 285bp sequence character ized amplified region (SCAR). A SCAR primer was found specific to R. solani . The detection sensitivity of R. solani was 0.5 ng for genomic DNA and 5 ng for DNA extracted from infected chickpea roots. Quantitative analysis using real time PCR assay clearly showed that the SCAR primer set was able to detect R. solani up to 1 pg genomic DNA obtained from infected chickpea roots. This is the first study on cultural, pathogenic behaviour, molecular characterization and development of SCAR marker for detection of chickpea isolates of R. solani |
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Date |
2016-06-24T14:00:31Z
2016-06-24T14:00:31Z 2013 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/67951
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Format |
application/pdf
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Publisher |
IARI, DIVISION OF PL ANT PATHOLOGY
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