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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.
 
Creator P. GANESHAMOORTHI
 
Contributor S. C. DUBEY
 
Subject packaging, flavouring, animal husbandry, productivity, application methods, fats, proteins, economic systems, storage, milk products
 
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
 
Date 2016-06-24T14:00:31Z
2016-06-24T14:00:31Z
2013
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/67951
 
Format application/pdf
 
Publisher IARI, DIVISION OF PL ANT PATHOLOGY