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Screening of mungbean and ricebean entries for Mungbean Yellow Mosaic Virus Resistance through Agroinfection. M. Sudha, Research Scholar. Department of Plant Molecular Biology and Biotechnology, Centre for Plant Molecular Biology, Tamil Nadu Agricultural University, Coimbatore. - PowerPoint PPT Presentation
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Screening of mungbean and ricebean entries for Mungbean Yellow Mosaic Virus Resistance through Agroinfection
Department of Plant Molecular Biology and Biotechnology,Centre for Plant Molecular Biology, Tamil Nadu Agricultural University,
Coimbatore.
M. Sudha,Research Scholar
- Dr. Norman Borlaug
White flies
Yellow mosaic disease in pulses
Agroinfection is an effective method by which infectious viral clones are introduced into plants using Agrobacterium tumefaciens
A partial tandem repeat of a virus is constructed and placed in the T-DNA of a binary vector and introduced into A. tumefaciens
Upon inoculation of plants with the Agrobacterium strains and transfer of the T-DNA, the viral genome is released in the plant cells from the partial tandem repeat of the virus
AgroinfectionAgroinfection
Infectivity analysis of two variable DNA B components of Mungbean yellow mosaic virus-Vigna in Vigna mungo and Vigna radiata
Mungbean yellow mosaic virus-Vi Agroinfection by codelivery of DNA A and DNA B from one Agrobacterium Strain
Analysis of an isolate of Mungbean yellow mosaic virus (MYMV) with a highly variable DNA B component
Infectivity analysis of a soybean isolate of Mungbean yellow mosaic Indiavirus by agroinoculation
Studies on Agroinfection in pulses
Agrobacterium strains
VA221 - (KA 22 DNA B + KA30 DNA A)
VA239 - (KA 27DNA B + KA30 DNA A)
These strains cloned from infected black gram were obtained from Dr. K. Veluthambi (Madurai Kamaraj University)
MYMV Symptom expression in Co 5 black gram for VA 221 and VA 239 strains
Susceptible reaction to MYMV through agroinfection in mungbean LM 122 (field susceptible)
Field screening of germplasm entries against MYMV
S.No Germplasm Number of entries
screened
Number of resistant entries
1. Mungbean 413 28
2. Ricebean 24 24
Agroinfection screening of mungbean entries against MYMVAgroinfection screening of mungbean entries against MYMV
ReactionReaction Number of entriesNumber of entries
Resistant to VA221 3Resistant to VA 239 1
Susceptible 24Total 28
S.No Entries S.No Entries S.No Entries
1 KMG 149 11 ML 131 21 PDM 12-1
2 PUSA 9071 12 ML 1108 22 COGG 917
3 PUSA 9030-BOK 13 EC 27087 23 KMG 189
4 SEL 2 14 ZKM 26 24 SP 84
5 PLS 3087 15 VGG 24 25 SU 4-147
6 PLS 303-B 16 SU 4-145 26 VGG 148
7 VBM-92-3-B 17 SU4-139 27 VGG 04-006
8 NL 5 18 SU4-148 28 ML 1165
9 PDM-84-1464 19 VGG04-013
10 COGG-924 20 ML 818
Resistant to VA221 strain Resistant to VA239 strain
List of field resistant mungbean entries subjected to agroinfectionList of field resistant mungbean entries subjected to agroinfection
MYMV Symptom(s) expression in mungbean – ML 1108 (VA221 resistant )
MYMV Symptom(s) expression in mungbean - ML 818 (VA239 resistant )
FOR VA221 Strain
703 bp
PCR amplification in MYMV agroinoculated mungbean entries
FOR VA239 Strain
Positive control
1-20 : Agroinoculated entriesPrimer used – Coat protein region of DNA A
100bp 1 2
1. CO 5 blackgram-VA2212. CO5 blackgram-VA239
100bp 1 2 3 4 5 6 7 8 9 10 11 12 13 14151617181920
100bp 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17181920
Agroinfection screening of ricebean entries against MYMV
Reaction Number of entries
Resistant 24Susceptible 0
Total 24
S.No Entries S.No Entries
1 V. umbellata TNAU red 13 LRB 89 green
2 LRB 41 14 LRB 11
3 LRB 297 15 LRB 29 l.green mottled
4 LRB 110 Yellow 16 LRB 35
5 LRB 113 original 17 TNAU Yellow
6 LRB 23 18 LRB 111
7 V. Umbellata TNAU green 19 LRB 104
8 LRB 113 black 20 LRB 282
9 LRB mottled green 21 LRB 5 mottled
10 V. Umbellata dark green 22 LRB 55
11 LRB tendrills 23 LRB 109 green
12 LRB 15 green 24 LRB 15 green
List of Rice bean entries subjected to agroinfection
Resistance reaction to MYMV through agroinfection in ricebean LRB 297
Resistant symptom expression in Ricebean TNAU RED along with positive check blackgram Co 5 through agroinfection
1000bp 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
703 bp
Lane1 : Uninoculated Control TNAU red Lane 2 : First dayLane 3 : Second dayLane 4 : Nineth dayLane 5 : Tenth dayLane 6 : Twelth dayLane 7 : Fifteenth day Lane 8 : Seventeenth dayLane 9 :Twenty-second day
Lane10 : Immediately after inoculation( embryos alone)Lane11 : Third day ( embryos alone)Lane12 : Fourth day (embryos alone)Lane13 : Nineth dayLane14 : Tenth dayLane15 :Twelth dayLane16 : Fifteenth day Lane17 : Seventeenth dayLane 18 : Eighteenth dayLane19 : Twentith dayLane 20 : Twenty-first day
PCR amplification of agroinoculated ricebean TNAU RED for PCR amplification of agroinoculated ricebean TNAU RED for VA221 strainVA221 strain
700 bp
800 bp
Lane 1 : 2 month old lower leavesLane 2 : 1 month old upper leavesLane 3 : 1 month old lower leavesLane 4 : Twenty four days old leavesLane 5 : Fifteenth days old leaves
100bp 1 2 3 4 5
703 bp700 bp
800 bp
PCR amplification of agroinoculated ricebean TNAU RED VA221 strain at different growth stages
Ongoing works…
Real-Time PCR assays for detection and quantification of MYMV viruses for the identified resistant mungbean entries which were strain specific
Identification of resistant proven donors for MYMV resistance through agroinfection which can be used as a genetic material