TY - JOUR
T1 - Integration of a hairpin RNA-encoding gene derived from Tobacco streak virus confers resistance in groundnut (Arachis hypogaea L.) against peanut stem necrosis disease
AU - Reddy, Madam Gurivi
AU - Senthilraja, Chinnaiah
AU - Adhithya, Rangasamy
AU - Satya, Vijayalakshmi Kothandaraman
AU - Kokiladevi, Easwaran
AU - Sudhakar, Durailagaraja
AU - Rabindran, Ramalingam
AU - Velazhahan, Rethinasamy
PY - 2016/10
Y1 - 2016/10
N2 - The feasibility of controlling peanut stem necrosis disease caused by Tobacco streak virus (TSV) in groundnut (Arachis hypogaea L.) was explored by expressing double-stranded RNA of the replicase (Rep) gene of TSV in groundnut through genetic engineering. A hairpin (hp) RNAi construct containing 535-bp sense and antisense TSV-Rep sequences flanking a 742-bp spacer sequence (Pdk intron) under the control of the constitutive Cauliflower mosaic virus 35S promoter was made in the binary vector pART27. This chimeric gene construct was then mobilized into Agrobacterium tumefaciens strain LBA4404 via triparental mating using pRK2013 as a helper. Cotyledon explants of groundnut cultivar TMV-7 were transformed with A. tumefaciens harboring the hpRNA cassette. The presence of the transgene in the transgenic plants was confirmed up to T3 generation by PCR amplification of the 535-bp fragment of TSV-Rep gene. The bioassay results indicated that necrotic lesions were observed on the leaves of the wild-type plants 7-9 days after inoculation with TSV and stem necrosis appeared 16-20 days after inoculation, whereas the transgenic plants did not develop symptoms until harvest. ELISA results indicated that the wild-type plants inoculated with TSV recorded the highest virus concentration as compared to the transgenic lines.
AB - The feasibility of controlling peanut stem necrosis disease caused by Tobacco streak virus (TSV) in groundnut (Arachis hypogaea L.) was explored by expressing double-stranded RNA of the replicase (Rep) gene of TSV in groundnut through genetic engineering. A hairpin (hp) RNAi construct containing 535-bp sense and antisense TSV-Rep sequences flanking a 742-bp spacer sequence (Pdk intron) under the control of the constitutive Cauliflower mosaic virus 35S promoter was made in the binary vector pART27. This chimeric gene construct was then mobilized into Agrobacterium tumefaciens strain LBA4404 via triparental mating using pRK2013 as a helper. Cotyledon explants of groundnut cultivar TMV-7 were transformed with A. tumefaciens harboring the hpRNA cassette. The presence of the transgene in the transgenic plants was confirmed up to T3 generation by PCR amplification of the 535-bp fragment of TSV-Rep gene. The bioassay results indicated that necrotic lesions were observed on the leaves of the wild-type plants 7-9 days after inoculation with TSV and stem necrosis appeared 16-20 days after inoculation, whereas the transgenic plants did not develop symptoms until harvest. ELISA results indicated that the wild-type plants inoculated with TSV recorded the highest virus concentration as compared to the transgenic lines.
KW - Agrobacterium tumefaciens
KW - Arachis hypogaea
KW - Genetic engineering
KW - Replicase
KW - Rnai
KW - Tobacco streak virus
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U2 - 10.1007/s41348-016-0039-7
DO - 10.1007/s41348-016-0039-7
M3 - Article
AN - SCOPUS:84986612933
SN - 1861-3829
VL - 123
SP - 205
EP - 214
JO - Journal of Plant Diseases and Protection
JF - Journal of Plant Diseases and Protection
IS - 5
ER -