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Amplicon-based RNA interference targeting V2 gene of cotton leaf curl kokhran virus- burewala strain can provide resistance in transgenic cotton plants

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Title Amplicon-based RNA interference targeting V2 gene of cotton leaf curl kokhran virus- burewala strain can provide resistance in transgenic cotton plants
 
Creator Akram, Faheem
 
Contributor Kiani, Sarfraz
Butt, Afshan
Qayyum Rao, Abdul
Ahmad, Aftab
Ahmad Nasir, Idrees
Husnain, Tayyab
Mansoor, Shahid
Amin, Imran
Aftab, Shaheen
Zubair, Muhammad
Tahir, Muhammad Nouman
Akhtar, Sohail
Scheffler, Jodi
Scheffler, Brian
 
Description The conserved coat or V2 gene of begomoviruses is responsible for viral movement in the plant cells. RNAi technology was used to silence V2 gene for resistance against these viruses in transgenic plants. The transformation of the RNAi-based gene construct targeting V2 gene of CLCuKoV-Bur, cloned under 35S promoter, was done in two elite cotton varieties MNH-786 and VH- 289 using shoot apex cut method of gene transformation.
The transformation efficiency was found to be 3.75 and 2.88 % in MNH-786 and VH-289, respectively. Confirmation of successful transformation was done through PCR in T0, T1, and T2 generations using gene-specific primers. Transgenic cotton plants were categorized on the basis of
the virus disease index in T1 generation. Copy number and transgene location were observed using FISH and karyotyping in T2 generation which confirmed random integration of V2 RNAi amplicon at chromosome 6 and 16. Real-time quantitative PCR analyses of promising transgenic lines showed low virus titer compared to wild-type control plants upon challenging them with viruliferous whiteflies in a contained environment. From the results, it was concluded that amplicon V2 RNAi construct was able to limit virus replication and can be used to control CLCuV
in the field.
 
Date 2016-10-15
2017-01-19T10:51:47Z
2017-01-19T10:51:47Z
 
Type Journal Article
 
Identifier https://mel.cgiar.org/reporting/download/hash/Fx4LibNj
Faheem Akram, Sarfraz Kiani, Afshan Butt, Abdul Qayyum Rao, Aftab Ahmad, Idrees Ahmad Nasir, Tayyab Husnain, Shahid Mansoor, Imran Amin, Shaheen Aftab, Muhammad Zubair, Muhammad Nouman Tahir, Sohail Akhtar, Jodi Scheffler, Brian Scheffler. (15/10/2016). Amplicon-based RNA interference targeting V2 gene of cotton leaf curl kokhran virus- burewala strain can provide resistance in transgenic cotton plants. Molecular Biotechnology, 58 (12), pp. 807-820.
https://hdl.handle.net/20.500.11766/5525
Open access
 
Language en
 
Rights CC-BY-NC-4.0
 
Format PDF
 
Publisher Humana Press
 
Source Molecular Biotechnology;58,(2016) Pagination 807-820