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. 2016 May;23(3):358-62.
doi: 10.1016/j.sjbs.2014.11.013. Epub 2014 Nov 20.

Development of Cotton leaf curl virus resistant transgenic cotton using antisense ßC1 gene

Sayed Sartaj Sohrab  1 , Mohammad A Kamal  1 , Abdul Ilah  2 , Azamal Husen  3 , P S Bhattacharya  4 , D Rana  4
Affiliations

Development of Cotton leaf curl virus resistant transgenic cotton using antisense ßC1 gene

Sayed Sartaj Sohrab et al. Saudi J Biol Sci. 2016 May.

Abstract

Cotton leaf curl virus (CLCuV) is a serious pathogen causing leaf curl disease and affecting the cotton production in major growing areas. The transgenic cotton (Gossypium hirsutum cv. Coker 310) plants were developed by using βC1 gene in antisense orientation gene driven by Cauliflower mosaic virus-35S promoter and nos (nopaline synthase) terminator and mediated by Agrobacterium tumefaciens transformation and somatic embryogenesis system. Molecular confirmation of the transformants was carried out by polymerase chain reaction (PCR) and Southern blot hybridization. The developed transgenic and inoculated plants remained symptomless till their growth period. In conclusion, the plants were observed as resistant to CLCuV.

Keywords: A. tumefaciens, Agrobacterium tumefaciens; CLCuV resistance; CLCuV, Cotton leaf curl virus; CP, coat protein; Gene transfer; MS, Murashige and Skoog’s; Molecular verification; PCR, polymerase chain reaction; PDR, pathogen derived resistance; Transgenic cotton plants; Virus resistance.

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Figures

Figure 1
Figure 1
Schematic representation of the binary vector pBI 121 carrying full length of 35S CaMV promoter, NPTII, NOS promotor and terminator and βC1 gene in antisense orientation. LB, Left border; RB, Right border.
Figure 2
Figure 2
Stage of transformed cotton regeneration and plant development. (A) Hypocotyls. (B) Somatic embryos. (C) Emerged cotton plants. (D) Elongated plants. (E) Rooted plants. (F) Hardened cotton plant.
Figure 3
Figure 3
Electrophoretic analysis of putative cotton transgenic showing PCR amplification of βC1 gene using specific primers. Lane 1–19. Putative plants; M: 1 kb ladder.
Figure 4
Figure 4
Southern blot analysis of transgenic cotton plants. Lane 1–3; Independent transgenic cotton lines Lane 4; Non-transformed cotton. 5: Positive control (Cloned DNA).
See this image and copyright information in PMC

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