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Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton
Published on: August 20, 2011
Cotton Mi-1.2-like Gene: A potential source of whitefly resistance
Muhammad Qasim Aslam1, Athar Hussain2, Afzal Akram1
1National Institute for Biotechnology and Genetic Engineering (NIBGE), college of Pakistan Institute of Engineering and Applied Sciences (PIEAS), Faisalabad, Pakistan.
Abstract:
Whitefly (Bemisia tabaci) inflicts tremendous yield losses to cotton crops in many parts of the world by sap-sucking and transmitting viral diseases. The tomato-associated Mi-1.2 gene has been successfully deployed in tomato cultivars to attain whitefly resistance. In the current study, putative Mi-1.2-like orthologs were identified in five whitefly hosts and functionally validated through virus-inducing gene silencing (VIGS) in cotton plants. The expression profiling and qPCR results depicted differential regulation of the Mi-1.2-like gene in various tissue types and under different biotic and abiotic stresses, especially in whitefly susceptible and resistant cotton plants. The upregulation of the Mi-1.2-like gene (Gadrp RPP-13 Like gene) was observed at 24 h and 48 h post-whitefly exposure (PWFE) in whitefly resistant (FDH-228) and tolerant (Mac7) cotton plants as compared to susceptible plants of Coker-312. However, delayed expression was recorded at 72 h of PWFE in Coker-312 plants. In TRV based gene silencing experiment, silencing of the Mi-1.2-like gene, significantly enhanced the whitefly infestation on both whitefly-resistant and susceptible cotton genotypes. Based on these results, we conducted the evolutionary analysis of Mi-1.2-like orthologs among cotton, cassava, tomato, papaya, and cucumber hosts. This indicated that cotton associated Mi-1.2 like gene has a close relation with cassava and tomato. These results suggested that Mi-1.2-like R genes could be the potential candidate for deriving whitefly resistance response in cotton plants.

