Transformation and Experssion of the Coat Protein genes of Potato virus Y and Potato virus S Iranian isolates in Solanum tubersum var Agria Lines

Document Type : research

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Abstract

In this project, infected plants with Potato virus Y (tobacco veinal necrosis strain: PVYN) and Potato virus S (PVS) from Lalezar in Kerman province were isolated by ELISA test. ELISA positive samples were inoculated on test plants. The coat protein (CP) genes from these two virus isolates were amplified with specific primers contained BamH1 and Sac1 restriction sites, cloned into pTZ57R/T plasmid separately and then transformed to E. coli competent cells. Two samples of each genes were sequenced and compared with the sequences available in the Gen Bank, and then cloned in pBI121 vector and transformed to E.coli competent cells. Accuracy of these constructs tested with digestion by BamH1 and Sac1 and PCR reaction with specific primers. These two constructs (pBI-PVS-CP and pBI-PVYN-CP) were transformed in Agrobacterium tumefacience (GV3850) including rifamepcin resistant gene and injected in to micro tuber disks. Transgenic plantlets were rooted and transferred to pots and tested by PCR and ELISA for transformation and expression of the CP genes. Out of 20 lines, only 6 and 4 lines were contained CP genes of PVYN-CP and PVS-CP, respectively. Moreover, in 2 lines both CP genes were identified.

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Agricultural Biotechnology
Volume 4, Issue 1 - Serial Number 1
February 2014
Pages 35-45

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