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Pak. J. Bot., 47(1): 211-216, 2015. |
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Updated: 24-02-15 | ||||
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AGROBACTERIUM MEDIATED TRANSFORMATION OF BRASSICA JUNCEA (L.) CZERN. WITH CHITINASE GENE CONFERRING RESISTANCE AGAINST FUNGAL INFECTIONS
BASHIR AHMAD1*, AMBREEN1, MUHAMMAD SAYYAR KHAN2 ALI HAIDER3 AND IBRAR KHAN1
Abstract: Brassica juncea (Czern & Coss., L.) is an important oilseed crop. Since it is attacked by several bacterial and fungal diseases, therefore, we developed an easy and simple protocol for the regeneration and transformation of B. juncea variety RAYA ANMOL to give rise to transgenic plants conferring resistance against various fungal diseases. The transformation was carried out using Agrobacterium with Chitinase gene. This gene was isolated from Streptomyces griseus HUT6037. We used two types of explants for transformation i.e. hypocotyls and cotyledons. Only hypocotyls explants showed good results regarding callus initiation. Different hormonal concentrations were applied i.e. BAP 2, 4 and 6 mgL-1 and NAA 0.1, 0.2 and 0.3 mgL-1. However, high transformation efficiency was observed by supplementing the medium with combination of 2 mgL-1 BAP and 0.2 mgL-1 for initiation of callus. Similarly 10 mgL-1 kanamycin and 200 mgL-1 cefotaxime also proved successful for the selection of transformed callus. In order to confirm the presence of transgenic callus Polymerase chainreaction was performed using specific primers for Chitinase gene.1Centre of Biotechnology and Microbiology, University of Peshawar, Khyber PakhtunKhwa, Pakistan 2Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, Pakistan. 3PGMI Lady Reading Hospital Peshawar, Pakistan *Corresponding author e-mail: * bashirdr2001@yahoo.com |
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