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Production of transgenic potato exhibiting enhanced resistance to fungal infections and herbicide applications
Authors:Raham Sher Khan  Rinaldi Sjahril  Ikuo Nakamura  Masahiro Mii
Affiliation:(1) Laboratory of Plant Cell Technology, Graduate School of Horticulture, Chiba University, 648 Matsudo, Matsudo Chiba, 271-8510, Japan;(2) Institute of Biotechnology and Genetic Engineering, NWFP Agricultural University, Peshawar, Pakistan;(3) Faculty of Agriculture and Forestry, Hasanuddin University, Jl. Perintis Kemerdekaan Km 10, Makassar, 90245, South Sulawesi, Indonesia
Abstract:Potato (Solanum tuberosum L.), one of the most important food crops, is susceptible to a number of devastating fungal pathogens in addition to bacterial and other pathogens. Producing disease-resistant cultivars has been an effective and useful strategy to combat the attack of pathogens. Potato was transformed with Agrobacterium tumefaciens strain EHA101 harboring chitinase, (ChiC) isolated from Streptomyces griseus strain HUT 6037 and bialaphos resistance (bar) genes in a binary plasmid vector, pEKH1. Polymerase chain reaction (PCR) analysis revealed that the ChiC and bar genes are integrated into the genome of transgenic plants. Different insertion sites of the transgenes (one to six sites for ChiC and three to seven for bar) were indicated by Southern blot analysis of genomic DNA from the transgenic plants. Expression of the ChiC gene at the messenger RNA (mRNA) level was confirmed by Northern blot analysis and that of the bar gene by herbicide resistance assay. The results obviously confirmed that the ChiC and bar genes are successfully integrated and expressed into the genome, resulting in the production of bialaphos-resistant transgenic plants. Disease-resistance assay of the in vitro and greenhouse-grown transgenic plants demonstrated enhanced resistance against the fungal pathogen Alternaria solani (causal agent of early blight).
Keywords:Solanum tuberosum   Transformation   Alternaria solani   Chitinase C   Bar gene
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