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首页> 外文期刊>African Journal of Biotechnology >An improved Agrobacterium mediated transformation in tomato using hygromycin as a selective agent
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An improved Agrobacterium mediated transformation in tomato using hygromycin as a selective agent

机译：以潮霉素为选择剂的改良农杆菌介导的番茄转化

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Bacterial wilt is a devastating disease of tomato crop throughout the world. This disease is very dangerous in hot and humid regions, where it spreads with the irrigation water to whole field within days, which resulted in severe decline in yield. Two varieties of tomato were used for developing bacterial wilt resistance. Riogrande responded more efficiently as compared to Roma. Regeneration frequency of 90.6% was achieved when leaf discs were used and 82.5% when hypocotyls were used as explant for tomato cv. Riogrande.? While 65.4% regeneration was achieved from hypocotyls and 72.6% regeneration was obtained when leaf discs were used as explant for cv. Roma. Explants were co-cultivated with?Agrobacterium?strain EHA101 containing a binary vector pTCL5, having hygromycin phosphotransferase (HPT) gene which confers resistance to hygromycin and β-glucuronidase (GUS) gene in addition to Xa21 gene.? Hygromycin (25 mg/l) was used as selectable marker while GUS is a reporter gene. Acetosyringone (50?mM) enhanced transformation effeciency. Preselection period of 7 days was found to be indispensable for successful transformation of tomato crop. Transformation efficiency of 24% was observed for Riogrande and 8% for Roma.? Molecular analysis of transgenic plants produced was carried out for hygromycin resistance gene. Transgenic plants contained the expected band of 670 bp.?? PCR analysis confirmed the foreign DNA in to the plant genome.
机译：细菌性枯萎病是全世界番茄作物的毁灭性疾病。这种疾病在炎热和潮湿的地区非常危险，在几天之内它随灌溉水扩散到整个田地，导致产量严重下降。番茄的两个变种被用于发展抗青枯病。与罗姆人相比，里奥格兰德的回应更为有效。当使用叶盘时，再生频率达到90.6％;当使用下胚轴作为番茄cv的外植体时，再生频率达到82.5％。里奥格兰德。当使用叶盘作为cv的外植体时，从下胚轴获得了65.4％的再生，并且获得了72.6％的再生。罗马。将外植体与含有二元载体pTCL5的农杆菌菌株EHA101共培养，该载体具有除Xa21基因外还具​​有潮霉素抗性的潮霉素磷酸转移酶（HPT）基因和β-葡糖醛酸糖苷酶（GUS）基因。潮霉素（25 mg / l）用作选择标记，而GUS是报告基因。乙酰丁香酮（50？mM）增强了转化效率。发现7天的预选期对于番茄作物的成功转化是必不可少的。 Riogrande的转化效率为24％，Roma的转化效率为8％。对产生的潮霉素抗性基因进行了转基因植物的分子分析。转基因植物的预期条带为670 bp。 PCR分析证实了外源DNA进入了植物基因组。

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