大豆β-1,3-葡聚糖酶基因转化烟草及抗病性的研究

Tobacco Transformation of Soybean Beta-1, 3-glucanase Gene and Disease Resistance Assay

构建了大豆β－１，３－葡聚糖酶（β－１，３－ｇｌｕｃａｎａｓｅ）基因的植物表达载体ｐＢＩ１２１－ｇｌｕ，并通过直接转化方法将其导入根癌农杆菌（Ａ．ｔｕｍｅｆａｃｉｅｎｓ）ＬＢＡ４４０４受体菌中，构建了用于植物遗传转化的工程菌株ＬＢＡ４４０４（ｐＢＩ１２１－ｇｌｕ），并以烟草为转化对象进行了遗传转儿，获得了大量再生的转基因烟草．ＰＣＲ，ＰＣＲ－Ｓｏｕｔｈｅｍ以及Ｓｏｕｔｈｅｍ杂交检测结果表明目的基因已整合到烟草基因组中．转基因植株苗期抗立枯病实验表明，部分转化β－１，３－葡聚糖酶基因的工程烟草对立枯丝核菌（Ｒｈｉｚｏｃｔｏｎｉａ ｓｏｌａｎｉ．）表明出不同程度的抗性提高．

In this paper, plant expression vector pBI121-glu is constructed by cloning the beta-1, 3-glucanase gene to pBI121 plasmid. Then, the engineered strain LBA4404 (pBI121 -glu) is constructed by conducting the pBI121-glu to A. tumefaciens LBA4404 and is used to transform tobacco. The PCR, PCR -Southern and Southern blotting analysis indicate that the soybean beta-1, 3-glucanase gene has been integrated into the genome of tobacco. The infection assay indicates that parts of beta-1, 3-glucanase transgenic tobacco exhibit enhanced resistance against fungal pathogen Rhizoctonia solani.