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基因枪法向小麦导入几丁质酶基因的研究
Studies on transferring chitinase gene into wheat via microprojectile bombardment
【摘要】 利用基因枪法,以菜豆几丁质酶基因转化小麦幼胚愈伤组织。在轰击压力1300psi,轰击距离6cm、100μg金粉/枪和轰击距离9cm、150μg金粉/枪的2种处理条件下,获得4株春小麦东农7742转化植株,转化频率分别为0.36%和0.56%。经PCR和PCR-Southern杂交分析,证实菜豆几丁质酶基因已整合到T0和T1小麦基因组中。采用氨基葡萄糖法测定几丁质酶活力,结果表明,转基因小麦的几丁质酶活力明显高于对照株;转基因植株对白粉病症状减缓,并获得一株赤霉菌接种未扩展的转基因T1植株。
【Abstract】 Using embryogenic calli from young embryos as explants,bean chitinase gene was transferred into spring wheat plants by microprojectile bombardment.Four transgenic plants were obtained under the conditions of 6 cm distance,100 μg of gold particles per bombardment and 9 cm,150 μg.Both bombardment pressures were 1 300 psi.Transformation frequency of the two treatments was 0.36% and 0.56% respectively.It was confirmed that the bean chitinase gene had been integrated into T0 and T1 wheat genomes by means of PCR and Southern analysis.The activity of chitinase were determined by measuring the amount of aminoglucose.The result indicated that activity of chitinase in transgenic plant was much higher than that of CK.The symptoms of some transgenic plants to powder mildew were weaker than that of CK.No spreading disease of wheat scab was observed in one transgenic T1 plant.
【Key words】 chitinase gene; transformation; wheat; microprojectile bombardment;
- 【文献出处】 西北植物学报 ,Acta Botanica Boreali-occidentalia Sinica , 编辑部邮箱 ,2003年01期
- 【分类号】S512.1
- 【被引频次】63
- 【下载频次】216