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小麦芽生长点的基因枪转化技术研究

DOI: 10.7668/hbnxb.2009.05.001, PP. 1-6

Keywords: 小麦,芽生长点,基因枪,转化

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Abstract:

为建立简易的不依赖组织培养和抗性筛选的小麦转基因技术,以小麦品种石4185为对象,以gus和badh/bar为外源基因,围绕适于基因枪转化的种子芽生长点受体的制备、基因枪轰击参数的优化等开展了试验,建立了“用解剖刀去除种子根、用镊子把芽鞘掰去、用镊尖扫去包盖生长点的未展开叶、将带生长点的部分与胚乳分离”为特点的受体制备方法;确定1100Psi+9cm为最佳轰击参数组合。对轰击后的小麦芽生长点(70个×3批)进行gus基因瞬时表达检测,以受体数统计时,转化率为22.9%~35.7%;以生长点数统计时,转化率为5.7%~8.6%。用含badh/bar的pABH9质粒转化1000个受体,发育成了802个T0植株,对0植株所结种子长成的穗行喷200mg/kg的PPT进行抗性筛选,获得了105个抗性株,究其来源为42个0株,由此计算抗性率为5.2%。用PCR检测1植株,6个呈阳性。初步建立了基因枪法转化小麦种子芽生长点的技术。

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