摘要
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1100 Psi＋9 cm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T0植株所结种子长成的穗行喷200 mg/kg的PPT进行抗性筛选,获得了105个抗性株,究其来源为42个T0株,由此计算抗性率为5.2%。用PCR检测T1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。
In order to set up a simple and easy method for genetic transformation of wheat,which do not depend on tissue culture and resistance selection,experiments of target-preparation of shoot apical point of buds for particle delivery and parameter-optimization of bombardment were conducted with a cultivar of Shi4185 as a recipient,and with gus,badh/bar as foreign genes.A suitable way was successfully founded to prepare shoot apical point targets from germinated seeds of wheat to bombardment,and the optimum parameters combination was determined as 1 100 Psi ＋ 9 cm. Statistics of gus detection of transient expression with 70 × 3 batches indicated that 22.9 % - 35.7 % of targets and 5.7 % - 8.6 % of shoot apical points were transformed. With the bombardment of pABH9 containing badh/bar, 802/1 000 targets devel oped to normal plants. 105 resistant Tl plants were obtained by spraying PPT of 200 ppm to the seedlings of seeds corresponding to 42 To plants with a resistance rate of 5.2 %. PCR analysis showed that 6 T1 plants were positive. Preliminarily, the technique to transform shoot apical point of wheat buds by particle bombardment was established.
出处
《华北农学报》
CSCD
北大核心
2009年第5期1-6,共6页
Acta Agriculturae Boreali-Sinica
基金
转基因生物新品种培育重大专项(2009ZX08010-011B)
关键词
小麦
芽生长点
基因枪
转化
Wheat
Shoot apical point
Particle bombardment
Transformation
