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。
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 target2preparation 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 optimumparameters combination was determined as 1100Psi+9cm.Statistics of gus detection of transient expression with 70×3 batches indicated that 2219%-3517% of targets and 5.7%-8.6% of shoot apical points were transformed.With the bombardment of pABH9 containing badh/bar,802/1000 targets developed to normal plants.105 resistant T1 plants were obtained by spraying PPT of 200 ppm to the seedlings of seeds corresponding to 42 T0 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.
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