基因枪法介导大白菜小孢子转基因技术研究初报

发布时间：2018-06-03 14:38

本文选题：大白菜 + 小孢子　；参考：《园艺学报》2017年01期

【摘要】：围绕基因枪微粒的制备、大白菜小孢子对基因枪轰击的耐受能力以及小孢子的胚胎发生能力开展研究,初步建立了以小孢子为外植体的遗传转化体系。高胚胎发生的大白菜材料‘CC-4’的小孢子在基因枪7.58×10~6 Pa爆破压力,9 cm的轰击距离下,利用直径0.6μm,亚精胺包裹的金粒子微弹以每次500μg的金粒子量连续轰击5次,小孢子活力不受影响,培养1 d后GUS基因瞬时表达检测,平均每皿可获得20个以上转化小孢子;小孢子培养15 d后成功获得胚状体,GUS基因瞬时表达检测,平均每皿最多获得4个遗传转化的胚状体,40个花蕾可获得20个以上转化胚状体。初步探讨基因枪法介导大白菜小孢子结果表明:大白菜小孢子数量庞大易收集,可耐受基因枪的多次轰击,并能继续保持胚胎发生的能力,作为新尝试的遗传转化受体,有可能是解决白菜类作物再生困难,提高转基因效率的有效手段。
[Abstract]:The microspore tolerance to bombardment and embryogenesis of microspores in Chinese cabbage were studied. A genetic transformation system with microspore as explant was established. Under the bombardment distance of 7.58 脳 10 ~ (6) Pa blasting pressure of a gene gun, the microspore of Chinese cabbage material CC-4 'with high embryogenesis was bombarded five times with the amount of gold particles of 500 渭 g each time using 0.6 渭 m in diameter. The activity of microspore was not affected, the transient expression of GUS gene was detected after 1 day culture, the average of transformed microspore was more than 20 per dish, and the transient expression of Gus gene in embryoid was detected successfully after 15 days of microspore culture. An average of 4 transformed embryoids were obtained per dish, and more than 20 transformed embryoids were obtained from 40 flower buds. The results of preliminary study on microspore mediated by gene gunshot in Chinese cabbage showed that the large number of microspores in Chinese cabbage was easy to collect and could withstand multiple bombardment by gene gun, and could continue to maintain the ability of embryogenesis as a new type of genetic transformation receptor. It may be an effective means to solve the difficulty of regeneration of cabbage crops and to improve the efficiency of transgenic plants.
【作者单位】：中国农业科学院蔬菜花卉研究所;中国农业大学农学与生物技术学院;
【基金】：国家重点研发计划项目(2016YFD0101701) 农业部园艺作物生物学与种质创制重点实验室项目
【分类号】：S634.1

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