陈凌娜1 ，曲延英1，王兰岚2*，张金文2，3*，刘桂珍2，陈正华2
（1.新疆农业大学农学院，中国新疆 乌鲁木齐 830052；2.甘肃亚盛集团博士后科研工作站北京分站，中国北京100101；3.甘肃农业大学农学院，中国甘肃 兰州 730070）
摘 要：以棉花感受态萌动种胚作为外源基因转化的受体，用激光微束穿刺法将β-1,3- 葡聚糖酶及几丁质酶双价基因导入棉花，所构建的植物表达载体pBIBGC携带有筛选标记npt-Ⅱ（新霉素磷酸转移酶）基因。激光转化处理的种胚经卡那霉素（Kan）筛选，已经获得了抗性小植株。本文还研究了微束激光转化法用于棉花感受态萌动胚转化预培养的时间、高渗液对材料的处理等。研究表明用微束激光转化处理种胚是一种操作简便、重复性好的转化方法，用该法可将外源基因导入植物感受态萌动胚，因而避开植株离体再生的困难。关键词：微束激光；β-1,3- 葡聚糖酶基因；几丁质酶基因；感受态萌动种胚；转化
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Studies of Introduction of Antifungal Genes into Cotton Plants
by Laser Microbeam Puncture
CHEN Ling-na1, QU Yan-ying1,WANG Lan-lan2*, ZHANG Jin-wen2.3*, LIU Gui-zhen2, CHEN Zheng-hua2
(1. Agronomy College, Xinjiang Agricultural University, Urumqi 830052, Xinjiang, China； 2. Postdoctoral Scientific Research Station of Gansu Yasheng Industrial (Group) Co. Ltd, Beijing Branch, Beijing 100101, China； 3. Agronomy Department, Gansu Agricultural University, Lanzhou 730070, Gansu, China)
Abstract: Using cotton susceptive seed-germinating embryos as transgenic receptor, the bivalent genes pBIBGC, which includes β-1,3-glucanase gene and chitinase gene, was introduced into cotton by Laser Microbeam Puncture Techniques (LMPT). There are npt-Ⅱ(neomycin phosphotransferase Ⅱ) gene as screening marker in the plant bivalent expression vector. From treated embryos by laser transformation, resistant plantlets have been obtained after kanamycin resistance screening. In this research, we have also studied some key factors that influenced LMPT, such as the duration of pretreatment of susceptive seed-germinating embryos and hypertonic treatment, and so on. The results indicated that the LMPT was easy to operate and repetitive. The foreign genes were introduced into susceptive seed-germinating embryos by this technique, which avoids difficulties of regeneration of plantlets.
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