芝麻人工构建雄性不育基因的转化研究初报

Transformation of Engineered Male Sterile Gene and Establishment of Transgenic Plants in Sesame (Sesamum indicum L.)

用基因枪将人工构建的植物雄性不育基因ＴＡ２９－Ｂａｒｎａｓｅ及其标记基因ｂａｒ导入芝麻豫芝４号的离体子叶，在ＭＳ＋５ｍｇ·Ｌ－１６－ＢＡ＋１ｍｇ·Ｌ－１ＩＡＡ＋１ｍｇ·Ｌ－１ＡＢＡ＋５ｍｇ·Ｌ－１ＡｇＮＯ３的培养基上交培养１２天，转入光、暗光替培养，诱导出不定芽。经添加２ｍｇ·Ｌ－１除草剂（ＢＡＳＴＡ）的上述培养基筛选，获得了１１个ＢＡＳＴＡ抗性株系，对其中３个株系进行Ｓｏｕｔｈｅｒｎ杂交鉴定分析，证实其中２个为转基因株系，转化率为１．４６％

The engineered male sterile gene barnase together with the mark gene bar were transformed to the isolated cotyledon of a sesane variety Yuzhi 4 by microprojectile bomebardment.The transformed cotyledon was cultured under darkness for 12 days then converted into dark 1ight alternative conditions on the culture media MS+5mg·L -1 6-BA+1mg·L -1 IAA+1mg·L -1 ABA+5mg·L - AgNO 3 Resistant calli were selected on herbicide Basta 2mg.L -1 11 anti Basta plants were obtained.Southern blot of 3 well developed green plants confirmed stable integration of both barnase and bar genes into nuclear DNA.The transfomation efficiency reached 1.46%.