用基因枪法获得转异天南星基因aha抗蚜虫小麦

凝集素是一类具有特异糖结合活性的蛋白，对蚜虫等害虫有很强的抗杀作用。利用异天南星凝集素基因aha(Arisaema heterophyllum agglutinin)以及水稻Rubisco小亚基启动子，构建了aha基因植物绿色组织特异性表达载体，并采用基因枪转化方法, 与携带bar基因的pAHC20载体共转化到小麦品种科农199中。经过愈伤诱导、再生和筛选以及PCR鉴定，获得aha转基因植株42株，平均转化效率为2.41%。对转aha基因植株后代PCR鉴定表明，T₁代转基因植株的分离比例基本符合孟德尔遗传规律。利用室内多目标综合判别法评定抗蚜虫特性，8个T₁代转基因株系中有高抗材料1份，低抗材料3份，占参试比例44.4%。本研究为获得抗蚜虫转基因小麦新材料奠定了基础。

Generation of Aphid Resistant Transgenic Wheat with aha from Arisaema heterophyllum by Particle Bombardment

Agglutinin is a class of mannose-binding protein,which has detrimental effect on aphids and other pests.In this study,the vector harboring aha cloned from Arisaema heterophyllum directed by rice Rubisco small subunit promoter(rbcS) was constructed.By co-transformed with pAHC20 that harbors bar selection gene,the aha gene was transferred into wheat variety Kenong 199 via bombardment.After induction,regeneration,two rounds of selection and with the conformation by PCR analysis,42 transgenic plants with foreign aha gene were obtained,average co-transformation efficiency of 2.41%.According to PCR analysis,the segregation of the T1 plants was basically consistent with Mendel’s separation.Aphid resistance bioassay was carried out using eight randomly selected transgenic lines by multiple discrimination method.One high-resistant and three low-resistant transgenic lines were identified,accounting for 44.4% of the tested materials.This study has laid a basis for application of aha gene in development of aphid resistant transgenic wheat.