淀粉分支酶sbeIIb基因RNA干涉片段转化玉米自交系的研究

玉米高直链淀粉育种是玉米分子育种的一个重要研究方向.本实验中,首先研究了不同诱导愈伤培养基对再生体系的影响,确定了LS+2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 %蔗糖为诱导培养基.同时,构建并验证了含有淀粉分支酶sbeIIb基因双干涉片段载体和胚乳特异性启动子的表达载体pCAMBIA 1301+Glu+1620,并转入根癌农杆菌EHA105,以农杆菌转化法转化玉米自交系178.通过PCR检测,5株转化株表现阳性,初步证明了干涉片段已整合入玉米基因组中.

A Study on Starch Branching sbeIIb Gene RNAi Fragments Transformation of Maize Inbred-lines

High amylose maize (Zea mays L.) breeding is an important field of molecular maize breeding. In this study, through research of the effect of inducing culture, the optimal inducing culture was fixed as LS+ 2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 % sucrose. Then, we constructed and detected the expression vector pCAMBIA 1 301+Glu+1 620 which carries endosperm specific promoter (Glu promoter from wheat glutenin) and two interferential fragment of the 16 and 20 exon in maize starch branching sbeIIb gene, transferred into Agrobacterium tumefaciens strain EHA105, and transformed the maize elite inbred line 178 with the Agrobacterium-protocol. Five transgenic plants which were detected by PCR reaction showed positive signal, indicating that interferential fragment was transformed preliminarily into genome of maize.