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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-D2.0mg/L+L-pro700mg/L+CH500mg/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pCAMBIA1301+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。
Maize high amylose breeding is an important research direction of maize molecular breeding. In this experiment, we first studied the effects of different callus induction medium on the regeneration system, and determined that LS + 2,4-D2.0mg / L + L-pro700mg / L + CH500mg / L + 3% sucrose as induction medium . At the same time, we constructed and verified the expression vector pCAMBIA1301 + Glu + 1620 containing double interfering fragment of starch breakout enzyme sbeIIb gene and endosperm-specific promoter and transformed it into Agrobacterium tumefaciens EHA105. 178. The 5 transformants were positive by PCR and initially showed that the interference fragment was integrated into the genome of maize.