转异质型ACCase复合基因对油菜含油量的影响

发布时间：2018-05-15 05:02

本文选题：ACCase + 农杆菌介导转化法　；参考：《分子植物育种》2017年03期

【摘要】：乙酰辅酶A羧化酶(ACCase)是脂肪酸生物合成的限速酶,在油料作物脂肪酸代谢和调控基因工程中具有重要的应用价值。本研究利用同尾酶反复酶切的方法,构建了拟南芥Rubisco SSU小亚基转运肽与大肠杆菌异质型ACCase各亚基基因的融合串联种子特异表达载体,在根癌农杆菌(Agrobacterium tumefaciens)介导的下胚轴转化甘蓝型油菜高油品种CY2,研究异质型ACCase转入油菜对含油量的影响。经过抗生素筛选获得11株转化再生植株,PCR、Southern分子检测证明四个目的基因已整合到T0代再生植株油菜基因组中。RT-PCR检测其中4株,说明在转基因株系中目标基因在籽粒中特异表达。对T2代转基因油菜含油量进行测定,研究表明转基因株系的含油量比对照提高了约5.7%。
[Abstract]:Acetyl-CoA carboxylase (ACCase) is a rate-limiting enzyme for fatty acid biosynthesis, which has important application value in oil crop fatty acid metabolism and gene engineering. In this study, we constructed the fusion series-specific expression vector of Arabidopsis thaliana Rubisco SSU small subunit transporter peptide and E. coli heterotype ACCase subunit gene by using the same tail enzyme digestion method. Hypocotyl transformation mediated by Agrobacterium tumefaciens (Agrobacterium tumefaciens) from Agrobacterium tumefaciens was used to study the effect of heterogenous ACCase on oil content of Brassica napus. 11 transformed regenerated plants were screened by antibiotics, and Southern analysis showed that the four target genes had been integrated into the genome of regenerated plants of T0 generation. RT-PCR was used to detect 4 of them, which indicated that the target genes were specifically expressed in the seeds of transgenic lines. The oil content of transgenic rapeseed in T2 generation was measured. The results showed that the oil content of transgenic lines was about 5.7% higher than that of control.
【作者单位】：浙江省农业科学院病毒学与生物技术所浙江省植物有害生物防控重点实验室-省部共建国家重点实验室培育基地;
【基金】：国家自然科学基金(31571700;31272114) 浙江省植物有害生物防控重点实验室-省部共建国家重点实验室培育基地(2010DS700124-KF1702;2010D700124-KF1609)共同资助
【分类号】：S565.4

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