农杆菌介导法将高赖氨酸蛋白基因sb401导入水稻

Introduction of Lysine-rich Protein Gene sb401 into Rice by Agrobacterium tumefactions-mediated Method

培育高赖氨酸的水稻品种是提高稻米营养品质的重要途径。本研究利用农杆菌介导法将水稻胚乳特异表达启动子PGlu驱动的马铃薯花粉特异水溶性蛋白基因sb401导入粳稻品种日本晴和籼稻恢复系501R的幼胚愈伤组织中,以期提高水稻胚乳中赖氨酸的含量。经PCR检测,从70株T0再生植株中筛选出18株转基因植株(日本晴13株,501R5株)。对转基因植株的Southernblotting分析表明,sb401基因已经转入并整合进了水稻基因组中。测定10株T0代转基因植株成熟种子的总蛋白含量和赖氨酸含量,结果表明,有8株的总蛋白含量,5株的赖氨酸含量皆提高了20%以上;其中,8号样品(日本晴转基因植株)的种子总蛋白含量和赖氨酸含量分别为10.5%和0.383%,与对照相比分别提高了32.74%和35.34%,表明sb401基因在大部分转基因植株中的翻译水平上得到了比较有效的表达。

The breeding of high lysine content variety is an effective path to increase the nutritional value of rice. The sb401 gene,which was from the potato species (Solanum berthaultii) encoding a pollen-specific protein with high lysine content and under the control of a rice endosperm-specific expresstion promoter, was successfully introduced into calli derived from immature embryos immature of japonica rice Nipponbare and indica rice 501R by Agrobacterium tumefaction in this study. 18 transgenic plants(13 Nipponbare and 5 501R, respectively) were identified by PCR from 70 T0 regenerated plants obtained by effective selection for hygromycin resistance. Southern blotting assay for the T0 transgenic plants indicaed that the sb401 gene has been successfully introduced and integrated into the genome of the receptors. The analysis of total protein and lysine content in mature seeds from 10 T0 transgenic plants showed that the content of total protein in 8 transgenic plants and lysine in 5 transgenic plants was increased more than 20% , and in No.8 sample, compared to the non-transgenic rice control,the total protein content was increased by 32.74% as well as the lysine content was increased by 35.34%. The results showed that the sb401 gene had been expressed effectively in translation level of great majority transgenic plants.