采用一种新型RNAi载体培育转基因高直链淀粉马铃薯.pdfVIP

作物学报 ACTA AGRONOMICA SINICA 2009, 35 5 : 809?815 /zwxb/ ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@ DOI: 10.3724/SP.J.1006.2009.00809 采用一种新型RNAi 载体培育转基因高直链淀粉马铃薯 郭志鸿1 王亚军1 张金文2 张玉宝1 王金牛1 谢忠奎1,* 陈正华3 1 , 730000; 2 , 730070; 3 , 100101 : PCR nos axi1 ubi.u4 , Sbe1 Sbe2 SIII, “ubi.u4 SIIInos axi1 nos ”3′UTR RNAi pCUSNI, 3 2 , 16 , 14 , 53.80%~85.33%; , RT-PCR , 80%Sbe1 Sbe2 mRNA , 3′UTR RNAi pCUSNI Sbe1 Sbe2 , RNAi pCUSNI , , pCUSNI BamH I Xba I SIII , , : RNAi; nos ; ; Development of Transgenic High-Amylose Potato Using a Novel RNAi Vector 1 1 2 1 1 1,* GUO Zhi-Hong , WANG Ya-Jun , ZHANG Jin-Wen , ZHANG Yu-Bao ,WANG Jin-Niu , XIE Zhong-Kui , 3 and CHEN Zheng-Hua 1 Cold and Arid Region’s Environmental and Engineering Institute, Chinese Academy of Sciences, Lanzhou 730000, China; 2 College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China; 3 Postdoctoral Scientific Research Station of Gansu Yasheng Industrial Group . Co. Ltd., Beijing Branch, Beijing 100101, China Abstract: Amylose from potato starch is of great advantage for applications in many fields because of its higher degree of polym- erization and lower gelling temperature compared with cereal starch. However, there is no natural mutant of high-amylose potato. RNAi technique is efficient and specific for plant gene silence but traditional RNAi vector is laborious to prepare. To design an easily-prepared RNAi vector and to develop transgenic high-amylose potato, PCR technique was employed to amplify the nos terminator, the tobacco axi1 and the tobacco ubi.u4 promoter, and to sub-clone a fused fragment SIII which is partially homolo- gous to potato Sbe1 and to Sbe2. Then, a newly designed vector pCUSNI containing “ubi.u4 promotor–antisense SIII–antisense nos terminator–axi1 gene intron–sense nos terminator” was generated and transformed into potato varieties Longshu 3, Gannong- shu 2, and Atlantic by Agrobacterium-mediated transformation. Sixteen t