[1]朱彩云,黄翠香,于 伟,等.利用RNA 干扰技术创制高直链淀粉马铃薯新材料[J].南京师大学报(自然科学版),2014,37(02):112.
 Zhu Caiyun,Huang Cuixiang,Yu Wei,et al.Using RNAi Technology to Produce High-Amylose Potato New Material[J].Journal of Nanjing Normal University(Natural Science Edition),2014,37(02):112.
点击复制

利用RNA 干扰技术创制高直链淀粉马铃薯新材料()
分享到:

《南京师大学报(自然科学版)》[ISSN:1001-4616/CN:32-1239/N]

卷:
第37卷
期数:
2014年02期
页码:
112
栏目:
生命科学
出版日期:
2014-06-30

文章信息/Info

Title:
Using RNAi Technology to Produce High-Amylose Potato New Material
作者:
朱彩云黄翠香于 伟王忠亚杨金金徐盼盼徐 春季 勤
淮阴师范学院生命科学学院,江苏省环洪泽湖生态农业生物技术重点实验室,江苏淮安223300
Author(s):
Zhu CaiyunHuang CuixiangYu WeiWang ZhongyaYang JinjinXu PanpanXu ChunJi Qin
School of Life Sciences,Huaiyin Normal College,Jiangsu Key Laboratory for Eco-Agricultural and Biotechnology Around Hongze Lake,Huaian 223300,China
关键词:
转基因马铃薯SBE 基因RNA 干扰直链淀粉
Keywords:
transgenic potatoSBE geneRNA interferenceamylose
分类号:
Q786
文献标志码:
A
摘要:
直链淀粉由于其独特的理化性质,在工业上有着广泛的应用. 郭志鸿[1] 等利用RNA 干扰技术抑制马铃薯中两类淀粉分支酶亚基基因Sbe1 和Sbe2 的表达,有效提高了马铃薯的直链淀粉含量. 本研究利用RNA 干扰技术,构建马铃薯SBEⅠ(the ndogenesis starch branching enzymeⅠ) 和SBEⅡ( the endogenesis starch branchingenzyme Ⅱ)单基因以及SBE玉和SBE域双基因的RNA 干扰双元表达质粒载体. 采用农杆菌介导法将重组基因
Abstract:
Amylose is extensively used in industrial applications because of it’s unique physicochemical property. Zhihong Guo inhibted expression of two potato starch branching enzyme subunit genes Sbe1 and Sbe2 by using RNA interference technology,effectively improved the amylase content in potato. To develop transgenic potato plants with high-amylose starch in tubers, the endogenesis starch branching enzyme Ⅰ( SBE Ⅰ) and Ⅱ( SBE Ⅱ) genes were, single or simutaneously,targeted for RNA mediated gene silencing by expression of RNA interference genes in potato. In this study,RNA interference was used for gene inhibition of the starch branching enzymeⅠ(SBEⅠ)and Ⅱ(SBEⅡ)genes. Three SBE-RNA interference plasmids were constructed and transformed into potato cultivar Désirée. Three transformed series were obtained and analyzed by PCR. The level of silencing of SBEⅠand SBEⅡin transgenic plants was investigated by analysis of apparent amylose content. The results showed that silencing of SBEⅠor SBEⅡalone had yielded a very low frequency of high-amylose lines, and even a reduction in amylose content in most of transgenic starches. However,production of high-amylose potato lines were be achieved by simultaneous inhibition of SBEⅠand SBEⅡ. In this transgenic series,more than 50% of the transgenic lines showed that amylose content was higher than thire control. The highest amylose content in one line was 49. 1%,increasing 20. 6% in comparison with untransformed starch ones(28. 5%).

参考文献/References:

[1]郭志鸿,张金文,王蒂,等. 用RNA 干扰技术创造高直链淀粉马铃薯材料[J]. 中国农业科学,2008,41(2):494-501.
[2] Englyst H N,Andersen V,Cummings J H. Starch and non鄄starch polysaccharides in some cereal foods[J]. J Sci Food Agric,1983,34:1 434-1 440.
[3] 宋同明. 发展我国特用玉米产业的意义潜力与前景[J]. 玉米科学,1996,5(4):6-11.
[4] Alan M M,Matthew K M,Steven G,et al. Progress toward understanding biosynthesis of the amylopectin crystal[ J]. Plant Physiology,2000,122(4):989-997.
[5] Fernie A R,Willmitzer L,Trethewey R N. Sucrose to starch:a transition in molecular plant physiology[ J]. Trends in Plant Science,2002,7(1):35-41.
[6] Abad Marta C,Binderup K,Rkos S J,et al. The X-ray crystallographic structure of Escherichia coli branching enzyme[ J].Journal of Biological Chemistry,2002,277:42 164-42 170.
[7] Waterhouse P M,Helliwell C A. Exploring plant genomes by RNA induced gene silencing[ J]. Nature Reviews Genetics,2002(4):29-38.
[8] Mattew L. RNAi for plant functional genomics[J]. Comparative and Functional Genomics,2004(5):240-244.
[9] Ogita S,Uefuji H,Yamguchi Y,et al. Producing decaffeinated coffee plants[J]. Nature,2003,423(6 942):823.
[10] Kusaba M,Miyahara K,Lida S,et al. Low lutenin content 1:a dominant mutation that suppresses the glutenin multigene family via RNA silencing in rice[J]. The Plant Cell,2003,15:1 455-1 467.
[11] Wang M B,Abbott D C,Waterhouse P M. A single copy of a virus derived transgene encoding hairpin RNA gives immunity to barley yellow dwarf virus[J]. Molecular Plant Pathology,2000,1(6):347-356.
[12] Ogita S,Hirotaka U,Masayuki M,et al. Application of RNAi to confirm theobromine as the major intermediate for caffeine bio- synthesis in coffee plants with potential for construction of decaffeinated varieties[J]. Plant Molecular Biology,2004,54:931- 941.
[13] Jobling S A,Schwall G P,Westcott R J,et al. A minor form of starch branching enzyme in potato( Solanum tuberosum L. )tubers has a major effect on starch structure:cloning and charac-terization of multiple forms of SBE A[J]. The Plant Journal,1999,18(2):163-171.
[14] Rydberg U,Andersson L,Andersson R,et al. Comparison of starch branching enzymeⅠand Ⅱfrom potato[ J]. European Journal of Biochemistry,2001,268(23):6 140-6 145.
[15] Larsson C T,Hofvander P,Khoshnood J,et al. Three isoforms of starch synthase and two isoforms of branching enzyme are present in potato tuber starch[J]. Plant Science,1996,117:9-16.
[16] Ji Q,Vincken J P,Suurs L C,et al. Microbial starch-binding domains as a tool for targeting proteins to granules during starch biosynthesis[J]. Plant Molecules Biological,2003,51:789-801.
[17] 黄翠香,季勤,张云峰,等. 不同启动子对禽流感病毒H5HA 基因在马铃薯体中表达的影响[J]. 南京师大学报:自然科学版,2012,35(1):75-79,83.
[18] Murray M G,Thompson W F. Rapid isolation of high molecular weight DNA[J]. Nucleic Acids Research,1980,8(19):4321-4325.
[19] Schwall G P,safford R,Westcott R J,et al. Production of very-high-amylose potato starch by inhibition of SBE A abd B[J].Nature Biotechnology,2000,18:551-554.
[20] Wesley S V,Helliwell C A,Smith N A,et al. Construct design for efficient,effective and high-throughput gene silencing in plants[J]. The Plant Journal,2001,27(6):581-590.

相似文献/References:

[1]黄翠香,季勤,张云峰,等.不同启动子对禽流感病毒H5HA基因在马铃薯体中表达的影响[J].南京师大学报(自然科学版),2012,35(01):75.
 Huang Cuixiang,Ji Qin,Zhang Yunfeng,et al.Effect of Promoters on Expression of H5HA Avian Influenza Virus Gene in Potato Plant[J].Journal of Nanjing Normal University(Natural Science Edition),2012,35(02):75.

备注/Memo

备注/Memo:
收稿日期:2013-03-28.
基金项目:国家自然科学基金(30771367).
通讯联系人:季勤,博士,教授,研究方向:分子生物学. E-mail:jiqin@hytc.edu.cn
更新日期/Last Update: 2014-06-30