[1]颜文飞,张启军,秦海龙,等.运用农杆菌介导的针刺法将外源基因转入水稻[J].江苏农业学报,2015,(04):718-724.[doi:10.3969/j.issn.1000-4440.2015.04.002]
　YAN Wen-fei,ZHANG Qi-jun,QIN Hai-long,et al.Transforming gene into rice through agrobacterium-mediation assisted by needle puncture[J].,2015,(04):718-724.[doi:10.3969/j.issn.1000-4440.2015.04.002]
点击复制

运用农杆菌介导的针刺法将外源基因转入水稻()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年04期
页码:
718-724
栏目:
遗传育种·生理生化
出版日期:
2015-08-31

文章信息/Info

Title:
Transforming gene into rice through agrobacterium-mediation assisted by needle puncture
作者:
颜文飞1; 张启军2; 秦海龙1; 廖慧敏1; 宗寿余2; 夏士健2; 吕川根2
(1.南京农业大学农学院，江苏南京210095；2.江苏省农业科学院粮食作物研究所/江苏省优质水稻工程技术研究中心，江苏南京210014)
Author(s):
YAN Wen-fei1; ZHANG Qi-jun2; QIN Hai-long1; LIAO Hui-min1; ZONG Shou-yu2; XIA Shi-jian2; L Chuan-gen2
(1.College of Agriculture, Nanjing Agricultural University, Nanjing 210095, China;2.Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu High Quality Rice R&D Center, Nanjing 210014, China)
关键词:
水稻; 转基因; 胚芽生长点; 针刺法
Keywords:
rice; transgene; apical meristem; needle puncture
分类号:
S511
DOI:
10.3969/j.issn.1000-4440.2015.04.002
文献标志码:
A
摘要:
选用含转化质粒pBI121-GUS的农杆菌为转化载体菌株，以水稻南粳45为转基因受体品种，用含有外源基因的农杆菌菌液浸蘸接种针，然后直接侵染受体水稻发芽种子的胚芽生长点，不进行愈伤组织培养，直接完成转基因操作。结果表明，生长成植株的48株中，16株经PCR检测和GUS组织化学检测均为阳性，说明外源基因不仅导入水稻基因组中，且已在蛋白质水平上表达。与传统的农杆菌介导法相比，本方法不需诱导愈伤组织以及之后的一系列组织培养过程，节省了大量的人力和财力，转化过程所需时间缩短，具有简单、快速、高效的优点，是一种改良的水稻转基因新技术。
Abstract:
The solution of agrobacterium EHA105 containing plasmid pBI121-GUS was used to infect the apical meristem of japonica rice variety, Nanjing45 through agrobacterium-mediation assisted by needle puncture to transform gus gene into rice without callus induction and a series of tissue culture. Among 48 survived seedlings, 16 positive plants were verified by PCR detection and GUS histochemical assays, indicating that the GUS gene has been transformed into rice genome and expressed on protein level. Compared to the traditional method of agrobacterium-mediated transformation, the present method does not need to induce or culture callus, and can save a large amount of labor, time and costs. It is a simple, quick and efficient way for rice gene transformation.

参考文献/References:


[1]杨长青,王凌健,毛颖波,等. 植物转基因技术[J].科学,2011,63(3):21-24.
[2]JAMES C. 2009年全球生物技术/转基因作物商业化发展态势[J].生物工程学报,2010, 30(2): 1-22.
[3]万建民. 我国转基因植物研发形势及发展战略[J].生命科学,2011,23(2):157-167.
[4]CHILTON M D, SAIKIR K. Highly conserved DNA of Ti plasmids overlaps T-DNA maintained plant tumors [J]. Nature, 1978, 275:147-149.
[5]BABA A, HASEZAWA S, SYONO K. Cultivation of rice protoplasts and their transformation mediated by Agrobacterium protoplasts [J].Plant Cell Physio,1986,27:463-468.
[6]郑腾,陆承平. 转基因技术对粮食生产的影响[J].生物学通报,2004,39(1):6-9.
[7]宁约瑟,刘雄伦. 根癌农杆菌介导水稻遗传转化研究进展及展望[J].中国农学通报,2007,23(3):47-51.
[8]李卫,郭光沁,郑锠.根癌农杆菌介导遗传转化研究的若干新进展[J].科学通报,2000,45(8):798-807.
[9]杜立群,李银心,麻密,等. 小麦生长点转化法初报[J].植物学报,1996,38(11):921-924.
[10]RAZZAQ A,张艳敏,杨帆,等.小麦茎生长点转化研究初报[J].华北农学报,2005,20(1):17-22.
[11]董福双,张艳敏,杨帆,等. 小麦芽生长点的基因枪转化技术研究[J]. 华北农学报, 2009,24(5):1-6.
[12]孙传波,李海华,郭嘉,等. 农杆菌介导法向玉米茎尖导入抗草甘膦EPSPS基因的研究[J]. 生物技术通报,2011（3）:91-93.
[13]林拥军. 农杆菌介导的水稻转基因研究[D].武汉:华中农业大学,2001. 
[14]PUTU S, TSUTOMU S, HIDENARI S, et al. Development of simple and efficient in plant-a transformation method for rice (Oryza sativa L.) using Agrobacterium tumefaciens [J]. Bioscience and Bioengineering, 2005, 100(4):391-397.
[15]HIEI Y, OHTA S, KOMARI T, et al. Efficient transformation of rice mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA [J]. Plant J,1994,6(2):271-282.
[16]殷丽青,张建军,王慧梅,等. 提高根癌农杆菌介导水稻遗传转化频率的研究[J]. 上海交通大学学报：农业科学版, 2003, 21（增）:43-47.
[17]刘元凤,刘彦卓,王金花,等. 根癌农杆菌介导籼稻遗传转化影响因素研究[J]. 分子植物育种, 2005,3(5):737-743.
[18]任永霞,季静,王罡,等. 植物遗传转化方法概述[J]. 河北北方学院学报：自然科学版, 2005, 21(6):38-42.
[19]胡丽华,吴慧敏,周泽民,等. 利用农杆菌介导法将柠檬酸合成酶基因（CS）导入籼稻品种明恢86[J]. 分子植物育种,2006,4(2):160-166.
[20]刘永巍,田红刚,李玉华,等. 根癌农杆菌的水稻遗传转化[J].牡丹江师范学院学报：自然科学版, 2008（1）:30-32.
[21]吴金平. 利用离体培养技术筛选魔芋软腐病抗源材料的研究[D]. 武汉:华中农业大学, 2004.〖ZK）〗〖FL）〗

相似文献/References:

[1]王士磊,丁正权,黄海祥.水稻隐性早熟突变体ref早熟性的遗传分析和基因定位[J].江苏农业学报,2016,(04):721.[doi:10.3969/j.issn.100-4440.2016.04.001]
　WANG Shi-lei,DING Zheng-quan,HUANG Hai-xiang.Inheritance and gene mapping of recessive earliness in rice (Oryza sativa L.)[J].,2016,(04):721.[doi:10.3969/j.issn.100-4440.2016.04.001]
[2]王在满,郑乐,张明华,等.不同播种方式对直播水稻倒伏指数和根系生长的影响[J].江苏农业学报,2016,(04):725.[doi:10.3969/j.issn.100-4440.2016.04.002]
　WANG Zai-man,ZHENG Le,ZHANG Ming-hua,et al.Effects of seeding manners on lodging index and root growth of directseeded rice[J].,2016,(04):725.[doi:10.3969/j.issn.100-4440.2016.04.002]
[3]易能,薛延丰,石志琦,等.微囊藻毒素对水稻种子萌发和幼苗生长的胁迫作用[J].江苏农业学报,2016,(04):729.[doi:10.3969/j.issn.100-4440.2016.04.003]
　YI Neng,XUE Yan-feng,SHI Zhi-qi,et al.Inhibitory effect of microcystins on seed germination and seedling growth of rice[J].,2016,(04):729.[doi:10.3969/j.issn.100-4440.2016.04.003]
[4]刘凯,王爱民,严国红,等.一个水稻显性矮秆突变体的遗传特性与降株高能力[J].江苏农业学报,2016,(05):968.[doi:10.3969/j.issn.1000-4440.2016.05.002]
　LIU Kai,WANG Ai-min,YAN Guo-hong,et al.Genetic analysis and plant height reduction of a dominant dwarf mutant of rice[J].,2016,(04):968.[doi:10.3969/j.issn.1000-4440.2016.05.002]
[5]王红,杨镇,裴文琪,等.功能性微生物制剂对镉胁迫下水稻生长及生理特性的影响[J].江苏农业学报,2016,(05):974.[doi:10.3969/j.issn.1000-4440.2016.05.003]
　WANG Hong,YANG Zhen,PEI Wen-qi,et al.Growth and physiological characteristics of cadmium-stressed rice influenced by functional microorganism agent[J].,2016,(04):974.[doi:10.3969/j.issn.1000-4440.2016.05.003]
[6]孙玲,单捷,毛良君,等.基于遥感和Moran's I指数的水稻面积变化空间自相关性研究[J].江苏农业学报,2016,(05):1060.[doi:10.3969/j.issn.1000-4440.2016.05.017]
　SUN Ling,SHAN Jie,MAO Liang-jun,et al.Spatial autocorrelation of changes in paddy rice area based on remote sensing and Moran’s I index[J].,2016,(04):1060.[doi:10.3969/j.issn.1000-4440.2016.05.017]
[7]张晓忆,李卫国,景元书,等.多种光谱指标构建决策树的水稻种植面积提取[J].江苏农业学报,2016,(05):1060.[doi:10.3969/j.issn.1000-4440.2016.05.018]
　ZHANG Xiao-yi,LI Wei-guo,JING Yuan-shu,et al.Extraction of paddy rice area by constructing the decision tree with multiple spectral indices[J].,2016,(04):1060.[doi:10.3969/j.issn.1000-4440.2016.05.018]
[8]裔传灯,李玮,王德荣,等.水稻GW5基因的1212-bp Indel变异对粒形的影响[J].江苏农业学报,2016,(06):1201.[doi:doi:10.3969/j.issn.1000-4440.2016.06.001]
　YI Chuan-deng,LI Wei,WANG De-rong,et al.Effect of 1212-bp Indel variation of gene GW5 on rice grain shape[J].,2016,(04):1201.[doi:doi:10.3969/j.issn.1000-4440.2016.06.001]
[9]刘红江,陈虞雯,张岳芳,等.不同播栽方式对水稻叶片光合特性及产量的影响[J].江苏农业学报,2016,(06):1206.[doi:doi:10.3969/j.issn.1000-4440.2016.06.002]
　LIU Hong-jiang,CHEN Yu-wen,ZHANG Yue-fang,et al.Effects of planting pattern on leaf photosynthetic characteristics and yield of rice[J].,2016,(04):1206.[doi:doi:10.3969/j.issn.1000-4440.2016.06.002]
[10]郭保卫,许轲,魏海燕,等.钵苗机插水稻茎秆的抗倒伏能力[J].江苏农业学报,2016,(06):1280.[doi:doi:10.3969/j.issn.1000-4440.2016.06.014]
　GUO Bao-wei,XU Ke,WEI Hai-yan,et al.Culm lodging resistance characteristics of bowl seedling mechanical-transplanting rice[J].,2016,(04):1280.[doi:doi:10.3969/j.issn.1000-4440.2016.06.014]

备注/Memo

备注/Memo:
收稿日期：2014-12-25 基金项目：国家转基因生物新品种培育重大专项（2014ZX08001-004） 作者简介：颜文飞，（1988- ），女，福建晋江人，硕士研究生，主要研究水稻遗传育种。（E-mail）2010801171@njau.edu.cn。张启军为共同第一作者。 通讯作者：吕川根，（E-mail）rb8@jaas.ac.cn
更新日期/Last Update: 2015-08-31