欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2012, Vol. 38 ›› Issue (05): 773-779.doi: 10.3724/SP.J.1006.2012.00773

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

抗纹枯病、根腐病的转SN1基因小麦的获得与鉴定

王金凤1,2，杜丽璞1，李钊1，黄素萍1,2，叶兴国1，冯斗2，张增艳1,*   

  1. 1中国农业科学院作物科学研究所 / 农作物基因资源与基因改良国家重大科学工程 / 农业部作物遗传育种重点实验室, 北京 100081; 2广西大学农学院, 广西南宁 530004
  • 收稿日期:2011-11-08 修回日期:2012-01-19 出版日期:2012-05-12 网络出版日期:2012-03-05
  • 通讯作者: 张增艳, E-mail: zhangzy@mail.caas.net.cn
  • 基金资助:

    本研究由国家转基因生物新品种培育科技重大专项(2008ZX08002-001, 2009ZX08002-006B)资助。

Development and Characterization of SN1 Transgenic Wheat Plants with Enhanced Resistance to Rhizoctonia cerealis and Bipolaris sorokiniana

WANG Jin-Feng1,2,DU Li-Pu1,LI Zhao1,HUANG Su-Ping1,2,YE Xing-Guo1,FENG Dou2,ZHANG Zeng-Yan1,*   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Crop Genetic and Breeding, Ministry of Agriculture / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 College of Agronomy, Guangxi University, Nanning 530004, China
  • Received:2011-11-08 Revised:2012-01-19 Published:2012-05-12 Published online:2012-03-05
  • Contact: 张增艳, E-mail: zhangzy@mail.caas.net.cn

PDF

1597

被引次数

10

摘要： SN1是源于马铃薯的一种抗菌肽, 可以抑制多种植物病原菌的生长。小麦纹枯病(主要病原菌为禾谷丝核菌Rhizoctonia cerealis)和根腐病(主要病原菌为平脐蠕孢菌Bipolaris sorokiniana)是小麦的主要土传真菌病害。本研究利用基因工程技术构建了SN1基因的单子叶植物表达载体pA25-SN1, 它受玉米泛素(ubiquitin)启动子的控制;采用基因枪法将pA25-SN1转化小麦推广品种扬麦18幼胚愈伤组织4 000块, 获得203株再生植株, 通过PCR检测出阳性植株55株, 转化率为1.38%。对转SN1基因小麦T0~T2代植株, 进行外源基因的PCR、Southern blot、RT-PCR、荧光定量RT-PCR(Q-RT-PCR)分析和小麦纹枯病菌与根腐病菌接种及其抗病性鉴定。结果表明, 转入的SN1基因已经整合到转基因小麦的基因组中, 能够在转基因小麦中遗传、转录与表达。SN1基因的表达提高了转基因植株对小麦纹枯病和根腐病的抗性, 其抗病性可以遗传。

关键词: SN1基因, 转基因小麦, 小麦纹枯病菌, 小麦根腐病菌, 抗性

Abstract: SN1 is an anti-microbial peptide in potato, and can inhibit the growth of important phytopathogens. In this study, the transformation vector of SN1 gene expressing highly in monocot plants, pA25-SN1, was correctly constructed, in which SN1 gene was driven by maize ubiquitinpromoter. Four thousand embryo calli of Yangmai 18 were bombarded by the particle containing pA25-SN1. Among the regenerated 203 plants, 55 positive transgenic individuals were identified by PCR assay in the T0 generation with a transformation frequency of 1.38%. The transgenic wheat plants from T0 to T2 generations were subjected to PCR, Southern blot, RT-PCR, and Q-RT-PCR assays. The disease responses of these positive transgenic plants were also evaluated by inoculating with Rhizoctonia cerealisand Bipolaris sorokiniana. The results showed that SN1 gene was integrated into these transgenic lines, inherited from T0 to T2, and expressed in the wheat background. The transgenic wheat plants expressing SN1 displayed anenhanced resistance to R. cerealis and B. sorokiniana compared with the untransformed Yangmai 18, and the enhanced resistance was inheritable.

Key words: SN1 gene, Transgenic wheat, Rhizoctonia cerealis, Bipolaris sorokiniana, Resistance

[l]Chen Y-X(陈延熙), Tang W-H(唐文华), Zhang D-H(张敦华), Jian X-Y(简小鹰). A preliminary study on etiology of sharp eye-spot of wheat in China. Acta Phytophylacica Sin (植物保护学报), 1986, 13(1): 39–44 (in Chinese with English abstract)

[2]Lu Y(路妍), Zhang Z-Y(张增艳), Ren L-J(任丽娟), Liu B-Y(刘宝业), Liao Y(廖勇), Xu H-J(徐惠君), Du L-P(杜丽璞), Ma H-X(马鸿翔), Ren Z-L(任正隆), Jing J-X(井金学), Xin Z-Y(辛志勇). Molecular analyses on Rs-AFP2 transgenic wheat plants and their resistance to Rhizoctonia cerealis. Acta Agron Sin (作物学报), 2009, 35(4): 640–646 (in Chinese with English abstract)

[3]Kumar J, Schafer P, Hückelhoven R, Langen G, Baltruschat H, Stein E, Nagarajan, S, Kogel K H. Bipolaris sorokiniana, a cereal pathogen of global concern: cytological and molecular approaches towards better control. Mol Plant Pathol, 2002, 3: 185–195

[4]Theis T, Stahl U. Antifungal proteins: targets, mechanisms and prospective applications. Cell Mol Life Sci, 2004, 61: 437–455

[5]Thevissen K, Terras F R G, Broekaert W F. Permeabilization of fungal membranes by plant defensins inhibits fungal growth. Appl Environ Microbiol, 1999, 65: 5451–5458

[6]Turrini A,  Sbrana C, Pitto L, Castiglione M R, Giorgetti L, Briganti R, Bracci T, Evangelista M, Nuti M P, Giovannetti M. The antifungal Dm-AMP1 protein from Dahlia merckii expressed in Solanum melongena is released in root exudates and differentially affects pathogenic fungi and mycorrhizal symbiosis. New Phytol, 2004, 163: 393–403

[7]Segura A, Moreno M, Madueño F, Molina A, García-Olmedo F. Snakin-1, a peptide from potato that is active against plant pathogens. Mol Plant-Microbe Interact, 1999, 12: 16–23

[8]Jiang W(蒋雯), Liu X(刘欣), Zhang Z-Y(张增艳). Cloning and overexpression of defensin SN1 in Escherichia coli and its antifungal assay. J Plant Genet Resour (植物遗传资源学报), 2011, 12(2): 286–290 (in Chinese with English abstract)

[9]Alnasia N, Bazzini A, Hopp H E, Vazquez-Rovere C. Overexpression of snakin-1 gene enhances resistance to Rhizoctonia solani and Erwinia carotovora in transgenic potato plants. Mol Plant Pathol, 2008, 9: 329–338

[10]Xu H-J(徐惠君), Pang J-L(庞俊兰), Ye X-G(叶兴国), Du L-P(杜丽璞), Li L-C(李连城), Xin Z-Y(辛志勇), Ma Y-Z(马有志), Chen J-P(陈剑平), Chen J(陈炯), Cheng S-H(程顺和), Wu H-Y(吴宏亚). Study on the gene transferring of Nib8 into wheat for its resistance to the Yellow mosaic virus by bombardment. Acta Agron Sin (作物学报), 2001, 27(6): 684–689 (in Chinese with English abstract)

[11]Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucl Acids Res, 1980, 8: 4321–4325 (in Chinese with English abstract)

[12]Sharp P J, Kries M, Shewry P R, Gale M D. Location of amylase sequences in wheat and its relatives. Theor Appl Genet, 1988, 75: 286–290

[13]Cai S-B(蔡士宾), Ren L-J(任丽娟), Yan W(颜伟), Wu J-Z(吴纪中), Chen H-G(陈怀谷), Wu X-Y(吴小有), Zhang X-Y(张仙义). Germplasm development and mapping of resistance to sharp eyespot (Rhizoctonia cerealis) in wheat. Sci Agric Sin (中国农业科学), 2006, 39(5): 928–934 (in Chinese with English abstract)

[14]Dong N, Liu X, Lu Y, Du L, Xu H, Liu H, Xin Z, Zhang Z. Overexpression of TaPIEP1, a pathogen-induced ERF gene of wheat, confers host-enhanced resistance to fungal pathogen Bipolaris sorokiniana. Func Integr Genomics, 2010, 10: 215–226

[15]Li Z, Zhou M, Zhang Z, Ren L, Du L, Zhang B, Xu H, Xin Z. Expression of a radish defensin in transgenic wheat confers increased resistance to Fusarium graminearum and Rhizoctonia cerealis. Func Integr Genomics, 2011, 11: 63–70

[16]Jha S, Tank H G, Prasad B D, Chattoo B B. Expression of Dm-AMP1 in rice confers resistance to Magnaporthe oryzae and Rhizoctonia solani. Transgenic Res, 2009, 18: 59–69
[1] 邓钊, 江南, 符辰建, 严天泽, 符星学, 胡小淳, 秦鹏, 刘珊珊, 王凯, 杨远柱. 隆两优与晶两优系列杂交稻的稻瘟病抗性基因分析[J]. 作物学报, 2022, 48(5): 1071-1080.
[2] 刘丹, 周彩娥, 王晓婷, 吴启蒙, 张旭, 王琪琳, 曾庆东, 康振生, 韩德俊, 吴建辉. 利用集群分离分析结合高密度芯片快速定位小麦成株期抗条锈病基因YrC271[J]. 作物学报, 2022, 48(3): 553-564.
[3] 杨昕, 林文忠, 陈思远, 杜振国, 林杰, 祁建民, 方平平, 陶爱芬, 张立武. 黄麻双生病毒CoYVV的分子鉴定和抗性种质筛选[J]. 作物学报, 2022, 48(3): 624-634.
[4] 张思梦, 倪文荣, 吕尊富, 林燕, 林力卓, 钟子毓, 崔鹏, 陆国权. 影响甘薯收获期软腐病发生的指标筛选[J]. 作物学报, 2021, 47(8): 1450-1459.
[5] 习玲, 王昱琦, 朱微, 王益, 陈国跃, 蒲宗君, 周永红, 康厚扬. 78份四川小麦育成品种(系)条锈病抗性鉴定与抗条锈病基因分子检测[J]. 作物学报, 2021, 47(7): 1309-1323.
[6] 左香君, 房朋朋, 李加纳, 钱伟, 梅家琴. 有毛野生甘蓝(Brassica incana)抗蚜虫特性研究[J]. 作物学报, 2021, 47(6): 1109-1113.
[7] 马燕斌, 王霞, 李换丽, 王平, 张建诚, 文晋, 王新胜, 宋梅芳, 吴霞, 杨建平. 玉米光敏色素A1基因(ZmPHYA1)在棉花中的转化及分子鉴定[J]. 作物学报, 2021, 47(6): 1197-1202.
[8] 蒋伟, 潘哲超, 包丽仙, 周福仙, 李燕山, 隋启君, 李先平. 马铃薯资源晚疫病抗性的全基因组关联分析[J]. 作物学报, 2021, 47(2): 245-261.
[9] 张荣跃, 王晓燕, 杨昆, 单红丽, 仓晓燕, 李婕, 王长秘, 尹炯, 罗志明, 李文凤, 黄应昆. 甘蔗新品种及主栽品种对褐锈病抗性与Bru1基因分子检测[J]. 作物学报, 2021, 47(2): 376-382.
[10] 仓晓燕, 夏红明, 李文凤, 王晓燕, 单红丽, 王长秘, 李婕, 张荣跃, 黄应昆. 甘蔗优良品种(系)对黑穗病的抗性评价[J]. 作物学报, 2021, 47(11): 2290-2296.
[11] 陈同睿, 罗艳君, 赵潘婷, 贾海燕, 马正强. 过表达TaJRL53基因提高了小麦赤霉病抗性[J]. 作物学报, 2021, 47(1): 19-29.
[12] 崔静, 王志城, 张新雨, 柯会锋, 吴立强, 王省芬, 张桂寅, 马峙英, 张艳. 棉花GbSTK基因调控开花和黄萎病抗性的功能研究[J]. 作物学报, 2021, 47(1): 30-41.
[13] 闻竞, 沈彦岐, 韩四平, 邢跃先, 张叶, 王梓钰, 李世界, 杨小红, 郝东云, 张艳. 玉米拟轮枝镰孢菌穗腐病抗性基因的挖掘[J]. 作物学报, 2020, 46(9): 1303-1311.
[14] 张雪翠,钟超,段灿星,孙素丽,朱振东. 大豆品种郑97196抗疫霉病基因RpsZheng精细定位[J]. 作物学报, 2020, 46(7): 997-1005.
[15] 李威涛,徐志军,蔡岩,郭建斌,喻博伦,黄莉,陈玉宁,周小静,罗怀勇,刘念,陈伟刚,任小平,姜慧芳. 抗青枯病兼大果和高出仁率的花生新种质创制[J]. 作物学报, 2020, 46(4): 484-490.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2