Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (9): 1790-1798.doi: 10.3864/j.issn.0578-1752.2013.09.006

• PLANT PROTECTION • Previous Articles     Next Articles

Molecular Characterization of T-DNA Integration of the Ustilaginoidea virens Mutant 5062

 YU  Mi-Na, HU  Jian-Kun, HUANG  Lei, YU  Jun-Jie, YIN  Xiao-Le, NIE  Ya-Feng, CHEN  Zhi-Yi, LIU  Yong-Feng   

  1. Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014
  • Received:2012-12-24 Online:2013-05-01 Published:2013-02-18

PDF

751

Cited

3

Abstract: 【Objective】 The objective of the experiment is to establish a method for cloning genes of the Ustilaginoidea virens mutant, and to shed light on the pathogenic mechanism of the U. virens, depending on a new mutant 5062, which has increased virulence. 【Method】With the wild type strain 70-22 as a control, phenotypic analysis, such as growth rate, colonial morphologies and sporulation, were analyzed. TAIL-PCR combined with RACE were used to identify the T-DNA integration site and the genes of flanking right site of the T-DNA. The expression levels of the genes were analyzed by RT-PCR as well. 【Result】Colonial morphologies and pigment on PSA and TB3 medium were no different between 70-22 and 5062. But on MM and water-agar medium, 5062 grew slower and produced more conidiophores. The flanking sequences of T-DNA were non-adjacent in the wild type. The expression of the gene flanking right site of the T-DNA was upregulated.【Conclusion】In 5062, both the T-DNA insertion and inversion of chromosomal fragment caused a mutation of the increased virulence.

Key words: Ustilaginoidea virens , virulence , T-DNA , mutant , chromosomal rearrangement

[1]杨健源, 曾列先, 陈深, 李传瑛, 韦靖鸾, 谭新球, 朱小源. 我国稻曲病研究进展. 广东农业科学, 2011(2): 77-79.

Yang J Y, Zeng X L, Chen S, Li C Y, Wei J L, Tan X Q, Zhu X Y. Research advance on rice false smut (Ustilaginoidea virens) in China. Guangdong Agricultural Sciences, 2011(2): 77-79.（in Chinese）

[2]李小娟, 刘二明, 肖启明, 刘年喜, 王金辉, 谭小平, 郑和斌. 水稻对稻曲病抗性的分级及相应级别的产量损失. 湖南农业大学学报: 自然科学版, 2011, 37(3): 275-279.

Li X J, Liu E M, Xiao Q M, Liu N X, Wang J H, Tan X P, Zheng H B. Rating of rice resistance against Ustilaginoidea virens and relationship between resistance and yield loss. Journal of Hunan Agricultural University: Natural Sciences, 2011, 37(3): 275-279. (in Chinese)

[3]尹小乐, 陈志谊, 刘永锋, 于俊杰, 李燕, 俞咪娜. 稻曲毒素A的相对含量分析及其与致病力的相关性. 中国农业科学, 2012, 45(22): 4720-4727.

Yin X L, Chen Z Y, Liu Y F, Yu J J, Li Y, Yu M N. Detection of the relative content of ustiloxin A in rice false smut balls and correlation analysis between pathogenicity and ustiloxin A production of Ustilaginoidea virens. Scientia Agricultura Sinica, 2012, 45(22): 4720-4727. (in Chinese)

[4]Li P X, Evans C D, Wu Y Z, Cao B, Hamel E, Joullié M M. Evolution of the total syntheses of ustiloxin natural products and their analogues. Journal of American Chemical Society, 2008, 130(7): 2351-2364.

[5]刘朋娟, 王政逸, 王秋华, 李德葆. 农杆菌介导的稻瘟病菌转化及致病缺陷突变体筛选. 中国水稻科学, 2006, 20(3): 231-237.

Liu P J, Wang Z Y, Wang Q H, Li D B. Agrobacterium tumefaciens- mediated transformation of Magnaporthe grisea and identification of pathogenicity defective mutant. Chinese Journal of Rice Science, 2006, 20(3): 231-237. (in Chinese)

[6]Chen X L, Yang J, Peng Y L. Large-scale insertional mutagenesis in Magnaporthe oryzae by Agrobacterium tumefaciens-mediated transformation. Methods in Molecular Biology, 2011, 722: 213-224.

[7]Maruthachalam K, Klosterman S J, Kang S, Hays R J, Subbarao K V. Identification of pathogenicity-related genes in the vascular wilt fungus Verticillium dahliae by Agrobacterium tumefaciens-mediated T-DNA insertional mutagenesis. Molecular Biotechnology, 2011, 49(3): 209-221.

[8]Chi M H, Park S Y, Kim S, Lee Y H. A novel pathogenicity gene is required in the rice blast fungus to suppress the basal defenses of the host. PLoS Pathogens, 2009, 5(4): e1000401.

[9]Li Y, Yan X, Wang H, Liang S, Ma W B, Fang M Y, Talbot N J, Wang A Y. MoRic8 is a novel component of G-protein signaling during plant infection by the rice blast fungus Magnaporthe oryzae. Molecular Plant Microbe Interactions, 2010, 23(3): 317-331.

[10]Goh J, Jeon J, Kim K S, Park J, Park S Y, Lee Y H. The PEX7-mediated peroxisomal import system is required for fungal development and pathogenicity in Magnaporthe oryzae. PLoS One, 2011, 6(12): e28220.

[11]Mullins E D, Chen X, Romaine P, Raina R, Geiser D M, Kang S. Agrobacterium-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer. Phytopathology, 2001, 91(2): 173-180.

[12]张震, 杜新法, 柴荣耀, 毛雪琴, 邱海萍, 王艳丽, 王教瑜, 孙国 昌. 根癌农杆菌介导遗传转化稻曲病菌. 中国水稻科学, 2006, 20(4): 440-442.

Zhang Z, Du X F, Chai R Y, Mao X Q, Qiu H P, Wang Y L, Wang J Y, Sun G C. Agrobacterium tumefaciens-mediated transformation of the pathogen of Ustilaginoidea virens. Chinese Journal of Rice Science, 2006, 20(4): 440-442. (in Chinese)

[13]张君成, 陈志谊, 张炳欣, 刘永锋, 陆凡. 稻曲病菌接种技术研究. 植物病理学报, 2004, 34(5): 463-467.

Zhang J C, Chen Z Y, Zhang B X, Liu Y F, Lu F. Study on biology of conidia of Ustilaginoidea virens. Acta Phytopathologica Sinica, 2004, 34(5): 463-467. (in Chinese)

[14]张君成, 张炳欣, 陈志谊, 刘永锋, 陆凡. 稻曲病菌分生孢子的生物学研究. 植物病理学报, 2003, 33(1): 44-47.

Zhang J C, Zhang B X, Chen Z Y, Liu Y F, Lu F. Study on biology of conidia of Ustilaginoidea virens. Acta Phytopathologica Sinica, 2003, 33(1): 44-47. (in Chinese)

[15]顾志敏, 丁正中, 陈析丰, 郭龙彪, 曾大力, 钱前, 马伯军. 实时荧光定量PCR筛选稻曲病菌内参基因. 中国水稻科学, 2012, 26(5): 615-618.

Gu Z M, Ding Z Z, Chen X F, Guo L B, Zeng D L, Qian Q, Ma B J. Reference genes selection of Ustilaginoidea virens by real-time quantitative polymerase chain reaction (PCR). Chinese Journal of Rice Science, 2012, 26(5): 615-618. (in Chinese)

[16]胡东维, 王疏. 稻曲病菌侵染机制研究现状与展望. 中国农业科 学, 2012, 45(22): 4604-4611.

Hu D W, Wang S. Progress and perspectives in infection mechanism of Ustilaginoidea virens. Scientia Agricultura Sinica, 2012, 45(22): 4604-4611. (in Chinese)

[17]刘见平, 唐涛, 张松柏, 郑和斌. 稻曲病初侵染源及病菌侵染适期初步研究. 杂交水稻, 2009, 24(1): 74-77.

Liu J P, Tang T, Zhang S B, Zheng H B. Preliminary studies on initial infection sources and pathogen-infecting favorable stage for rice false smut. Hybrid Rice, 2009, 24(1): 74-77. (in Chinese)

[18]Tang Y X, Jin J, Hu D W, Yong M L, Xu Y, He L P.  Elucidation of the infection process of Ustilaginoidea virens (teleomorph: Villosiclava virens) in rice spikelets. Plant Pathology, 2013, 62: 1-8.

[19]李燕, 于俊杰, 刘永锋, 尹小乐, 张荣胜, 俞咪娜, 陈志谊. 稻曲病菌产孢能力及致病力测定. 中国农业科学, 2012, 45(20): 4166-4177.

Li Y, Yu J J, Liu Y F, Yin X L, Zhang R S, Yu M N, Chen Z Y. Determination of sporulation and pathogenicity of Ustilaginoidea virens. Scientia Agricultura Sinica, 2012, 45(20): 4166-4177. (in Chinese)

[20]Guo M, Chen Y, Du Y, Dong Y H, Guo W, Zhai S, Zhang H F, Dong S M, Zhang Z G, Wang Y C, Wang P, Zheng X B. The bZIP transcription factor MoAP1 mediates the oxidative stress response and is critical for pathogenicity of the rice blast fungus Magnaporthe oryzae. PLoS Pathogens, 2011, 7(2): e1001302.

[21]刘连盟, 王玲, 黄雯雯, 刘恩勇, 黄世文. 水稻稻曲病菌G蛋白β亚基基因的克隆、表达与序列分析. 中国水稻科学, 2010, 24(4): 353-359.

Liu L M, Wang L, Huang W W, Liu E Y, Huang S W. Cloning, expression and sequence analysis of G protein β subunit gene of rice false smut pathogen Ustilaginoidea virens. Chinese Journal of Rice Science, 2010, 24(4): 353-359. (in Chinese)

[22]张震, 杜新法, 柴荣耀, 王教瑜, 邱海萍, 毛雪琴, 孙国昌. 稻曲病菌PMK1类同源基因克隆及在稻瘟病菌遗传互补中的功能验证. 微生物学报, 2008, 48(11): 1473-1478.

Zhang Z, Du X F, Chai R Y, Wang J Y, Qiu H P, Mao X Q, Sun G C. Cloning of a homologous gene of Magnaporthe grisea PMK1 Ttype MAPK from Ustilaginoidea virens and functional identification by complement in Magnaporthe grisea corresponding mutant. Acta Microbiologica Sinica, 2008, 48(11): 1473-1478. (in Chinese)

[23]Tanaka E, Kumagawa T, Tanaka C, Koga H. Simple transformation of the rice false smut fungus Villosiclava virens by electroporation of intact conidia. Mycoscience, 2011, 52: 344-348.

[24]Zhu Q H, Ramm K, Eamens A L, Dennis E S, Upadhyaya N M. Transgene structures suggest that multiple mechanisms are involved in T-DNA integration in plants. Plant Science, 2006, 171: 308-322.

[25]Müller A E, Atkinson R G, Sandoval R B, Jorgensen R A. Microhomologies between T-DNA ends and target sites often occur in inverted orientation and may be responsible for the high frequency of T-DNA-associated inversions. Plant Cell Reports, 2007, 26: 617-630.

[26]Zhu C F, Wu J H, He C Z. Induction of chromosomal inversion by integration of T-DNA in the rice genome. Journal of Genetics and Genomics, 2010, 37: 189-196.

[27]Li G H, Zhou Z Z, Liu G F, Zheng F C, He C Z. Characterization of T-DNA insertion patterns in the genome of rice blast fungus Magnaporthe oryzae. Current Genetics, 2007, 51: 233-243.
[1] LIU RUI, ZHAO YuHan, FU ZhongJu, GU XinYi, WANG YanXia, JIN XueHui, YANG Ying, WU WeiHuai, ZHANG YaLing. Distribution and Variation of PWL Gene Family in Rice Magnaporthe oryzae from Heilongjiang Province and Hainan Province [J]. Scientia Agricultura Sinica, 2023, 56(2): 264-274.
[2] WANG WenJuan,SU Jing,CHEN Shen,YANG JianYuan,CHEN KaiLing,FENG AiQing,WANG CongYing,FENG JinQi,CHEN Bing,ZHU XiaoYuan. Pathogenicity and Avirulence Genes Variation of Magnaporthe oryzae from a Rice Variety Meixiangzhan 2 in Guangdong Province [J]. Scientia Agricultura Sinica, 2022, 55(7): 1346-1358.
[3] TANG ZiYun,HU JianXin,CHEN Jin,LU YiXing,KONG LingLi,DIAO Lu,ZHANG FaFu,XIONG WenGuang,ZENG ZhenLing. Relationship Between Biofilm Formation and Molecular Typing of Staphylococcus aureus from Animal Origin [J]. Scientia Agricultura Sinica, 2022, 55(3): 602-612.
[4] JIANG QiQi,XU JianJian,SU Yue,ZHANG Qi,CAO Peng,SONG ChenHu,LI ZhongAn,SONG Zhen. Construction and Application of Infectious Clone of Citrus Yellow Mosaic Virus [J]. Scientia Agricultura Sinica, 2022, 55(24): 4840-4850.
[5] MA XueMeng,YU ChengMin,SAI XiaoLing,LIU Zhen,SANG HaiYang,CUI BaiMing. PSORA: A Strategy Based on High-Throughput Sequence for Analysis of T-DNA Insertion Sites [J]. Scientia Agricultura Sinica, 2022, 55(15): 2875-2882.
[6] YUAN JingLi,ZHENG HongLi,LIANG XianLi,MEI Jun,YU DongLiang,SUN YuQiang,KE LiPing. Influence of Anthocyanin Biosynthesis on Leaf and Fiber Color of Gossypium hirsutum L. [J]. Scientia Agricultura Sinica, 2021, 54(9): 1846-1855.
[7] CHEN ChaoXi,LI YuHan,TAN Min,WANG Lu,HUANG ZhiHong. Biofilm-Forming Phenotype, Antibacterial Resistance Genes, Integrase Genes and Virulence Genes Detection of Escherichia coli Isolated from Yaks and Tibetan Pigs in Northwest Sichuan Plateau [J]. Scientia Agricultura Sinica, 2021, 54(23): 5144-5162.
[8] CAI Ni,YAN DuoZi,NONG XiangQun,WANG GuangJun,TU XiongBing,ZHANG ZeHua. Adhesin Gene mad2 Knockout and Functional Effects on Biological Characteristics and Inducing Plant Responses in Metarhizium anisopliae [J]. Scientia Agricultura Sinica, 2021, 54(22): 4800-4812.
[9] ZHENG Wei,SHI Zheng,LONG Mei,LIAO YunCheng. Photosynthetic and Physiological Characteristics Analysis of Yellow- Green Leaf Mutant in Wheat of Jimai5265yg [J]. Scientia Agricultura Sinica, 2021, 54(21): 4539-4551.
[10] WANG ChengLi,YIN ZhiYuan,NIE JiaJun,LIN YongHui,HUANG LiLi. Identification and Virulence Analysis of CAP Superfamily Genes in Valsa mali [J]. Scientia Agricultura Sinica, 2021, 54(16): 3440-3450.
[11] ZHANG Shuo,ZHI Hui,TANG ChanJuan,LUO MingZhao,TANG Sha,JIA GuanQing,JIA YanChao,DIAO XianMin. Cytological Characters Analysis and Low-Resolution Mapping of Stripe-Leaf MutantA36-S in Foxtail Millet [J]. Scientia Agricultura Sinica, 2021, 54(14): 2952-2964.
[12] LI FeiFei,WANG BeiBei,LAI YingFang,YANG FeiYing,YOU MinSheng,HE WeiYi. Knockout of Single Allele of fl(2)d Significantly Decreases the Fecundity and Fertility inPlutella xylostella [J]. Scientia Agricultura Sinica, 2021, 54(14): 3029-3042.
[13] LI ZhaoWei,LING DongLan,SUN CongYing,ZENG HuiLing,LIU KaiJi,LAN YingShan,FAN Kai,LIN WenXiong. CRISPR/Cas9 Targeted Editing of OsIAA11 in Rice [J]. Scientia Agricultura Sinica, 2021, 54(13): 2699-2709.
[14] ZHAO Xue,WANG Feng,WANG WenJing,LIU XiaoFeng,BIAN ShiQuan,LIU YanHua,LIU XinMin,DU YongMei,ZHANG ZhongFeng,ZHANG HongBo. Splicing Property Analyses of the NRSE1 Element from Tobacco PR3b mRNA After Fusion Expression with GUS Gene [J]. Scientia Agricultura Sinica, 2020, 53(8): 1524-1531.
[15] KunNeng ZHOU,JiaFa XIA,Peng YUN,YuanLei WANG,TingChen MA,CaiJuan ZHANG,ZeFu LI. Transcriptome Research of Erect and Short Panicle Mutant esp in Rice [J]. Scientia Agricultura Sinica, 2020, 53(6): 1081-1094.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd