中国水稻科学
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  2010, Vol. 24 Issue (6): 617-622     DOI: 10.3969/j.issn.1001-7216.2010.06.009
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Establishment of Agrobacterium tumefaciensMediated Transformation System for Rhizoctonia solani AG-1 ⅠA, the Causal Agent of Rice Sheath Blight
YANG Ying-qing#, YANG Mei#, LI Ming-hai, LI Yong, HE Xiao-xia, ZHOU Er-xun*
Department of Plant Pathology, South China Agricultural University, Guangzhou 510642, China; #These authors contributed equally to this paper; *Corresponding author, E-mail: exzhou@scau.edu.cn
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Abstract  In order to construct the TDNA insertional mutagenesis transformation system for rice sheath blight pathogen Rhizoctonia solani AG1 ⅠA, the virulent isolate GD118 of this pathogen was selected as the initial isolate for transformation. The conditions for the transformation of GD118 were optimized in 5 aspects, i.e. preinduction time, coculture time, acetosyringone concentration at coculture stage, coculture temperature and pH values of solid induction medium (SIM) at coculture phase. Finally, an Agrobacterium tumefaciensmediated transformation (ATMT) system for R. solani AG1 ⅠA was established successfully. The optimal conditions for this ATMT system are as follows: the concentration of hygromycin B at 30 μg/mL for transformant screening, 8 h of preinduction time, 20 h of coculture time, 200 μmol/L of acetosyringone in SIM at coculture stage, coculture temperature at 25℃ and pH 5.6 to 5.8 of SIM at coculture phase. The transformants still showed high resistance to hygromycin B after 5 generations’ subcultures. Ten transformants were randomly picked out for PCR verification using the specific primers designed from the hph gene, and the results revealed that an expected band of 500 bp was amplified from all of the 10 transformants. Meantime, PCR amplification for these 10 transformants was carried out using specific primers designed from the Vir gene of A. tumefaciens, with 4 strains of A. tumefaciens as positive controls for eliminating the falsepositive caused by A. tumefaciens′s contamination, and the results showed that an expected band of 730 bp could be amplified from the 4 strains of A. tumefaciens, whereas no corresponding DNA band could be amplified from the 10 transformants. The results of above two PCR amplifications clearly showed that TDNA was indeed inserted into the targeted isolate GD118.
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Articles by authors
YANG Ying-qing#
YANG Mei#
LI Ming-hai
LI Yong
HE Xiao-xia
ZHOU Er-xun*
Key wordsrice sheath blight   Rhizoctonia solani   Agrobacterium tumefaciensmediated transformation   TDNA insertional mutagenesis   methodology     
Received: 1900-01-01;
Cite this article:   
YANG Ying-qing#,YANG Mei#,LI Ming-hai et al. Establishment of Agrobacterium tumefaciensMediated Transformation System for Rhizoctonia solani AG-1 ⅠA, the Causal Agent of Rice Sheath Blight[J]. , 2010, 24(6): 617-622 .
 
[1] Lee F N, Rush M C. Rice sheath blight: A major rice disease. Plant Dis, 1983, 67(7): 829-832.
[2] 周而勋, 曹菊香, 杨媚, 等. 我国南方六省 (区)水稻纹枯病菌遗传多样性的研究. 南京农业大学学报, 2002, 25(3): 36-40.
[3] 周而勋, 杨媚, 陈友林. 土壤环境因素对水稻纹枯病菌腐生定殖能力的影响. 植物病理学报, 2002, 32(3): 214-218.
[4] 黄世文, 王玲, 陈惠哲, 等. 氮肥施用量和施用方法对超级杂交稻纹枯病发生的影响. 植物病理学报, 2009, 39(1): 104-109.
[5] 肖勇, 刘明伟, 李刚, 等. 四川省水稻立枯丝核菌的遗传分化与致病力. 中国水稻科学, 2008, 22(1): 87-92. 浏览
[6] 黄江华, 杨媚, 周而勋, 等. 13种植物丝核菌对水稻、甜玉米、黄瓜和甘蓝的交互致病性. 华中农业大学学报, 2008, 27(2): 198-203.
[7] Cubeta M A, Vilgalys R. Population biology of the Rhizoctonia solani complex. Phytopathology, 1997, 87: 480-484.
[8] Ogoshi A. Ecology and pathogenicity of anastomosis and intraspecific groups of Rhizoctonia solani Kühn. Ann Rev Phytopathol, 1987, 25: 125-143.
[9] Stodart B J, Harvey P R, Neate S M, et al. Genetic variation and pathogenicity of anastomosis group 2 isolates of Rhizoctonia solani in Australia. Mycol Res, 2007, 111: 891-900.
[10] 王政逸, 李德葆. 限制酶介导的遗传转化及其在丝状真菌中的应用. 菌物系统, 2001, 20(1): 142-147.
[11] Mullins E D, Kang S. Transformation: A tool for studying fungal pathogens of plants. Cell Mol Life Sci, 2001, 58: 2043-2052.
[12] Combier J P, Melayah D, Raffier C, et al. Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in the symbiotic ectomycorrhizal fungus Hebeloma cylindrosporum. FEMS Microbiol Lett, 2003, 220(1): 141-148.
[13] Michielse C B, Hooykaas P J J, van den Hondel C A, et al. Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet, 2005, 48(1): 1-17.
[14] 李敏慧, 张荣, 姜大刚, 等. 根癌农杆菌介导的香蕉枯萎病菌4号生理小种的转化. 植物病理学报, 2009, 39(4): 405-412.
[15] Wu J, O′Brien P A. Stable transformation of Rhizoctonia solani with a modified hygromycin resistance gene. Australas Plant Pathol, 2009, 38: 79-84.
[16] Robinson H L, Deacon J W. Protoplast preparation and transient transformation of Rhizoctonia solani. Mycol Res, 2001, 105(11): 1295-1303.
[17] Sawada H, Ieki H, Matsuda I. PCR detection of Ti and Ri plasmids from phytopathogenic Agrobacterium strains. Appl Environ Microbiol, 1995, 61(2): 828-831.
[18] Tsuji G, Fujii S, Fujihara N, et al. Agrobacterium tumefaciens-mediated transformation for random insertional mutagenesis in Colletotrichum lagenarium. J General Plant Pathol, 2003, 69(4): 230-239.
[19] Meyer V, Mueller D, Strowig T, et al. Comparison of different transformation methods for Aspergillus giganteus. Curr Genet, 2003, 43(5): 371-377.
[20] Takahara H, Tsuji G, Kubo Y, et al. Agrobacterium tumefaciens-mediated transformation as a tool for random mutagenesis of Colletotrichum trifolii. J General Plant Pathol, 2004, 70(2): 93-96.
[21] Rogers C W, Challen M P, Green J R, et al. Use of REMI and Agrobacterium-mediated transformation to identify pathogenicity mutants of the biocontrol fungus, Coniothyrium minitans. FEMS Microbiol Lett, 2004, 241(2): 207-214.
[22] 迟彦, 周东坡, 平文祥, 等. 根癌农杆菌介导的真菌遗传转化及其应用.菌物学报, 2005, 24(4): 612-619.
[23] Samils N, Elfstrand M, Czederpiltz D L, et al. Development of a rapid and simple Agrobacterium tumefaciens mediated transformation system for the fungal pathogen Heterobasidion annosum. FEMS Microbiol Lett, 2006, 255(1): 82-88.
[24] Turk S C H J, Melchers L S, Dulk-Ras H D, et al. Environmental conditions differentially affect vir gene induction in different Agrobacterium strains: Role of the Vir A sensor protein. Plant Mol Biol, 1991, 16: 1051-1059.
[25] Rogowsky P M , Close T J, Chimera J A, et al. Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTi C58. J Bacteriol, 1987, 169(11): 5101-5112.
[26] Stachel S E, Zambryski P C. Agrobacterium tumefaciens and the susceptible plant cell: A novel adaptation of extracellular recognition and DNA conjugation. Cell, 1986, 47(2): 155-157.
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[2] XU De-jin,GU Zhong-yan*,XU Guang-chun,XU Xiao-long,FAN Peng. Effects of Solution Surface Tension and Spray Methods on Deposition of Droplets on Rice Plants[J]. , 2011, 25(2): 213-218 .
[3] JI Ying-hua,GAO Rui-zhen,ZHANG Ye,CHENG Zhao-bang,ZHOU Tong,FAN Yong-jian,ZHOU Yi-jun. A Simplified Method for Quick Detection of Rice BlackStreaked Dwarf Virus and Southern Rice BlackStreaked Dwarf Virus
[J]. , 2011, 25(1): 91-94 .
[4] WANG Ling,HUANG Wen-wen,HUANG Shi-wen,LIU Lian-meng,LIU En-yong,. Pathogenicity Differentiation of Rice Sheath Blight Pathogen Rhizoctonia solani AG-1 ⅠA Isolates from Anhui and Hubei Provinces, China[J]. , 2010, 24(6): 623-629 .
[5] SUN Chuan,CHEN Gang,RAO Yu-chun,ZHANG Guang-heng,GAO Zhen-yu,LIU Jian,JU Pei-na,HU Jiang,GUO Long-biao,QIAN Qian,ZENG Da-li . A Simple Method for Rapid Preparation of Rice Genomic DNA[J]. , 2010, 24(6): 677-680 .
[6] ZHOU Tong#,WU Li-juan#,WANG Ying,CHENG Zhao-bang,JI Ying-hua,FAN Yong-jian,ZHOU Yi-jun*. Preliminary Report on the Transmission of Rice BlackStreaked Dwarf Virus from Frozen Infected Leaves to Rice by Insect Vector Small Brown Planthopper (Laodelphax striatellus[J]. , 2010, 24(4): 425-428 .
[7] CHEN Tao, ZHANG Zhen, CHAI Rong-yao,WANG Jiao-yu,MAO Xue-qin,QIU Hai-ping,DU Xin-fa,JIANG Hua, WANG Li-an,WANG Yan-li .

Genetic Diversity and Pathogenicity Variation of Different Rhizoctonia solani Isolates in Rice from Zhejiang Province, China

[J]. , 2010, 24(1): 67-72 .
[8] XIAO Yong,LI Shuang-cheng,CHU Ming-guang,ZHOU Peng,GUAN Peng,LIU Li,WANG Ling-xia,ZHENG Ai-ping,LI Ping. Cloning,Expression and Characterization of G Protein β-subunit Gene in Rhizoctonia solani from Rice [J]. , 2008, 22(5): 541-544 .
[9] AO Yan,HU Zhi-qiu,TANG Zai-xiang,WANG Xue-feng,XU Chen-wu*. A General Method for QTL Mapping in Multiple Related Populations Derived from Multiple Parents[J]. , 2008, 22(4): 427-431 .
[10] XIAO Yong ,LIU Ming-wei ,LI Gang ,ZHOU Er-xun ,WANG Ling-xia ,TANG Jie ,TAN Fu-rong ,ZHENG Ai-ping ,LI ping. Genetic Diversity and Pathogenicity Variation of Different Rhizoctonia solani Isolates in Rice from Sichuan Province, China[J]. , 2008, 22(1): 87-92 .
[11] WEN Li,LIU Gai,LI Shao-qing,LI Guo-min,TAO Jun,ZHU Ying-guo. Establishment and Application of Two-Dimensional Gel Electrophoresis Analysis of Proteins from Rice Pollen [J]. , 2007, 21(1): 13-19 .
[12] YU Chu-lang ,YAN Shun-ping ,SUN Wei-ning ,YANG Ling. High-Resolution Two-Dimensional Electrophoresis for Total Proteins in Rice Roots, Leaves and Suspension Cells[J]. , 2006, 20(5): 549-552 .
[13] SUN Cheng-xiao ,DUAN Bin-wu ,XIE Li-hong ,CHEN Neng. Determination of Several Quality Characteristics of Brown Rice by Near Infrared Transmission Spectroscopy[J]. , 2006, 20(4): 451-454 .
[14] LIU Peng-juan ,WANG Zheng-yi ,WANG Qiu-hua ,LI De-bao . Agrobacterium tumefaciens-Mediated Transformation of Magnaporthe grisea and Identification of Pathogenicity Defective Mutant[J]. , 2006, 20(3): 231-237 .
[15] DUAN Gui-fang ,ZHANG Jian-ping ,ZHOU Yong-jun ,YU Liu-qing ,YUAN Qin-sheng . Biocontrol of Rice Sheath Blight by Helminthosporium gramineum Rabenh f. sp. echinochloae and Its Metabolite Ophiobolin A[J]. , 2006, 20(3): 337-339 .
[16] SUN Shu-bin ,LI Bao-zhen ,HU Jiang ,XU Guo-hua. Establishment and Application of a Real-Time Fluorescence Quantitative PCR for Detecting Transcripts of Low Abundance Gene, OsAMT1;3,in Rice[J]. , 2006, 20(1): 8-12 .
[17] DAI Wei-min ,ZHANG Ke-qin ,DUAN Bin-Wu ,SUN Cheng-xiao ,ZHENG Kang-le ,CAI Run ,ZHUANG Jie-yun . Rapid Determination of Silicon Content in Rice(Oryza sativa)[J]. , 2005, 19(5): 460-462 .
[18] MAO Xing-xue ,XIAO Xin ,CHEN Jian-wei ,LUO Wen-yong ,LIU Yan-zhuo ,LI Xiao-fang.

One-Step PCR Method for Detecting the First Base of Splice Donor of Wx Intron 1 in Rice

[J]. , 2005, 19(3): 285-287 .
[19] ZHANG Hai-yan ,DONG Hai-tao ,YAO Hai-gen ,XIANG Yue-wu ,TAN Xue-lin ,FANG Yong-qi ,Deng Ye ,JIN Qing-chao ,Cabezas DANIEL ,JIANG Yu-xin ,DAI Cheng-en ,YAN Wen-zhao ,LI De-bao. Isolation of Total RNA from Rice Endosperm and Expression of Important Genes Related to Grain Quality in Rice (Oryza sativa)[J]. , 2005, 19(2): 105-110 .
[20] TANG Shao-qing,SHI Chun-hai,JIAO Gui-ai,HU Pei-song,WANG Hai-lian,WAN Jian-min. Analysis of Fat Content in Milled Rice by Near Infrared Reflectance Spectroscopy[J]. , 2004, 18(6): 563-566 .
[21] CHENG Fang*,YING Yi-bin. Vision Inspection Analysis of Seed Characteristics in Hybrid Rice and Construction of Image Information Base [J]. , 2004, 18(5): 461-465 .
[22] SHANG Yan-fen,HOU Cai-yun,CHANG Guo-hua,ZHOU Xiao-feng. Automatic Detection of Head Rice[J]. , 2004, 18(5): 466-468 .
[23] WANG Jian-fei,CHEN Hong-you,YANG Qing-li,YAO Ming-zhe,ZHOU Guo-an,ZHANG Hong-sheng. Effects of Salt Concentration and Temperature on the Screening of Salt-Tolerance in Rice[J]. , 2004, 18(5): 449-454 .
[24] SHEN Bo ,PING Xiao-fei ,TANG Fu-bing ,YAN Hong-lan ,WANG Xi . Determination of Cytokinins in Rice Root Exudates by High Performance Liquid Chromatography(HPLC)[J]. , 2004, 18(4): 362-364 .
[25] LI Xiang-min ,HU Bai-shi ,XU Zhi-gang ,MEW T.W. . Threshold Population Sizes of Bacillus subtilis B5423-R to Suppress the Occurrence of Rice Sheath Blight [J]. , 2003, 17(4): 360-364 .
[26] LU De-zhao ,ZHU Shui-jin ,QIAN Qian ,WANG Hui-zhong ,YAN Mei-xian ,HUANG Da-nian . Research on the Methods for Transforming BADH Gene into Rice[J]. , 2003, 17(4): 323-327 .
[27] SHEN Wen-biao ,YU Wei-wei ,WANG Ren ,JIANG Ling ,ZHAI Hu-qu ,CHENG Shi-hua ,WAN Jian-min . A Rapid Method for Screening Storable Rice Cultivars by Carotene Bleaching[J]. , 2003, 17(4): 387-389 .
[28] HU Pei-song ,ZHAI Hu-qu ,TANG Shao-qing ,WAN Jian-min . A Simplified Method for Evaluating Gelatinization Temperature and Amylose Content of Rice[J]. , 2003, 17(3): 284-286 .
[29] XIAO Xin,CHEN Yi,LUO Wen-yong,LIU Yan-zhuo,MAO Xing-xue,LI Xiao-fang * . Determination of Amylose Content in Single Rice Grain by Near Infrared Transmittance Spectroscopy(NITS)[J]. , 2003, 17(3): 287-290 .
[30] ZHENG Ai-ping,LI Ping,WANG Shi-quan,WANG Ling-xia,MA Bing-tian. Screening,Taxonomy of Antagonistic Strain B34 Against Rhizoctonia solani and Acquirement of the Antiseptic Protein[J]. , 2002, 16(4): 356-360 .
[31] YU Xue-fang ,XIE Guan-lin ,J. SWINGS ,T. W. MEW . Relationships of Different Cultural Patterns of a Predominant Japonica Rice Variety and Population Variation of Antagonistic Bacteria Against Rice Sheath Blight and the Bacterial Species[J]. , 2002, 16(4): 351-355 .
[32] HE Qing-fang,CHEN Wei-liang,MA Zhi-chao. Purification and Properties of Antagonistic Peptide Produced by Bacillus subtilis A30[J]. , 2002, 16(4): 361-365 .
[33] LIU Ze-wen,HAN Zhao-jun,ZHANG Ling-chun,WANG Yin-chang. Methods for Insect Raising and Insecticide Resistance Selection with Rice Planthoppers[J]. , 2002, 16(2): 167-170 .
[34] TAN Gen-jia,WANG Zi-ying. Temporal and Spatial Niches of Rice Sheath Blight[J]. , 2002, 16(2): 182-184 .
[35] TANG Jia-bin,MA Bing-tian,WANG Ling-xia,LI Ping,ZHENG Ai-ping,CHEN Hong . Biological Control of Rice Sheath Blight with Trichoderma and Thichoderma-like[J]. , 2002, 16(1): 63-66 .
[36] WANG Xiu-feng,YANG Jian-bo,XIANG Tai-he,LI Li,NI Da-hu . A New Method for PCR Reaction from Alkali-Treated Rice Leaf Tissues[J]. , 2002, 16(1): 67-70 .
[37] CHEN Zong-xiang,ZOU Jun-huang,HAN Yue-peng,XU Jing-you,TONG Yun-hui,YU Heng-xiu,ZHANG Ya-fang,PAN Xue-biaoonclick="TPI_openwindow('/GRID/knowledgenet.aspx?sdb=CJFD&,sfield=%e%b%b%e%%%e%bd%c%e%%&,skey=PAN+Xue-biao++%ef%bc%Agricultural+College','knet','status=yes,scrollbars=yes,width=,height=')">. An Innovated Method for Research on Inheritance of Resistance to Rice Sheath Blight and Its Verification[J]. , 2002, 16(1): 74-76 .
[38] SU Zu-fang,ZHOU Pei-nan,XU Nai-xia,ZHANG Ya-jie. Effects of Nitrogen and Planting Density on N-Absorption and Yield of Rice[J]. , 2001, 15(4): 281-286 .
[39] XIAO Lang-tao ,LI Dong-hui ,LIN Wan-huang ,HONG Bin ,HONG Ya-hui . An Objective Method to Measure Chalkiness of Rice Grain [J]. , 2001, 15(3): 206-208 .
[40] TANG Jian,PING Xiao-fei,TANG Fu-bing,YANG Bao-jun. Determination of Juvenile Hormone in Brown Planthopper (Nilaparvata lugens) by GC/MS [J]. , 2001, 15(2): 142-144 .
[41] BAO Jin-song,SHU Qing-yao,WU Dian-xing,XIA Ying-wu. Comparative Study on the RVA Profiles from the Milled and Brown Rice Flour[J]. , 2001, 15(2): 145-146 .
[42] WEI Xing-hua,TANG Sheng-xiang,YU Han-yong,JIANG Yun-zhu,QIU Zong-en. Studies on Methods of Developing a Core Collection for China Traditional Japonica Rice Germplasm[J]. , 2000, 14(4): 237-240 .
[43] CHENG Shi-hua. Current Status and Prospect in the Development of Breeding Materials and Breeding Methodology of Hybrid Rice[J]. , 2000, 14(3): 165-169 .
[44] LI Li ,YANG Jian-bo ,D.J.MACKILL ,P.M.COLOWIT . Comparison of Different Detection Methods for Rice SSR Analysis[J]. , 2000, 14(3): 185-188 .
[45] WU Wei-ming,SONG Xiang-fu,ZHOU Guo-yan. Application of Floating Culture Method on Natural Waters for Rice Root Research[J]. , 2000, 14(3): 189-192 .
[46] CHEN Zhi-yi ,XU Zhi-gang ,GAO Tai-dong ,NI Shou-kun ,YAN Da-fu ,LU Fan ,LIU Yong-feng . Evaluation and Utilization of Antagonistic Bacteria against Rice Sheath Blight[J]. , 2000, 14(2): 98-102 .
[47] Fan Longjiang,Hu Binming,Xu Dexin. A Method for Selecting Locations in Rice Cultivar Trials[J]. , 2000, 14(1): 58-60 .
[48] Sun Yansong,Shang Shuhua,Zhang Yunjiang. Selection Efficiency of Rice Anther Culture Breeding in Cold Region[J]. , 1999, 13(3): 176-178 .
[49] Dai Luyuan,Ye Changrong,Xiong Jianhua,Wang Huiyi. Methods for Evaluating Cold Tolerance in Rice[J]. , 1999, 13(1): 62-62 .
[50] Chen Zhiyi,T. W. Mew. The Distribution and Species of Antagonistic Bacteria Against Sheath Blight in Rice Ecosystem (in English)[J]. , 1998, 12(1): 35-39 .
[51] Qing Houguo,Ye Zhengxiang,Li Hua. Studies on Estimation Methods of Larva Density of Hydrellia sasakii[J]. , 1996, 10(3): 185-188 .
[52] Wu Lianghua,Wu Ping,Fang Ping,Tao Qinnan. Optimal Regulation Technique in Soil Moisture at Paddy Field (in English)[J]. , 1995, 9(4): 230-234 .
[53] Yang Wei,Bi Xuezhi,Huang Danian,Xiao Yuhua. A Method for the Two-dimensional Gel Electrophoresis of Rice Proteins[J]. , 1993, 7(1): 43-47 .
[54] Lu Yangjiang,Zheng Kangle. A Simple Method for Isolation of Rice DNA[J]. , 1992, 6(1): 47-48 .
[55] Pan Jun,Qiu Lingcang,Duan Binwu. Studies on the Simultaneous Analysis of Elements Content in Rice by ICP-AES with Microwave Oven Digesting[J]. , 1991, 5(2): 87-90 .
[56] Zhu Zhiwei,Lin Ronghui. Alkaline Digestion and Oxidation for the Speed Determination of the Silica in Rice Hull [J]. , 1990, 4(2): 89-91 .
[57] Xie Shaoping,Ni Jinshan. Correlation between K+ Influx and H+ Efflux by Roots of Intact Seedlings of indica Rice Varieties: A Method for Screening Efficient K+-uptaking Lines[J]. , 1990, 4(1): 22-26 .
[58] Zhang Zhitao,E A Heinrichs,F G Medrano. Screening for Resistance in Rice to the Brown Planthopper,Nilaparvata lugens(Stal),Using the Modified Seedbox Screening Technique[J]. , 1986, 1(1): 74-79 .
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