分离自棉花的轮枝菌“种”的鉴定

doi:10.3864/j.issn.0578-1752.2013.10.008

摘要/Abstract

摘要： 【目的】已知引起植物萎蔫病的轮枝菌有5个“种”，本研究旨在澄清在中国引起棉花黄萎病的轮枝菌的“种”类。【方法】从中国12个省84个县分离获得310个轮枝菌菌株，采用生物学性状观察和分子鉴定相结合的方法，对其进行“种”的鉴定。【结果】298株均产生黑色微菌核和轮枝状分生孢子梗，为典型的大丽轮枝菌，另有12个菌株培养性状较为特殊（不产生微菌核，出现黑色菌丝或厚壁孢子），经温度敏感试验、PCR特异扩增及ITS序列进一步鉴定，2株被确定为变黑轮枝菌，其它10株被确定为大丽轮枝菌，没有出现黑白轮枝菌。【结论】在中国三大棉区，棉花黄萎病主要是由大丽轮枝菌引起的。但在长江流域和黄河流域棉区均分离到1株变黑轮枝菌，其在棉花黄萎病发生过程中的作用及其与大丽轮枝菌的互作还有待进一步研究。

关键词: 大丽轮枝菌 , 变黑轮枝菌 , 黑白轮枝菌 , 棉花黄萎病

Abstract: 【Objective】In the genus of Verticillium, there are five ‘species’ as plant pathogens which could lead to plant wilt symptom. The aim of this study was to identify the specific ‘species’ caused Verticillium wilt of cotton.【Method】A total of 310 Verticillium isolates were obtained from 84 counties, 12 provinces of China. To clarify the ‘species’ of them, both observation on biological characteristics and molecular identification were used.【Result】A total of 298 isolates were typical V. dahliae producing black microsclerotia and verticillate conidiophores identified by specific PCR amplifications. However, 12 special isolates producing non-microsclerotia, black mycelia or chlamydospore were tested by temperature sensitivity, specific PCR amplification and ITS sequencing. Except for 2 V. nigrescens isolates, the rest 10 isolates were identified as V. dahliae, no V. albo-atrum was found. 【Conclusion】 In 3 main cotton growing regions of China, the causal agent of cotton Verticillium wilt is V. dahliae. And V. nigrescens also presented in Yangzi River and Yellow River basins. In cotton Verticillium wilt happening process, the interaction between V. dahliae and V. nigrescens needs to be further studied.

Key words: Verticillium dahlia , Verticillium nigrescens , Verticillium albo-atrum , cotton Verticillium wilt

朱荷琴, 冯自力, 李志芳, 师勇强, 赵丽红, 李彩红, 刘义杰, 王玲飞. 分离自棉花的轮枝菌“种”的鉴定[J]. 中国农业科学, 2013, 46(10): 2032-2040.

ZHU He-Qin, FENG Zi-Li, LI Zhi-Fang, SHI Yong-Qiang, ZHAO Li-Hong, LI Cai-Hong, LIU Yi-Jie, WANG Ling-Fei. Identification of Isolates of Verticillium Species from Cotton[J]. Scientia Agricultura Sinica, 2013, 46(10): 2032-2040.

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参考文献

[1]陈旭升, 陈永萱, 黄骏麒. 棉花黄萎病菌鉴定技术进展. 棉花学报, 1997, 9(2): 64-67.

Chen X S, ChenY X, Huang J Q. The developmental trend of Verticillium identification technique. Acta Gossypii Sinica, 1997, 9(2): 64-67. (in Chinese)

[2]Pegg G F, Brady B L. Verticillium Wilts. New York: CABI Publishing, 2002: 142-167.

[3]沈其益. 棉花病害—基础研究与防治. 北京: 科学出版社, 1992: 128-151.

Shen Q Y. Cotton Diseases -Basic Research and Prevention. Beijing: Science Press, 1992: 128-151. (in Chinese)

[4]张绪振, 张树琴, 陈吉隶, 李庆基, 陈壁, 姚跃文. 我国棉花黄萎病菌种的鉴定. 植物病理学报, 1981, 11(3): 13-18.

Zhang X Z, Zhang S Q, Chen J L, Li Q J, Chen B, Yao Y W. Identification of Verticillium wilt pathogen of cotton in China. Acta Phytopathologica Sinica, 1981, 11(3): 13-18. (in Chinese)

[5]简桂良, 马存, 石磊岩. 1995年北方棉区黄萎病大发生及综合防治措施. 植物保护, 1996, 22(3): 37-38.

Jian G L, Ma C, Shi L Y. The outbreak and integral control of Verticillium wilt in northern cotton producing region in 1995. Plant Protection, 1996, 22(3): 37-38. (in Chinese)

[6]陈婧, 白应文, 杨继娟, 陈卫民, 张莉, 胡小平. 苜蓿黄萎病菌中国菌株生物学特性研究. 草地学报, 2010, 18(2): 274-279.

Chen J, Bai Y W, Yang J J, Chen W M, Zhang L, Hu X P. Biological characteristics of Chinese strain of Verticillium albo-atrum from alfalf. Acta Agrestia Sinica, 2010, 18(2): 274-279. (in Chinese)

[7]薛光华, 范伟功, 张祥林. 新疆截获进境植物疫情分析及检疫监管措施. 新疆农业科学, 2004, 41(5): 263-267.

Xue G H, Fan W G, Zhang X L. Analysis on the intercepted situation of the entry plant pests and the quarantine & supervisory measures in Xinjiang. Xinjiang Agricultural Sciences, 2004, 41(5): 263-267. (in Chinese)

[8]吴翠萍, 李彬, 粟寒, 陈贯源, 戴忠军, 安榆林. 进境美国苜蓿草中苜蓿黄萎病菌的检疫鉴定. 植物检疫, 2011, 25(1): 42-46.

Wu C P, Li B, Su H, Chen G Y, Dai Z J, An Y L. Detection of Verticillium albo-atrum in alfalfa samples imported from the United States. Plant Quarantine, 2011, 25(1): 42-46. (in Chinese)

[9]黄胜光, 卢兆山, 邱世明, 奚国华, 陆道, 钟恒禄. 广西防城港首次截获苜蓿黄萎病菌. 植物保护, 2012, 38(1): 180-183.

Huang S G, Lu Z S, Qiu S M, Xi G H, Lu D, Zhong H L. First interception of Verticillium albo-atrum in Fangchenggang of Guangxi. Plant Protection, 2012, 38(1): 180-183. (in Chinese)

[10]段维军, 郭立新, 张慧丽, 刘培源, 夏侯阳, 陈先锋. 进境台湾芹菜上一株变黑轮枝菌的鉴定. 植物检疫, 2012, 26(4): 30-34.

Duan W J, Guo L X, Zhang H L, Liu P Y, Xia H Y, Chen X F. Identification of Verticillium nigrescens isolated in celery from Taiwan. Plant Quarantine, 2012, 26(4): 30-34. (in Chinese)

[11]刘学堂, 宋晓轩, 郭金城. 棉花黄萎病菌的研究及最新进展. 棉花学报, 1998, 10(1):6-13.

Liu X T, Song X X, Guo J C. Studies and advances on cotton Verticillium wilt. Acta Gossypii Sinica, 1998, 10(1): 6-13. (in Chinese)

[12]宋晓轩, 朱荷琴, 郭金城. 棉花黄萎病 (Verticillium dahliae Kleb.) 安阳菌系致病力分化研究. 中国农业科学, 1997, 30(1): 13-18.

Song X X, Zhu H Q, Guo J C. Study on pathogenicity differentiation of Verticillium dahliae Kleb. strains from Anyang China. Scientia Agricultura Sinica, 1997, 30(1): 13-18. (in Chinese)

[13]毛岚, 宋培玲, 杨家荣. 陕西棉花黄萎病菌致病力分化及其遗传多样性. 植物保护学报, 2009, 36(1): 27-31.

Mao L, Song P L, Yang J R. Pathogenicity differentiation and genetic diversity of Verticillium dahliae of cotton in Shaanxi. Acta Phytophylacica Sinica, 2009, 36(1): 27-31. (in Chinese)

[14]金利容, 万鹏, 孔令甲, 喻大昭, 黄薇, 黄民松, 汪莉. 湖北省棉花黄萎病病菌致病力分化研究. 棉花学报, 2011, 23(6): 566-572.

Jin L R, Wan P, Kong L J, Yu D Z, Huang W, Huang M S, Wang L. The study on the pathogenicity differentiation of Verticillium dahliae in Hubei province. Cotton Science, 2011, 23(6): 566-572. (in Chinese)

[15]林玲, 章如意, 张昕, 邓晟, 徐敬友, 周益军. 江苏省棉花黄萎病菌的培养特性及致病力分化监测. 棉花学报, 2012, 24(3): 199-206.

Lin L, Zhang R Y, Zhang X, Deng S, Xu J Y, Zhou Y J. Cultural characteristics and pathogenicity differentiation among strains of Verticillium dahliae from cotton in Jiangsu Province. Cotton Science, 2012, 24(3): 199-206. (in Chinese)

[16]Li K N, Rouse D I, Eyestone E J, German T L. The generation of specific DNA primers using random amplified polymorphic DNA and its application to Verticillium dahliae. Mycological Research, 1999, 103(11): 1361-1368.

[17]Nazar R N, Hu X, Schmidt J, Culham D, Robb J. Potential use of PCR-amplified ribosomal intergenic sequences in the detection and differentiation of Verticillium wilt pathogens. Physiological and Molecular Plant Pathology, 1991, 39(1): 1-11.

[18]朱荷琴, 冯自力, 尹志新, 李志芳, 赵丽红, 师勇强, 杨家荣. 我国棉花黄萎病菌致病力分化及ISSR指纹分析. 植物病理学报, 2012, 42(3): 225-235.

Zhu H Q, Feng Z L, Yin Z X, Li Z F, Zhao L H, Shi Y Q, Yang J R. Pathogenicity differentiation and ISSR fingerprint analysis of cotton Verticillium dahliae in China. Acta Phytopathologica Sinica, 2012, 42(3): 225-235. (in Chinese)

[19]White T J, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Genetics Selection Evolution, 1990, 38: 315-322.

[20]Isaac I. A comparative study of pathogenic isolates of Verticillium. Transactions of the British Mycological Society, 1949, 32: 137-157.

[21]Klosterman S J, Atallah Z K, Vallad G E, Subbarao K V. Diversity, pathogenicity, and management of Verticillium species. Annual Review of Phytopathology, 2009, 47: 39-62.

[22]Smith H C. The morphology of Verticillium albo-atrum, V. dahliae, and V. tricorpus. New Zealand Journal of Agricultural Research, 1965, 8(3): 450-478.

[23]Barbara D J, Clewes E. Plant pathogenic Verticillium species: how many of them are there? Molecular Plant Pathology, 2003, 4(4): 297-305.

[24]Carrell J C, Gordon T R, McCain A H. Vegetative compatibility and pathogenicity of Verticillium albo-utrum. Phytopathology, 1988, 78(8): 1017-1021.

[25]Hu X P, Wang M X, Hu D F. First report of wilt on alfalfa in China caused by Verticillium nigrescens. Plant Disease, 2011, 95(12): 1591.

[26]王克荣, 陆家云. 变黑轮枝菌种的鉴定及其致病力的测定. 南京农业大学学报, 1987(2): 59-62.

Wang K R, Lu J Y. Identification and pathogenicity determination of isolates of Verticillium Nigrescens Pethybr. Journal of Nanjing Agricultural University, 1987(2): 59-62. (in Chinese)

[27]金利容, 万鹏, 黄民松, 黄薇, 杨绍丽, 喻大昭. 棉花变黑轮枝菌的鉴定及致病性的测定. 湖北大学学报: 自然科学版, 2011, 33(4): 413-417.

Jin L R, Wan P, Huang M S, Huang W, Yang S L, Yu D Z. Identification and pathogenicity determination of isolates of Verticillium Nigrescens Pethybr. Journal of Hubei University: Natural Science, 2011, 33(4): 413-417. (in Chinese)

[28]Zhu H Q, Feng Z L, Li Z F, Shi Y Q, Zhao L H, Yang J R. Characterization of two fungal isolates from cotton and evaluation of their potential for biocontrol of Verticillium wilt of cotton. Journal of Phytopathology, 2013, 161: 70-77.