北疆棉区棉花黄萎病菌培养性状及致病力分化
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摘要
【目的】研究新疆北疆棉区棉花黄萎病菌的致病类型、培养特性及致病力分化。【方法】从北疆主要植棉区采集棉花黄萎病病株,利用大丽轮枝菌特异引物VDS-F/VDS-R、落叶型菌系特异性引物D1/D2、非落叶型菌系特异性引物ND1/ND2对所有分离的菌株进行检测;根据落叶型和非落叶型菌株在单个条田中所占的比例对田间地块进行分析;选取18个代表菌株对其在PDA上的培养特性及对棉花的致病力分析。【结果】从北疆棉田病株上分离到644个大丽轮枝菌菌株,67.5%的菌株为落叶型菌系,27.8%为非落叶型菌系,4.7%菌株不能归类,供试菌株的致病类型存在分化,其中以落叶型菌系为主要致病类型。对田间地块进行分析,单一落叶型菌株发生的地块最多,其次是落叶型和非落叶型菌株混合发生地块,田间以落叶型菌株为优势菌株的地块占70.1%,落叶型菌系为田间地块的主要致病类型。18个代表性菌株形成菌核型、菌丝型和中间型3种菌落类型,菌核型为主要的培养类型,并进一步分化形成3种不同的类型。对棉花感病品种的致病力进行测定,落叶型菌株引起的病指普遍高于非落叶型菌株引起的病指,非落叶型菌株中也有病指很高的菌株,北疆棉花黄萎病菌的致病力存在明显分化。【结论】北疆棉田采集的棉花黄萎病菌,其致病类型、培养特性及致病力均存在明显分化。
【Objective】 This research focuses on determining the pathotypes, cultural characteristics and pathogenicity differentiation of Verticillium dahliae from cottons in Northern Xinjiang. 【Method】All strains isolated from infected cotton plants were detected with specific primer pair VDS-F and VDS-R for V. dahliae, with specific primer pair D1 and D2 for defoliating strain, with specific primer pair ND1 and ND2 for non-defoliating strain, respectively. Field plots were analyzed according to the proportion of the defoliating and non-defoliating strains in every plot. 18 representative strains of V. dahliae were selected to investigate their cultural characteristics on PDA media and virulence to cotton seedling. 【Result】 All 644 strains isolated from infected cotton plants were determined to be V. dahliae, and the defoliating and non-defoliating strains accounted for 67.5% and 27.8%, respectively, 4.7% strains could not be classified into defoliating or non-defoliating strains, indicating that the pathogenic types of the tested strains were differentiated, among which defoliating strain was the main pathotype. Analysis of field plots showed that the plots only with defoliating strains were the most, and then mixed plots with defoliating strains and non-defoliating strains. 70.1% of the field plots were dominated by defoliating strains, suggesting that defoliating strains had become the main pathogenic type in the fields. The results of cultural characteristics on PDA media showed that 18 representative strains produced three colony morphologies: sclerotium type, hyphal type and intermediate type. Sclerotium type colony was the main culture type and further differentiated into three different colony types. The infection assays displayed that the average disease index of cotton seedling infected with defoliating strains was higher than that of non-defoliating strains, and there were strains with high disease index among the non-defoliating strains. 【Conclusion】The differentiation of pathogenic types, culture characteristics and virulence exist in strains of V. dahliae from cotton fields in northern Xinjiang.
【Objective】 This research focuses on determining the pathotypes, cultural characteristics and pathogenicity differentiation of Verticillium dahliae from cottons in Northern Xinjiang. 【Method】All strains isolated from infected cotton plants were detected with specific primer pair VDS-F and VDS-R for V. dahliae, with specific primer pair D1 and D2 for defoliating strain, with specific primer pair ND1 and ND2 for non-defoliating strain, respectively. Field plots were analyzed according to the proportion of the defoliating and non-defoliating strains in every plot. 18 representative strains of V. dahliae were selected to investigate their cultural characteristics on PDA media and virulence to cotton seedling. 【Result】 All 644 strains isolated from infected cotton plants were determined to be V. dahliae, and the defoliating and non-defoliating strains accounted for 67.5% and 27.8%, respectively, 4.7% strains could not be classified into defoliating or non-defoliating strains, indicating that the pathogenic types of the tested strains were differentiated, among which defoliating strain was the main pathotype. Analysis of field plots showed that the plots only with defoliating strains were the most, and then mixed plots with defoliating strains and non-defoliating strains. 70.1% of the field plots were dominated by defoliating strains, suggesting that defoliating strains had become the main pathogenic type in the fields. The results of cultural characteristics on PDA media showed that 18 representative strains produced three colony morphologies: sclerotium type, hyphal type and intermediate type. Sclerotium type colony was the main culture type and further differentiated into three different colony types. The infection assays displayed that the average disease index of cotton seedling infected with defoliating strains was higher than that of non-defoliating strains, and there were strains with high disease index among the non-defoliating strains. 【Conclusion】The differentiation of pathogenic types, culture characteristics and virulence exist in strains of V. dahliae from cotton fields in northern Xinjiang.
引文
[1]段维军,李国英,张莉,等.新疆棉花黄萎病菌致病性分化监测研究[J].新疆农业科学,2004,(5):324-328.DUAN Wei-jun,LI Guo-ying,ZHANG Li,et al.(2004).Monitoring of pathogenic differentiation of Verticillium dahliae in Xinjiang [J].Xinjiang Agricultural Sciences,(5):324-328.(in Chinese)
[2]Schnathorst,W.C.,& Mathre,D.E.(1966).Host range and differentiation of a severe form of verticillium albo-atrum in cotton.Phytopathology,56(10):1,155-1,161.
[3]姚耀文,傅翠真,王文录,等.棉花黄萎病菌生理型鉴定的初步研究[J].植物保护学报,1982,(3):145-148.YAO Yao-wen,FU Cui-zhen,WANG Wen-lu,et al.(1982).Preliminary study on physiological identification of Verticillium dahliae [J].Journal of Plant Protection,(3):145-148.(in Chinese)
[4]朱荷琴,冯自力,赵丽红,等.我国棉花黄萎病菌的致病力分化及落叶型菌株的分布[C]//.中国植物病理学会学术年会,2010.ZHU He-qin,FENG Zi-li,ZHAO Li-hong,et al.(2010).Pathogenicity differentiation of Verticillium dahliae in China and distribution of defoliated strains [C]//.Academic Annual Meeting of the Chinese Society of Plant Pathology.(in Chinese)
[5]韩宏伟,刘培源,高峰,等.新疆北部棉区黄萎病菌种群致病性分化及变异[J].植物保护学报,2011,38(2):121-126.HAN Hong-wei,LIU Pei-yuan,GAO Feng,et al.(2011).Pathogenic differentiation and variation of Verticillium dahlia population in northern cotton area of Xinjiang [J].Journal of Plant Protection,38 (2):121-126.(in Chinese)
[6]李国英,张新全,宋玉萍,等.北疆棉区棉花黄萎病发生趋势、抗性研究[J].新疆农业科学,2015,52(1):185-190.LI Guo-ying,ZHANG Xin-quan,SONG Yu-ping,et al.(2015).Study on the occurrence trend and resistance of cotton Verticillium in cotton areas in northern Xinjiang [J].Xinjiang Agricultural Sciences,52(1):185-190.(in Chinese)
[7]刘海洋,王伟,张仁福,等.新疆主要棉区棉花黄萎病发生概况[J].植物保护,2015,41(3):138-142.LIU Hai-yang,WANG Wei,ZHANG Ren-fu,et al.(2015).The occurrence of cotton verticillium in the main cotton areas of Xinjiang [J].Plant Protection,41(3):138-142.(in Chinese)
[8]Jonge,R.D.,Esse,H.P.V.,Maruthachalam,K.,Bolton,M.D.,& Thomma,B.P.H.J.(2012).Tomato immune receptor ve1 recognizes effector of multiple fungal pathogens uncovered by genome and RNA sequencing.Proceedings of the National Academy of Sciences,109(13):5,110-5,115.
[9]朱荷琴,冯自力,尹志新,等.我国棉花黄萎病菌致病力分化及ISSR指纹分析[J].植物病理学报,2012,42(3):225-235.ZHU He-qin,FENG Zi-li,YIN Zhi-xin,et al.(2012).Pathogenicity differentiation and ISSR fingerprint analysis of Verticillium dahliae in China [J].Journal of Plant Pathology,42(3):225-235.(in Chinese)
[10]石磊岩,孙文姬.棉花抗黄萎病苗期鉴定方法[J].植物保护,1987,(1):42.SHI Lei-yan,SUN Wen-ji.(1987).Identification method of cotton against Verticillium wilt disease seedling stage [J].Plant Protection,(1):42.(in Chinese)
[11]庞莉,李梅,孙青,等.棉花黄萎病病原菌大丽轮枝菌的快速分子检测[J].植物保护学报,2016,43(6):892-899.PANG Li,LI Mei,SUN Qing,et al.(2016).Rapid molecular detection of Verticillium dahliae,a pathogen of cotton Verticillium wilt [J].Journal of Plant Protection,43 (6):892-899.(in Chinese)
[12]Short,D.P.G.,Gurung,S.,Maruthachalam,K.,Atallah,Z.K.,& Short,D.P.G.(2014).Verticillium dahliae race 2-specific pcr reveals a high frequency of race 2 strains in commercial spinach seed lots and delineates race structure.Phytopathology,104(7):799-795.
[13]段维军.新疆棉花黄萎病菌致病性分化及其快速检测技术研究[D].石河子:石河子大学硕士论文,2005.DUAN Wei-jun.(2005).Study on pathogenic differentiation and rapid detection technology of Verticillium dahliae in Xinjiang [D].Master Dissertation.Shihezi University,Shihezi.(in Chinese)
[14]彭姗,吕学莲,高峰,等.一种新的棉花黄、枯萎病快速接种方法的研究[J].棉花学报,2008,20(3):174-178.PENG Shan,Lü Xue-lian,GAO Feng,et al.(2008).A new method for rapid inoculation of cotton yellow and wilt disease [J].Cotton Journal,20(3):174-178.(in Chinese)
[15]石磊岩,王波,文学.我国棉花黄萎病菌类型分化及培养特性研究[J].植物保护学报,1993,20(3):247-252.SHI Lei-yan,WANG Bo,WEN Xue.(1993).Study on the differentiation and culture characteristics of Verticillium dahliae in China [J].Journal of Plant Protection,20(3):247-252.(in Chinese)
[16]刘琳琳,张文文,周易,等.棉花与番茄抗棉花黄萎病不依赖于Ve1[J].中国科学:生命科学,2014,(8):803-814.LIU Lin-lin,ZHANG Wen-wen,ZHOU Yi,et al.(2014).Cotton and tomato resistant to cotton verticillium does not depend on Ve1 [J].Chinese Science:Life Sciences,(8):803-814.(in Chinese)
[17]喻秀秀,王卿,张昕,等.江苏省大丽轮枝菌的培养、遗传及致病特性分析[J].植物病理学报,2018,48(3):378-388.YU Xiu-xiu,WANG Qing,ZHANG Xin,et al.(2018).Analysis of culture,genetic and pathogenic characteristics of Verticillium dahliae in Jiangsu Province [J].Journal of Plant Pathology,48(3):378-388.(in Chinese)
[18]刘廷利,惠慧,阚家亮,等.新疆北部棉花黄萎病菌培养特性、致病型、致病性分化及ISSR遗传变异研究[J].棉花学报,2017,(6):51-59.LIU Ting-li,HUI Hui,KAN Jia-liang,et al.(2017).Culture characteristics,pathogenicity,pathogenic differentiation and ISSR genetic variation of Verticillium dahliae in northern Xinjiang [J].Cotton Journal,(6):51-59.(in Chinese)
[19]袁洪波,艾尼江,赵建军,等.棉花黄萎病菌致病力分化与 ISSR 遗传变异分析[J].华北农学报,2013,28(5):84-89.YUAN Hong-bo,AI Ni-jiang,ZHAO Jian-jun,et al.(2013).Pathogenicity differentiation of Verticillium dahliae and ISSR genetic variation analysis [J].North China Agricultural Journal,28(5):84-89.(in Chinese)
[20]朱荷琴,李志芳,冯自力,等.我国棉花黄萎病研究十年回顾及展望[J].棉花学报,2017,(S1):37-50.ZHU He-qin,LI Zhi-fang,FENG Zi-li,et al.(2017).Review and prospect of research on cotton Verticillium in China [J].Cotton Journal,(S1):37-50.(in Chinese)
[21]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[J].中国农业科学,1997,30(1):13-18.(in Chinese)SONG Xiao-xuan,ZHU He-qin,GUO Jin-cheng.(1997).Study on pathogenicity differentiation of Anyang strain of Verticillium dahliae Kleb.[J].Chinese Agricultural Sciences,30(1):13-18.(in Chinese)
[22]田秀明.山西棉花黄萎病菌致病力分化与其类型和生理的研究[J].植物保护,1995,21(3):8-10.TIAN Xiu-ming.(1995).Study on pathogenicity differentiation and type and physiology of Verticillium dahliae in Shanxi [J].Plant Protection,21(3):8 -10.(in Chinese)
[2]Schnathorst,W.C.,& Mathre,D.E.(1966).Host range and differentiation of a severe form of verticillium albo-atrum in cotton.Phytopathology,56(10):1,155-1,161.
[3]姚耀文,傅翠真,王文录,等.棉花黄萎病菌生理型鉴定的初步研究[J].植物保护学报,1982,(3):145-148.YAO Yao-wen,FU Cui-zhen,WANG Wen-lu,et al.(1982).Preliminary study on physiological identification of Verticillium dahliae [J].Journal of Plant Protection,(3):145-148.(in Chinese)
[4]朱荷琴,冯自力,赵丽红,等.我国棉花黄萎病菌的致病力分化及落叶型菌株的分布[C]//.中国植物病理学会学术年会,2010.ZHU He-qin,FENG Zi-li,ZHAO Li-hong,et al.(2010).Pathogenicity differentiation of Verticillium dahliae in China and distribution of defoliated strains [C]//.Academic Annual Meeting of the Chinese Society of Plant Pathology.(in Chinese)
[5]韩宏伟,刘培源,高峰,等.新疆北部棉区黄萎病菌种群致病性分化及变异[J].植物保护学报,2011,38(2):121-126.HAN Hong-wei,LIU Pei-yuan,GAO Feng,et al.(2011).Pathogenic differentiation and variation of Verticillium dahlia population in northern cotton area of Xinjiang [J].Journal of Plant Protection,38 (2):121-126.(in Chinese)
[6]李国英,张新全,宋玉萍,等.北疆棉区棉花黄萎病发生趋势、抗性研究[J].新疆农业科学,2015,52(1):185-190.LI Guo-ying,ZHANG Xin-quan,SONG Yu-ping,et al.(2015).Study on the occurrence trend and resistance of cotton Verticillium in cotton areas in northern Xinjiang [J].Xinjiang Agricultural Sciences,52(1):185-190.(in Chinese)
[7]刘海洋,王伟,张仁福,等.新疆主要棉区棉花黄萎病发生概况[J].植物保护,2015,41(3):138-142.LIU Hai-yang,WANG Wei,ZHANG Ren-fu,et al.(2015).The occurrence of cotton verticillium in the main cotton areas of Xinjiang [J].Plant Protection,41(3):138-142.(in Chinese)
[8]Jonge,R.D.,Esse,H.P.V.,Maruthachalam,K.,Bolton,M.D.,& Thomma,B.P.H.J.(2012).Tomato immune receptor ve1 recognizes effector of multiple fungal pathogens uncovered by genome and RNA sequencing.Proceedings of the National Academy of Sciences,109(13):5,110-5,115.
[9]朱荷琴,冯自力,尹志新,等.我国棉花黄萎病菌致病力分化及ISSR指纹分析[J].植物病理学报,2012,42(3):225-235.ZHU He-qin,FENG Zi-li,YIN Zhi-xin,et al.(2012).Pathogenicity differentiation and ISSR fingerprint analysis of Verticillium dahliae in China [J].Journal of Plant Pathology,42(3):225-235.(in Chinese)
[10]石磊岩,孙文姬.棉花抗黄萎病苗期鉴定方法[J].植物保护,1987,(1):42.SHI Lei-yan,SUN Wen-ji.(1987).Identification method of cotton against Verticillium wilt disease seedling stage [J].Plant Protection,(1):42.(in Chinese)
[11]庞莉,李梅,孙青,等.棉花黄萎病病原菌大丽轮枝菌的快速分子检测[J].植物保护学报,2016,43(6):892-899.PANG Li,LI Mei,SUN Qing,et al.(2016).Rapid molecular detection of Verticillium dahliae,a pathogen of cotton Verticillium wilt [J].Journal of Plant Protection,43 (6):892-899.(in Chinese)
[12]Short,D.P.G.,Gurung,S.,Maruthachalam,K.,Atallah,Z.K.,& Short,D.P.G.(2014).Verticillium dahliae race 2-specific pcr reveals a high frequency of race 2 strains in commercial spinach seed lots and delineates race structure.Phytopathology,104(7):799-795.
[13]段维军.新疆棉花黄萎病菌致病性分化及其快速检测技术研究[D].石河子:石河子大学硕士论文,2005.DUAN Wei-jun.(2005).Study on pathogenic differentiation and rapid detection technology of Verticillium dahliae in Xinjiang [D].Master Dissertation.Shihezi University,Shihezi.(in Chinese)
[14]彭姗,吕学莲,高峰,等.一种新的棉花黄、枯萎病快速接种方法的研究[J].棉花学报,2008,20(3):174-178.PENG Shan,Lü Xue-lian,GAO Feng,et al.(2008).A new method for rapid inoculation of cotton yellow and wilt disease [J].Cotton Journal,20(3):174-178.(in Chinese)
[15]石磊岩,王波,文学.我国棉花黄萎病菌类型分化及培养特性研究[J].植物保护学报,1993,20(3):247-252.SHI Lei-yan,WANG Bo,WEN Xue.(1993).Study on the differentiation and culture characteristics of Verticillium dahliae in China [J].Journal of Plant Protection,20(3):247-252.(in Chinese)
[16]刘琳琳,张文文,周易,等.棉花与番茄抗棉花黄萎病不依赖于Ve1[J].中国科学:生命科学,2014,(8):803-814.LIU Lin-lin,ZHANG Wen-wen,ZHOU Yi,et al.(2014).Cotton and tomato resistant to cotton verticillium does not depend on Ve1 [J].Chinese Science:Life Sciences,(8):803-814.(in Chinese)
[17]喻秀秀,王卿,张昕,等.江苏省大丽轮枝菌的培养、遗传及致病特性分析[J].植物病理学报,2018,48(3):378-388.YU Xiu-xiu,WANG Qing,ZHANG Xin,et al.(2018).Analysis of culture,genetic and pathogenic characteristics of Verticillium dahliae in Jiangsu Province [J].Journal of Plant Pathology,48(3):378-388.(in Chinese)
[18]刘廷利,惠慧,阚家亮,等.新疆北部棉花黄萎病菌培养特性、致病型、致病性分化及ISSR遗传变异研究[J].棉花学报,2017,(6):51-59.LIU Ting-li,HUI Hui,KAN Jia-liang,et al.(2017).Culture characteristics,pathogenicity,pathogenic differentiation and ISSR genetic variation of Verticillium dahliae in northern Xinjiang [J].Cotton Journal,(6):51-59.(in Chinese)
[19]袁洪波,艾尼江,赵建军,等.棉花黄萎病菌致病力分化与 ISSR 遗传变异分析[J].华北农学报,2013,28(5):84-89.YUAN Hong-bo,AI Ni-jiang,ZHAO Jian-jun,et al.(2013).Pathogenicity differentiation of Verticillium dahliae and ISSR genetic variation analysis [J].North China Agricultural Journal,28(5):84-89.(in Chinese)
[20]朱荷琴,李志芳,冯自力,等.我国棉花黄萎病研究十年回顾及展望[J].棉花学报,2017,(S1):37-50.ZHU He-qin,LI Zhi-fang,FENG Zi-li,et al.(2017).Review and prospect of research on cotton Verticillium in China [J].Cotton Journal,(S1):37-50.(in Chinese)
[21]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[J].中国农业科学,1997,30(1):13-18.(in Chinese)SONG Xiao-xuan,ZHU He-qin,GUO Jin-cheng.(1997).Study on pathogenicity differentiation of Anyang strain of Verticillium dahliae Kleb.[J].Chinese Agricultural Sciences,30(1):13-18.(in Chinese)
[22]田秀明.山西棉花黄萎病菌致病力分化与其类型和生理的研究[J].植物保护,1995,21(3):8-10.TIAN Xiu-ming.(1995).Study on pathogenicity differentiation and type and physiology of Verticillium dahliae in Shanxi [J].Plant Protection,21(3):8 -10.(in Chinese)