广东西番莲茎基腐病病原的分离及鉴定
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摘要
【目的】明确引起广东西番莲茎基腐病的病原菌,为有效防治该病害提供理论依据。【方法】采用组织分离法对广东番莲茎基腐病的病原进行分离,利用柯赫氏法则测定病原菌的致病性,光学显微镜观察病原菌分生孢子的形态学特征,测序分析病原菌的rDNA ITS序列。【结果】基于柯赫氏法则的致病性测定,证实从发病西番莲茎基部主干分离获得的病原菌对西番莲具有致病性。BLAST在线比对分析结果显示,病原菌r DNA ITS序列与多个腐皮镰孢菌菌株的相似性在99%以上。结合病原菌形态学特征、致病性测定及rDNA ITS序列分析结果,可确定引起广东西番莲茎基腐病的病原菌为腐皮镰孢菌[Fusarium solani(Mart)Sacc)]。【结论】近年来引起广东西番莲茎基腐病的病原菌为腐皮镰孢菌。防治西番莲茎基腐病需注重早期预防,提早防治;合理规划种植,加强栽培;做好冬季清园,合理疏剪。
【Objective】The aim of the study was to clarify the pathogen causing stem rot of Passiflora Linn.,and provide a theoretical basis for effective prevention and treatment of the disease.【Method】The pathogen was obtained by tissue separation method,and the pathogenicity of the pathogen was determined by Koch's rule. The morphological characteristics of the conidia of the pathogen were observed by optical microscope,and the r DNA-ITS sequence of the pathogen was analyzed by online BLAST.【Result】Based on the pathogenicity measurement of Koch's rule,it was confirmed that the isolated bacteria from the stem stalk was pathogenic to Passiflora Linn.. The BLAST online alignment showed that the similarity between the pathogen ITS sequence and those of multiple Fusarium solani strains was over 99%. Combined with the pathological characteristics of pathogen,the pathogenicity determination and sequence analysis of ITS,the pathogen causing stem rot of Passiflora Linn. was identified as Fusarium solani(Mart)Sacc.【Conclusion】F. solani is the pathogen that can infect the base stem of Passiflora Linn. Controlling the stem rot of Passiflora Linn. requires early prevention,planning planting,strengthening cultivation,clearing garden and rationally cutting.
【Objective】The aim of the study was to clarify the pathogen causing stem rot of Passiflora Linn.,and provide a theoretical basis for effective prevention and treatment of the disease.【Method】The pathogen was obtained by tissue separation method,and the pathogenicity of the pathogen was determined by Koch's rule. The morphological characteristics of the conidia of the pathogen were observed by optical microscope,and the r DNA-ITS sequence of the pathogen was analyzed by online BLAST.【Result】Based on the pathogenicity measurement of Koch's rule,it was confirmed that the isolated bacteria from the stem stalk was pathogenic to Passiflora Linn.. The BLAST online alignment showed that the similarity between the pathogen ITS sequence and those of multiple Fusarium solani strains was over 99%. Combined with the pathological characteristics of pathogen,the pathogenicity determination and sequence analysis of ITS,the pathogen causing stem rot of Passiflora Linn. was identified as Fusarium solani(Mart)Sacc.【Conclusion】F. solani is the pathogen that can infect the base stem of Passiflora Linn. Controlling the stem rot of Passiflora Linn. requires early prevention,planning planting,strengthening cultivation,clearing garden and rationally cutting.
引文
曹雪梅,李生兵,张惠玲,胡小平.2014.甘草根腐病病原菌鉴定[J].植物病理学报,44(2):213-216.[Cao X M,Li SB,Zhang H L,Hu X P.2014.Identification of the pathogens causing root rot of Glycyrrhiza uralensis Fisch[J].Acta Phytopathologica Sinica,44(2):213-216.]
陈企村,朱有勇,李振岐,唐永生,康振生.2009.不同品种混种对小麦产量及条锈病的影响[J].中国生态农业学报,17(1):29-33.[Chen Q C,Zhu Y Y,Li Z Q,Tang Y S,Kang Z S.2009.Effect of wheat cultivar mixtures on wheat yield and stripe rust[J].Chinese Journal of EcoAgriculture,17(1):29-33.]
陈星,高淑梅,李迎宾,张治萍,蒋娜,罗来鑫,李健强.2017.广西防城港西番莲茎基腐病发生为害调查与病原鉴定[C]//中国植物病理学会2017年学术年会论文集.北京:中国农业科学技术出版社:147.[Chen X,Gao S M,Li Y B,Zhang Z P,Jiang N,Luo L X,Li J Q.2017.Investigation and pathogen identification of stem rot of Passiflora in Fangchenggang,Guangxi[C]//Proceedings of the 2017 Annual Meeting of the Chinese Society of Plant Pathology.Beijing:China Agricultural Science and Technology Press:147.]
杜宾.2017.镰刀菌的分类和研究现状[J].太原学院学报(自然科学版),35(4):68-70.[Du B.2017.The classification and research status of Fusarium[J].Journal of Taiyuan University(Natural Science Edition),35(4):68-70.]
方中达.1998.植病研究方法[M].第3版.北京:中国农业出版社:122-145.[Fang Z D.1998.Plant Disease Research Methods[M].The 3rdEdition.Beijing:China Agriculture Press:122-145.]
高芬,岳换弟,秦雪梅,雷振宏,王梦亮.2018.植物致病镰刀菌细胞壁降解酶的研究进展[J].江苏农业学报,34(4):955-960.[Gao F,Yue H D,Qin X M,Lei Z H,Wang ML.2018.Research advances on cell wall degrading enzymes produced by pathogenic Fusarium causing plant diseases[J].Jiangsu Journal of Agricultural Sciences,34(4):955-960.]
黄冲,孙振宇,马占鸿.2010.影响品种混种模式下植物病害流行时空动态的各因素的随机模拟分析[C]//第二次全国植物抗病虫和病害流行与控制学术研讨会论文集.北京:中国植物保护学会:6.[Huang C,Sun Z Y,Ma Z H.2010.Stochastic simulation analysis of the factors affecting the temporal and spatial dynamics of plant diseases in mixed breeding modes[C]//Proceedings of the Second National Symposium on Plant Disease Resistance and Disease Epidemiology and Control.Beijing:China Society of Plant Protection:6.]
黄志巧,韦应贤,李德越,陆秋萍.2017.4种药剂防治西番莲茎基腐病药效试验初报[J].广西植保,(3):36-37.[Huang Z Q,Wei Y X,Li D Y,Lu Q P.2017.Preliminary report on the efficacy of four kinds of pesticides against the stem rot of Passiflora[J].Guangxi Plant Protection,(3):36-37.]
霍丹群,蒋兰,马璐璐,侯长军,杨平.2012.百香果功能研究及其开发进展[J].食品工业科技,33(19):391-395.[Huo D Q,Jiang L,Ma L L,Hou C J,Yang P.2012.Function of Passiflor and its comprehensive processing utility[J].Science and Technology of Food Industry,33(19):391-395.]
姜延涛,许韬,段霞瑜,周益林.2015.品种混种控制小麦白粉病及其对小麦产量和蛋白质含量的影响[J].作物学报,41(2):276-285.[Jiang Y T,Xu T,Duan X Y,Zhou Y L.2015.Effect of variety mixure planting on powdery mildew controlling as well as yield and protein contents in common wheat[J].Acta Agronomica Sinica,41(2):276-285.]
雷娅红,况卫刚,郑春生,李秀璋,高文娜,李春杰.2016.基于DNA条形码技术对镰刀菌属的检测鉴定[J].植物保护学报,43(4):544-551.[Lei Y H,Kuang W G,Zheng C S,Li X Z,Gao W N,Li C J.2016.Detection and identification of the genus Fusarium by DNA barcoding[J].Journal of Plant Protection,43(4):544-551.]
李德福,杨佳琪,张晓燕,孙龙芬.1993.福建省西番莲茎基腐病病原鉴定[J].植物病理学报,23(4):86.[Li D F,Yang J Q,Zhang X Y,Sun L F.1993.Identification of the pathogen causing stem rot disease of Passiflora in Fujian[J].Acta Phytopathologica Sinica,23(4):86.]
林泗海,黄添基,傅成业.2015.西番莲茎基腐病的发生特点及防治技术[J].现代园艺,(7):128.[Lin S H,Huang TJ,Fu C Y.2015.Occurrence characteristics and control techniques of stem rot of Passiflora[J].Xiandai Horticulture,(7):128.]
卢桂玲,黄英晴.2017.百香果根腐病的综合防治技术[J].江西农业,(19):23.[Lu G L,Huang Y Q.2017.Integrated control technology of root rot disease of Passiflora[J].Jiangxi Agriculture,(19):23.]
王桂清,马迪.2017.5种不同镰刀菌的酶学特性分析[J].河南农业科学,46(4):84-88.[Wang G Q,Ma D.2017.Enzymatic properties of five different Fusarium species[J].Journal of Henan Agricultural Sciences,46(4):84-88.]
王世伟,王卿惠,李小鹏,张其法,马兆魁,唐海平.2018.镰刀菌分子鉴定与重要应用的研究进展[J].微生物学通报,45(4):907-919.[Wang S W,Wang Q H,Li X P,Zhang Q F,Ma Z K,Tang H P.2018.Progress in molecular identification in the genus Fusarium and its important applications[J].Microbiology China,45(4):907-919.]
王秀荣,段安安,许玉兰.2003.国内西番莲引种栽培现状及改良思路[J].西南林学院学报,23(2):88-91.[Wang XR,Duan A A,Xu Y L.2003.A review on introduction and cultivation and improvement strategy of Passiflora fruit in China[J].Journal of Southwest Forestry College,23(2):88-91.]
许韬,徐志,徐媛,段霞瑜.2014.小麦品种混种对白粉病的调控及其对部分产量性状及蛋白质含量的影响[J].植物保护,40(3):87-93.[Xu T,Xu Z,Xu Y,Duang X Y.2014.Effects of wheat cultivar mixtures on powdery mildew,wheat yield and grain protein content[J].Plant Protection,40(3):87-93.]
严佳文,袁启凤,彭志军,王丽娟,解璞,陈楠,马玉华.2018.西番莲病毒病害研究进展[J].热带农业科学,38(4):85-94.[Yan J W,Yuan Q F,Peng Z J,Wang L J,Xie P,Chen N,Ma Y H.2018.Research progress on viruses of passionfruit[J].Chinese Journal of Tropical Agriculture,38(4):85-94.]
詹儒林,郑服丛,H H Ho.2003.海南西番莲茎腐病病原的分离与鉴定[J].热带作物学报,24(4):39-42.[Zhan R L,Zheng F C,Ho H H.2003.Isolation and identification of stalk butt rot of passion fruit in Hainan[J].Chinese Journal of Tropical Crops,24(4):39-42.]
郑继华.2000.西双版纳黄果西番莲茎基腐病调查[J].云南热作科技,23(3):40.[Zheng J H.2000.Investigation on stem rot of Passiflora edulis in Xishuangbanna[J].Journal of Yunnan Tropical Crops Science&Technology,23(3):40.]
郑加协,黄盈,梁渭州,张景文.1992.福建西番莲茎基腐病的病原菌鉴定[J].福建省农科院学报,7(2):65-68.[Zheng J X,Huang Y,Liang W Z,Zhang J W.1992.Identification of the pathogen causing base rot disease of passionfruit in Fujian[J].Journal of Fujian Academy of Agricultural Sciences,7(2):65-68.]
Antognoni F,Zheng S P,Pagnucco C,Baraldi R,Poli F,Biondi S.2007.Induction of flavonoid production by UV-Bradiation in Passiflora quadrangularis callus cultures[J].Fitoterapia,78(5):345-352.
Hebert P D N,Stoeckle M Y,Zemlak T S,Francis C M.2004.Identification of birds through DNA barcodes[J].PLoS Biology,2(10):e312.
Zeraik M L,Yariwake J H.2010.Quantification of isoorientin and total flavonoids in Passiflora edulis fruit pulp by HPLC-UV/DAD[J].Microchemical Journal,96(1):86-91.
陈企村,朱有勇,李振岐,唐永生,康振生.2009.不同品种混种对小麦产量及条锈病的影响[J].中国生态农业学报,17(1):29-33.[Chen Q C,Zhu Y Y,Li Z Q,Tang Y S,Kang Z S.2009.Effect of wheat cultivar mixtures on wheat yield and stripe rust[J].Chinese Journal of EcoAgriculture,17(1):29-33.]
陈星,高淑梅,李迎宾,张治萍,蒋娜,罗来鑫,李健强.2017.广西防城港西番莲茎基腐病发生为害调查与病原鉴定[C]//中国植物病理学会2017年学术年会论文集.北京:中国农业科学技术出版社:147.[Chen X,Gao S M,Li Y B,Zhang Z P,Jiang N,Luo L X,Li J Q.2017.Investigation and pathogen identification of stem rot of Passiflora in Fangchenggang,Guangxi[C]//Proceedings of the 2017 Annual Meeting of the Chinese Society of Plant Pathology.Beijing:China Agricultural Science and Technology Press:147.]
杜宾.2017.镰刀菌的分类和研究现状[J].太原学院学报(自然科学版),35(4):68-70.[Du B.2017.The classification and research status of Fusarium[J].Journal of Taiyuan University(Natural Science Edition),35(4):68-70.]
方中达.1998.植病研究方法[M].第3版.北京:中国农业出版社:122-145.[Fang Z D.1998.Plant Disease Research Methods[M].The 3rdEdition.Beijing:China Agriculture Press:122-145.]
高芬,岳换弟,秦雪梅,雷振宏,王梦亮.2018.植物致病镰刀菌细胞壁降解酶的研究进展[J].江苏农业学报,34(4):955-960.[Gao F,Yue H D,Qin X M,Lei Z H,Wang ML.2018.Research advances on cell wall degrading enzymes produced by pathogenic Fusarium causing plant diseases[J].Jiangsu Journal of Agricultural Sciences,34(4):955-960.]
黄冲,孙振宇,马占鸿.2010.影响品种混种模式下植物病害流行时空动态的各因素的随机模拟分析[C]//第二次全国植物抗病虫和病害流行与控制学术研讨会论文集.北京:中国植物保护学会:6.[Huang C,Sun Z Y,Ma Z H.2010.Stochastic simulation analysis of the factors affecting the temporal and spatial dynamics of plant diseases in mixed breeding modes[C]//Proceedings of the Second National Symposium on Plant Disease Resistance and Disease Epidemiology and Control.Beijing:China Society of Plant Protection:6.]
黄志巧,韦应贤,李德越,陆秋萍.2017.4种药剂防治西番莲茎基腐病药效试验初报[J].广西植保,(3):36-37.[Huang Z Q,Wei Y X,Li D Y,Lu Q P.2017.Preliminary report on the efficacy of four kinds of pesticides against the stem rot of Passiflora[J].Guangxi Plant Protection,(3):36-37.]
霍丹群,蒋兰,马璐璐,侯长军,杨平.2012.百香果功能研究及其开发进展[J].食品工业科技,33(19):391-395.[Huo D Q,Jiang L,Ma L L,Hou C J,Yang P.2012.Function of Passiflor and its comprehensive processing utility[J].Science and Technology of Food Industry,33(19):391-395.]
姜延涛,许韬,段霞瑜,周益林.2015.品种混种控制小麦白粉病及其对小麦产量和蛋白质含量的影响[J].作物学报,41(2):276-285.[Jiang Y T,Xu T,Duan X Y,Zhou Y L.2015.Effect of variety mixure planting on powdery mildew controlling as well as yield and protein contents in common wheat[J].Acta Agronomica Sinica,41(2):276-285.]
雷娅红,况卫刚,郑春生,李秀璋,高文娜,李春杰.2016.基于DNA条形码技术对镰刀菌属的检测鉴定[J].植物保护学报,43(4):544-551.[Lei Y H,Kuang W G,Zheng C S,Li X Z,Gao W N,Li C J.2016.Detection and identification of the genus Fusarium by DNA barcoding[J].Journal of Plant Protection,43(4):544-551.]
李德福,杨佳琪,张晓燕,孙龙芬.1993.福建省西番莲茎基腐病病原鉴定[J].植物病理学报,23(4):86.[Li D F,Yang J Q,Zhang X Y,Sun L F.1993.Identification of the pathogen causing stem rot disease of Passiflora in Fujian[J].Acta Phytopathologica Sinica,23(4):86.]
林泗海,黄添基,傅成业.2015.西番莲茎基腐病的发生特点及防治技术[J].现代园艺,(7):128.[Lin S H,Huang TJ,Fu C Y.2015.Occurrence characteristics and control techniques of stem rot of Passiflora[J].Xiandai Horticulture,(7):128.]
卢桂玲,黄英晴.2017.百香果根腐病的综合防治技术[J].江西农业,(19):23.[Lu G L,Huang Y Q.2017.Integrated control technology of root rot disease of Passiflora[J].Jiangxi Agriculture,(19):23.]
王桂清,马迪.2017.5种不同镰刀菌的酶学特性分析[J].河南农业科学,46(4):84-88.[Wang G Q,Ma D.2017.Enzymatic properties of five different Fusarium species[J].Journal of Henan Agricultural Sciences,46(4):84-88.]
王世伟,王卿惠,李小鹏,张其法,马兆魁,唐海平.2018.镰刀菌分子鉴定与重要应用的研究进展[J].微生物学通报,45(4):907-919.[Wang S W,Wang Q H,Li X P,Zhang Q F,Ma Z K,Tang H P.2018.Progress in molecular identification in the genus Fusarium and its important applications[J].Microbiology China,45(4):907-919.]
王秀荣,段安安,许玉兰.2003.国内西番莲引种栽培现状及改良思路[J].西南林学院学报,23(2):88-91.[Wang XR,Duan A A,Xu Y L.2003.A review on introduction and cultivation and improvement strategy of Passiflora fruit in China[J].Journal of Southwest Forestry College,23(2):88-91.]
许韬,徐志,徐媛,段霞瑜.2014.小麦品种混种对白粉病的调控及其对部分产量性状及蛋白质含量的影响[J].植物保护,40(3):87-93.[Xu T,Xu Z,Xu Y,Duang X Y.2014.Effects of wheat cultivar mixtures on powdery mildew,wheat yield and grain protein content[J].Plant Protection,40(3):87-93.]
严佳文,袁启凤,彭志军,王丽娟,解璞,陈楠,马玉华.2018.西番莲病毒病害研究进展[J].热带农业科学,38(4):85-94.[Yan J W,Yuan Q F,Peng Z J,Wang L J,Xie P,Chen N,Ma Y H.2018.Research progress on viruses of passionfruit[J].Chinese Journal of Tropical Agriculture,38(4):85-94.]
詹儒林,郑服丛,H H Ho.2003.海南西番莲茎腐病病原的分离与鉴定[J].热带作物学报,24(4):39-42.[Zhan R L,Zheng F C,Ho H H.2003.Isolation and identification of stalk butt rot of passion fruit in Hainan[J].Chinese Journal of Tropical Crops,24(4):39-42.]
郑继华.2000.西双版纳黄果西番莲茎基腐病调查[J].云南热作科技,23(3):40.[Zheng J H.2000.Investigation on stem rot of Passiflora edulis in Xishuangbanna[J].Journal of Yunnan Tropical Crops Science&Technology,23(3):40.]
郑加协,黄盈,梁渭州,张景文.1992.福建西番莲茎基腐病的病原菌鉴定[J].福建省农科院学报,7(2):65-68.[Zheng J X,Huang Y,Liang W Z,Zhang J W.1992.Identification of the pathogen causing base rot disease of passionfruit in Fujian[J].Journal of Fujian Academy of Agricultural Sciences,7(2):65-68.]
Antognoni F,Zheng S P,Pagnucco C,Baraldi R,Poli F,Biondi S.2007.Induction of flavonoid production by UV-Bradiation in Passiflora quadrangularis callus cultures[J].Fitoterapia,78(5):345-352.
Hebert P D N,Stoeckle M Y,Zemlak T S,Francis C M.2004.Identification of birds through DNA barcodes[J].PLoS Biology,2(10):e312.
Zeraik M L,Yariwake J H.2010.Quantification of isoorientin and total flavonoids in Passiflora edulis fruit pulp by HPLC-UV/DAD[J].Microchemical Journal,96(1):86-91.