玉屏油茶炭疽病病原菌鉴定及同源性分析
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摘要
油茶是贵州玉屏脱贫致富的重要产业,但近年来,炭疽病危害日益严重。为了防治油茶炭疽病,本研究采用林间病情观察、科赫氏法则验证、形态学鉴定及多基因分子鉴定的方法,对分离的3株炭疽病病原菌进行了系统鉴定。在ITS-CAL-GADPH 3基因所构建的系统发育树中,3株病原菌与ICMP10643*、ICMP-10646*两炭疽属模式菌株聚为一个独立的进化枝且置信度为99%,结合其形态特征,将其鉴定为山茶炭疽菌(Colletotrichum camelliae)。依据林间观察结果与玉屏油茶的生长发育规律,提出了防治油茶炭疽病的关键时间。玉屏油茶炭疽病种类的分子鉴定可为抗病种质的分子育种及该病的防治提供科学依据。
Camellia oleifera is an important industry of Yuping Dong Autonomous county, Guizhou Province to get rid of poverty and become better off. However, in recent years, the damage caused by anthrax has become increasingly serious. In order to prevent and control anthrax in C. oleifera, three isolated pathogen strains were systematically identified based on field observation, Koch's postulation, morphological characteristics, and multi-gene molecular identification. The multi-gene phylogenetic tree system based on ITS-CAL-GADPH showed that the 3 pathogenic strains and the 2 type strains of ICMP10643*, ICMP10646* were clustered into one clade,with the confidence level of 99%. Combined with its morphological characteristics, the 3 pathogenic strains were identified as Colletotrichum camelliae. According to the observations in the fields and growth habit of C. oleifera in Yuping county, the optimal time of controlling the anthracnose was suggested. The molecular identification of pathogens of C. oleifera could provide a scientific basis for the molecular breeding of germplasm with resistance to anthracnose and the prevention and control of this disease.
Camellia oleifera is an important industry of Yuping Dong Autonomous county, Guizhou Province to get rid of poverty and become better off. However, in recent years, the damage caused by anthrax has become increasingly serious. In order to prevent and control anthrax in C. oleifera, three isolated pathogen strains were systematically identified based on field observation, Koch's postulation, morphological characteristics, and multi-gene molecular identification. The multi-gene phylogenetic tree system based on ITS-CAL-GADPH showed that the 3 pathogenic strains and the 2 type strains of ICMP10643*, ICMP10646* were clustered into one clade,with the confidence level of 99%. Combined with its morphological characteristics, the 3 pathogenic strains were identified as Colletotrichum camelliae. According to the observations in the fields and growth habit of C. oleifera in Yuping county, the optimal time of controlling the anthracnose was suggested. The molecular identification of pathogens of C. oleifera could provide a scientific basis for the molecular breeding of germplasm with resistance to anthracnose and the prevention and control of this disease.
引文
Cheng J.J.,Liu H.Y.,Fan H.,Cheng S.Y.,Zhang S.M.,Chen Y.L.,and Liu Z.,2016,Comparative bioinformatics analysis of candidate genes resistant to yellow dwarf disease in Setaria italica,Hordeum vulgare and Sorghum bicolor,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(7):1767-1779(程金金,刘红云,范会,程思远,张书敏,陈宇龙,刘征,2016,谷子、大麦、高粱抗黄矮病候选基因的生物信息学比较分析,基因组学与应用生物学,35(7):1767-1779)
Crouch J.A.,Clarke B.B.,and Hillman B.I.,2009a,What is the value of ITS sequence data in Colletotrichum systematics and species diagnosis?A case study using the falcate-spored graminicolous Colletotrichum group,Mycologia,101(5):648-656
Crouch J.A.,Clarke B.B.,White J.J.F.,and Hillman B.I.,2009b Systematic analysis of the falcate-spored graminicolous Colletotrichum and a description of six new species from warm-season grasses,Mycologia,101(5):717-732
Damm U.,Cannon P.F.,Woudenberg J.H.C.,and Crous P.W.2012,The Colletotrichum acutatum species complex,Stud Mycol.,73(1):37-113
Ding G.J.,Ruan C.J.,Zhao Y.,Chen W.C.,Yang B.,and Liu SH.,2016,Correlation and principal component analysis of fruit characteristics in fine Camellia oleifera germplasm in Yuping,Jingjilin Yanjiu(Nonwood Forest Research),34(1)26-32(丁桂娇,阮成江,赵月,陈文昌,杨波,刘四黑,2016玉屏油茶良种果实性状的相关性和主成分分析,经济林研究,34(1):26-32)
Li H.,Li Y.,Jiang S.Q.,Liu J.A.,and Zhou G.Y.,2017,Pathogen of oil-tea trees anthracnose caused by Colletotrichum spp.in Hu'nan Province,Linye Kexue(Scientia Silvae Sinicae),53(8):43-53(李河,李杨,蒋仕强,刘君昂,周国英,2017,湖南省油茶炭疽病病原鉴定,林业科学,53(8):43-53)
Liu X.Q.,Qi J.M.,Chen Y.S.,Chen M.C.,Chen M.X.,Liu W.Fang P.P.,Lin L.H.,and Tao A.F.,2012,Identification and r DNA-ITS sequence analysis of the pathogen of kenaf anthracnose,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(17):3515-3521(刘晓倩,祁建民,陈玉森,陈绵才,陈美霞,刘伟,方平平,林荔辉,陶爱芬,2012,中国红麻炭疽病病原菌的分离鉴定及r DNA-ITS序列分析,中国农业科学,45(17):3515-3521)
Moriwaki J.,Tsukiboshi T.,and Sato T.,2002,Grouping of Colletotrichum species in Japan based on r DNA sequences,JGen.Plant Pathol.,68(4):307-320
Sharma G.,Kumar N.,Weir B.S.,Hyde K.D.,and Shenoy B.D.,2013,The ApMat marker can resolve Colletotrichum species:a case study with Mangifera indica,Fungal Divers.,61(1):117-138
Smith B.J.,and Black L.L.,1990,Morphological,cultural and pathogenic variation among Colletotrichum species isolated from strawberry,Plant Dis.,74(1):69-76
Sreenivasaprasad S.,Mills P.R.,Meehan B.M.,and Brown A.E.,1996,Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences,Genome,39(3):499-512
Tang Y.L.,Zhou G.Y.,Li H.,Zhong W.B.,Gong H.E.,and Wang L.Y.,2015,Identification of a new anthracnose of Camellia oleifera based on multiple-gene phylogeny,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),36(5):972-977(汤铱泠,周国英,李河,钟文斌,龚洪恩,王丽云,2015,多基因序列鉴定油茶炭疽病原Colletotrichum boninense新种,热带作物学报,36(5):972-977)
Wang Y.C.,Hao X.Y.,Huang Y.T.,Yue C.,Wang B.,Cao H.L.,Wang L.,Wang X.C.,Yang Y.J.,and Xiao B.,2015,Phylogenetic study of Colletotrichum species associated with Camellia sinensis from the major tea areas in China,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(24):4924-4935(王玉春,郝心愿,黄玉婷,岳川,王博,曹红利,王璐,王新超,杨亚军,肖斌,2015,中国主要茶区茶树炭疽菌系统发育学,中国农业科学,48(24):4924-4935)
Weir B.S.,Johnston P.R.,and Damm U.,2012,The Colletotrichum gloeosporioides species complex,Stud.Mycol.,73(1):115-180
Yu H.Q.,Liu Y.,Xu M.L.,and Li Y.P.,2015,Identification of resistance to black shank of new tobacco germplasm,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(5):1027-1033(于海芹,刘勇,许美玲,李永平,2015,新引进烟草种质资源的黑胫病抗性鉴定,基因组学与应用生物学,34(5):1027-1033)
Zhu Y.Z.,Liao W.J.,Zou D.X.,Wu Y.J.,and Deng Y.,2015,I-dentification and biological characteristics of the pathogen from Camellia oleifera anthracnose in Guangxi,Zhiwu Baohu Xuebao(Journal of Plant Protection),42(3):382-389(朱英芝,廖旺姣,邹东霞,吴耀军,邓艳,2015,广西油茶炭疽病病原菌鉴定及生物学特性,植物保护学报,42(3):382-389)
Crouch J.A.,Clarke B.B.,and Hillman B.I.,2009a,What is the value of ITS sequence data in Colletotrichum systematics and species diagnosis?A case study using the falcate-spored graminicolous Colletotrichum group,Mycologia,101(5):648-656
Crouch J.A.,Clarke B.B.,White J.J.F.,and Hillman B.I.,2009b Systematic analysis of the falcate-spored graminicolous Colletotrichum and a description of six new species from warm-season grasses,Mycologia,101(5):717-732
Damm U.,Cannon P.F.,Woudenberg J.H.C.,and Crous P.W.2012,The Colletotrichum acutatum species complex,Stud Mycol.,73(1):37-113
Ding G.J.,Ruan C.J.,Zhao Y.,Chen W.C.,Yang B.,and Liu SH.,2016,Correlation and principal component analysis of fruit characteristics in fine Camellia oleifera germplasm in Yuping,Jingjilin Yanjiu(Nonwood Forest Research),34(1)26-32(丁桂娇,阮成江,赵月,陈文昌,杨波,刘四黑,2016玉屏油茶良种果实性状的相关性和主成分分析,经济林研究,34(1):26-32)
Li H.,Li Y.,Jiang S.Q.,Liu J.A.,and Zhou G.Y.,2017,Pathogen of oil-tea trees anthracnose caused by Colletotrichum spp.in Hu'nan Province,Linye Kexue(Scientia Silvae Sinicae),53(8):43-53(李河,李杨,蒋仕强,刘君昂,周国英,2017,湖南省油茶炭疽病病原鉴定,林业科学,53(8):43-53)
Liu X.Q.,Qi J.M.,Chen Y.S.,Chen M.C.,Chen M.X.,Liu W.Fang P.P.,Lin L.H.,and Tao A.F.,2012,Identification and r DNA-ITS sequence analysis of the pathogen of kenaf anthracnose,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(17):3515-3521(刘晓倩,祁建民,陈玉森,陈绵才,陈美霞,刘伟,方平平,林荔辉,陶爱芬,2012,中国红麻炭疽病病原菌的分离鉴定及r DNA-ITS序列分析,中国农业科学,45(17):3515-3521)
Moriwaki J.,Tsukiboshi T.,and Sato T.,2002,Grouping of Colletotrichum species in Japan based on r DNA sequences,JGen.Plant Pathol.,68(4):307-320
Sharma G.,Kumar N.,Weir B.S.,Hyde K.D.,and Shenoy B.D.,2013,The ApMat marker can resolve Colletotrichum species:a case study with Mangifera indica,Fungal Divers.,61(1):117-138
Smith B.J.,and Black L.L.,1990,Morphological,cultural and pathogenic variation among Colletotrichum species isolated from strawberry,Plant Dis.,74(1):69-76
Sreenivasaprasad S.,Mills P.R.,Meehan B.M.,and Brown A.E.,1996,Phylogeny and systematics of 18 Colletotrichum species based on ribosomal DNA spacer sequences,Genome,39(3):499-512
Tang Y.L.,Zhou G.Y.,Li H.,Zhong W.B.,Gong H.E.,and Wang L.Y.,2015,Identification of a new anthracnose of Camellia oleifera based on multiple-gene phylogeny,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),36(5):972-977(汤铱泠,周国英,李河,钟文斌,龚洪恩,王丽云,2015,多基因序列鉴定油茶炭疽病原Colletotrichum boninense新种,热带作物学报,36(5):972-977)
Wang Y.C.,Hao X.Y.,Huang Y.T.,Yue C.,Wang B.,Cao H.L.,Wang L.,Wang X.C.,Yang Y.J.,and Xiao B.,2015,Phylogenetic study of Colletotrichum species associated with Camellia sinensis from the major tea areas in China,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(24):4924-4935(王玉春,郝心愿,黄玉婷,岳川,王博,曹红利,王璐,王新超,杨亚军,肖斌,2015,中国主要茶区茶树炭疽菌系统发育学,中国农业科学,48(24):4924-4935)
Weir B.S.,Johnston P.R.,and Damm U.,2012,The Colletotrichum gloeosporioides species complex,Stud.Mycol.,73(1):115-180
Yu H.Q.,Liu Y.,Xu M.L.,and Li Y.P.,2015,Identification of resistance to black shank of new tobacco germplasm,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(5):1027-1033(于海芹,刘勇,许美玲,李永平,2015,新引进烟草种质资源的黑胫病抗性鉴定,基因组学与应用生物学,34(5):1027-1033)
Zhu Y.Z.,Liao W.J.,Zou D.X.,Wu Y.J.,and Deng Y.,2015,I-dentification and biological characteristics of the pathogen from Camellia oleifera anthracnose in Guangxi,Zhiwu Baohu Xuebao(Journal of Plant Protection),42(3):382-389(朱英芝,廖旺姣,邹东霞,吴耀军,邓艳,2015,广西油茶炭疽病病原菌鉴定及生物学特性,植物保护学报,42(3):382-389)