海南菠萝一种叶斑病病原菌的分离与鉴定及多基因序列分析  

Isolation,Identification and Analysis of Multiple Gene Sequences of a Pathogen of Leaf Spot Disease on Pineapple in Hainan

作　　者：唐中发 秦春秀[1] 缪卫国[1] 林春花[1] 郑服丛[1] 刘文波[1] Tang Zhongfa;Qin Chunxiu;Miao Weiguo;Lin Chunhua;Zheng Fucong;Liu Wenbo(Key Laboratory of Ministry of Education for Green Prevention and Control of Tropical Plant Diseases and Pests,College of Plant Protection,Hainan University,Haikou,570228)

机构地区：[1]海南大学植物保护学院,热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《基因组学与应用生物学》2021年第3期1219-1226,共8页Genomics and Applied Biology

基　　金：海南省重大科技项目(ZDKJ201817-23);国家自然科学基金(31560495,31760499,31660033);国家重点研发计划(2018YFD0201105);海南大学青年基金项目(hdkyxj201708)共同资助。

摘　　要：为明确菠萝叶斑病病原菌可可毛色二孢菌在海南省各市县的亲缘关系及遗传差异,从海南省的海口、澄迈、儋州、三亚、保亭等16个市县进行病样采集和病样分离,根据科赫氏法则鉴定,获得42株病原菌。观察形态特征,并基于多基因联合序列分析其遗传多样性。结果表明,通过形态学鉴定和ITS与TUB2基因序列联合进化树分析发现,其中来自海口、澄迈、儋州、三亚、保亭等16个市县的16株代表病原菌均鉴定为可可毛色二孢菌(Lasiodiplodia theobromae),其分生孢子平均大小为(22.06~31.07)μm×(11.77~16.48)μm;通过ITS、TUB2、EF-1α、GAPDH、CHS-1和ACT基因拼接序列聚类分析,结果分为3个类群,海南岛的中部(儋州,昌江,白沙,五指山,万宁,琼海等地)聚为一个类群,中北部(屯昌,临高)聚为一个类群,西南部(东方,乐东,三亚等地)聚为一个类群。该结果说明来源于不同产地不同菠萝品种上的可可毛色二孢菌遗传多样性丰富,在海南省16个市县菠萝叶斑病菌中可可毛色二孢菌L.theobromae为优势种。In order to clarify the genetic diversity and relationship of pathogens of Lasiodiplodia theobromae associated with leaf spot of Ananas comosus from cities and counties in Hainan,the diseased pineapple leaf samples from 16 cities and counties such as Haikou,Chengmai,Danzhou,Sanya and Baoting in Hainan province were used to isolate pathogens.According to the Koch's rule,42 pathogens were obtained,then the morphological characteristics were observed,and the genetic diversity was analyzed based on the multiple gene combination sequence.The results showed that 16 pathogens among them were identified as Lasiodiplodia theobromae by morphological characteristics and analysis of phylogenetic tree based on ITS-TUB2 gene combined sequence.The average size of conidia was(22.06~31.07)μm×(11.77~16.48)μm.Then phylogenetic tree based on combined sequences of ITS,TUB-2,ITS,TUB2,EF-1α,GAPDH,CHS-1 and ACT showed that the 16 pathogens were divided into three groups:one mainly concentrated in the middle part of Hainan Island(Danzhou,Changjiang,Baisha,Wuzhishan,Wanning,Qionghai,etc);one at the north-central part of Hainan Island(Tunchang,Lingao);the last mainly in the southwest ofHainan Island(Dongfang,Ledong,Sanya,etc).The results indicated that the genetic diversity of L.theobromae from different pineapple varieties in different producing areas was rich,and it was also clear that L.theobromae was the dominant species in the leaf spot pathogens of pineapple in 16 cities and counties in Hainan province.

关 键 词：菠萝 叶斑病 可可毛色二孢菌 多基因系统发育 遗传多样性 

分 类 号：S436.68[农业科学—农业昆虫与害虫防治]