海南省油茶及其他寄主植物果生刺盘孢菌群体遗传结构分析

doi:10.11707/j.1001-7488.20161010

摘要/Abstract

摘要： [目的] 研究海南省油茶树及其他寄主植物炭疽病菌果生刺盘孢菌群体遗传结构、群体遗传多样性及单倍型地理分布，探索海南地区不同寄主植物炭疽病菌的分子遗传变异规律。[方法] 利用单孢分离法对采集自海南省南圣镇、通什镇、枫木林场和澄迈林场4个地区9种不同寄主植物的炭疽病病叶进行炭疽菌分离，并对每个菌株ITS，CAL和GAPDH 3个基因进行PCR扩增和测序，按顺序对3个基因的序列进行拼接，然后采用GenALEx6，DnaSP 5.0，TCS1.21和MEGA6.0软件分别对数据进行分析。[结果] 分离、鉴定共获得12个病菌种群的108个果生刺盘孢菌病菌样品，可推导出18种单倍体型（haplotype），其中单倍型Haplotype11共70个菌株，为优势单倍型，在每个病菌种群中均有分布；单倍型网络结构分析表明，病原菌以单倍型Haplotype11为辐射中心，呈现出1个比较典型星状发散图，推测Haplotype11为原始单倍型；108株病菌表现出丰富的遗传多样性；AMOVA分析显示，种群间和种群内的遗传变异分别占总变异的3%和97%，变异主要存在种群内，而种群间的差异小，说明不同的寄主植物未对病菌交流造成隔离障碍；遗传分化和基因流分析发现，12个病菌种群间遗传分化小，种群间基因交流频繁；另外，油茶种群之间以及油茶与鲫鱼胆、葛根和山矾等植物种群之间的果生刺盘孢菌存在遗传重组现象；系统发育分析表明，来自同一个果生刺盘孢菌种群的单倍型未能聚在一起，而是散乱的分布在系统树中，没有显示出明显的寄主植物分布族群；核苷酸不配对分析表明病菌种群历史上并没有发生显著的种群扩张。[结论] 海南省果生刺盘孢菌遗传多样性丰富，不同地区和不同寄主植物病菌种群之间存在有效基因流，揭示该病菌可以在油茶树及其他寄主植物之间迁移，这为整体考虑海南省油茶及其他寄主植物制定可持续控制油茶炭疽病策略提供重要的理论依据。

关键词: 油茶, 寄主植物, 果生刺盘孢菌, 群体遗传结构

Abstract: [Objective] The population genetic structure genetic diversity and the geographical distribution of haplotype patterns of Colletotrichum fructicola from Oil-tea and other host plants in Hainan province were analysised to explore the molecular genetic variation pattern of the pathogen in different hosts.[Method] C.fructicola associated with anthracnose diseases of oil-tea and other native plants were isolated using single spore isolation and confirmed by DNA sequencing of the partial sequences of rDNA-ITS, CAL and GAPDH region. GenALEx6, DnaSP 5.0,TCS1.21 and MEGA6.0 software was used to analyze the data of ITS-CAL-GAPDH sequences.[Result] A total of 108 strains was isolated, and 18 haplotypes were identified. The most common sequence type, haplotype 11, was found in 70 samples and distributed across all populations. AMOVA test showed that 3% of genetic differentiation occurred among populations and 97% within population. There was a very low genetic divergence in Fst and Nm values among the populations and the high level of gene flow was among populations. There existed genetic recombination among some populations of C. fructicola from Oil-tea camellia and the other host plants. [Conclusion] Results of this research illustrates that there exists high degree of genetic intermixing among natural populations of C. fructicola. Our study provides a foundation that a more holistic approach would be needed to take into account of the life histories of different hosts in order to develop effective strategies for controlling anthracnose.

Key words: Camellia oleifera, host plant, Colletotrichum fructicola, population genetic structure

中图分类号:

S763.1

李河, 李杨, 徐建平, 周国英. 海南省油茶及其他寄主植物果生刺盘孢菌群体遗传结构分析[J]. 林业科学, 2016, 52(10): 80-88.

Li He, Li Yang, Xu Jianping, Zhou Guoying. Population Genetic Structure of Colletotrichum fructicola from Oil-Tea and Other Host Plants in Hainan province[J]. Scientia Silvae Sinicae, 2016, 52(10): 80-88.

参考文献

柏云爱,宋大海,张富强,等.2008. 油茶籽油与橄榄油营养价值的比较. 中国油脂, 33(3):39-41.
(Bo Y A, Song D H, Zhang F Q, et al. 2008. The comparison of nutritional value between tea seed oil and palm oil. China Oils and Fats, 33(3):39-41.[in Chinese])
陈守常,田泽君,郭隆錫.1965.油茶炭疽病的发生与蔓延. 植物保护学报, 4(3):207-218.
(Chen S C,Tian Z J,Guo L X.1965. Development and spread of the anthracnose of Camellia oleifera (Lour.) rehd.Acta Phytophylacica Sinica, 4(3):207-218.[in Chinese])
陈守常,田泽君,陈大昌.1964.油茶炭疽病菌有性世代的形态特征、出现频率、致病力及其产生条件. 植物病理学报,3(1):45-52.
(Chen S C, Tian Z J, Chen D C.1964. Studies on the perithecial stage of the anthracnose fungus on Camellia oleifera (Lour.) rehd. Acta Phytopathologica Sinica, 3(1):45-52.[in Chinese])
方中达.1998.植病研究法.3版. 北京:中国农业出版社, 122-145.
(Fang Z D. Methods in plant pathology.3rd edition. Beijing:China Agriculture Press,122-145.[in Chinese])
黄翠莉,吴苏喜,刘瑞兴,等.2011. 油茶籽油对大鼠降血脂和预防脂肪肝的影响.食品科学, 32(13):332-335.
(Huang C L, Wu S X, Liu R X, et al. 2011.Blood lipid-lowering and fatty liver-preventing effects of tea seed (Camellia oleifera Abel.) oil in rats. Food Science, 32(13):332-335.[in Chinese])
梁子超,岑炳沾,戴品初.1964.油茶炭疽病菌的有性世代、越冬方式和侵入途径. 植物保护学报, 3(3):316-317.
(Liang Z C,Chen B Z,Dai P C. 1964. Studies on anthracnose of Camellia oleifera (Lour.) rehd.I. The perfect stage,mode of over wintering and ingress of the pathogen. Acta Phytophylacica Sinica, 3(3):316-317.[in Chinese])
刘威,叶乃兴,陈玉森,等. 2014.茶树炭疽菌Colletotrichum fructicola的鉴定及系统发育分析.茶叶科学, 34(1):95-104.
(Liu W, Ye N X, Chen Y S, et al. 2014. Identification and phylogenetic analysis of anthracnose pathogen Colletotrichum fructicola Isolated from Camellia sinensis. Journal of Tea Science, 34(1):95-104.[in Chinese])
李河,周国英,徐建平,等.2014.一种油茶新炭疽病原的多基因系统发育分析鉴定. 植物保护学报, 41(5):602-607.
(Li H, Zhou G Y, Xu J P, et al. 2014. Pathogen identification of a new anthracnose of Camellia oleifera in China based on multiple-gene phylogeny. Acta Phytophylacica Sinica,41(5):602-607.[in Chinese])
李河, 李杨, 蒋仕强, 等. 2016. 海南省不同寄主植物炭疽病菌侵染油茶研究. 植物保护, 42(4):64-70.
(Li H, Li Y, Jiang S Q, et al. 2016. The initial inoculums source of oil tea anthracnose in Hainan Province. Plant Protection, 42(4):64-70.[in Chinese])
龙正海,王道平.2008. 油茶籽油与橄榄油化学成分研究.中国粮油学报, 23(2):121-123.
(Long Z H,Wang D P. 2008. Chemical constituents of olive oil and from Camellia oleifera seed oil. Journal of the Chinese Cereals and Oils Association, 23(2):121-123.[in Chinese])
曲若竹,侯林,吕红丽,等.2004. 群体遗传结构中的基因流.遗传,26(3):377-382.
(Qu R Z, Hou L, Lü H L, et al. 2004. The gene flow of population genetic structure. Hereditas, 26(3):377-382.[in Chinese])
汤富彬,沈丹玉,刘毅华,等. 2013.油茶籽油和橄榄油中主要化学成分分析. 中国粮油学报,28(7):108-113.
(Tang F B, Shen D Y, Liu Y H, et al.2013. Analysis of main chemical components in Camellia oil and Olive oil. Journal of the Chinese Cereals and Oils Association, 28(7):108-113.[in Chinese])
王义勋,刘伟,陈京元,等.2013.油茶炭疽病菌潜伏越冬场所、菌态及菌量研究. 湖北林业科技,1:32-33.
(Wang Y X, Liu W, Chen J Y, et al. 2013. Overwintering sites, various stages and amount of Colletotrichum gloeosporioides on Camellia oleifera. Hubei Forestry Science and Technology,1:32-33.[in Chinese])
杨涛,关天舒,白金铠.2000. 灰霉菌准性生殖现象及其生物学意义.辽宁农业科学, 6:24-26.
(Yang T, Guan T S, Bai J K, et al. 2000. The parasexuality of Botrytis cinerea and its biological significance. Liaoning Agricultural Sciences, 6:24-26.[in Chinese])
杨友联.2010. 中国贵州、云南、广西炭疽菌属真菌多基因分子系统学研究. 武汉:华中农业大学博士学位论文.
(Yang Y L. 2010. Multi-locus phylogeny of Colletotrichum species in guizhou,yunnan and guanxi,China. Wuhang:PhD thesis of Huazhong Agricultural University.[in Chinese])
袁嗣令,张能唐,翁月霞,等.1963. 油茶炭疽病的研究. 植物保护学报,2(3):253-262.
(Yuan S L, Zhang N T, Weng Y X,et al.1963. The anthracnose disease of Camellia oleosa (Lour.) rehd. Acta Phytophylacica Sinica, 2(3):253-262.[in Chinese])
郑道君,潘孝忠,谢良商,等.2015. 海南省油茶产业发展现状调查与分析.经济林研究,33(1):131-135.
(Zheng DJ,Pan XZ,Xie LS,et al.2015. Investigation and analysis of Camellia oleifera industry development status in Hainan Province.Nonwood Forest Research,33(1):131-135.[in Chinese])
祝雯,詹家绥.2012. 植物病原物的群体遗传学.遗传,34(2):157-166.
(Zhu W, Zhan J S. 2012. Population genetics of plant pathogens. Hereditas, 34(2):157-166.[in Chinese])
Clement M, Posada D, Crandall K A. 2000. TCS:a computer program to estimate gene genealogies. Molecular Ecology, 9(10):1657-1659.
Excoffier L, Smouse P E, Quattro J M. 1992. Analysis of molecular variance inferred from metric distances among DNA haplotypes:application to human mitochondrial DNA restriction data. Genetics,131(2):479-491.
Librado P, Rozas J. 2009. DnaSP v5:A software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25(11):1451-1452.
Li H, Zhou G Y, Liu J A, et al. 2016. Population genetic analyses of the fungal pathogen Colletotrichum fructicola on tea-oil trees in China. PLoS ONE, 11(6):1-24.
Maisa C G, Cristina B A, Pereira S. 2014. Recent Introduction and Recombination in Colletotrichum acutatum populations associated with Citrus postbloom fruit drop epidemics in São Paulo, Brazil. Population Biology,104(7):769-778.
Peakall R, Smouse P E. 2006. GENALEX 6:genetic analysis in excel. Population genetic software for teaching and research. Molecular Ecology Notes, 6(1):288-295.
Prihastuti H,Cai L,Chen H,Hyde K D.2009. Characterization of Colletotrichum species associated with coffee berries in northern Thailand. Fungal Diversity, 39:89-109.
Peng L J, Sun T, Yang Y L, et al. 2013. Colletotrichum species on grape in Guizhou and Yunnan provinces, China. Mycoscience, 54(1):29-41.
Souza E A, Jr C, Pinto J M A. 2010. Sexual recombination in Colletotrichum lindemuthianum occurs on a fine scale. Genet Mol Res, 9(3):1759-1769.
Souza E A, Fávaro L C, Casela C R. 2003. Genetic tecombination in Colletotrichum sublineolum. Phytopathology, 151(6):329-334.
Tamura K, Stecher G, Peterson D, et al. 2013. MEGA6:molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30(12):2725-2729.
Wright S. 1931. Evolution in mendelian populations. Genetics, 16(2):97-159.
Weir B S, Johnston P R, Damm U. 2012. The Colletotrichum gloeosporioides species complex. Studies in mycology, 73(1):115-180.

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