浙江大盘山华顶杜鹃菌根真菌的分离和鉴定
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摘要
于2016年5月在浙江省大盘山国家级自然保护区的3处采样地,每处采集5株华顶杜鹃Rhododendron huadingense的1～2支侧根,培养分离华顶杜鹃的菌根真菌,然后结合菌落形态特征和r DNAITS序列分析进行鉴定。结果表明:华顶杜鹃根系内共分离到165个菌株,分为22个菌落形态类型;结合r DNA ITS序列分析,最终分为20个不同的菌种类型,其中19个菌种类型属于子囊菌门Ascomycota,包括8个属于粪壳菌纲Sordariomycetes和6个属于锤舌菌纲Leotiomycetes;3个优势菌种分别为Phialocephala fortinii,Ilyonectria radicicola和某粪壳菌纲真菌的近缘种,分别占菌株总数的41.82%,22.42%和10.91%;其余17个菌种类型合计只占总数的24.85%。
Three sample plots were established in Dapanshan National Nature Reserve, Zhejiang province in May of 2016. 5 Rhododendron huadingense were selected for 1-2 lateral roots each. Ericoid mycorrhiza(ERM) was cultured from collected roots and studied by morphological characteristics and molecular identification of r DNA ITS sequence. The result showed that 165 mycorrhizal fungal strains were isolated which were classified into 22 types based on colony characteristics. r DNA ITS demonstrated that these mycorrhizal fungi could be identified into 20 species,among them, 19 species belonging to Ascomycota, including 8 Sordariomycetes, 6 Leotiomycetes. The dominant 3 species were Phialocephala fortinii, Ilyonectria radicicola and Sordariomycetes sp., respectively accounting for 41.82 %, 22.42% and 10.91% of the total strains. The lest 17 species occupied only 24.9%.
Three sample plots were established in Dapanshan National Nature Reserve, Zhejiang province in May of 2016. 5 Rhododendron huadingense were selected for 1-2 lateral roots each. Ericoid mycorrhiza(ERM) was cultured from collected roots and studied by morphological characteristics and molecular identification of r DNA ITS sequence. The result showed that 165 mycorrhizal fungal strains were isolated which were classified into 22 types based on colony characteristics. r DNA ITS demonstrated that these mycorrhizal fungi could be identified into 20 species,among them, 19 species belonging to Ascomycota, including 8 Sordariomycetes, 6 Leotiomycetes. The dominant 3 species were Phialocephala fortinii, Ilyonectria radicicola and Sordariomycetes sp., respectively accounting for 41.82 %, 22.42% and 10.91% of the total strains. The lest 17 species occupied only 24.9%.
引文
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[20]魏杰,杨秀丽.大兴安岭4种杜鹃花科植物的DSE真菌分布[J].内蒙古林业科技,2014,40(4):43-46.
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[22]KOHOUT P,TAMAEA T,ROY M,et al.A diverse fungal community associated with Pseudorchis albida(Orchidaceae)roots[J].Fung Ecol,2013,6(1):50-64.
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[24]NEWSHAM K K.A meta-analysis of plant responses to dark septate root endophytes[J].New Phytol,2011,190(3):783-793.
[25]BRUZONE M C,FONTENLA S B,VOHNíK M.Is the prominent ericoid mycorrhizal fungus Rhizoscyphus ericae absent in the Southern Hemisphere’s Ericaceae?A case study on the diversity of root mycobionts in Gaultheria spp.from northwest Patagonia,Argentina[J].Mycorrhiza,2015,25(1):25-40.
[26]张新军.极小种群野生植物大树杜鹃的形态解剖特征及内生真菌多样性研究[D].昆明:云南大学,2015,32-42.
[27]CARLOS A B,GRAMAJE D,JOSE G J,et al.Detection of black-foot disease pathogens in the grapevine nursery;propagation process in Spain[J].Europ J Plant Pathol,2013,137(1):103-112.
[28]VITALE A,AIELLO D,GUARNACCIA V,et al.First Report of Root Rot Caused by Ilyonectria(Neonectria)macrodidyma on Avocado(Persea americana)in Italy[J].J Phytopathol,2012,160(3):156-159.
[29]KOGEL K H,FRANKEN P,HüCKELHOVEN R.Endophyte or parasite-what decides?[J].Curr Opin Plant Biol,2006,9(4):358-363.
[2]徐丽娟,刁志凯,李岩,等.菌根真菌的生理生态功能[J].应用生态学报,2012,23(1):288-295.
[3]PELLETT N E,ROWAN N,ALEONG J.Cold Hardiness of Various Provenances of Flame,Roseshell,and Swamp Azaleas[J].J Am Soc Hort,1991,116(1):23-26.
[4]FUKUCHI S,OBASE K,TAMAI Y,et al.Vegetation and colonization status of mycorrhizal and endophytic fungi in plant species on acidic barren at crater basin of volcano Esan in Hokkaido,Japan[J].Eurasian J For Res,2011,14(1):1-11.
[5]CAIRNEY J W G,MEHARG A A.Ericoid mycorrhiza:a partnership that exploits harsh edaphic conditions[J].Europ J Soil Sci,2010,54(4):735-740.
[6]DIAZ A,GREEN I,BENVENUTO M,et al.Are Ericoid Mycorrhizas a factor in the success of Calluna vulgaris heathland restoration?[J].Restor Ecol,2010,14(2):187-195.
[7]丁炳扬,方云亿.浙江杜鹃花属一新种[J].植物研究,1990,10(1):31-33.
[8]杨旭,丁炳扬,胡仁勇.浙江杜鹃花属植物资源开发及利用[J].中国野生植物资源,2005,24(2):22-25.
[9]周莹莹,孙莉,陈子林,等.珍稀特有植物华顶杜鹃的群落基本特征分析[J].杭州师范大学学报:自然科学版,2012,11(3):211-216.
[10]褚文珂,周莹莹,陈子林,等.珍稀植物华顶杜鹃群落分类和物种多样性研究[J].杭州师范大学学报:自然科学版,2013,12(3):240-244,257.
[11]陈珍慧.珍稀特有植物华顶杜鹃的种群特征和保护遗传学研究[D].杭州:杭州师范大学,2016,40-47.
[12]陈亚琳,杨碧烟.大盘山志[M].杭州:浙江科学技术出版社,2016:2-8.
[13]张春英,尹丽娟,王胤,等.一种简捷的分离杜鹃花类菌根真菌的方法[J].生物技术,2007,17(6):28-32.
[14]刘仁阳,欧静,李冠楠,等.梵净山雷山杜鹃根部真菌分离与鉴定[J].西北农业学报,2014,23(4):178-185.
[15]杨兵,张春英,王献,等.杜鹃花根系内生菌资源调查及多样性分析[J].河南农业大学学报,2010,44(3):290-294.
[16]陈真,杨兵,张春英,等.锦绣杜鹃菌根真菌r DNA ITS序列分析及接种效应研究[J].菌物学报,2011,30(5):729-737.
[17]庄彩云,李潞滨,胡陶,等.适用于r DNA ITS分析的兰属菌根真菌培养及DNA提取方法[J].北京农学院学报,2007,22(3):4-6,31.
[18]张丽娜,荣昌鹤,何远,等.常用系统发育树构建算法和软件鸟瞰[J].动物学研究,2013,34(6):640-650.
[19]LEOPOLD D R.Ericoid fungal diversity:Challenges and opportunities for mycorrhizal research[J].Fung Ecol,2016,24(Part B):114-123.
[20]魏杰,杨秀丽.大兴安岭4种杜鹃花科植物的DSE真菌分布[J].内蒙古林业科技,2014,40(4):43-46.
[21]徐风美,王春燕,褚洪龙,等.松萎蔫病发生区和未发生区油松根部真菌群落研究[J].西北植物学报,2014,34(8):1627-1634.
[22]KOHOUT P,TAMAEA T,ROY M,et al.A diverse fungal community associated with Pseudorchis albida(Orchidaceae)roots[J].Fung Ecol,2013,6(1):50-64.
[23]孙茜,薛子可,解琳琳,等.沙冬青及其伴生植物深色有隔内生真菌物种多样性[J].植物生态学报,2017,41(7):729-737.
[24]NEWSHAM K K.A meta-analysis of plant responses to dark septate root endophytes[J].New Phytol,2011,190(3):783-793.
[25]BRUZONE M C,FONTENLA S B,VOHNíK M.Is the prominent ericoid mycorrhizal fungus Rhizoscyphus ericae absent in the Southern Hemisphere’s Ericaceae?A case study on the diversity of root mycobionts in Gaultheria spp.from northwest Patagonia,Argentina[J].Mycorrhiza,2015,25(1):25-40.
[26]张新军.极小种群野生植物大树杜鹃的形态解剖特征及内生真菌多样性研究[D].昆明:云南大学,2015,32-42.
[27]CARLOS A B,GRAMAJE D,JOSE G J,et al.Detection of black-foot disease pathogens in the grapevine nursery;propagation process in Spain[J].Europ J Plant Pathol,2013,137(1):103-112.
[28]VITALE A,AIELLO D,GUARNACCIA V,et al.First Report of Root Rot Caused by Ilyonectria(Neonectria)macrodidyma on Avocado(Persea americana)in Italy[J].J Phytopathol,2012,160(3):156-159.
[29]KOGEL K H,FRANKEN P,HüCKELHOVEN R.Endophyte or parasite-what decides?[J].Curr Opin Plant Biol,2006,9(4):358-363.