橡胶树臭根病菌的鉴定及其生物学特性研究
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摘要
利用海南省橡胶树臭根病病株上采集的病根样本,采用菌索组织分离法获得菌落后,进行子实体诱导产生分生孢子,再经单孢分离纯化获得菌株HNDZ001,通过柯赫氏法则验证、形态学特征观察及分子生物学技术对该菌株进行鉴定,确定菌株HNDZ001为引起橡胶树臭根病的匐灿球赤壳菌(Sphaerostilbe repens Berk.&Br.)。生物学特性测定结果表明,菌株HNDZ001在PDA培养基上菌落近圆形,表面黄白色,致密,背面呈深褐色至浅黄色,后期会产生黄褐色菌索和孢梗束;该菌菌丝生长的适宜条件为温度25~34℃、p H 7~9,最适生长条件是温度28℃、pH值为8、甘露醇为碳源、牛肉浸膏为氮源、完全黑暗,且在黄瓜汁培养基上菌丝生长最好。
Utilizing the diseased root samples collected from the stinking root rot of rubber tree in Hainan Province, colonies were obtained using the method of rhizomorph tissue isolation, and then the sporocarp was induced to produce conidia. The strain HNDZ001 was obtained by single spore isolation and purification. Based on the Koch's postulates, morphological characteristics and molecular biological technology, the strain HNDZ001 was identified as Sphaerostilbe repens Berk. & Br., and It was the pathogen causing stinking root rot of rubber tree. The biological characteristics of the strain showed that the colonies were nearly round, yellowish-white and dense on the surface, dark brown to light yellow on the back, and produced yellow brown rhizomorph synnema on the PDA medium; The optimum conditions for mycelium growth were 25-34 ℃, p H 7-9. The optimum growth conditions were 28 ℃, p H 8, mannitol as carbon source, beef extract as nitrogen source, completely dark, and mycelium grew best in cucumber juice agar medium.
Utilizing the diseased root samples collected from the stinking root rot of rubber tree in Hainan Province, colonies were obtained using the method of rhizomorph tissue isolation, and then the sporocarp was induced to produce conidia. The strain HNDZ001 was obtained by single spore isolation and purification. Based on the Koch's postulates, morphological characteristics and molecular biological technology, the strain HNDZ001 was identified as Sphaerostilbe repens Berk. & Br., and It was the pathogen causing stinking root rot of rubber tree. The biological characteristics of the strain showed that the colonies were nearly round, yellowish-white and dense on the surface, dark brown to light yellow on the back, and produced yellow brown rhizomorph synnema on the PDA medium; The optimum conditions for mycelium growth were 25-34 ℃, p H 7-9. The optimum growth conditions were 28 ℃, p H 8, mannitol as carbon source, beef extract as nitrogen source, completely dark, and mycelium grew best in cucumber juice agar medium.
引文
[1]黄蔚寒,王金林.橡胶树根病及其防治[C]//中国天然橡胶发展大会.2010.
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[11]de Oliveira E J,de Oliveira S A S,Boas S A V,et al.Selection of cassava accessions with multiple resistance to pathogens associated with root rot disease[J].Euphytica,2017,213(8):185.
[12]Boas S A V,Oliveira S A S D,Bragan?a C A D,et al.Survey of fungi associated with cassava root rot from different producing regions in Brazil[J].Scientia Agricola,2017,74(1):60-67.
[13]Zinsou V A,Afouda L A C,Ahohuendo B C,et al.Importance of fungal root rot pathogens of cassava in Benin[J].Pakistan Journal of Botany,2017,49(5):2 023-2 028.
[14]Roger L.Phytopathologies des pays chauds[C]//Roger L.Encyclopédie Mycologique.Paris:Paul Lechevalier,1951:1116-1 126.
[15]Botton B.Morphogenesis of coremia and rhizomorphs in the Ascomycete Sphaerostilbe repens[J].Protoplasma,1983,116(2-3):99-114.
[16]Guillaumin J.Geographical distribution and ecology of the Armillaria species in Western Europe;European journal of forest pathology[J].Forest Pathology,1993,23(6-7):321-341.
[2]John K P.Minor root diseases--a reappraisal[J].Rubber Research Institute of Malaya.1964,75:244-248.
[3]George,Weber F,黄朝豪.橡胶树的次要病害[J].世界热带农业信息,1973(6):32-36.
[4]John K P.Spore dissemination of root disease[J].World Tropical Agriculture Information,1965(3):13-15.
[5]方中达.植病研究方法[M].北京:中国农业出版社,1998:1-427.
[6]孙广宇,宗兆锋.植物病理学实验技术[M].北京:中国农业出版社,2002.
[7]White T J,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[C]/Innis M,Gel-fand J,Sainsky J,et al.PCR protocols:a guide to methods and applications.San Diego:Academic Press.1990:315-322.
[8]李增平,罗大全.橡胶树病虫害诊断图谱[M].北京:中国农业出版社,2007.
[9]Doilom M,Taylor J E,Bhat D J,et al.Checklist of fungi on teak[J].Mycosphere,2016,7(5):656-678.
[10]Azza M K A,Mohamed S Y.Peach canker caused by Nectria mauritiicola and its control in Egypt[J].International Journalof Scientific&Engineering Research,2015,6(5):1 353-1 360.
[11]de Oliveira E J,de Oliveira S A S,Boas S A V,et al.Selection of cassava accessions with multiple resistance to pathogens associated with root rot disease[J].Euphytica,2017,213(8):185.
[12]Boas S A V,Oliveira S A S D,Bragan?a C A D,et al.Survey of fungi associated with cassava root rot from different producing regions in Brazil[J].Scientia Agricola,2017,74(1):60-67.
[13]Zinsou V A,Afouda L A C,Ahohuendo B C,et al.Importance of fungal root rot pathogens of cassava in Benin[J].Pakistan Journal of Botany,2017,49(5):2 023-2 028.
[14]Roger L.Phytopathologies des pays chauds[C]//Roger L.Encyclopédie Mycologique.Paris:Paul Lechevalier,1951:1116-1 126.
[15]Botton B.Morphogenesis of coremia and rhizomorphs in the Ascomycete Sphaerostilbe repens[J].Protoplasma,1983,116(2-3):99-114.
[16]Guillaumin J.Geographical distribution and ecology of the Armillaria species in Western Europe;European journal of forest pathology[J].Forest Pathology,1993,23(6-7):321-341.