학술논문
蘘荷叶枯病病原菌的分离鉴定与生物学特性 / Isolation,identification and biological characteristics of the pathogen causing leaf blight of Zingiber mioga
Document Type
Academic Journal
Author
王小秋; 葛礼姣; 仇亮; 翟彩娇; 王广达; 宋益民; 程玉静; WANG Xiaoqiu; GE Lijiao; QIU Liang; ZHAI Caijiao; WANG Guangda; SONG Yimin; CHENG Yujing
Source
湖南农业大学学报（自然科学版） / Journal of Hunan Agricultural University(Natural Sciences). 49(6):689-736
Subject
Language
Chinese
ISSN
1007-1032
Abstract
从发病严重田块采集蘘荷叶枯病病叶,采用组织分离法进行病原菌分离、纯化,获得代表性致病菌株RH-1,对RH-1进行鉴定,并测定其生物学特性.结合病原菌形态学特征和分子生物学鉴定,构建真菌内转录间隔区(ITS)、β-微管蛋白基因(β-Tubulin)和TEF-1α系统发育树,鉴定该菌为尖孢镰刀菌(Fusarium oxysporum).分别在温度(5、10、15、20、25、30、35℃)、pH(4、5、6、7、8、9、10)和光照时间(连续光照、连续黑暗、12 h光照/12 h黑暗、24 h光照/24 h黑暗)条件下用PDA培养基培养RH-1,结果在 30℃、pH 9和连续光照下菌丝生长较快.分别用葡萄糖、α-乳糖和麦芽糖以及酵母膏、牛肉膏、(NH4)2SO4、NH4Cl、蛋白胨和NH2CH2COOH等量替代 Czapek 基础培养基中的碳源和氮源,发现菌丝生长的较适碳源为葡萄糖或麦芽糖,最适氮源为牛肉膏或蛋白胨.将RH-1分别接种于红蘘荷和绿蘘荷,发现绿蘘荷的抗病性强于红蘘荷的.
Diseased leaves from Zingiber mioga with leaf blight were collected from severely affected fields,and pathogenic bacteria were isolated and purified using tissue isolation method.The representative pathogenic strain RH-1 was obtained,further identification of strain RH-1 was conducted and biological characteristics of the strain were determined.Combining the results of morphological characteristics,molecular and biological identification,constructed fungal internal transcriptional spacer regions(ITS),phylogenetic tree based on β-Tubulin and TEF-1α,the pathogen was identified as Fusarium oxysporum.Strain RH-1 was cultured on PDA medium with different temperatures(5,10,15,20,25,30,35℃),pHs(4,5,6,7,8,9,10),and light durations(continuous light,continuous darkness,12 hours of light/12 hours of darkness,and 24 hours of light/24 hours of darkness).The results showed that the mycelium grew faster under 30℃,pH9,and continuous light conditions.Using glucose α-lactose and maltose as carbon sources,and yeast extract,beef extract,(NH4)2SO4,NH4Cl,peptone,and NH2CH2COOH as nitrogen sources to replace the carbon and nitrogen sources in Czapek basic culture medium in equal amounts.mycelium growth showed that the optimal carbon source was glucose or maltose,and the optimal nitrogen source was beef extract or peptone.Strain RH-1 was inoculated into red and green Zingiber mioga respectively,and it was found that green Zingiber mioga has stronger disease resistance.
Diseased leaves from Zingiber mioga with leaf blight were collected from severely affected fields,and pathogenic bacteria were isolated and purified using tissue isolation method.The representative pathogenic strain RH-1 was obtained,further identification of strain RH-1 was conducted and biological characteristics of the strain were determined.Combining the results of morphological characteristics,molecular and biological identification,constructed fungal internal transcriptional spacer regions(ITS),phylogenetic tree based on β-Tubulin and TEF-1α,the pathogen was identified as Fusarium oxysporum.Strain RH-1 was cultured on PDA medium with different temperatures(5,10,15,20,25,30,35℃),pHs(4,5,6,7,8,9,10),and light durations(continuous light,continuous darkness,12 hours of light/12 hours of darkness,and 24 hours of light/24 hours of darkness).The results showed that the mycelium grew faster under 30℃,pH9,and continuous light conditions.Using glucose α-lactose and maltose as carbon sources,and yeast extract,beef extract,(NH4)2SO4,NH4Cl,peptone,and NH2CH2COOH as nitrogen sources to replace the carbon and nitrogen sources in Czapek basic culture medium in equal amounts.mycelium growth showed that the optimal carbon source was glucose or maltose,and the optimal nitrogen source was beef extract or peptone.Strain RH-1 was inoculated into red and green Zingiber mioga respectively,and it was found that green Zingiber mioga has stronger disease resistance.