甜瓜镰刀菌果腐病新病原菌Fusarium incarnatum的鉴定及生物学特性
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摘要
采用常规组织分离法对采自山西省太谷县疑似镰孢刀菌引起的甜瓜果实新病害进行病原菌分离,根据柯赫氏法则明确其致病性。基于形态特征以及核糖体内转录间隔区(rDNA-ITS)、转录延伸因子(EF-1α)、组蛋白(Histone 3)和线粒体中核糖体小亚基(mtSSU)序列分析,鉴定该病原菌为变红镰刀菌(Fusarium incarnatum)。病原菌的生物学特性测定结果表明,菌丝生长和产孢最适培养基均为马铃薯蔗糖琼脂培养基(PSA);温度为20～35℃菌丝生长较快,20～25℃产孢量最大;羧甲基纤维素钠、淀粉和木糖等碳源适合菌丝生长,产孢的最佳碳源为乳糖;病原菌在多种氮源上均能较好生长,产孢的最佳氮源为酵母粉;pH为8时菌丝生长最快,不同pH处理的产孢量间差异不显著;病原菌在全光照和全黑暗条件下能较好生长,不同光照条件下产孢量差异不显著;菌丝致死温度为61℃(10min),分生孢子致死温度为56℃(10 min)。
A novel disease of suspected Fusarium fruit rot was observed on muskmelon in Taigu County,Shanxi Province. The isolate was obtained from diseased sample by conventional tissue isolation method. The pathogenicity was confirmed by Koch's postulates. The pathogen was identified as Fusarium incarnatum based on morphological characteristics and the sequence analysis of rDNA-ITS,EF-1α,histone 3 and mtSSU. The results of biological characteristics showed that PSA medium was most conducive to mycelium growth and sporulation;the ideal temperature conditions for mycelium growth and sporulation were in the range of 20–35 ℃ and 20–25 ℃,respectively;the suitable carbon sources formycelium growth were CMC-Na,starch and xylose,the optimal carbon source for sporulation was lactose. The pathogen could grow fastly on the medium with kinds of nitrogen sources,the optimal nitrogen source for sporulation was yeast powder. The optimal pH for mycelium growth was 8. The full illumination and full darkness was suitable for mycelium growth. The sporulation under pH and light conditions had not significant difference. The mycelial lethal temperature was at 61 ℃ for 10 min,and the spore lethal temperature was at 56 ℃ for 10 min.
A novel disease of suspected Fusarium fruit rot was observed on muskmelon in Taigu County,Shanxi Province. The isolate was obtained from diseased sample by conventional tissue isolation method. The pathogenicity was confirmed by Koch's postulates. The pathogen was identified as Fusarium incarnatum based on morphological characteristics and the sequence analysis of rDNA-ITS,EF-1α,histone 3 and mtSSU. The results of biological characteristics showed that PSA medium was most conducive to mycelium growth and sporulation;the ideal temperature conditions for mycelium growth and sporulation were in the range of 20–35 ℃ and 20–25 ℃,respectively;the suitable carbon sources formycelium growth were CMC-Na,starch and xylose,the optimal carbon source for sporulation was lactose. The pathogen could grow fastly on the medium with kinds of nitrogen sources,the optimal nitrogen source for sporulation was yeast powder. The optimal pH for mycelium growth was 8. The full illumination and full darkness was suitable for mycelium growth. The sporulation under pH and light conditions had not significant difference. The mycelial lethal temperature was at 61 ℃ for 10 min,and the spore lethal temperature was at 56 ℃ for 10 min.
引文
Bi Shu-juan,Liu Yu-xia,Wang Fei,Liu Hong-yan,Dong Cheng-ming.2012.Biological characteristics of two pathogenic Fusarium species from honeysuchle.Journal of Henan Agricultural Science,41(6):115-118.(in Chinese)毕淑娟,刘玉霞,王飞,刘红彦,董诚明.2012.2种忍冬致病镰刀菌的生物学特性研究.河南农业科学,41(6):115-118.
Chai Zhao-xiang,Li Jin-hua,Li Min-quan,Hao Feng-xia.2004.Isolation of the pathogens of muskmelon fruit rot and toxin bioassay of the predominant pathogen.Chinese Journal and Applied Environmental Biology,10(5):671-674.(in Chinese)柴兆祥,李金花,李敏权,郝风霞.2004.白兰瓜果腐病病原菌的分离鉴定及优势病菌的毒性测定.应用与环境生物学报,10(5):671-674.
Chai Zhao-xiang,Li Jin-hua,Li Min-quan,Fan Xiao-jun.2005.Identification of the superior pathogen of muskmelon fruit rot and producing condition of its conidium.Journal of Fruit Science,22(1):40-43.(in Chinese)柴兆祥,李金花,李敏权,樊小军.2005.白兰瓜果腐病优势病菌鉴定及其产孢条件研究.果树学报,22(1):40-43.
Deng J J,Bi Y,Zhang Z K,Xie D F,Ge Y H,Li W H,Wang J J.2015.Postharvest oxalic acid treatment induces resistance against pink rot by priming in muskmelon(Cucumis melo L.)fruit.Postharvest Biology and Technology,106:53-61.
Ding Z L,Wu J P,Yang C Z,Zhou J,Jiao Z B,Guo F L.2018.First report of Fusarium meridionale and Fusarium incarnatum causing dry rot of Konjac in China.Plant Disease,102(1):247-247.
Fang Zhong-da.1998.Plant pathology research methods.Beijing:China Agriculture Press.(in Chinese)方中达.1998.植病研究方法.北京:中国农业出版社.
Fei Nuo-ya.2018.Identification and biological characteristics of root rot pathogen on blueberry[M.D.Dissertation].Shenyang:Shenyang Agricultural University.(in Chinese)费诺亚.2018.蓝莓根腐病病原菌鉴定及生物学特性研究[硕士论文].沈阳:沈阳农业大学.
Fogliata G M,Martínez C V,Acosta M E,Mu?oz M L,Ploper L D.2013.First report of Fusarium rot caused by Fusarium oxysporum on lemon in Tucumán,Argentina.Plant Disease,97(7):989.
Gai X T,Yang R X,Pan X J,Yuan Y,Wang S N,Liang B B,Gao Z G.2016.First report of Fusarium incarnatum causing stalk rot on maize in China.Plant Disease,100(5):1010.
Glass N L,Donaldson G C.1995.Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes.Applied and Environmental Microbiology,61(4):1323-1330.
Guo K F,Sun J,Zhao S F,He L.2016.Black spot disease of Chinese jujube(Ziziphus jujuba)caused by Fusarium incarnatum in China.Plant Disease,100(2):529.
Jacobs A,Mojela L,Summerell B,Venter E.2018.Characterisation of members of the Fusarium incarnatum-equiseti species complex from undisturbed soils in South Africa.Antonie van Leeuwenhoek,111(11):1999-2008.
Jiang Xian-quan,Wang Wei,Tang Jian-hui,Guo Yu-ren.2007.Characteristics and biocontrol of phomopsis rot on postharvest rock melon.Acta Phytophylacica Sinica,34(2):129-135.(in Chinese)蒋贤权,王伟,唐建辉,郭玉人.2007.甜瓜采后拟茎点霉菌腐烂病及其生物防治.植物保护学报,34(2):129-135.
Lei Ya-hong,Kuang Wei-gang,Zheng Chun-sheng,Li Xiu-zhang,Gao Wen-na,Li Chun-jie.2016.Detection and identification of the genus Fusarium by DNA barcoding.Journal of Plant Protection,43(4):544-551.(in Chinese)雷娅红,况卫刚,郑春生,李秀璋,高文娜,李春杰.2016.基于DNA条形码技术对镰刀菌属的检测鉴定.植物保护学报,43(4):544-551.
Leslie J F,Summerell B A.2006.The Fusarium laboratory manual.Ames:Blackwell Publishing.
Leyva-Madrigal K Y,Larralde-Corona C P,Apodaca-Sánchez M A,Quiroz-Figueroa F R,Mexia-Bola?os P A,Portillo-Valenzuela S,Ordaz-Ochoa J,Maldonado-Mendoza I E.2015.Fusarium species from the Fusarium fujikuroi species complex involved in mixed infections 1 of maize in Northern Sinaloa,Mexico.Journal of Phytopathology,163(6):486-497.
Li Min-quan,Xu Bing-liang,Xu Peng,Xue Li.2001.Perliminary identification of causal organisms of muskmelon pastharvest fruit rot disease.Journal of Gansu Agricultural University,36(4):405-408.(in Chinese)李敏权,徐秉良,徐鹏,薛莉.2001.厚皮甜瓜郁金香采后腐烂病害的病原分离鉴定.甘肃农业大学学报,36(4):405-408.
Li Xin-feng,Wang Jian-ming,Zhang Zuo-gang,Hao Xiao-juan,Zhang Zu-wei,Tian Hong-xian.2015.Analysis of genetic diversity and cross-species transferability based on Fusarium oxysporum EST-SSR markers.Journal of Plant Protection,42(5):777-786.(in Chinese)李新凤,王建明,张作刚,郝晓娟,张祖维,田宏先.2015.尖镰孢菌EST-SSR遗传多样性分析及通用性评价.植物保护学报,42(5):777-786.
Liu Zhi-heng,Hou Yue,Hu Ji-xiang,Teng Xiao-fei,Huang Xin-yang,Wang Shi-wei,Zhao Ting-chang.2013.Identification and biological characteristics of muskmelon fruit rot pathogen in Liaoning Province.Acta Horticulturae Sinica,40(1):89-97.(in Chinese)刘志恒,侯悦,胡积祥,滕晓菲,黄欣阳,王世维,赵廷昌.2013.辽宁省甜瓜果腐病病原菌鉴定及生物学特性初探.园艺学报,40(1):89-97.
Mckeen C D,Thorpe H J.2011.A pythium root rot of muskmelon.Canadian Journal of Botany,46(10):1165-1171.
Palmore T N,Shea Y R,Childs R W,Sherry R M,Walsh T J.2010.Fusarium proliferatum soft tissue infection at the site of a puncture by a plant:recovery,isolation,and direct molecular identification.Journal of Clinical Microbiology,48(1):338-342.
Ramdial H,Latchoo R K,Hosein F N,Rampersad S N.2017.Phylogeny and haplotype analysis of fungi within the Fusarium incarnatum-equiseti species complex.Phytopathology,107(1):109-120.
Seta S,Gonzalez M,Lori G.2004.First report of walnut canker caused by Fusarium incarnatum in Argentina.Plant Pathology,53(2):248-248.
Singh B,Kalha C S,Razdan V K,Verma V S.2011.First report of walnut canker caused by Fusarium incarnatum from India.Plant Disease,95(12):1587.
Stenglein S A,Rodriguero M S,Chandler E,Jennings P,Salerno G L,Nicholson P.2010.Phylogenetic relationships of Fusarium poae based on EF-1αand mtSSU sequences.Fungal Biology,114(1):96-106.
Wang Lin,Li Xin-feng,Xu Yu-mei,Chang Yin-dong,Wang Jian-ming.2017.Analysis of population distribution and genetic variation of plant pathogenic Fusarium in Shanxi Province.Scientia Agricultura Sinica,50(10):1802-1816.(in Chinese)王琳,李新凤,徐玉梅,畅引东,王建明.2017.山西省植物病原镰孢菌种群分布及遗传变异分析.中国农业科学,50(10):1802-1816.
Xiao Min,Ji Xun-cong,Wang Yun-qin,Chen Mian-cai,Rui Kai,Zhang Rong-yi.2011.Research of chemical control on muskmelon Fusarium rot in Hainan Island.Journal of Changjiang Vegetables,(2):63-65.(in Chinese)肖敏,吉训聪,王运勤,陈绵才,芮凯,张荣意.2011.海南岛甜瓜镰刀菌果腐病药剂防治研究.长江蔬菜,(2):63-65.
Zhang You-lin,Zhang Run-guang,Sun Gang,Chen Ting.2010.Study on postharvest physiology,storage diseases and quality changes of Jiashi muskmelon.Scientia Agricultura Sinica,43(6):1220-1228.(in Chinese)张有林,张润光,孙刚,陈婷.2010.伽师瓜采后生理、贮期病害及贮藏保鲜技术.中国农业科学,43(6):1220-1228.
Zhou Jie,Ding Zi-li,Yang Chao-zhu,Wen Liang,Jiao Zhen-biao,Wu Jin-ping.2018.Pathogen identification of dry rot of Amorphophallus konjac in storage period and indoor fungicide-screening.China Plant Protection,38(2):11-16.(in Chinese)周洁,丁自立,杨朝柱,温亮,矫振彪,吴金平.2018.魔芋贮藏期干腐病病原菌鉴定及室内药效初报.中国植保导刊,38(2):11-16.
Chai Zhao-xiang,Li Jin-hua,Li Min-quan,Hao Feng-xia.2004.Isolation of the pathogens of muskmelon fruit rot and toxin bioassay of the predominant pathogen.Chinese Journal and Applied Environmental Biology,10(5):671-674.(in Chinese)柴兆祥,李金花,李敏权,郝风霞.2004.白兰瓜果腐病病原菌的分离鉴定及优势病菌的毒性测定.应用与环境生物学报,10(5):671-674.
Chai Zhao-xiang,Li Jin-hua,Li Min-quan,Fan Xiao-jun.2005.Identification of the superior pathogen of muskmelon fruit rot and producing condition of its conidium.Journal of Fruit Science,22(1):40-43.(in Chinese)柴兆祥,李金花,李敏权,樊小军.2005.白兰瓜果腐病优势病菌鉴定及其产孢条件研究.果树学报,22(1):40-43.
Deng J J,Bi Y,Zhang Z K,Xie D F,Ge Y H,Li W H,Wang J J.2015.Postharvest oxalic acid treatment induces resistance against pink rot by priming in muskmelon(Cucumis melo L.)fruit.Postharvest Biology and Technology,106:53-61.
Ding Z L,Wu J P,Yang C Z,Zhou J,Jiao Z B,Guo F L.2018.First report of Fusarium meridionale and Fusarium incarnatum causing dry rot of Konjac in China.Plant Disease,102(1):247-247.
Fang Zhong-da.1998.Plant pathology research methods.Beijing:China Agriculture Press.(in Chinese)方中达.1998.植病研究方法.北京:中国农业出版社.
Fei Nuo-ya.2018.Identification and biological characteristics of root rot pathogen on blueberry[M.D.Dissertation].Shenyang:Shenyang Agricultural University.(in Chinese)费诺亚.2018.蓝莓根腐病病原菌鉴定及生物学特性研究[硕士论文].沈阳:沈阳农业大学.
Fogliata G M,Martínez C V,Acosta M E,Mu?oz M L,Ploper L D.2013.First report of Fusarium rot caused by Fusarium oxysporum on lemon in Tucumán,Argentina.Plant Disease,97(7):989.
Gai X T,Yang R X,Pan X J,Yuan Y,Wang S N,Liang B B,Gao Z G.2016.First report of Fusarium incarnatum causing stalk rot on maize in China.Plant Disease,100(5):1010.
Glass N L,Donaldson G C.1995.Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes.Applied and Environmental Microbiology,61(4):1323-1330.
Guo K F,Sun J,Zhao S F,He L.2016.Black spot disease of Chinese jujube(Ziziphus jujuba)caused by Fusarium incarnatum in China.Plant Disease,100(2):529.
Jacobs A,Mojela L,Summerell B,Venter E.2018.Characterisation of members of the Fusarium incarnatum-equiseti species complex from undisturbed soils in South Africa.Antonie van Leeuwenhoek,111(11):1999-2008.
Jiang Xian-quan,Wang Wei,Tang Jian-hui,Guo Yu-ren.2007.Characteristics and biocontrol of phomopsis rot on postharvest rock melon.Acta Phytophylacica Sinica,34(2):129-135.(in Chinese)蒋贤权,王伟,唐建辉,郭玉人.2007.甜瓜采后拟茎点霉菌腐烂病及其生物防治.植物保护学报,34(2):129-135.
Lei Ya-hong,Kuang Wei-gang,Zheng Chun-sheng,Li Xiu-zhang,Gao Wen-na,Li Chun-jie.2016.Detection and identification of the genus Fusarium by DNA barcoding.Journal of Plant Protection,43(4):544-551.(in Chinese)雷娅红,况卫刚,郑春生,李秀璋,高文娜,李春杰.2016.基于DNA条形码技术对镰刀菌属的检测鉴定.植物保护学报,43(4):544-551.
Leslie J F,Summerell B A.2006.The Fusarium laboratory manual.Ames:Blackwell Publishing.
Leyva-Madrigal K Y,Larralde-Corona C P,Apodaca-Sánchez M A,Quiroz-Figueroa F R,Mexia-Bola?os P A,Portillo-Valenzuela S,Ordaz-Ochoa J,Maldonado-Mendoza I E.2015.Fusarium species from the Fusarium fujikuroi species complex involved in mixed infections 1 of maize in Northern Sinaloa,Mexico.Journal of Phytopathology,163(6):486-497.
Li Min-quan,Xu Bing-liang,Xu Peng,Xue Li.2001.Perliminary identification of causal organisms of muskmelon pastharvest fruit rot disease.Journal of Gansu Agricultural University,36(4):405-408.(in Chinese)李敏权,徐秉良,徐鹏,薛莉.2001.厚皮甜瓜郁金香采后腐烂病害的病原分离鉴定.甘肃农业大学学报,36(4):405-408.
Li Xin-feng,Wang Jian-ming,Zhang Zuo-gang,Hao Xiao-juan,Zhang Zu-wei,Tian Hong-xian.2015.Analysis of genetic diversity and cross-species transferability based on Fusarium oxysporum EST-SSR markers.Journal of Plant Protection,42(5):777-786.(in Chinese)李新凤,王建明,张作刚,郝晓娟,张祖维,田宏先.2015.尖镰孢菌EST-SSR遗传多样性分析及通用性评价.植物保护学报,42(5):777-786.
Liu Zhi-heng,Hou Yue,Hu Ji-xiang,Teng Xiao-fei,Huang Xin-yang,Wang Shi-wei,Zhao Ting-chang.2013.Identification and biological characteristics of muskmelon fruit rot pathogen in Liaoning Province.Acta Horticulturae Sinica,40(1):89-97.(in Chinese)刘志恒,侯悦,胡积祥,滕晓菲,黄欣阳,王世维,赵廷昌.2013.辽宁省甜瓜果腐病病原菌鉴定及生物学特性初探.园艺学报,40(1):89-97.
Mckeen C D,Thorpe H J.2011.A pythium root rot of muskmelon.Canadian Journal of Botany,46(10):1165-1171.
Palmore T N,Shea Y R,Childs R W,Sherry R M,Walsh T J.2010.Fusarium proliferatum soft tissue infection at the site of a puncture by a plant:recovery,isolation,and direct molecular identification.Journal of Clinical Microbiology,48(1):338-342.
Ramdial H,Latchoo R K,Hosein F N,Rampersad S N.2017.Phylogeny and haplotype analysis of fungi within the Fusarium incarnatum-equiseti species complex.Phytopathology,107(1):109-120.
Seta S,Gonzalez M,Lori G.2004.First report of walnut canker caused by Fusarium incarnatum in Argentina.Plant Pathology,53(2):248-248.
Singh B,Kalha C S,Razdan V K,Verma V S.2011.First report of walnut canker caused by Fusarium incarnatum from India.Plant Disease,95(12):1587.
Stenglein S A,Rodriguero M S,Chandler E,Jennings P,Salerno G L,Nicholson P.2010.Phylogenetic relationships of Fusarium poae based on EF-1αand mtSSU sequences.Fungal Biology,114(1):96-106.
Wang Lin,Li Xin-feng,Xu Yu-mei,Chang Yin-dong,Wang Jian-ming.2017.Analysis of population distribution and genetic variation of plant pathogenic Fusarium in Shanxi Province.Scientia Agricultura Sinica,50(10):1802-1816.(in Chinese)王琳,李新凤,徐玉梅,畅引东,王建明.2017.山西省植物病原镰孢菌种群分布及遗传变异分析.中国农业科学,50(10):1802-1816.
Xiao Min,Ji Xun-cong,Wang Yun-qin,Chen Mian-cai,Rui Kai,Zhang Rong-yi.2011.Research of chemical control on muskmelon Fusarium rot in Hainan Island.Journal of Changjiang Vegetables,(2):63-65.(in Chinese)肖敏,吉训聪,王运勤,陈绵才,芮凯,张荣意.2011.海南岛甜瓜镰刀菌果腐病药剂防治研究.长江蔬菜,(2):63-65.
Zhang You-lin,Zhang Run-guang,Sun Gang,Chen Ting.2010.Study on postharvest physiology,storage diseases and quality changes of Jiashi muskmelon.Scientia Agricultura Sinica,43(6):1220-1228.(in Chinese)张有林,张润光,孙刚,陈婷.2010.伽师瓜采后生理、贮期病害及贮藏保鲜技术.中国农业科学,43(6):1220-1228.
Zhou Jie,Ding Zi-li,Yang Chao-zhu,Wen Liang,Jiao Zhen-biao,Wu Jin-ping.2018.Pathogen identification of dry rot of Amorphophallus konjac in storage period and indoor fungicide-screening.China Plant Protection,38(2):11-16.(in Chinese)周洁,丁自立,杨朝柱,温亮,矫振彪,吴金平.2018.魔芋贮藏期干腐病病原菌鉴定及室内药效初报.中国植保导刊,38(2):11-16.