澳洲坚果叶斑病新病原及生物学特性
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摘要
在澳洲坚果园发现一种叶部病害,为明确该病害的病原菌种类,将采集病叶进行病原菌分离,并运用柯赫法则测定致病性,根据病原菌的形态学特征和rDNA-ITS序列分析对病原菌进行鉴定。结果表明,分离菌株的菌落形态和分生孢子形态都与小孢拟盘多毛孢Pestalotiopsis microspora一致,经分子生物学鉴定,其ITS序列与小孢拟盘多毛孢的ITS序列的同源性为99%,将引起澳洲坚果叶斑病的病原菌鉴定为小孢拟盘多毛孢。PDA培养基最有利于该病原菌生长,培养3d后,菌落直径达62.9mm,最佳碳源为葡萄糖,最佳氮源为硫酸铵;病原菌在15~30℃均能生长,最适生长温度28℃;病原菌在pH值4~11范围内均可生长,pH值7生长最佳,全光照有利于病原菌生长。首次报道Pestalotiopsis microspora引起澳洲坚果叶斑病。
A macadamia leaf disease was found in macadamiaplantation.In order to identify the pathogen,isolates were obtained by isolation and purification.The pathogenicity was tested using the Koch's Postulates,pathogenicity of isolates from the symptomatic leaf.Identification of pathogen was conducted according to the morphological characteristics and ITS sequence.The pathogen isolated from the macadamia is Pestalotiopsis microspora.Biological analysis showed that the PDA was the most conducive medium for the growth of pathogen.The colony diameters were up to 62.9 mm after incubation at 25℃ in dark for three days.The best carbon source for bacteria growth was dextrose,and the best nitrogen source was ammonia sulfate.Pathogenic bacteria could grow at 15~30 ℃,and the most suitable temperature was 28 ℃.Pathogenic bacteria could grow at pH value ranging from 4 to 11,and the most suitable pH value was 7.Continuous illumination was the most favorable for the growth of mycelium.This is the first report of Pestalotiopsis microspora causing diseases on macadamia.
A macadamia leaf disease was found in macadamiaplantation.In order to identify the pathogen,isolates were obtained by isolation and purification.The pathogenicity was tested using the Koch's Postulates,pathogenicity of isolates from the symptomatic leaf.Identification of pathogen was conducted according to the morphological characteristics and ITS sequence.The pathogen isolated from the macadamia is Pestalotiopsis microspora.Biological analysis showed that the PDA was the most conducive medium for the growth of pathogen.The colony diameters were up to 62.9 mm after incubation at 25℃ in dark for three days.The best carbon source for bacteria growth was dextrose,and the best nitrogen source was ammonia sulfate.Pathogenic bacteria could grow at 15~30 ℃,and the most suitable temperature was 28 ℃.Pathogenic bacteria could grow at pH value ranging from 4 to 11,and the most suitable pH value was 7.Continuous illumination was the most favorable for the growth of mycelium.This is the first report of Pestalotiopsis microspora causing diseases on macadamia.
引文
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[6]陈剑,纪烨斌,张能,等.短穗鱼尾葵灰斑病菌的鉴定及生物学特性[J].植物保护,2011,37(4):48-51.
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[14]LEE S,CROUS P W,WINGFIELD M J.Pestalotioid fungi from Restionaceae in the Cape Floral Kingdom[J].Studies in Mycology,2006,55(1):175
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[17]张小媛,何红,胡汉桥,等.红海榄赤斑病病原菌鉴定及其生物学特性研究[J].植物病理学报,2009,39(6):584-592
[18]KEITH L M,VELASQUEZ M E,ZEE F T.Identification and characterization of Pestalotiopsisspp.causing scab disease of guava,Psidium guajava,in Hawaii[J].Plant Disease,2006,90(1):16-23
[19]METZ A M,HADDAD A,WORAPONG J,et al.Induction of the sexual stage of Pestalotiopsis microspora,a taxol-producing fungus[J].Microbiology,2000,146(8):2079-2089
[20]李建宏,谢昌平,王延丽,等.散尾葵拟盘多毛孢叶斑病菌的鉴定[J].热带农业科学,2013,33(2):62-64
[21]管斌,徐超,张红岩,等.红叶石楠叶斑病病原菌分离鉴定及致病性测定研究[J].西部林业科学,2013,42(2):56-61
[22]肖仲久,李小霞.白术叶枯病病原菌鉴定及其生物学特性研究[J].热带作物学报,2013,34(5):973-977
[23]崔朝宇,王园秀,蒋军喜,等.美国红枫褐斑病病原菌鉴定[J].林业科学,2015,51(10):142-147
[24]ZHANG C Q,LIU Y H,WU H M,et al.Baseline sensitivity of Pestalotiopsis microspora,which causes black spot disease on Chinese hickory(Carya cathayensis),to pyraclostrobin[J].Crop Protection,2012,42:256-259
[25]王璇.山核桃干腐病病原菌的鉴定及其系统发育分析[D].杭州:浙江农林大学,2013
[26]JEON Y H,CHEON W.First report of leaf blight of Japanese yew caused by Pestalotiopsis microspora in Korea[J].Plant Disease,2014,98(5):691
[2]杨为海,张明楷,邹明宏,等.澳洲坚果不同种质果仁粗脂肪及脂肪酸成分的研究[J].热带作物学报,2012,33(7):1297-1302
[3]张剑,董晔欣,张金林,等.一株具有高除草活性的真菌菌株[J].菌物学报,2008,27(5):645-651
[4]肖仲久,李小霞,段帅,等.麻风树叶枯病菌的鉴定(英文)[J].菌物学报,2010,29(6):874-878
[5]张传清,徐志宏,孙品雷,等.新病害——山核桃果实黑斑病病原菌的鉴定[J].植物保护,2010,36(4):160-162
[6]陈剑,纪烨斌,张能,等.短穗鱼尾葵灰斑病菌的鉴定及生物学特性[J].植物保护,2011,37(4):48-51.
[7]方中达.植病研究方法[M].第3版.北京:中国农业出版社,1998
[8]王志龙,王胜,谭志文,等.无患子枝干新病害——溃疡病病原鉴定[J].植物病理学报,2014,44(2):208-212
[9]张剑.小孢拟盘多毛孢的鉴定与除草活性物质分析[D].保定:河北农业大学,2008
[10]牛晓庆,陈良秋,付登强,等.油茶叶枯病菌的鉴定及生物学特性研究[J].热带作物学报,2013,34(2):352-357
[11]ZHANG H T,LEE Y K,ZHANG W,et al.Culturable actinobacteria from the marine sponge Hymeniacidon perleve:isolation and phylogenetic diversity by 16S rRNA gene-RFLP analysis[J].Antonie Van Leeuwenhoek,2006,90(2):159-169
[12]BARR M E.Pestalosphaeria,a new genus in the Amphisphaeriaceae[J].Mycologia,1975,67(1):187-194
[13]KANG J C,HYDE K D,KONG R Y C.Studies on Amphisphaeriales:the Amphisphaeriaceae(sensu stricto)[J].Mycological Research,1999,103(1):53-64
[14]LEE S,CROUS P W,WINGFIELD M J.Pestalotioid fungi from Restionaceae in the Cape Floral Kingdom[J].Studies in Mycology,2006,55(1):175
[15]丁榕,王延丽,李博勋,等.杧果拟盘多毛孢叶枯病菌鉴定及其生物学特性研究[J].中国南方果树,2010,39(4):20-24
[16]韦继光,徐同,潘秀湖,等.拟盘多毛孢属的分类学研究进展[J].广西农业生物科学,2006,25(1):78-85
[17]张小媛,何红,胡汉桥,等.红海榄赤斑病病原菌鉴定及其生物学特性研究[J].植物病理学报,2009,39(6):584-592
[18]KEITH L M,VELASQUEZ M E,ZEE F T.Identification and characterization of Pestalotiopsisspp.causing scab disease of guava,Psidium guajava,in Hawaii[J].Plant Disease,2006,90(1):16-23
[19]METZ A M,HADDAD A,WORAPONG J,et al.Induction of the sexual stage of Pestalotiopsis microspora,a taxol-producing fungus[J].Microbiology,2000,146(8):2079-2089
[20]李建宏,谢昌平,王延丽,等.散尾葵拟盘多毛孢叶斑病菌的鉴定[J].热带农业科学,2013,33(2):62-64
[21]管斌,徐超,张红岩,等.红叶石楠叶斑病病原菌分离鉴定及致病性测定研究[J].西部林业科学,2013,42(2):56-61
[22]肖仲久,李小霞.白术叶枯病病原菌鉴定及其生物学特性研究[J].热带作物学报,2013,34(5):973-977
[23]崔朝宇,王园秀,蒋军喜,等.美国红枫褐斑病病原菌鉴定[J].林业科学,2015,51(10):142-147
[24]ZHANG C Q,LIU Y H,WU H M,et al.Baseline sensitivity of Pestalotiopsis microspora,which causes black spot disease on Chinese hickory(Carya cathayensis),to pyraclostrobin[J].Crop Protection,2012,42:256-259
[25]王璇.山核桃干腐病病原菌的鉴定及其系统发育分析[D].杭州:浙江农林大学,2013
[26]JEON Y H,CHEON W.First report of leaf blight of Japanese yew caused by Pestalotiopsis microspora in Korea[J].Plant Disease,2014,98(5):691