进境美国苜蓿草中值得关注的真菌帚状刺壳霉
|
摘要
从进境美国苜蓿干草中发现带有褐色至黑色斑点的苜蓿草茎杆,经保湿培养,在病部表面形成分生孢子器,球形或近球形,褐色至黑色,孔口边缘着生刚毛。分生孢子透明,光滑,单细胞,近圆柱形至卵圆形。在PDA上,菌落初为黄褐色,后转为橄榄绿色,气生菌丝体稀疏,边缘不规则,表面呈绒状,有褶皱,背面呈橄榄绿色。培养5 d后开始产生分生孢子器。培养过程中有厚垣孢子产生和微循环产孢现象,菌丝束浅棕色。Blast分析表明,从分离物基因组中扩增到的ITS基因与Gen Bank中已知的帚状刺壳霉(Chaetopyrena penicillata(Fuckel)Hhn.1918)菌株的ITS序列同源性达99%~100%,基于ITS基因序列的系统发育分析显示分离物与帚状刺壳霉(C.penicillata)聚在同一簇。综合形态学特征、培养性状及ITS序列分析,将分离物鉴定为帚状刺壳霉(Chaetopyrena penicillata(Fuckel)Hhn.1918),此为我国口岸首次截获。迄今为止,除伊朗和南非外,世界其他国家和地区尚未有该病菌在苜蓿草上发生为害的报道。
Stems with brown to black spots were found on Alfalfa hay imported from the United States.Pycnidia formed on the disease area of stems under the condition of moisturizing. Pycnidia superficial,globose to subglobose,brown to black,1 ostiole well-necked with setae around. Conidia hyaline,smooth,1-celled,sub-cylindrical to ovoid. Colonies on PDA were yellowish brown at first,then olive green,aerial mycelium sparse,margins irregular,front velvet with fold and reverse olive green. Pycnidia formed within5 days on PDA. In culture chlamydospores and microcyclic conidiation can be seen,aerial mycelium bundled hyphae,pale brown. Blast analysis revealed ITS amplified from the genome DNA of isolates displayed 99%-100% sequence similarity to the known ITS of Chaetopyrena penicillata strains in Gen Bank.Based on the phylogenetic studies of ITS gene sequence,the isolates clustered together with C. penicillata. On the basis of morphological characteristics,culture patterns,ITS sequence analysis,the isolates were identified as Chaetopyrena penicillata(Fuckel) Hhn. 1918 which was the first time intercepted in the port of China. To our knowledge,C. penicillata has been reported only in Iran and South Africa on Alfalfa.
Stems with brown to black spots were found on Alfalfa hay imported from the United States.Pycnidia formed on the disease area of stems under the condition of moisturizing. Pycnidia superficial,globose to subglobose,brown to black,1 ostiole well-necked with setae around. Conidia hyaline,smooth,1-celled,sub-cylindrical to ovoid. Colonies on PDA were yellowish brown at first,then olive green,aerial mycelium sparse,margins irregular,front velvet with fold and reverse olive green. Pycnidia formed within5 days on PDA. In culture chlamydospores and microcyclic conidiation can be seen,aerial mycelium bundled hyphae,pale brown. Blast analysis revealed ITS amplified from the genome DNA of isolates displayed 99%-100% sequence similarity to the known ITS of Chaetopyrena penicillata strains in Gen Bank.Based on the phylogenetic studies of ITS gene sequence,the isolates clustered together with C. penicillata. On the basis of morphological characteristics,culture patterns,ITS sequence analysis,the isolates were identified as Chaetopyrena penicillata(Fuckel) Hhn. 1918 which was the first time intercepted in the port of China. To our knowledge,C. penicillata has been reported only in Iran and South Africa on Alfalfa.
引文
[1]环球网.宁波口岸从西班牙苜蓿草中截获鳞球茎茎线虫活体[EB/OL].(2017-02-21)[2017-09-20].http://china.huanqiu.com/hot/2016-10/9521341.html.
[2]Parhizi N,Zafari D,Soleimani M J,et al.First report of leaf spot caused by Chaetopyrena penicillata on Medicago sativa in Iran.Journal of Plant Pathology,2017,99:2.
[3]Gruyter J D,Woudenberg J H C,Aveskamp M M,et al.Systematic reappraisal of species in Phoma section Paraphoma,Pyrenochaeta and Pleurophoma.Mycologia,2010,102(5):1066.
[4]White T J,Brunns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics//Innis M A,Gelfand D H,Sninsky J J,et al.PCR protocols:a guide to methods and applications.San Diego,California:Academic Press,1990:315-322.
[5]Crous P W,Gams W,Stalpers J A,et al.Myco Bank:an online initiative to launch mycology into the 21st century.Studies in Mycology,2004,50(2004):19-22.
[6]Arzanlou M,Khodaei S.Phenotypic and molecular characterization of Chaetopyrena penicillata from Iran with description of a hyphomycete synanomorph.Agricultural&Biological Chemistry,2012,3(1):73-77.
[7]Wang Y,Jin L,Lin L,et al.New hosts for Bartalinia and Chaetopyrena in China.Mycotaxon,2016,131(1):1-6.
[8]郑欢,张芝元,韩燕峰,等.刺槐树洞悬土可培养真菌群落组成及其多样性分析.菌物学报,2017,36(5):625-632.
[2]Parhizi N,Zafari D,Soleimani M J,et al.First report of leaf spot caused by Chaetopyrena penicillata on Medicago sativa in Iran.Journal of Plant Pathology,2017,99:2.
[3]Gruyter J D,Woudenberg J H C,Aveskamp M M,et al.Systematic reappraisal of species in Phoma section Paraphoma,Pyrenochaeta and Pleurophoma.Mycologia,2010,102(5):1066.
[4]White T J,Brunns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics//Innis M A,Gelfand D H,Sninsky J J,et al.PCR protocols:a guide to methods and applications.San Diego,California:Academic Press,1990:315-322.
[5]Crous P W,Gams W,Stalpers J A,et al.Myco Bank:an online initiative to launch mycology into the 21st century.Studies in Mycology,2004,50(2004):19-22.
[6]Arzanlou M,Khodaei S.Phenotypic and molecular characterization of Chaetopyrena penicillata from Iran with description of a hyphomycete synanomorph.Agricultural&Biological Chemistry,2012,3(1):73-77.
[7]Wang Y,Jin L,Lin L,et al.New hosts for Bartalinia and Chaetopyrena in China.Mycotaxon,2016,131(1):1-6.
[8]郑欢,张芝元,韩燕峰,等.刺槐树洞悬土可培养真菌群落组成及其多样性分析.菌物学报,2017,36(5):625-632.