一株耐盐植物内生真菌的筛选与鉴定
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摘要
[目的]研究盐田原生态系统中的耐盐植物内生真菌。[方法]对38株从海南省莺歌海盐场和东方盐场分离的菌株进行耐盐生长测试。[结果]发现了1株在8%Na Cl浓度下生长较好的真菌,并根据其形态特征,ITS扩增序列聚类进化树分析,显示其分类地位为黑酵母菌(Hortaea werneckii)。经过高效液相色谱分析其代谢产物,发现该真菌在0、2%、4%、8%Na Cl浓度下的发酵产物均有所不同。[结论]盐田生境中存在特殊的植物内生真菌区系,其中黑酵母菌Hortaea werneckii具有一定代表性,有深入研究的价值。
[Objective]To study the endophytic fungi of halotolerant in the original ecosystem of saltern. [Method]Salt stress tests were conducted on the endophytic fungi isolated from Yingehai Saltern and Dongfang Saltern in Hainan Province,to discover the halotolerant fungi among them. [Result]One isolate was further identified according to morphological and ITS sequence-based phylogenic analyses,which suggested it belongs to Hortaea werneckii,a black yeast species,in taxonomy. The ferment substance under 0,2%,4%,8% of sodium chloride of this isolate was studied with HPLC finger printing approach,and differential metabolites according to salt concentrations were observed. [Conclusion]The solar salterns may be occupied by idiographic microflora of endophytes,included Hortaea werneckii observed in this study,which maybe representative of further studies.
[Objective]To study the endophytic fungi of halotolerant in the original ecosystem of saltern. [Method]Salt stress tests were conducted on the endophytic fungi isolated from Yingehai Saltern and Dongfang Saltern in Hainan Province,to discover the halotolerant fungi among them. [Result]One isolate was further identified according to morphological and ITS sequence-based phylogenic analyses,which suggested it belongs to Hortaea werneckii,a black yeast species,in taxonomy. The ferment substance under 0,2%,4%,8% of sodium chloride of this isolate was studied with HPLC finger printing approach,and differential metabolites according to salt concentrations were observed. [Conclusion]The solar salterns may be occupied by idiographic microflora of endophytes,included Hortaea werneckii observed in this study,which maybe representative of further studies.
引文
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[10]李炳学,李颖.产黑色素酵母状真菌[J].微生物学通报,2008,35(11):1791-1796.
[11]PLEMENITAA,GUNDE-CIMERMAN N.Cellular responses in the halophilic black yeast Hortaea werneckii to high environmental salinity[M]//GUNDE-CIMERMAN N,OREN A,PLEMENITAA.Adaptation to life at high salt concentrations in Archaea,Bacteria,and Eukarya.Dordrecht:Springer Verlag,2005.
[12]TURK M,MJANELLE L,ENTJURC M,et al.Salt-induced changes in lipid composition and membrane fluidity of halophilic yeast-like melanized fungi[J].Extremophiles,2004,8(1):53-61.
[13]CHEN J,XING X K,ZHANG L C,et al.Identification of Hortaea werneckii isolated from mangrove plant Aegiceras comiculatum based on morphology and r DNA sequences[J].Mycopathologia,2012,174:
[2]DOYLE J J,DOYLE J L.A rapid DNA isolation procedure for small quantities of fresh leaf tissue[J].Phytochem Bull,1987,19:11-15.
[3]HALL T A.Bio Edit:A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[J].Nucl Acids Symp Ser,1999,41:95-98
[4]DARRIBA D,TABOADA G L,DOALLO R,et al.j Model Test 2:More models,new heuristics and parallel computing[J].Nat Methods,2012,9(8):772.
[5]SWOFFORD D L.PAUP*.Phylogenetic analysis using parsimony(*and other methods)[M].Version 4.Sunderland,MA:Sinauer Associates,2000.
[6]HUELSENBECK J P,RONQUIST F.MRBAYES:Bayesian inference of phylogeny[J].Bioinformatics,2001,17:754-755.
[7]陈惠琴,朱剑梅,王佩,等.红树植物角果木内生真菌Penicillium sp.J54的次生代谢产物研究[J].中国抗生素杂志,2017,42(4):341-345.
[8]YURLOVA N A,DE HOOG S,VAN DEN ENDE B G.Taxonomy of Aureobasidium and allied genera[J].Studies in mycology,1999,43:63-69.
[9]NISHIMURA K,MIYAJI M.Further studies on the phylogenesis of the genus Exophiala and Hortaea[J].Mycophthologia,1985,92:101-109.
[10]李炳学,李颖.产黑色素酵母状真菌[J].微生物学通报,2008,35(11):1791-1796.
[11]PLEMENITAA,GUNDE-CIMERMAN N.Cellular responses in the halophilic black yeast Hortaea werneckii to high environmental salinity[M]//GUNDE-CIMERMAN N,OREN A,PLEMENITAA.Adaptation to life at high salt concentrations in Archaea,Bacteria,and Eukarya.Dordrecht:Springer Verlag,2005.
[12]TURK M,MJANELLE L,ENTJURC M,et al.Salt-induced changes in lipid composition and membrane fluidity of halophilic yeast-like melanized fungi[J].Extremophiles,2004,8(1):53-61.
[13]CHEN J,XING X K,ZHANG L C,et al.Identification of Hortaea werneckii isolated from mangrove plant Aegiceras comiculatum based on morphology and r DNA sequences[J].Mycopathologia,2012,174: