促进滇重楼皂苷类活性成分积累的内生真菌筛选
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摘要
【目的】植物内生菌(Endophyte)是近年来深受重视的一种新型的微生物能源,具有固氮、促进植物生长、增强宿主植物抗逆性等作用,目前研究表明:内生菌可增强宿主植物合成活性物质的能力,甚至它们本身也能产生与宿主植物相同或相近的生物活性物质。【方法】以传统药用植物滇重楼(Pairs polyphylla var.yunnanensis)为研究对象,从滇重楼地下块茎中分离得到23株内生菌,对其进行分子鉴定,之后与滇重楼组培苗进行共生培养,一段时间后利用高效液相色谱法检测组培苗的皂苷含量。【结果】最终筛选出2株对滇重楼组培苗皂苷含量有促进作用的菌株。【结论】人参生柱隔孢和木霉菌能促进滇重楼皂苷类活性成分的积累。
[Purpose]Endophyte have got much attention as a novel microbial energy source in recent years. Its functions include nitrogen fixation, plant growth promotion and stress resistance of host plants. It is thought that endophyte can produce the same or similarity substances as host plant. Furthermore, some endophytes can also help host plant synthesis more active ingredients.[Method]In this study, 23 strains of endophytic fungi were isolated from the tuberous tissue of Paris polyphylla var. yunnanensis, and then cultured with the tissue culture plant of P. polyphylla var. yunnanensis,after culturing for a certain time.[Results]Two strains were screened for their increase of saponin content in tissue culture seedlings.[Conclusion]Cylindrocarpon sp. and Trichoderma viride can promote the accumulation of active constituent of P. polyphylla var. yunnanensis.
[Purpose]Endophyte have got much attention as a novel microbial energy source in recent years. Its functions include nitrogen fixation, plant growth promotion and stress resistance of host plants. It is thought that endophyte can produce the same or similarity substances as host plant. Furthermore, some endophytes can also help host plant synthesis more active ingredients.[Method]In this study, 23 strains of endophytic fungi were isolated from the tuberous tissue of Paris polyphylla var. yunnanensis, and then cultured with the tissue culture plant of P. polyphylla var. yunnanensis,after culturing for a certain time.[Results]Two strains were screened for their increase of saponin content in tissue culture seedlings.[Conclusion]Cylindrocarpon sp. and Trichoderma viride can promote the accumulation of active constituent of P. polyphylla var. yunnanensis.
引文
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[18]孙静贤,高玉红,杨海英,等.滇重楼内生真菌抗菌活性的筛选和菌株鉴定[J].广东农业科学,2013,40(9):74.
[19]刘舒,陈春黎,刘敏,等.两种内生真菌对大花蕙兰的共生效应比较[J].华中农业大学学报,2016,35(1):43.
[20]DIJK E.Effects of mycorrhizal fungi on in-vitro nitrogen response of juvenile orchids[J].Agriculture Ecosystems&Environment,1990,29(1/4):91.
[21]金有亮,杨福孙,宋希强,等.文心兰组培苗与野生兰科植物内生真菌共培养的生长效应研究[J].热带作物学报,2011,32(8):1483.DOI:10.3969/j.issn.1000-2561.2011.08.020.
[22]张烨,吕霜霜,周浓,等.不同生长年限滇重楼中4种重楼皂苷的含量比较[J].中国药房,2011,22(43):4081.
[23]AINHOAMARTINEZ-MEDINA A.Phytohormore profiles induced by Trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants[J].Journal of Chemical Ecology,2014,7(15):804.
[24]何秀丽,丁嫚嫚,叶方,等.重楼植物中内生菌研究进展[J].中国药师,2016,19(5):970.DOI:10.3969/j.issn.1008-049X.2016.05.047.
[25]严雪瑞.人参西洋参锈腐病菌比较生物学研究[D].沈阳:沈阳农业大学,2002.
[26]杨玲玲.滇重楼根茎细菌多样性与根瘤菌系统发育基因组学研究[D].昆明:云南大学,2016.
[2]SONG Y L,WU P,LI Y F,et al.Effect of endophytic fungi on the host plant growth,expression of expansin gene and flavonoid content in Tetrastigma hemsleyanumDiels&Gilg ex Diels[J].Plant and Soil,2017,417(1/2):393 .DOI:10.1007/s11104-017-3266-1.
[3]SHI Y A,XIE H R,CAO L X,et al.Effects of Cd and Pb-resistant endophytic fungi on growth and phytoextraction of Brassica napus in metal-contaminated soils[J].Environmental Science and Pollution Research International,2017,24(1):417.DOI:10.1007/s11356-016-7693-y.
[4]文才艺,吴元华,田秀玲.植物内生菌研究进展及其存在的问题[J].生态学杂志,2004,23(2):86.DOI:10.3321/j.issn:1000-4890.2004.02.019.
[5]周敏.内生真菌及其诱导子与长春花悬浮细胞生物碱合成代谢相关性研究[D].长沙:湖南农业大学,2005.
[6]NAMDEO A,PATIL S,FULZELE D P.Influence of fungal elicitors of production of ajmalicine by cell cultures of Catharanthus roseus[J].Biotechnology Progress,2002,18(1):159.DOI:10.1021/bp0101280.
[7]张朝阳,赵庭周.重楼资源再生策略及其关键技术环节探讨[J].中草药,2009,40(2):319.DOI:10.3321/j.issn:0253-2670.2009.02.049.
[8]国家药典委员会.中华人民共和国药典:2015年版:1部[M].北京:中国医药科技出版社,2015.
[9]NAMBU T,HUANG X,SHU Y,et al.Chronotropic effect of the methanolic extracts of the plants of the Paris species and steroidal glycosides isolated from P.vietnamensis on spontaneous beating of myocardial cells[J].Planta Medica,1989,55(6):501.DOI:10.1055/s-2006-962080.
[10]季申,周坛树,张锦哲.中药重楼和云南白药中抗肿瘤细胞毒活性物质Gracillin的测定[J].中成药,2001,23(3):212.DOI:10.3969/j.issn.1001-1528.2001.03.022.
[11]杨丽云,陈翠,吕丽芬,等.云南重楼的组织培养与植株再生[J].植物生理学通讯,2008,44(5):947.
[12]毛玉东,梁社往,何忠俊,等.土壤pH对滇重楼生长、养分含量和总皂甙含量的影响[J].西南农业学报,2011,24(3):985.DOI:10.3969/j.issn.1001-4829.2011.03.031.
[13]何忠俊,曾波,韦建荣,等.钙对滇重楼生长和总皂甙含量的影响[J].云南农业大学学报(自然科学),2010,25(5):664.DOI:10.3969/j.issn.1004-390X.2010.05.013.
[14]张晓洁,汤丽丽,王一丁.产甾体皂甙华重楼内生真菌、放线菌的分离与筛选[J].现代生物医学进展,2007,7(3):358.DOI:10.3969/j.issn.1673-6273.2007.03.013.
[15]王茜,申仕康,张爱丽,等.滇重楼内生真菌分离与多样性研究[J].中国中药杂志,2013,38(22):3838.DOI:10.4268/cjcmm20132209.
[16]GUO L D,HYDE K D,LIEW E C.Detection and taxonomic placement of endophytic fungi within frond tissues of Livistona chinensis based on rDNA sequences[J].Molecular Phylogenetics and Evolution,2001,20(1):1.DOI:10.1006/mpev.2001.0942.
[17]宣群,张才军,张玲琪,等.1株滇重楼内生真菌的研究-抑真菌活性及分离鉴定[J].昆明医学院学报,2008,29(6):41.DOI:10.3969/j.issn.1003-4706.2008.06.010.
[18]孙静贤,高玉红,杨海英,等.滇重楼内生真菌抗菌活性的筛选和菌株鉴定[J].广东农业科学,2013,40(9):74.
[19]刘舒,陈春黎,刘敏,等.两种内生真菌对大花蕙兰的共生效应比较[J].华中农业大学学报,2016,35(1):43.
[20]DIJK E.Effects of mycorrhizal fungi on in-vitro nitrogen response of juvenile orchids[J].Agriculture Ecosystems&Environment,1990,29(1/4):91.
[21]金有亮,杨福孙,宋希强,等.文心兰组培苗与野生兰科植物内生真菌共培养的生长效应研究[J].热带作物学报,2011,32(8):1483.DOI:10.3969/j.issn.1000-2561.2011.08.020.
[22]张烨,吕霜霜,周浓,等.不同生长年限滇重楼中4种重楼皂苷的含量比较[J].中国药房,2011,22(43):4081.
[23]AINHOAMARTINEZ-MEDINA A.Phytohormore profiles induced by Trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants[J].Journal of Chemical Ecology,2014,7(15):804.
[24]何秀丽,丁嫚嫚,叶方,等.重楼植物中内生菌研究进展[J].中国药师,2016,19(5):970.DOI:10.3969/j.issn.1008-049X.2016.05.047.
[25]严雪瑞.人参西洋参锈腐病菌比较生物学研究[D].沈阳:沈阳农业大学,2002.
[26]杨玲玲.滇重楼根茎细菌多样性与根瘤菌系统发育基因组学研究[D].昆明:云南大学,2016.