不同生长年限霍山石斛内生菌的多样性与差异性
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摘要
为探索霍山石斛内生菌的多样性与差异性,该文采用宏基因组学的方法对不同生长年限5组30份霍山石斛样品内生细菌及真菌的菌群进行了分析。研究结果显示,从霍山石斛中鉴定出3 540个细菌OTUs,5组样品共有OTUs为138个;鉴定出2 168个真菌OTUs,5组样品共有OTUs为143个。霍山石斛优势细菌菌群为Sphingomonas sp.,Acinetobacter sp.,Burkholderia sp.,Methylobacterium sp.,Enterococcus sp.,Bacillus sp.,差异菌群Oceanobacillusd sp.,Actinomycetospora sp.,Paenibacillus sp.,霍山石斛优势真菌群为Zasmidium sp.,Zymoseptoria sp.,Alternaria sp.,Cladosporium sp.,Fusarium sp.,差异菌群为Cyphellophore sp.,Fusarium sp.,聚类结果显示无论是内生细菌还是内生真菌,ⅢY2,ⅢY3均独立聚为一类,ⅡY1,ⅢY1也独立聚为一类。研究结果丰富了霍山石斛内生细菌和内生真菌的种类与资源,也为霍山石斛合理采收提供了理论参考。
In order to explore endophytes diversity and difference in Dendrobium huoshanense,in this paper,the metagenomics method was used to analyze the endophytic bacteria and fungi community of 5 groups include 30 samples in different growth years. The results indicate that 3 540 bacterial OTUs were identified from D. huoshanense,and there are 138 OTUs in 5 groups simultaneously;2 168 fungal OTUs were identified,and 143 OTUs exist in 5 groups simultaneously. The dominate endophytic bacteria community are Sphingomonas sp.,Acinetobacter sp.,Burkholderia sp.,Methylobacterium sp.,Enterococcus sp.,Bacillus sp.,the difference endophytic bacteria community are Oceanobacillusd sp.,Actinomycetospora sp.,Paenibacillus sp.. The dominate endophytic fungi community are Zasmidium sp.,Zymoseptoria sp.,Alternaria sp.,Cladosporium sp.,Fusarium sp.,the difference endophytic fungi community are Cyphellophore sp.,Fusarium sp.. The results of clustering revealed that both the endophytic bacteria and the endophytic fungi,ⅢY2 and ⅢY3 are complete clustered,and ⅡY1 and ⅢY1 are also cluster completely. These enriched the species and resources of endophytic bacteria and fungi in D. huoshanense,and provided a theoretical reference for the reasonable harvest of D. huoshanense.
In order to explore endophytes diversity and difference in Dendrobium huoshanense,in this paper,the metagenomics method was used to analyze the endophytic bacteria and fungi community of 5 groups include 30 samples in different growth years. The results indicate that 3 540 bacterial OTUs were identified from D. huoshanense,and there are 138 OTUs in 5 groups simultaneously;2 168 fungal OTUs were identified,and 143 OTUs exist in 5 groups simultaneously. The dominate endophytic bacteria community are Sphingomonas sp.,Acinetobacter sp.,Burkholderia sp.,Methylobacterium sp.,Enterococcus sp.,Bacillus sp.,the difference endophytic bacteria community are Oceanobacillusd sp.,Actinomycetospora sp.,Paenibacillus sp.. The dominate endophytic fungi community are Zasmidium sp.,Zymoseptoria sp.,Alternaria sp.,Cladosporium sp.,Fusarium sp.,the difference endophytic fungi community are Cyphellophore sp.,Fusarium sp.. The results of clustering revealed that both the endophytic bacteria and the endophytic fungi,ⅢY2 and ⅢY3 are complete clustered,and ⅡY1 and ⅢY1 are also cluster completely. These enriched the species and resources of endophytic bacteria and fungi in D. huoshanense,and provided a theoretical reference for the reasonable harvest of D. huoshanense.
引文
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[25]杨绍周,吴毅歆,邵德林,等.鼓槌石斛内生细菌分离、鉴定及功能分析[J].中国农学通报,2014,30(25):171.
[26]杨安明.金钗石斛内生菌的分离鉴定及活性物质发现研究[D].贵阳:贵州大学,2017.
[27]梁益敏,郁阳,麻兵继,等.一种人工栽培霍山石斛内生真菌的分离鉴定及其组织分布[J].安徽中医药大学学报,2015,34(6):75.
[28]闵长莉,闵运江,徐慧欣,等.霍山石斛内生真菌的分离及其及活性菌株的鉴定[J].天然产物研究与开发,2018,30(6):968.
[29]刘刚,俞年军,韩荣春,等.不同生长年限霍山石斛高效液相指纹图谱研究[J].中草药,2018,49(6):1424.
[2]王长锁,王凯,孟欣,等.霍山石斛对小鼠肝脏细胞色素P450酶表达和活性的影响[J].中国中药杂志,2018,43(21):4323.
[3]Lin J,Chang Y J,Yang W B,et al.The multifaceted effects of polysaccharides isolated from Dendrobium Huoshanense on immune functions with the induction of interleukin-1 receptor antagonist(IL-1ra)in monocytes[J].PLoS ONE,2014,9(4):e94040.
[4]徐云燕,王令仪,黄彬,等.不同生长期金钗石斛和铁皮石斛中总生物碱及多糖的比较[J].华西药学杂志,2014,29(3):288.
[5]胡克兴.石斛属药用植物内生真菌多样性研究[D].北京:中国协和医科大学,2008.
[6]郁阳.不同栽培环境下霍山石斛内生真菌生物多样性的研究[D].合肥:安徽中医药大学,2017.
[7]邹文欣,谭仁祥.植物内生菌研究新进展[J].植物学报,2001,43(9):881.
[8]Stierle A,Strobel G,Stierle D.Taxol and taxane production by Taxomyces andreanae,an endophytic fungus of Pacific yew[J].Science,1993,260(5105):214.
[9]Zhou Y,Sun W,Chen J,et al.SmMYC2a and SmMYC2b played similar but irreplaceable roles in regulating the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza[J].Sci Rep-UK,2016,6:22852.
[10]侯晓强,郭顺星.铁皮石斛促生长内生真菌的筛选与鉴定[J].中国中药杂志,2014,39(17):3232.
[11]朱波,刘京晶,斯金平,等.铁皮石斛内生真菌对宿主组培苗生长与代谢成分的影响[J].中国中药杂志,2016,41(9):1602.
[12]Xing X,Ma X,Men J,et al.Phylogenetic constrains on mycorrhizal specificity in eight Dendrobium(Orchidaceae)species[J].Sci China Life Sci,2017,60(5):536.
[13]Logares R,Sunagawa S,Salazar G,et al.Metagenomic 16Sr DNA Illumina tags are a powerful alternative to amplicon sequencing to explore diversity and structure of microbial communities[J].Environ Microbiol,2014,16(9):2659.
[14]Wang Y,Sheng H F,He Y,et al.Comparison of the levels of bacterial diversity in freshwater,intertidal wetland,and marine sediments by using millions of Illumina Tags[J].Apply Environ Microb,2012,78(23):8264.
[15]王营,李浩华,谭国慧,等.广藿香内生真菌类群分析及其抗菌活性研究[J].中国中药杂志,2017,42(4):657.
[16]Contreras M,Magris M,Hidalgo G,et al.Human gut microbiome viewed across age and geography[J].Nature,2012,486(7402):222.
[17]Bai Y,Daniel B Müller,Srinivas G,et al.Functional overlap of the Arabidopsis leaf and root microbiota[J].Nature,2015,528(7582):364.
[18]Zhang J,Zhang N,Liu Y X,et al.Root microbiota shift in rice correlates with resident time in the field and developmental stage[J].Sci Chin Life Sci,2018,61(6):1.
[19]Colemanderr D,Desgarennes D,Fonsecagarcia C,et al.Plant compartment and biogeography affect microbiome composition in cultivated and native Agave species[J].New Phytol,2016,209(2):798.
[20]Walters W A,Jin Z,Youngblut N,et al.Large-scale replicated field study of maize rhizosphere identifies heritable microbes[J].Proc Natl Acad Sci USA,2018,115(28):7368.
[21]Timsina B,Rokaya M B,MünzbergováZ,et al.Diversity,distribution and host-species associations of epiphytic orchids in Nepal[J].Biodivers Conserv,2016,25(13):1.
[22]Adhikari Y P,Fischer H S,Fischer A.Host tree utilization by epiphytic orchids in different land-use intensities in Kathmandu Valley,Nepal[J].Plant Ecol,2012,213(9):1393.
[23]董丹丹,汪志平.石斛属植物内生菌的研究进展[J].药物分析杂志,2012,32(3):550.
[24]严亮,杨瑞娟,王桥美.云南铁皮石斛内生菌的分离与鉴定[J].云南农业大学学报:自然科学版,2015,30(5):760.
[25]杨绍周,吴毅歆,邵德林,等.鼓槌石斛内生细菌分离、鉴定及功能分析[J].中国农学通报,2014,30(25):171.
[26]杨安明.金钗石斛内生菌的分离鉴定及活性物质发现研究[D].贵阳:贵州大学,2017.
[27]梁益敏,郁阳,麻兵继,等.一种人工栽培霍山石斛内生真菌的分离鉴定及其组织分布[J].安徽中医药大学学报,2015,34(6):75.
[28]闵长莉,闵运江,徐慧欣,等.霍山石斛内生真菌的分离及其及活性菌株的鉴定[J].天然产物研究与开发,2018,30(6):968.
[29]刘刚,俞年军,韩荣春,等.不同生长年限霍山石斛高效液相指纹图谱研究[J].中草药,2018,49(6):1424.