叶底珠抑菌活性内生真菌的筛选及鉴定
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摘要
内生真菌存在于植物中,代谢产物十分丰富,具有多种生物活性,是新化合物、新药源的潜在资源。叶底珠通常根部入药,用于治疗风湿腰痛、四肢麻木和小儿疳积等疾病,对神经衰弱、急性肝损伤等也有一定的疗效。本文主要从叶底珠中分离筛选具有抑菌活性的内生真菌,并对其进行鉴定。采用平板培养法从叶底珠中分离内生真菌,通过滤纸片扩散法考察这些内生真菌的抑菌活性。对具有良好抑菌效果的内生真菌进行培养观察和ITS序列测定,鉴定其归属。从健康植株中分离到41株内生真菌,其中2个菌株的提取液对白色念珠菌的抑制效果最好,并发现其对另外7种病原菌具有不同程度的抑制作用。通过鉴定,确定菌株G11为球毛壳菌(Chaetomium globosum),菌株D11为枝细枝孢菌(Cladosporium ramotenellum)。从叶底珠中分离到2株具有较好抑菌作用的内生真菌,这些具有抑菌作用的内生真菌可用于生物制药和农业产业,值得进一步研究。
Endophytic fungi with antibacterial activity were isolated,screened from Securinega suffruticosa,and identified.These fungi were isolated from S.suffruticosa by plate culture method,and their antibacterial activity was examined by filter paper diffusion.The endophytic fungi with good antimicrobial activity were sequenced by ITS,and their corresponding genera were identified.A total of 41 endophytic fungi were isolated from healthy plants.Among these isolates,the extracts of 2 strains had the best inhibition effect on Candida albicans,and they had different inhibitory effects against the other seven pathogens.Strain G11 was identified as Chaetomium globosum,whereas strain D11 was identified as Cladosporium ramotenellum.Two endophytic fungi with good antimicrobial activity were isolated and screened from S.suffruticosa.These endophytic fungi can be used in biopharmaceutical and agricultural industries and merit further study.
Endophytic fungi with antibacterial activity were isolated,screened from Securinega suffruticosa,and identified.These fungi were isolated from S.suffruticosa by plate culture method,and their antibacterial activity was examined by filter paper diffusion.The endophytic fungi with good antimicrobial activity were sequenced by ITS,and their corresponding genera were identified.A total of 41 endophytic fungi were isolated from healthy plants.Among these isolates,the extracts of 2 strains had the best inhibition effect on Candida albicans,and they had different inhibitory effects against the other seven pathogens.Strain G11 was identified as Chaetomium globosum,whereas strain D11 was identified as Cladosporium ramotenellum.Two endophytic fungi with good antimicrobial activity were isolated and screened from S.suffruticosa.These endophytic fungi can be used in biopharmaceutical and agricultural industries and merit further study.
引文
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[32] 崔岩,王佩,孔凡栋,等.角果木内生真菌Cladosporium cladosporioides JG-12 次生代谢产物及其生物活性[J]. 热带生物学报,2017,8(1):29-36.
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[34] 王毅,王娟,王四海,等.蒜头果的内生真菌多样性分析[J].基因组学与应用生物学,2017(9):1-14.
[2] 吕海花,李雪晴,王冉冉,等.叶底珠根皮的生药学研究[J].山东中医杂志,2015,34(6):463-464.
[3] 孙亮.叶底珠化学成分及抗氧化活性的研究[D].延边:延边大学,2012:1-45.
[4] 刘毅,岳志华,张娜,等.一叶萩碱的研究进展[J].中国药事,2009,23(8):817-818,828.
[5] Qin S,Liang J Y,Gu Y C,et al.Suffruticosine,a novel octacyclic alkaloid with an unprecedented skeleton from Securinega suffruticosa (Pall.) Rehd[J]. Tetrahedron Letters,2008,49:7066-7069.
[6] 张刚,陈冬云,程静,等.左旋一叶萩碱诱导人非小细胞肺癌A549细胞自噬机制的研究[J].中国临床药理学与治疗学,2014,19(8):851-855.
[7] 贲亮,郭伟康,袁红艳,等.叶底珠枝叶水提物对小鼠急性肝损伤的保护作用[J].四川中医,2010,28(2):25-26.
[8] Zhang J,Yao J,Peng S,et al.Securinine disturbs redox homeostasis and elicits oxidative stress-mediated apoptosis via targeting thioredoxin reductase[J].Biochim Biophys Acta,2017,1863(1):129-138.
[9] Han S,Yang X,Pan Y,et al.L-securinine inhibits the proliferation of A549 lung cancer cells and promotes DKK1 promoter methylation[J].Oncol Lett,2017,14(4):4243-4248.
[10] Leonoudakis D,Rane A,Angeli S,et al.Anti-inflammatory and neuroprotective role of natural product securinine in activated glial cells:implications for Parkinson's Disease[J].Mediators Inflamm,2017,2017:8302636.
[11] Jia M,Chen L,Xin H L,et al.A friendly relationship between endophytic fungi and medicinal plants:a systematic review[J].Front Microbiol,2016,7:906.
[12] Khan A L,Hussain J,AI-Harrasi A,et al.Endophytic fungi:resource for gibberellins and crop abiotic stress resistance[J].Critical Reviews in Biotechnology,2013,35(1):62-74.
[13] Ibrahim SRM,Mohamed G A,Ross S A.Integracides F and G:new tetracyclic triterpenoids from the endophytic fungus Fusarium sp[J].Phytochemistry Letters,2016,15:125-130.
[14] Akone S H,Mándi A,Kurtán T,et al.Inducing secondary metabolite production by the endophytic fungus Chaetomium sp[J].through fungal-bacterial co-culture and epigenetic modification.Tetrahedron,2016,72(41):6340-6347.
[15] 李艳玲,王德才,史仁玖,等.泰山黄精内生真菌的分离鉴定及抑菌活性研究[J].中草药,2013,44(11):1490-1494.
[16] 汪友明,樊美珍,李增智.安徽黄山红豆杉内生真菌多样性与生态分布[J].菌物学报,2016,35(3):360-363.
[17] 刘宏玉,金慧清,王佳莹,等.烟草内生真菌多样性和种群结构[J].菌物学报,2015,34(6):1058‐1067.
[18] 刘雷,潘峰,杨远兵,等.川麦冬内生真菌分离和鉴定及抑菌活性初步研究[J].中草药,2016,47(8):1382-1391.
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[20] Herrera J,Khidir H H,Eudy D M,et al.Shifting fungal endophyte communities colonize Bouteloua gracilis:effect of host tissue and geographical distribution[J].Mycologia,2010,102(5):1012-1026.
[21] 李冬霞,武海燕,张猛,等.枣树内生真菌的分离、鉴定及其多样性分析[J].果树学报,2010,27(6):975-979.
[22] 徐全智,孙牧笛,李帆,等.宁夏枸杞内生真菌的分离及多样性分析[J].北方园艺,2017,(10):103-109.
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[25] 胡美玲,郑勇,孙翔,等.内生真菌促进玉米幼苗的抗旱性研究[J].菌物学报,2017,36(9):1-10.
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[27] 魏中琦.两株共生菌的次生代谢产物研究[D]. 扬州:扬州大学,2014:1-2.
[28] 张文豪,国政,魏少鹏,等.侧柏内生真菌Chaetomium globosum ZH-32 抗菌活性成分研究[J].农药学学报,2014,16(5):605-609.
[29] Li W,Yang X,Yang Y,et al.Anti-phytopathogen,multi-target acetylcholinesterase inhibitory and antioxidant activities of metabolites from endophytic Chaetomium globosum[J].Nat Prod Res,2016,8:1-4.
[30] 姚裕群.越南槐内生真菌多样性、抑菌活性及其次生代谢产物研究[D].南宁:广西大学,2017:94-96.
[31] 张昌飞.云南五种花卉植物内生真菌多样性及其对植物生长的影响研究[D]. 昆明:昆明理工大学,2013:48-49.
[32] 崔岩,王佩,孔凡栋,等.角果木内生真菌Cladosporium cladosporioides JG-12 次生代谢产物及其生物活性[J]. 热带生物学报,2017,8(1):29-36.
[33] 游玲,林娜,郭华,等.山苍子内生真菌多样性研究[J]. 西南师范大学学报(自然科学版),2013,38(1):121-126.
[34] 王毅,王娟,王四海,等.蒜头果的内生真菌多样性分析[J].基因组学与应用生物学,2017(9):1-14.