广西师范大学学报（自然科学版） ›› 2023, Vol. 41 ›› Issue (1): 155-163.doi: 10.16088/j.issn.1001-6600.2021102801

• 研究论文 • 上一篇    下一篇

柑橘内生真菌LJZ-Y-11次生代谢产物抑菌活性研究

李奇聪1,2,3,4, 陈洁萍1,2,3,4, 欧艳绍1,2,3,4, 杨树贤1,2,3,4, 邓志勇1,2,3,4, 骆海玉1,2,3,4*, 邓业成1,2,3,4*   

  1. 1.珍稀濒危动植物生态与环境保护教育部重点实验室(广西师范大学), 广西 桂林 541006;
    2.广西漓江流域景观资源保育与可持续利用重点实验室(广西师范大学), 广西 桂林 541006;
    3.广西珍稀濒危动物生态学重点实验室(广西师范大学), 广西 桂林 541006;
    4.广西师范大学可持续发展创新研究院, 广西 桂林 541006
  • 收稿日期:2021-10-28 修回日期:2021-12-04 出版日期:2023-01-25 发布日期:2023-03-07
  • 通讯作者: 骆海玉(1985—),女,广西桂林人,广西师范大学讲师,博士。E-mail:luohaiyu69@163.com; 邓业成(1965—),男,广西全州人,广西师范大学教授,博士。E-mail:dyecheng@163.com
  • 基金资助:
    广西自然科学基金(2018GXNSFAA281013); 广西重点研发计划(桂科AB21196065); 广西创新驱动发展专项(桂科AA20161002-1); 桂林市科学研究与技术开发计划(20190205); 广西师范大学珍稀濒危动植物生态与环境保护教育部重点实验室主任基金(ERESEP2021Z05)

Antimicrobial Activity of the Secondary Metabolites from Citrus Endophytic Fungus Nemania sp. LJZ-Y-11

LI Qicong1,2,3,4, CHEN Jieping1,2,3,4, OU Yanshao1,2,3,4, YANG Shuxian1,2,3,4, DENG Zhiyong1,2,3,4, LUO Haiyu1,2,3,4 *, DENG Yecheng1,2,3,4*   

  1. 1. Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Guangxi Normal University), Ministry of Education, Guilin Guangxi 541006, China;
    2. Guangxi Key Laboratory of Landscape Resources Conservation and Sustainable Utilization in Lijiang River Basin (Guangxi Normal University), Guilin Guangxi 541006, China;
    3. Guangxi Key Laboratory of Rare and Endangered Animal Ecology, Guilin Guangxi 541006, China;
    4. Institute for Sustainable Development and Innovation (Guangxi Normal University), Guilin Guangxi 541006, China
  • Received:2021-10-28 Revised:2021-12-04 Online:2023-01-25 Published:2023-03-07

PDF (PC)

72

摘要： 本文选取柑橘内生真菌Nemania sp. LJZ-Y-11进行发酵培养,提取制备其乙酸乙酯提取物并测定抑菌活性,旨在为其开发利用奠定基础。结果表明,其乙酸乙酯提取物对9种植物病原真菌和10种动物病原菌均具有显著抑制作用。处理浓度为2.5 g/L时,对9种植物病原真菌的抑制活性均超过50%,除对贡柑链格孢菌的抑制率为56.52%外,对其余8种病原菌的抑菌率均为100%,EC50值在0.004 8~0.154 4 g/L。其中,对玉米大斑病菌、金橘砂皮病菌和烟草黑胫病菌抑菌活性最为显著,EC50值分别为0.004 8、0.005 3和0.006 7 g/L,均小于0.01 g/L;其次为辣椒炭疽病菌、茶轮斑病菌和甘蔗凤梨病菌,EC50值分别为0.022 7、0.035 6和0.067 9 g/L,均小于0.1 g/L;对甘蓝黑斑病菌和水稻胡麻叶斑病菌也有较好的抑制活性,EC50值分别为0.101 3和0.154 4 g/L。对10种动物病原菌均具有不同程度的抑制效果,MIC值为0.078 1~2.500 0 g/L。其中,对枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、蜡样芽孢杆菌、炭疽杆菌、白色念珠菌具有较强的抑菌活性,MIC值均小于0.2 g/L,其中对枯草芽孢杆菌的抑菌活性最强,MIC值为0.078 1 g/L;对巨大芽孢杆菌和溶壁微球菌也有较好的抑菌活性,MIC值均为0.625 g/L;对产气肠杆菌和伤寒沙门氏菌的抑制活性相对较弱,MIC值仅为2.5 g/L。本文报道的柑橘内生真菌Nemania sp. LJZ-Y-11代谢产物具有广谱、显著抑菌活性,研究结果可为其次生代谢产物作为生物源抗菌剂开发利用提供依据。

关键词: 柑橘内生真菌, Nemania sp., 乙酸乙酯提取物, 抑菌活性

Abstract: Based on previous studies, citrus endophytic fungus Nemania sp. LJZ-Y-11 was selected for fermentation, extract preparation and evaluation of its antimicrobial activity, aiming to lay a foundation for the development and utilization of its metabolites. The antifungal and antibacterial activities of the ethyl acetate extract were determined by using a mycelium growth rate method and toxic medium coating method, respectively. The results showed that the ethyl acetate extract of LJZ-Y-11 exhibited an excellent antifungal activity against most of tested plant pathogenic fungi, and had an antibacterial activity against 10 animal pathogens to some extent. The inhibitory rate against 9 plant pathogenic fungi were all more than 50% at 2.5 g/L of the extract. Moreover, except that the inhibitory rate against Alternaria citri was 56.52%, the others were 100%. The toxicity of extract against 8 plant pathogenic fungi was determined furtherly, with EC50 values ranging from 0.004 8 g/L to 0.154 4 g/L. There was the highest inhibitory activity against Exserohilum turcicum, Diaporthe citri and Phytophthora parasitica var. nicotianae with EC50 values of 0.004 8, 0.005 3 and 0.006 7 g/L, respectively, less than 0.01 g/L, following by Colletotrichum capsici, Pestalotiopsis theae and Ceratocystis paradoxa with EC50 values of 0.022 7, 0.035 6 and 0.067 9 g/L, respectively, less than 0.1 g/L; and also had a significant inhibitory activity against Alternaria oleracea and Cochliobolus miyabeanus with EC50 values of 0.101 3 g/L and 0.154 4 g/L, respectively. In addition, the extract also showed significant antibacterial activities against 10 animal pathogens with MIC values of 0.078 1-2.5 g/L. There were high inhibitory activities against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Bacillus cereus, Bacillus anthraci and Monilia albican with MIC values of less than 0.2 g/L, and had the highest inhibitory activity against Bacillus subtilis with a MIC value 0.078 1 g/L. There were also strong inhibitory effects against Bacillus megaterium and Micrococcus lysodeikticus, with a MIC value of 0.625 g/L. The inhibition activity against Enterobacter aerogenes and Salmonella typhi was relatively weaker, with MIC values of 2.5 g/L. The results in this study reveal that the secondary metabolites of LJZ-Y-11 has broad-spectrum and significant antimicrobial activity against the common pathogens, providing a useful experiment basis for the development and utilization of the secondary metabolites as biological antimicrobial agents.

Key words: citrus endophytic fungi, Nemania sp. ethyl acetate extract, antimicrobial activity

中图分类号: 

  • TQ455
[1] HUMEERA N,AZRA N K,IRSHAD A N,et al. Fungal endophytes as prolific source of phytochemicals and other bioactive natural products:a review[J]. Microbial. Pathogenesis,2015,82:50-59. DOI:10.1016/j.micpath.2015.04.001.
[2]ANDRS M F,DIAZ C E,GIMENZ C,et al. Endophytic fungi as novel sources of biopesticides:the Macaronesian Laurel forest,a case study[J]. Phytochemistry Reviews,2017,16(5):1009-1022. DOI:10.1007/s11101-017-9514-4.
[3]SAIKKONEN K,FAETH S H,HELANDER M,et al. FUNGAL ENDOPHYTES:a continuum of interactions with host plants[J]. Annual Review of Ecology and Systematics,1998,29:319-343. DOI:10.1146/ANNUREV.ECOLSYS.29.1.319.
[4]LARRIBA E,JAIME M D L A,NISLOW C,et al. Endophytic colonization of barley(Hordeum vulgare) roots by the nematophagous fungus Pochonia chlamydosporia reveals plant growth promotion and a general defense and stress transcriptomic response[J]. Journal of Plant Research, 2015,128(4):665-678. DOI:10.1007/s10265-015-0731-x.
[5]SINGH A,SINGH D K,KHARWAR R N,et al. Fungal endophytes as efficient sources of plant-derived bioactive compounds and their prospective applications in natural product drug discovery:insights,avenues,and challenges[J]. Microorganisms,2021,9(1):197. DOI:10.3390/microorganisms9010197.
[6]WU H Y,YAN Z L,DENG Y C,et al. Endophytic fungi from the root tubers of medicinal plant Stephania dielsiana and their antimicrobial activity[J]. Acta Ecologica Sinica,2020,40(5):383-387. DOI:10.1016/j.chnaes.2020.02.008.
[7]SAAD M M G,GHAREEB R Y,SAEED A A. The potential of endophytic fungi as bio-control agents against the cotton leafworm,Spodoptera littoralis(Boisd.)(Lepidoptera:Noctuidae)[J]. Egyptian Journal of Biological Pest Control,2019,29(1):7. DOI:10.1186/s41938-019-0108-x.
[8]RATNAWEERA P B,JAYASUNDARA J M N,HERATH H H M S D,et al. Antifeedant,contact toxicity and oviposition deterrent effects of phyllostine acetate and phyllostine isolated from the endophytic fungus Diaporthe miriciae against Plutella xylostella larvae[J]. Pest Management Science,2020,76(4):1541-1548. DOI:10.1002/ps.5673.
[9]AMEEN F,STEPHENSON S L,ALNADHARI S,et al. Isolation,identification and bioactivity analysis of an endophytic fungus isolated from Aloe vera collected from Asir desert,Saudi Arabia[J]. Bioprocess and Biosystems Engineering,2021,44(6):1063-1070. DOI:10.1007/s00449-020-02507-1.
[10]甘琪,徐旭,张晓敏,等. 红树林植物内生真菌Stachybotrys chartarum HDN16-358次级代谢产物研究[J]. 中国海洋药物,2019,38(4):48-52. DOI:10.13400/j.cnki.cjmd.2019.04.007.
[11]黄燕,曾东强,唐文伟,等. 植物内生真菌生物活性成分研究进展[J]. 天然产物研究与开发,2020,32(8):1419-1437. DOI:10.16333/j.1001-6880.2020.8.019.
[12]崔迎,张志强,赵昕,等. 产西贝碱内生真菌的复壮[J]. 菌物研究,2010,8(1):41-43. DOI:10.13341/j.jfr.2010.01.002.
[13]VENUGOPALAN A,SRIVASTAVA Jr S. Endophytes as in vitro production platforms of high value plant secondary metabolites[J]. Biotechnology Advances,2015,33(6,Part 1):873-887. DOI:10.1016/j.biotechadv.2015.07.004.
[14]ARAU＇JO W L,MACCHERONI W Jr,AGUILAR-VILDOSO C I,et al. Variability and interactions between endophytic bacteria and fungi isolated from leaf tissues of citrus rootstocks[J]. Canadian Journal of Microbiology,2001,47(3):229-236. DOI:10.1139/w00-146.
[15]BAAYEN R P,BONANTS P J M,VERKLEY G,et al. Nonpathogenic isolates of the citrus black spot fungus,Guignardia citricarpa,identified as a cosmopolitan endophyte of woody plants,G. mangiferae (Phyllosticta capitalensis)[J]. Phytopathology,2002,92(5):464-477. DOI:10.1094/PHYTO.2002.92.5.464.
[16]GLIENKE-BLANCO C,AGUILAR-VILDOSO C I,VIEIRA M L C,et al. Genetic variability in the endophytic fungus Guignardia citricarpa isolated from citrus plants[J]. Genetics and Molecular Biology,2002,25(2):251-255. DOI:10.1590/S1415-47572002000200021.
[17]RODRIGUES K F,SIEBER T N,GRNIG C R,et al. Characterization of Guignardia mangiferae isolated from tropical plants based on morphology,ISSR-PCR amplifications and ITS1-5.8S-ITS2 sequences[J]. Mycological Research,2004,108(1):45-52. DOI:10.1017/S0953756203008840.
[18]DURN E L,PLOPER L D,RAMALLO J C,et al. The foliar fungal endophytes of Citrus limon in Argentina[J]. Canadian Journal of Botany,2005,83(4):350-355. DOI:10.1139/b05-009.
[19]罗永兰,张志元,冉国华. 柑橘内生真菌的分离与鉴定[J]. 湖南农业大学学报(自然科学版),2005,31(4):418-421.DOI:10.3321/j.issn:1007-1032.2005.04.019.
[20]赵昌会,黄芳,丁芳. 柑橘内生真菌的分离及抗菌活性的初步研究[J]. 湖南科技学院学报,2008,29(12):72-73.
[21]SUN Y L,HAN S H,PARK Y C,et al. Identification and taxonomy of the endophytic fungi and potential pathogens in Citrus plants[J]. Journal of Pure and Applied Microbiology,2013,7:611-619.
[22]HUANG F,UDAYANGA D,WANG X H,et al. Endophytic Diaporthe associated with Citrus:a phylogenetic reassessment with seven new species from China[J]. Fungal Biology,2015,119(5):331-347. DOI:10.1016/j.funbio.2015.02.006.
[23]DOUANLA-MELI C,UNGER J G. Phylogenetic study of the Colletotrichum species on imported citrus fruits uncovers a low diversity and a new species in the Colletotrichum gigasporum complex[J]. Fungal Biology,2017,121(10):858-868. DOI:10.1016/j.funbio.2017.06.003.
[24]GUARNACCIA V,GROENEWALD J Z,LI H,et al. First report of Phyllosticta citricarpa and description of two new species,P. paracapitalensis and P. paracitricarpa,from citrus in Europe[J]. Studies in Mycology,2017,87:161-185. DOI:10.1016/j.simyco.2017.05.003.
[25]PENA L C,JUNG L F,SAVI D C,et al. A Muscodor strain isolated from Citrus sinensis and its production of volatile organic compounds inhibiting Phyllosticta citricarpa growth[J]. Journal of Plant Diseases and Protection,2017,124(4):349-360. DOI:10.1007/s41348-016-0065-5.
[26]WACULICZ-ANDRADE C E,SAVI D C,BINI A P,et al. Colletotrichum gloeosporioides sensu stricto:an endophytic species or citrus pathogen in Brazil?[J]. Australasian Plant Pathology,2017,46(2):191-203. DOI:10.1007/s13313-017-0476-1.
[27]郭东升,原晨虹,翟颖妍,等. 柑橘内生燕麦镰刀菌Gds-1对柑橘青霉病的防治研究[J]. 农药学学报,2020,22(5):782-790. DOI:10.16801/j.issn.1008-7303.2020.0085.
[28]颜桢灵,陈洁萍,农小霞,等. 柑橘内生真菌的分离鉴定及其发酵产物对柑橘溃疡病菌的抑制活性[J]. 广西植物,2021,41(7):1196-1208. DOI:10.11931/guihaia.gxzw202002021.
[29]LUO H Y,ZHOU Q Y,DENG Y C,et al. Antifungal activity of the extract and the active substances of endophytic Nigrospora sp. from the traditional Chinese medicinal plant Stephania kwangsiensis[J]. Natural Product Communications,2017,12(12):1889-1892. DOI:10.1177/1934578X1701201219.
[30]DENG Y C, YU Y Z, LUO H Y, et al. Antimicrobial activity of extract and two alkaloids from traditional Chinese medicinal plant Stephania dielsiana[J]. Food Chemistry, 2011, 124(4): 1556-1560. DOI:10.1016/j.foodchem.2010.08.011.
[31]周秋艳,卿朕,骆海玉,等. 两株广西地不容内生真菌的抑菌活性研究[J]. 广东农业科学,2016,43(4):111-116. DOI:10.16768/j.issn.1004-874X.2016.04.022.
[32]林伟,骆海玉,邓业成,等. 血散薯块根内生真菌Periconia igniaria Stdif10发酵产物的生物活性研究[J]. 河南农业科学,2020,49(1):75-81. DOI:10.15933/j.cnki.1004-3268.2020.01.010.
[33]王慧芳,程叶新,白金虎,等. 艾灰黄酮超声提取工艺的优化及其抑菌活性[J]. 中成药,2021,43(9):2286-2292. DOI:10.3969/j.issn.1001-1528.2021.09.002.
[34]吴永祥,程满怀,江海涛,等. 白及萃取物的抑菌活性及其二氯甲烷萃取物化学成分分析[J]. 食品与机械,2017,33(12):76-79. DOI:10.13652/j.issn.1003-5788.2017.12.015.
[35]李雯霞,于司达,诸葛斌,等. 芍药花提取物对果蔬腐败菌的抑菌活性及其在樱桃番茄保鲜中的应用[J]. 食品与发酵工业,2018,44(2):228-232. DOI:10.13995/j.cnki.11-1802/ts.015384.
[36]赵国栋. 凤丹种荚提取物抗菌活性研究[J]. 现代农业科技,2019(13):200-201. DOI:10.3969/j.issn.1007-5739.2019.13.111.
[37]陆春雷,赵雨菡,姜金源,等. 樟芝深层发酵液萃取物对食源性致病菌的抑制效果[J]. 中国食用菌,2021,40(6):69-73,79. DOI:10.13629/j.cnki.53-1054.2021.06.014.
[38]李欣,乔家驹,冯汉青,等. 杜仲雄花乙酸乙酯提取物的抑菌活性研究[J]. 食品工业科技,2015,36(11):62-65. DOI:10.13386/j.issn1002-0306.2015.11.004.
[39]邵玲莉,钱思,施晓秦,等. 无患子皂苷的提取及抑菌性研究[J]. 安徽医药,2021,25(10):1943-1945. DOI:10.3969/j.issn.1009-6469.2021.10.008.
[40]陈炳智,杨立志,郑丽珠,等. 长裙竹荪抑菌活性物质的分离纯化及其抑菌效果[J]. 菌物学报,2020,39(8):1568-1579. DOI:10.13346/j.mycosystema.200017.
[41]KORNSAKULKARN J, SAEPUA S, SUVANNAKAD R,et al. Cytotoxic tropolones from the fungus Nemania sp. BCC 30850[J]. Tetrahedron,2017,73(25):3505-3512.DOI:10.1016/j.tet.2017.05.030.
[42] IBRAHIM A, SRENSEN D, JENKINS H A, et al. Epoxynemanione A, nemanifuranones A-F, and nemanilactones A-C, from Nemania serpens, an endophytic fungus isolated from Riesling grapevines[J]. Phytochemistry, 2017, 140: 16-26. DOI: 10.1016/j.phytochem.2017.04.009.
[1] 杜丽波, 李金玉, 张晓, 李永红, 潘卫东. 毛红椿皮的化学成分及生物活性研究[J]. 广西师范大学学报（自然科学版）, 2022, 40(4): 162-172.
[2] 刘汉甫, 马献力, 陈素一, 沈雪松, 黄富平. 锌三元配合物的合成及其抑菌活性研究[J]. 广西师范大学学报（自然科学版）, 2015, 33(4): 103-107.
[3] 邓业成, 宁蕾. 4种植物精油的抑菌活性及应用[J]. 广西师范大学学报（自然科学版）, 2012, 30(3): 288-294.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 卢其西, 林勇, 宾石玉, 唐章生, 彭婷. 罗非鱼6个家系的低温耐寒测定分析[J]. 广西师范大学学报（自然科学版）, 2011, 29(2): 104 -109 .
[2] 程瑞, 何明先, 钟春英, 罗树毅, 武正军. 野生与人工繁育鳄蜥游泳能力比较[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 79 -86 .
[3] 陈丹妮, 陈志林, 周善义. 中国蚁科昆虫名录——切叶蚁亚科(补遗)[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 87 -97 .
[4] 李爽, 刘上丽, 裴思玉, 唐绍清. 基于单拷贝核基因PAL研究贵州金花茶遗传多样性[J]. 广西师范大学学报（自然科学版）, 2021, 39(1): 128 -135 .
[5] 姚金洋, 胡颖, 王金华. 广义Howell设计GHD(n+5,3n)的存在性[J]. 广西师范大学学报（自然科学版）, 2021, 39(6): 119 -129 .
[6] 付美子, 林炳清. 临床测量中定量数据Bland-Altman一致性评价[J]. 广西师范大学学报（自然科学版）, 2022, 40(1): 125 -138 .
[7] 孙岩松, 杨亮, 林鸿飞. 基于多粒度的分词消歧和语义增强的情景剧幽默识别[J]. 广西师范大学学报（自然科学版）, 2022, 40(3): 57 -65 .
[8] 郝雅茹, 董力, 许可, 李先贤. 预训练语言模型的可解释性研究进展[J]. 广西师范大学学报（自然科学版）, 2022, 40(5): 59 -71 .
[9] 王喜敏, 袁杰, 寇巧媛. 一种基于多策略的改进黏菌算法[J]. 广西师范大学学报（自然科学版）, 2022, 40(6): 98 -108 .
[10] 张春梅, 闫芳, 宋海, 张喜峰, 陈叶. 祁连山黄参叶片转录组测序及生物信息学分析[J]. 广西师范大学学报（自然科学版）, 2022, 40(6): 247 -256 .
版权所有 © 广西师范大学学报（自然科学版）编辑部
地址：广西桂林市三里店育才路15号　邮编：541004
电话：0773-5857325　E-mail: gxsdzkb@mailbox.gxnu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发