[1]倪维鞠,张放,王瑞,等.蓝莓内生真菌BB01抗菌活性的研究[J].北方园艺,2021,(07):26-32.[doi:10.11937/bfyy.20202436]
　NI Weiju,ZHANG Fang,WANG Rui,et al.Antimicrobial Activity of Endophytic Fungi BB01 Derived From Blueberry[J].Northern Horticulture,2021,(07):26-32.[doi:10.11937/bfyy.20202436]
点击复制

蓝莓内生真菌BB01抗菌活性的研究

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 期数: 2021年07 页码: 26-32 栏目: 出版日期: 2021-04-15
Title:
Antimicrobial Activity of Endophytic Fungi BB01 Derived From Blueberry
作者:
倪维鞠; 张放; 王瑞; 雷霁卿
（贵阳学院 食品与制药工程学院，贵州 贵阳 550005）
Author(s):
NI Weiju; ZHANG Fang; WANG Rui; LEI Jiqing
(College of Food and Pharmaceutical Engineering，Guiyang University，Guiyang，Guizhou 550005)
关键词:
蓝莓; 内生真菌; 抗菌活性; 次级代谢产物
Keywords:
blueberry; endophytic fungi; antibacterial activity; secondary metabolism
DOI:
10.11937/bfyy.20202436
文献标志码:
A
摘要:
以贵州省麻江县蓝莓种植基地的“粉蓝”蓝莓为试材，通过分离纯化、生物学鉴定、对峙试验、菌丝生长试验及刃天青微量平板稀释试验等方法，研究了蓝莓内生真菌BB01的抗菌活性，以期为开发绿色生物保鲜剂提供参考依据。结果表明：蓝莓健康组织（茎、叶、果实）中分离纯化出41株内生真菌，其中BB01、BL22、BF27、BB01及BF41可抑制全部4种病害菌生长；发酵培养BB01获得肉汤乙酸乙酯粗提取物（broth ethyl acetate extract，BE）、菌丝体正己烷萃取物（cell hexane，CH）和菌丝体乙酸乙酯萃取物（cell ethyl acetate,CE）。提取物CH对青霉孢子抑制效果最好，最小抑菌浓度为64 μg?mL-1；提取物BE对枝孢菌、尖小丛壳菌及葡萄孢菌孢子抑制效果最好，最小抑菌浓度分别为16、256、128 μg?mL-1，同时，提取物BE终浓度500 μg?mL-1时，可有效抑制4种病害菌菌丝生长（抑菌率分别为98.067 302%±3.813 437%、92.187 892%±0.828 473%、94.251 883%±0.368 539%、96.909 029%±5.769 104%）。采用PCR扩增内生真菌基因组rDNA-ITS序列构建系统发育树鉴定BB01为枝孢菌属。
Abstract:
‘Pink blue’ blueberry collected from Majiang cultivation base in Guizhou was used as the test material.The isolation and purification experiment,biological identification,confrontation test ,mycelial growth test and resazurin microtitre plate antibacterial assay were performed to study the antibacterial activity of blueberry endophytic fungus BB01,in order to provide a research foundation for the development of green biological preservatives.The results showed that 41 strains endophytic fungi were isolated from the healthy tissue (stem,leaf,and fruit),among which BB01,BL22,BF27,BB01 and BF41 could inhibit the growth of all 4 pathogens.Endophytic fungus BB01 was undergone the process of liquid fermentation to obtain broth ethyl acetate extract (BE),it was found that CE exhibited the best inhibition against Penicillium,with minimum inhibitory concentrations (MIC) values of 64 μg?mL-1 ,and BE exhibited the best inhibition against Cladosporium，Glomerella acutata and Botrytis cinerea,with MIC values of 16 μg?mL-1，256 μg?mL-1 and 128 μg?mL-1,respectively.Moreover BE also exhibited 98.067 302%±3.813 437%,92.187 892%±0.828 473%,94.251 883%±0.368 539%,96.909 029%±5.769 104% growth inhibiton against 4 pathogens respectively at 500 μg?mL-1.PCR was used to amplify the genome rDNA-ITS sequence for phylogenetic tree.The results showed that BB01 was Cladosporium sp.

参考文献/References:

[1]苑兆和.世界蓝莓生产历史与发展趋势[J].落叶果树，2003，35(1)：49-52.[2]胡雅馨，李京，惠伯棣.蓝莓果实中主要营养及花青素成分的研究[J].食品科学，2006，27(10)：600-603.[3]任春光，杨丰，黄振兴，等.贵州省蓝莓主栽区真菌病害调查初报[J].中国森林病虫，2019，37(2)：22-27.[4]RODRIGUEZ R J，WHITE JR J F，ARNOLD A E，et al.Fungal endophytes：Diversity and functional roles[J].New Phytologist，2009，182(2)：314-330.[5]MALHADAS C，MALHEIRO R，PEREIRA J A，et al.Antimicrobial activity of endophytic fungi from olive tree leaves[J].World Journal of Microbiology and Biotechnology，2017，33(3)：46.[6]BADOTTI F，FONSECA P L C，TOM L M R，et al.ITS and secondary biomarkers in fungi：Review on the evolution of their use based on scientific publications[J].Brazilian Journal of Botany，2018，41(2)：471-479.[7]雷霁卿，吴文能，刘颖，等.贵州六盘水地区 “红阳” 猕猴桃软腐病病原菌分离鉴定及致病力差异测定[J].北方园艺，2019(4)：31-38.[8]张燕，张阳，张博，等.生防假单胞菌 2P24 不同碳源对抗生素 2，4-二乙酰基间苯三酚的影响[J].中国植物病理学会 2018 年学术年会论文集，2018.[9]SANTOS I P，SILVA L C N，SILVA M V，et al.Antibacterial activity of endophytic fungi from leaves of Indigofera suffruticosa Miller (Fabaceae)[J].Frontiers in Microbiology，2015(6)：350.[10]BALOUIRI M，SADIKI M，IBNSOUDA S K.Methods for in vitro evaluating antimicrobial activity：A review[J].Journal of Pharmaceutical Analysis，2016，6(2)：71-79.[11]CAMPANILE G，RUSCELLI A，LUISI N.Antagonistic activity of endophytic fungi towards Diplodia corticola assessed by in vitro and in planta tests[J].European Journal of Plant Pathology，2007，117(3)：237-246.[12]SARKER S D，NAHAR L，KUMARASAMY Y.Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth，and its application in the in vitro antibacterial screening of phytochemicals[J].Methods，2007，42(4):321-324.[13]SCHULZ B，BOYLE C，DRAEGER S，et al.Endophytic fungi：A source of novel biologically active secondary metabolites[J].Mycological Research，2002，106(9)：996-1004.[14]HYDE K D，SOYTONG K.The fungal endophyte dilemma[J].Fungal Divers，2008(33):163-173.[15]RODRIGUEZ R J，WHITE JR J F，ARNOLD A E，et al.Fungal endophytes：Diversity and functional roles[J].New Phytologist，2009，182(2)：314-330.[16]JI L L，SONG Y C，TAN R X.A potent feed preservative candidate produced by Calcarisporium sp.，an endophyte residing in stargrass (Cynodon dactylon)[J].Journal of Applied Microbiology，2004，96(2)：352-358.[17]YIN C，LIU H，SHAN Y，et al.Cytosporone B as a biological preservative：Purification，fungicidal activity and mechanism of action against Geotrichum citri-aurantii[J].Biomolecules，2019，9(4)：125.[18]SCOTT P M，WALBEEK W V，MACLEAN W M.Cladosporin，a new antifungal metabolite from cladosporium cladosporioides[J].The Journal of Antibiotics，1971，24(11):747-755.[19]BALAJI D S，BASAVARAJA S，DESHPANDE R，et al.Extracellular biosynthesis of functionalized silver nanoparticles by strains of Cladosporium cladosporioides fungus[J].Colloids & Surfaces B:Biointerfaces，2009，68(1):88-92.[20]PULIT J，BANACH M，SZCZYGOWSKA R，et al.Nanosilver against fungi.Silver nanoparticles as an effective biocidal factor[J].Acta Biochimica Polonica，2013，60(4):795-798.[21]WANG X，RADWAN M M，TARWNEH，AMER H，et al.Antifungal activity against plant pathogens of metabolites from the endophytic fungus cladosporium cladosporioides[J].Journal of Agricultural & Food Chemistry，2013，61(19):4551-4555.[22]HAN X，BAO X F，WANG C X，et al.Cladosporine A，a new indole diterpenoid alkaloid with antimicrobial activities from Cladosporium sp.[J].Natural Product Research，2019(12):1-7.[23]LINDER D，SCHOBERT R.Synthesis of the fungus metabolite cladosin C[J].Org Biomol Chem,2017,15(36):7672-7677.[24]BAI M，ZHENG C，TANG D，et al.Two new secondary metabolites from a mangrove-derived fungus Cladosporium sp.[J].JS1-2.J Antibiot，2019，72：779-782.

相似文献/References:

[1]王明洁,吴雨蹊,段亚东,等.蓝莓栽培土壤改良技术研究[J].北方园艺,2014,38(14):179.
　WANG Ming-jie,WU Yu-xi,DUAN Ya-dong,et al.Improvement Research of Technique on Cultivation Soil of Blueberry[J].Northern Horticulture,2014,38(07):179.
[2]王贺春.蓝莓冬季修剪技术[J].北方园艺,2013,37(13):58.
　WANG He-chun.Winter Pruning Techniqne of Blueberry[J].Northern Horticulture,2013,37(07):58.
[3]鲁海菊,董梅,崔同敏,等.枇杷内生真菌多样性初步研究[J].北方园艺,2014,38(04):100.
　LU Hai-ju,DONG Mei,CUI Tong-min,et al.Preliminary Study on Diversity of Endophytic Fungi From Eriobotrya japonica[J].Northern Horticulture,2014,38(07):100.
[4]聂飞,张玉春.我国蓝莓产业发展的机遇与挑战[J].北方园艺,2014,38(04):165.
　NIE Fei,ZHANG Yu-chun.Opportunities and Challenges for the Development of Blueberry Industry in China[J].Northern Horticulture,2014,38(07):165.
[5]杨芩,任永权,廖优江,等.五个兔眼蓝莓品种有效可授期研究[J].北方园艺,2013,37(14):5.
　YANG Qin,REN Yong-quan,LIAO You-jiang,et al.Study on Effective Pollination Period of Five Cultivars of Rabbiteye Blueberry[J].Northern Horticulture,2013,37(07):5.
[6]傅 冰,洪 震,刘跃钧,等.蓝莓SRAP-PCR反应体系的建立和优化及其遗传多样性分析[J].北方园艺,2013,37(17):95.
　FU Bing,HONG Zhen,LIU Yue-jun,et al.Optimization and Establishment of SRAP-PCR Reaction System of Vaccinium ssp. and Analysis of the Genetic Diversity[J].Northern Horticulture,2013,37(07):95.
[7]张文英,周 婧.低糖蓝莓番茄复合果酱的研制[J].北方园艺,2013,37(17):142.
　Brewing Technology Engineering Research Center,Department of Food Science,Jilin Agricultural Science and Technology College,et al.Preparation of Low-sugar Jam with Blueberry and Tomato[J].Northern Horticulture,2013,37(07):142.
[8]鲁海菊,郝小燕,崔同敏,等.抗枇杷根腐病病菌的枇杷主干内生真菌生物学特性研究[J].北方园艺,2014,38(08):108.
　LU Hai-ju,HAO Xiao-yan,CUI Tong-min,et al.Study on Biological Characteristics of Endophytic Fungal Resistant to Loquat Root Rot From Eriobotrya japonica Stem[J].Northern Horticulture,2014,38(07):108.
[9]龚 娜,肖 军,杨 镇,等.不同菌根真菌对蓝莓根际土壤微生物数量的影响[J].北方园艺,2013,37(17):175.
　GONG Na,XIAO Jun,YANG Zhen,et al.Effects of Different Mycorrhizal Fungi on Soil Microbial Number in the Blueberry Rhizosphere[J].Northern Horticulture,2013,37(07):175.
[10]王贺春.适于丹东地区温室栽培的蓝莓引种与筛选[J].北方园艺,2013,37(12):33.
　WANG He-chun.Introduction and Selection of Blueberry Varieties for Greenhouse That Suitable for Dandong Region[J].Northern Horticulture,2013,37(07):33.

备注/Memo

第一作者简介：倪维鞠（1996-），女，硕士研究生，研究方向为药理学。E-mail:962413636@qq.com.责任作者：雷霁卿（1985-），女，硕士，副教授，现主要从事微生物与食品安全等研究工作。E-mail:2012444896@qq.com.基金项目：贵州省教育厅自然科学研究资助项目（黔教合KY字[2014]308号）；贵州省2017年大学生创新创业训练计划资助项目（201710976074）；贵阳市财政支持贵阳学院学科建设与研究生教育资助项目(SY-2020)；贵阳学院科研资金资助项目（GYU-KY-[2021])；贵州省科技计划资助项目（黔科合成果[2019]4221号）。收稿日期：2020-06-11

更新日期/Last Update: 2021-06-10