蓝莓内生真菌BB01抗菌活性的研究

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64μg·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、128μg·mL^-1,同时,提取物BE终浓度500μg·mL^-1时,可有效抑制4种病害菌菌丝生长(抑菌率分别为98.067302%±3.813437%、92.187892%±0.828473%、94.251883%±0.368539%、96.909029%±5.769104%)。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。

Antimicrobial Activity of Endophytic Fungi BB01 Derived From Blueberry

‘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 64μg·mL^-1,and BE exhibited the best inhibition against Cladosporium,Glomerella acutata and Botrytis cinerea,with MIC values of 16μg·mL^-1,256μg·mL^-1 and 128μg·mL^-1,respectively.Moreover BE also exhibited 98.067302%±3.813437%,92.187892%±0.828473%,94.251883%±0.368539%,96.909029%±5.769104%growth inhibiton against 4 pathogens respectively at 500μg·mL^-1.PCR was used to amplify the genome rDNA-ITS sequence for phylogenetic tree.The results showed that BB01 was Cladosporium sp.