Food and Fermentation Industries >

2024 , Vol. 50 >Issue 18: 154 - 160

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.037588

Screening of endophytic bacteria from Cypripedium macranthum and antifungal activity against Aspergillus flavus

  • FU Yajuan ,
  • XU Xiaoqing ,
  • FAN Jianing ,
  • FAN Shuanghu ,
  • Han Hui ,
  • HOU Xiaoqiang
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  • 1(College of Life Science, Langfang Normal University, Langfang 065000, China)
    2(Technical Innovation Center for Utilization of Edible and Medicinal Fungi in Hebei Province, Langfang 065000, China)
    3(Key Laboratory of Biological Sample Analysis and Pesticide Residue Detection, Langfang 065000, China)

Received date: 2023-10-09

  Revised date: 2023-10-30

  Online published: 2024-10-14

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Abstract

To obtain the endophytic bacteria of Cypripedium macranthum with efficient antagonistic effect on Aspergillus flavus, 31 endophytic bacteria were screened against A.flavus through plate confrontation test and mycelia growth rate method.A series of methodologies were carried out as follows:The stability of antifungal substances test was measured by using mycelium growth rate method;the lipopeptide related and polyketide synthase genes from the genomic DNA of strain CmR-1n were amplified by PCR;the antifungal activity of the crude lipopeptide extracted from CmR-1n fermentation broth against A.flavus was evaluated by using cup and saucer method.The results showed that 3 endophytic bacteria strains (CmR-1n, CmR-3a and CmR-4b) possessed better antagonistic effects on A.flavus.Briefly, Bacillus sp. CmR-1n had the best antagonistic effect on A.flavus with the inhibition rate of 57.88%, while the inhibition rate of CmR-1n sterile fermentation broth against A.flavus reached 63.31%.Antifungal substances in CmR-1n fermentation broth had significant heat, acid, alkali, and UV resistance and storage stability.Besides, the CmR-1n genome contained gene fragments of iturin (Itu), surfactin (Srf), fengycin (fenB and fenD).Moreover, the crude lipopeptides obtained from CmR-1n fermentation broth in vitro inhibited A.flavus mycelium growth with the inhibition rate up to 62.58%.Therefore, it is speculated that lipopeptides from Bacillus sp. CmR-1n may be active substance against A.flavus.Bacillus sp. CmR-1n has certain application potential in biological prevention and control of A.flavus.

Key words： Cypripedium macranthum; endophytic Bacillus spp.; Aspergillus flavus; crude lipopeptides; antifungal activity

Cite this article

FU Yajuan , XU Xiaoqing , FAN Jianing , FAN Shuanghu , Han Hui , HOU Xiaoqiang . Screening of endophytic bacteria from Cypripedium macranthum and antifungal activity against Aspergillus flavus[J]. Food and Fermentation Industries, 2024 , 50(18) : 154 -160 . DOI: 10.13995/j.cnki.11-1802/ts.037588

References

[1] RUSHING B R, SELIM M I.Aflatoxin B1:A review on metabolism, toxicity, occurrence in food, occupational exposure, and detoxification methods[J].Food and Chemical Toxicology,2019, 124:81-100.
[2] ELAHI E, CUI W J, ZHANG H M, et al.Agricultural intensification and damages to human health in relation to agrochemicals:Application of artificial intelligence[J].Land Use Policy, 2019, 83:461-474.
[3] ZHAO S S, HAO X M, YANG F Y, et al.Antifungal activity of Lactobacillus plantarum ZZUA493 and its application to extend the shelf life of Chinese steamed buns[J].Foods, 2022, 11(2):195.
[4] CHEN L, SHI H, HENG J Y, et al.Antimicrobial, plant growth-promoting and genomic properties of the peanut endophyte Bacillus velezensis LDO2[J].Microbiological Research, 2019, 218:41-48.
[5] PRASERTSAN P, SAWAI B.Potential use of biofumigant from Streptomyces mycarofaciens for protection of maize seed contaminated by aflatoxin producing fungi[J].New Biotechnology, 2018, 44:S151.
[6] GONG A D, WU N N, KONG X W, et al.Inhibitory effect of volatiles emitted from Alcaligenes faecalis N1-4 on Aspergillus flavus and aflatoxins in storage[J].Frontiers in Microbiology, 2019, 10:1419.
[7] GONG A D, LEI Y Y, HE W J, et al.The inhibitory effect of Pseudomonas stutzeri YM6 on Aspergillus flavus growth and aflatoxins production by the production of volatile dimethyl trisulfide[J].Toxins, 2022, 14(11):788.
[8] NOGUEIRA M F, PEREIRA L, JENULL S, et al.Klebsiella pneumoniae prevents spore germination and hyphal development of Aspergillus species[J].Scientific Reports, 2019, 9(1):218.
[9] JIA J Y, FORD E, HOBBS S M, et al.Occidiofungin is the key metabolite for antifungal activity of the endophytic bacterium Burkholderia sp.MS455 against Aspergillus flavus[J].Phytopathology, 2022, 112(3):481-491.
[10] GONG A D, WANG G Z, SUN Y K, et al.Dual activity of Serratia marcescens Pt-3 in phosphate solubilizing and production of antifungal volatiles[J].BMC Microbiology, 2022, 22(1):26.
[11] LI S J, XU X G, ZHAO T Y, et al.Screening of Bacillus velezensis E2 and the inhibitory effect of its antifungal substances on Aspergillus flavus [J].Foods, 2022, 11(2):140.
[12] 姚彦坡, 丁丹, 张友青, 等.玉米黄曲霉毒素污染生防菌筛选及菌株B42-3抗菌活性研究[J].中国粮油学报, 2018, 33(3):84-88.
YAO Y P, DING D, ZHANG Y Q, et al.Screening of maize endophytic antagonists against Aspergillus flavus and activities research of antagonistic strain B42-3[J].Journal of the Chinese Cereals and Oils Association, 2018, 33(3):84-88.
[13] CHEN G J, FANG Q A, LIAO Z L, et al.Detoxification of aflatoxin B1 by a potential probiotic Bacillus amyloliquefaciens WF2020[J].Frontiers in Microbiology, 2022, 13:891091.
[14] 刘端木, 吴怿, 刘沄, 等.一株产毒曲霉拮抗细菌的筛选、鉴定及抑菌活性研究[J].生物技术通报, 2019, 35(8):42-50.
LIU D M, WU Y, LIU Y, et al.Screening, identification and antifungal properties of a bacterium with antagonistic activities against mycotoxin-producing Aspergillus spp.[J].Biotechnology Buttetin, 2019, 35(8):42-50.
[15] 潘虹余, 金玮鋆, 张晓蒙, 等.解淀粉芽孢杆菌B15抑菌物质对葡萄灰霉病灰葡萄孢的抑菌机理[J].微生物学报, 2018, 58(7):1245-1254.
PAN H Y, JIN Y J, ZHANG X M, et al.Inhibition of antifungal substances from Bacillus amyloquefaciens B15 against Botrytis Cinerea—the agent of “gray mold” of grape[J].Acta Microbiologica Sinica, 2018, 58(7):1245-1254.
[16] 杜宾, 李娜, 李燕.花椒内生细菌Y-6抗菌蛋白抑菌活性及稳定性研究[J].河南农业科学, 2017, 46(6):67-73.
DU B, LI N, LI Y.Antagonistic activity and stability of antifungal protein of endophytic bacterium Y-6 from Zanthoxylum bungeanum against plant pathogenic fungi[J].Journal of Henan Agricultural Sciences, 2017, 46(6):67-73.
[17] MA Z W, ZHANG S Y, ZHANG S H, et al.Isolation and characterization of a new cyclic lipopeptide surfactin from a marine-derived Bacillus velezensis SH-B74[J].The Journal of Antibiotics, 2020, 73(12):863-867.
[18] MARCHESE S, POLO A, ARIANO A, et al.Aflatoxin B1 and M1:Biological properties and their involvement in cancer development[J].Toxins, 2018, 10(6):214.
[19] FIRA D, DIMKIĆ I, BERIĆ T, et al.Biological control of plant pathogens by Bacillus species[J].Journal of Biotechnology, 2018, 285:44-55.
[20] ZHU J X, TAN T M, SHEN A R, et al.Biocontrol potential of Bacillus subtilis IBFCBF-4 against Fusarium wilt of watermelon[J].Journal of Plant Pathology, 2020, 102(2):433-441.
[21] YOU W J, GE C H, JIANG Z C, et al.Screening of a broad-spectrum antagonist Bacillus siamensis, and its possible mechanisms to control postharvest disease in tropical fruits[J].Biological Control, 2021, 157:104584.
[22] LAIRD M, PICCOLI D, WESELOWSKI B, et al.Surfactin-producing Bacillus velezensis 1B-23 and Bacillus sp.1D-12 protect tomato against bacterial canker caused by Clavibacter michiganensis subsp. Michiganensis [J].Journal of Plant Pathology, 2020, 102(2):451-458.
[23] FARZANEH M, SHI Z Q, AHMADZADEH M, et al.Inhibition of the Aspergillus flavus growth and aflatoxin B1 contamination on pistachio nut by fengycin and surfactin-producing Bacillus subtilis UTBSP1[J].The Plant Pathology Journal, 2016, 32(3):209-215.
[24] YUAN S L, WU Y J, JIN J, et al.Biocontrol capabilities of Bacillus subtilis E11 against Aspergillus flavus in vitro and for dried red chili (Capsicum annuum L.)[J].Toxins, 2023, 15(5):308.
[25] 胡忠亮, 郑催云, 田兴一, 等.解淀粉芽孢杆菌HZM9菌株发酵液的抑菌谱及稳定性测定[J].南京林业大学学报(自然科学版), 2017, 41(3):65-70.
HU Z L, ZHENG C Y, TIAN X Y, et al.Determination of the inhibitory spectrum and stability of culture filtrate from Bacillus amyloliquefaciens HZM9[J].Journal of Nanjing Forestry University (Natural Sciences Edition), 2017, 41(3):65-70.
[26] 孙力军, 王雅玲, 刘唤明, 等.抗菌豆豉发酵菌株的筛选及其脂肽组分鉴定和特性研究[J].中国生物工程杂志, 2013, 33(7):50-56.
SUN L J, WANG Y L, LIU H M, et al.Identification of antimicrobial lipopetides component produced by isolate from Douchi and its antimicrobial properties[J].China Biotechnology, 2013, 33(7):50-56.
[27] 丁飞鸿, 耿云龙, 樊士德, 等.抑制食源性致病菌脂肽的分离鉴定及生物学特性分析[J].食品与发酵工业, 2022, 48(6):195-203.
DING F H, GENG Y L, FAN S D, et al.Isolation and identification of lipopeptides from Bacillus pumilus inhibiting foodborne pathogens and analysis of its biological characteristics[J].Food and Fermentation Industries, 2022, 48(6):195-203.
[28] HASSAN Z U, AL THANI R, ALNAIMI H, et al.Investigation and application of Bacillus licheniformis volatile compounds for the biological control of toxigenic Aspergillus and Penicillium spp.[J].ACS Omega, 2019, 4(17):17186-17193.
[29] YANG M G, LU L F, PANG J, et al.Biocontrol activity of volatile organic compounds from Streptomyces alboflavus TD-1 against Aspergillus flavus growth and aflatoxin production[J].Journal of Microbiology, 2019, 57(5):396-404.
[30] BOUKAEW S, PRASERTSAN P.Efficacy of volatile compounds from Streptomyces philanthi RL-1-178 as a biofumigant for controlling growth and aflatoxin production of the two aflatoxin-producing fungi on stored soybean seeds[J].Journal of Applied Microbiology, 2020, 129(3):652-664.
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