{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T23:34:08Z","timestamp":1792884848619,"version":"4.5.0"},"reference-count":45,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T00:00:00Z","timestamp":1787443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011789","name":"Jilin Province Science and Technology Department","doi-asserted-by":"crossref","award":["20230402028GH"],"award-info":[{"award-number":["20230402028GH"]}],"id":[{"id":"10.13039\/501100011789","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100011789","name":"Jilin Province Science and Technology Department","doi-asserted-by":"crossref","award":["YDZJ202501ZYTS482"],"award-info":[{"award-number":["YDZJ202501ZYTS482"]}],"id":[{"id":"10.13039\/501100011789","id-type":"DOI","asserted-by":"crossref"}]},{"award":["20230402028GH"],"award-info":[{"award-number":["20230402028GH"]}],"id":[{"id":"https:\/\/ror.org\/049x38272","id-type":"ROR","asserted-by":"publisher"}]},{"award":["YDZJ202501ZYTS482"],"award-info":[{"award-number":["YDZJ202501ZYTS482"]}],"id":[{"id":"https:\/\/ror.org\/049x38272","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Panax ginseng, a high-value medicinal plant, faces substantial yield losses due to black spot disease, while conventional chemical controls cause pesticide residues and soil ecological damage, necessitating green biocontrol strategies. Here, two antagonistic strains, Bacillus sp. WY17 and WY26, were isolated from the surface-sterilized internal root tissues of 10-year-old ginseng. Through systematic optimization of carbon\/nitrogen sources, inorganic salts, and fermentation parameters (temperature, pH, agitation, inoculum size, and duration), the optimal culture conditions were established. The optimal consortium consisted of WY17 and WY26 in a 2:1 ratio (WY17:WY26 = 2:1), which achieved an antifungal inhibition rate of 84.94% against the pathogen compared to the untreated control group (pathogen only). Mechanistic investigations revealed that the crude protein extract exerted its antifungal effect by compromising the integrity of the pathogen\u2019s cell membrane, leading to increased permeability and leakage of intra-cellular contents, and produced cell wall-degrading enzymes (chitinase and \u03b2-1,3-glucanase), thereby inhibiting mycelial growth and spore germination. In vitro efficacy tests demonstrated that this crude protein extract performed comparably to the chemical fungicide 70% mancozeb, with no statistically significant difference observed between them (p &gt; 0.05). These findings identify a promising compound biocontrol agent derived from indigenous Bacillus strains, offering an effective and environmentally friendly alternative for managing ginseng black spot disease.<\/jats:p>","DOI":"10.3390\/microorganisms14091871","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T08:03:41Z","timestamp":1787558621000},"page":"1871","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Optimized Fermentation of Endophytic Bacillus sp. WY17 and WY26 Consortium for Biocontrol of Ginseng Black Spot Disease and Its Antifungal Activity via Crude Protein Extract"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-2810-1352","authenticated-orcid":false,"given":"Qiuyu","family":"Wang","sequence":"first","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihao","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchi","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajing","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyan","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunshi","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Sun","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiwei","family":"Dong","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenxiu","family":"Ji","sequence":"additional","affiliation":[{"name":"College of Agriculture, Yanbian University, Yanji 133002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1002\/med.21953","article-title":"A comprehensive and systemic review of ginseng-based nanomaterials: Synthesis, targeted delivery, and biomedical applications","volume":"43","author":"Balusamy","year":"2023","journal-title":"Med. Res. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/s11418-023-01747-1","article-title":"Genetic diversity of Panax ginseng cultivated in Japan and its relation with some plant characteristics","volume":"78","author":"Ito","year":"2024","journal-title":"J. Nat. Med."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Lan, J., Mei, S., Du, Y., Chi, M., Yang, J., Guo, S., Chu, M., He, R., and Gao, J. (2025). ApWD40a, a member of the WD40-repeat protein family, is crucial for fungal development, toxin synthesis, and pathogenicity in the ginseng alternaria leaf blight fungus Alternaria panax. J. Fungi, 11.","DOI":"10.3390\/jof11010059"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1080\/21501203.2023.2265662","article-title":"Mycotoxins from Alternaria Panax, the specific plant pathogen of Panax ginseng","volume":"14","author":"Chen","year":"2023","journal-title":"Mycology"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1007\/s13258-022-01241-0","article-title":"Transcriptome analysis reveals putative pathogenesis genes in Alternaria panax during infecting Panax notoginseng leaves","volume":"44","author":"Shah","year":"2022","journal-title":"Genes Genom."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Jia, Q., Fan, Y., Duan, S., Qin, Q., Ding, Y., Yang, M., Wang, Y., Liu, F., and Wang, C. (2023). Effects of Bacillus amyloliquefaciens XJ-BV2007 on growth of Alternaria alternata and production of tenuazonic acid. Toxins, 15.","DOI":"10.3390\/toxins15010053"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Feng, S., Zhao, Y., Wang, Q., Zhang, J., Liang, X., Fu, Z., Li, Y., Dong, W., and Ji, W. (2024). Biocontrol of rusted root rot in Panax ginseng by a combination of extracts from Bacillus amyloliquefaciens YY8 crude protein and Enterobacteriaceae YY115 ethyl acetate. BMC Microbiol., 24.","DOI":"10.1186\/s12866-024-03475-2"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Li, X., Liu, Q., Gao, Y., Zang, P., and Zheng, T. (2024). Effects of a co-bacterial agent on the growth, disease control, and quality of ginseng based on rhizosphere microbial diversity. BMC Plant Biol., 24.","DOI":"10.1186\/s12870-024-05347-3"},{"key":"ref_9","first-page":"75","article-title":"Evaluation of Bacillus isolates as a biological control agents against soilborne phytopathogenic fungi","volume":"28","author":"Eltem","year":"2025","journal-title":"Int. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"20929","DOI":"10.1038\/s41598-023-48171-9","article-title":"Whole genome sequencing provides evidence for Bacillus velezensis SH-1471 as a beneficial rhizosphere bacterium in plants","volume":"13","author":"Shen","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1007\/s00284-023-03470-2","article-title":"Induced resistance mechanism of Bacillus velezensis S3-1 against pepper wilt","volume":"80","author":"Fan","year":"2023","journal-title":"Curr. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zou, Q., Zhang, Y., Niu, X., Yang, H., Chu, M., Wang, N., Bao, H., Zhan, F., Yang, R., and Lou, K. (2024). Antifungal activity of rhizosphere Bacillus isolated from Ziziphus jujuba Against Alternaria alternata. Microorganisms, 12.","DOI":"10.20944\/preprints202410.0957.v1"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2250","DOI":"10.1111\/1751-7915.14348","article-title":"Biocontrol mechanisms of Bacillus: Improving the efficiency of green agriculture","volume":"16","author":"Zhang","year":"2023","journal-title":"Microb. Biotechnol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"148669","DOI":"10.1016\/j.gene.2024.148669","article-title":"Unlocking the growth-promoting and antagonistic power: A comprehensive whole genome study on Bacillus velezensisstrains","volume":"927","author":"Ercole","year":"2024","journal-title":"Gene"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4110","DOI":"10.1002\/ps.8118","article-title":"Bacillus-derived consortium enhances Ginkgo biloba\u2019s health and resistance to Alternaria tenuissima","volume":"80","author":"Ji","year":"2024","journal-title":"Pest Manag. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Li, Q., Yan, N., Miao, X., Zhan, Y., and Chen, C. (2022). The potential of novel bacterial isolates from healthy ginseng for the control of ginseng root rot disease (Fusarium oxysporum). PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0277191"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e17424","DOI":"10.7717\/peerj.17424","article-title":"The unexpected effect of the compound microbial agent NP-M2 on microbial community dynamics in a nonylphenol-contaminated soil: The self-stability of soil ecosystem","volume":"12","author":"Chen","year":"2024","journal-title":"PeerJ"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1007\/s00284-024-03652-6","article-title":"Evaluation of antifungal activity of endophytic Bacillus spp. and identification of secondary metabolites produced against the phytopathogenic fungi","volume":"81","author":"Patel","year":"2024","journal-title":"Curr. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/j.jgr.2023.08.005","article-title":"Botrytis cinerea hypovirulent strain \u25b3BcSpd1 induced Panax ginseng defense","volume":"47","author":"Zhang","year":"2023","journal-title":"J. Ginseng. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1007\/s00294-024-01288-4","article-title":"Comprehensive genomic analysis of Burkholderia arboris PN-1 reveals its biocontrol potential against Fusarium solani-induced root rot in Panax notoginseng","volume":"70","author":"Yang","year":"2024","journal-title":"Curr. Genet."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1007\/s00203-022-03136-w","article-title":"Fungal disease suppression and growth promotion potential of endophytic bacteria from ethnomedicinal plants","volume":"204","author":"Khunjamayum","year":"2022","journal-title":"Arch. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6071","DOI":"10.1007\/s00253-023-12642-w","article-title":"Synergistic biocontrol of Bacillus subtilis and Pseudomonas fluorescens against early blight disease in tomato","volume":"107","author":"Jia","year":"2023","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1007\/s10534-022-00440-2","article-title":"Bacillus sp. extract used to fabricate ZnO nanoparticles for their antagonist effect against phytopathogens","volume":"35","author":"Vinay","year":"2022","journal-title":"Biometals"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Harish, B.N., Nagesha, S.N., Ramesh, B.N., Shyamalamma, S., Nagaraj, M.S., Girish, H.C., Pradeep, C., Shiva Kumar, K.S., Tharun Kumar, K.S., and Pavan, S.N. (2023). Molecular characterization and antifungal activity of lipopeptides produced from Bacillus subtilis against plant fungal pathogen Alternaria alternata. BMC Microbiol., 23.","DOI":"10.1186\/s12866-023-02922-w"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Li, X., Wang, J., Shen, H., Xing, C., Kong, L., Song, Y., Hou, W., Gao, J., Jiang, Y., and Chen, C. (2024). Biocontrol and growth promotion potential of Bacillus velezensis NT35 on Panax ginseng based on the multifunctional effect. Front. Microbiol., 15.","DOI":"10.3389\/fmicb.2024.1447488"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1002\/jobm.202200530","article-title":"Characterization of lipopeptide produced by Bacillus altitudinis Q7 and inhibitory effect on Alternaria alternata","volume":"63","author":"Guo","year":"2023","journal-title":"J. Basic Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1007\/s00248-024-02450-8","article-title":"Paenibacillus as a biocontrol agent for fungal phytopathogens: Is P. polymyxa the only one worth attention?","volume":"87","year":"2024","journal-title":"Microb. Ecol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Li, E., Liu, K., Yang, S., Li, L., Ran, K., Sun, X., Qu, J., Zhao, L., Xin, Y., and Zhu, F. (2024). Analysis of the complete genome sequence of Paenibacillus sp. lzh-N1 reveals its antagonistic ability. BMC Genom., 25.","DOI":"10.1186\/s12864-024-10206-4"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e202402744","DOI":"10.1002\/cbdv.202402744","article-title":"Two chroman type polyketides from Trichoderma sp. YUD24002 associated with Aconitum","volume":"22","author":"Zi","year":"2025","journal-title":"Chem. Biodivers."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Feng, S., Wang, C., Xu, Z., Dou, B., Wang, X., Yang, L., Lu, B., and Gao, J. (2024). Evaluation of the Antifungal and Biochemical Activities of Fungicides and Biological Agents against Ginseng Sclerotinia Root Rot Caused by Sclerotinia nivalis. Microorganisms, 12.","DOI":"10.3390\/microorganisms12091761"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e0131122","DOI":"10.1128\/mra.01311-22","article-title":"Genome sequence resource of Bacillus velezensis strain ML61, a potential biocontrol bacterium isolated from the rhizosphere of bok choy (Brassica rapa var. chinensis)","volume":"12","author":"Ma","year":"2023","journal-title":"Microbiol. Resour. Announc."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Palacios-Rodriguez, A.P., Espinoza-Culup\u00fa, A., Dur\u00e1n, Y., and S\u00e1nchez-Rojas, T. (2024). Antimicrobial Activity of Bacillus amyloliquefaciens BS4 against gram-negative pathogenic bacteria. Antibiotics, 13.","DOI":"10.3390\/antibiotics13040304"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Wang, K., Wang, Z., Ding, Y., Yu, Y., Wang, Y., Geng, Y., Li, Y., and Wen, X. (2023). Optimization of heterotrophic culture conditions for the algae Graesiella emersonii WBG-1 to produce proteins. Plants, 12.","DOI":"10.3390\/plants12122255"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1007\/s12223-025-01247-7","article-title":"Goan mangrove yeast: A source of therapeutic enzyme L-asparaginase","volume":"70","author":"Marathe","year":"2025","journal-title":"Folia Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"31011","DOI":"10.1038\/s41598-024-82167-3","article-title":"Upregulation of ACC deaminase gene in Bacillus velezensis UTB96 improved yield and shelf Life of Agaricus bisporus","volume":"14","author":"Dousti","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Zhou, Y., Yang, X., Li, Q., Peng, Z., Li, J., and Zhang, J. (2023). Optimization of fermentation conditions for surfactin production by B. subtilis YPS-32. BMC Microbiol., 23.","DOI":"10.1186\/s12866-023-02838-5"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"105025","DOI":"10.1016\/j.psj.2025.105025","article-title":"Optimization of the enrichment medium for recombinant ChIL-4-ChIL-2 in Lactococcus lactis through response surface methodology","volume":"104","author":"Zhang","year":"2025","journal-title":"Poult. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"110580","DOI":"10.1016\/j.ijfoodmicro.2024.110580","article-title":"Bacillus velezensis RC218 and emerging biocontrol agents against Fusarium graminearum and Fusarium poae in barley: In vitro, greenhouse and field conditions","volume":"413","author":"Zanon","year":"2024","journal-title":"Int. J. Food Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1186\/s12934-024-02372-7","article-title":"Research advances in the identification of regulatory mechanisms of surfactin production by Bacillus: A review","volume":"23","author":"Qiao","year":"2024","journal-title":"Microb. Cell Fact."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1007\/s00203-023-03582-0","article-title":"Biochemical and molecular depictions to develop ech42 gene-specific SCAR markers for recognition of chitinolytic Trichoderma inhibiting Macrophomina phaseolina (Maubl.) Ashby","volume":"205","author":"Gajera","year":"2023","journal-title":"Arch. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Zhang, L., Bao, L., Li, S., Liu, Y., and Liu, H. (2023). Active substances of myxobacteria against plant diseases and their action mechanisms. Front. Microbiol., 14, Erratum in Front. Microbiol. 2024, 15, 1392109.","DOI":"10.3389\/fmicb.2023.1294854"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"126021","DOI":"10.1016\/j.ijbiomac.2023.126021","article-title":"Chitinases as key virulence factors in microbial pathogens: Understanding their role and potential as therapeutic targets","volume":"249","author":"Jeong","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1002\/ps.7872","article-title":"Biological control of tomato early blight in Pakistan using local rhizobacteria","volume":"80","author":"Riaz","year":"2024","journal-title":"Pest Manag. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Kumari, R., Kumar, V., Arukha, A.P., Rabbee, M.F., Ameen, F., and Koul, B. (2024). Screening of the biocontrol efficacy of Potent Trichoderma strains against Fusarium oxysporum f.sp. ciceri and Scelrotium rolfsii causing wilt and collar rot in chickpea. Microorganisms, 12.","DOI":"10.3390\/microorganisms12071280"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"135084","DOI":"10.1016\/j.jhazmat.2024.135084","article-title":"One stone two birds: Endophytes alleviating trace elements accumulation and suppressing soilborne pathogen by stimulating plant growth, photosynthetic potential and defense related gene expression","volume":"476","author":"Imran","year":"2024","journal-title":"J. Hazard. Mater."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/9\/1871\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T04:33:24Z","timestamp":1787632404000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/9\/1871"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,23]]},"references-count":45,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["microorganisms14091871"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14091871","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,23]]}}}