Identification of Endogenous Antagonistic Bacteria in Grape Leaves and Its Inhibitory Effect on Postharvest <i>Aspergillus niger</i> in Thompson Seedless Grapes

ZHANG Miaomiao; YANG Yizhong; ZHANG Yuchen; MA Jie; XU Zhenzhen; WANG Wei; ZHANG Lijuan; WANG Ning; FAN Yingying; WANG Cheng; HUANG Wei; LIU Fengjuan

doi:10.13386/j.issn1002-0306.2024070065

Turn off MathJax

Article Contents

Abstract

References

ZHANG Miaomiao, YANG Yizhong, ZHANG Yuchen, et al. Identification of Endogenous Antagonistic Bacteria in Grape Leaves and Its Inhibitory Effect on Postharvest Aspergillus niger in Thompson Seedless Grapes[J]. Science and Technology of Food Industry, 2025, 46(10): 1−10. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024070065.

Citation:

PDF (6163 KB)

Identification of Endogenous Antagonistic Bacteria in Grape Leaves and Its Inhibitory Effect on Postharvest Aspergillus niger in Thompson Seedless Grapes

1.
College of Food Science and Pharmacy, Xinjiang Agricultural University, Urumqi 830052, China
2.
Key Laboratory of Agro-products Quality and Safety of Xinjiang, Laboratory of Quality and Safety Risk Assessment for Agri-products (Urumqi), Key Laboratory of Functional Nutrition and Health of Characteristic Agricultural Products in Desert Oasis Ecological Region (Co-construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Institute of Quality Standards & Testing Technology for Agri-products, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China
3.
Microbial Application Institute, Xinjiang Academy of Agricultural Sciences, Urumqi 830000, China
4.
Institute of Quality Standard & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, China

More Information

Received Date: July 07, 2024
Available Online: March 20, 2025

Graphical Abstract

Abstract

Abstract

To screen endophytic strains with strong antibacterial activity from grape leaves and assess their biocontrol potential, providing a theoretical basis for the development of new biological control strategies against grape Aspergillus rot disease. Using the plate confrontation method, antagonistic strains displaying notable inhibitory effects on Aspergillus niger were isolated from the leaves of ‘Thompson Seedless Grapes’. Morphological, physiological, biochemical, and molecular-biological analyses identified the antagonistic strains to species-level resolution. Lipopeptide extracts from the fermentation supernatant were analyzed by UPLC-Vion-IMS-Q-TOF mass spectrometry, revealing their chemical composition. In vivo experiments further confirmed the efficacy of the antagonistic fermentation broth in reducing postharvest Aspergillus rot on 'Thompson Seedless Grapes'. From the leaves of ‘Thompson Seedless’ grapes, 25 endophytic bacterial strains with inhibitory effects on A. niger were isolated. Among these, strain TLF4-3 demonstrated the strongest antagonistic activity, exhibiting an inhibition zone diameter of 27.27 ± 0.05 mm and an inhibitory titer of 18.27 ± 0.53 cm³/mL. UPLC-Vion-IMS-Q-TOF analysis identified the primary antimicrobial compounds produced by this strain as Bacillomycin D (C15) and Iturin C (C14). Treatments with TLF4-3 fermentation broth at 3 days postharvest achieved the best control effect, reducing lesion diameter by 2.3 mm and achieving an antibacterial rate of 68.26%±0.91%. These results offer a theoretical framework and pragmatic direction for the control and avoidance of Aspergillus rot in postharvest grapes.
- antagonizing endogenous bacteria,
- lipopeptides,
- A. niger,
- B. velezensis,
- biological control

FullText(HTML)

References (34)

References

[1]	刘凤之. 我国葡萄产业现状与高质量发展方向[J]. 农业知识,2023,75(10):10−14. [LIU F Z. Current situation and high quality development direction of grape industry in China[J]. Agricultural Knowledge,2023,75(10):10−14.] LIU F Z. Current situation and high quality development direction of grape industry in China[J]. Agricultural Knowledge, 2023, 75（10）: 10−14.
[2]	张苗苗, 张雨晨, 杨怡中, 等. 拮抗菌防治葡萄采后病害研究进展[J]. 食品工业科技,2024,45(9):410−418. [ZHANG M M, ZHANG Y C, YAANG Y Z, et al. Research progress on antagonistic antimicrobial prevention and control of grape postharvest diseases[J]. Food Industry Science and Technology,2024,45(9):410−418.] ZHANG M M, ZHANG Y C, YAANG Y Z, et al. Research progress on antagonistic antimicrobial prevention and control of grape postharvest diseases[J]. Food Industry Science and Technology, 2024, 45（9）: 410−418.
[3]	李小红, 李运景, 马晓青, 等. 我国葡萄产业发展现状与展望[J]. 中国南方果树,2021,50(5):161−166. [LI X H, LI Y J, MA X Q, et al. Current situation and prospect of grape industry development in China[J]. Fruit Trees of Southern China,2021,50(5):161−166.] LI X H, LI Y J, MA X Q, et al. Current situation and prospect of grape industry development in China[J]. Fruit Trees of Southern China, 2021, 50（5）: 161−166.
[4]	ASSAF L, PEDROZO L P, NALLY M C, et al. Use of yeasts from different environments for the control of Penicillium expansum on table grapes at storage temperature[J]. International Journal of Food Microbiology,2020,37(320):108520.
[5]	孟创鸽, 曹红霞, 韩峪, 等. 葡萄贮藏保鲜技术研究进展[J]. 黑龙江农业科学,2022,45(5):102−106. [MENG C G, CAO H X, HAN Y, et al. Research progress of grape storage and fresh-keeping technology[J]. Agricultural Sciences in Heilongjiang,2022,45(5):102−106.] MENG C G, CAO H X, HAN Y, et al. Research progress of grape storage and fresh-keeping technology[J]. Agricultural Sciences in Heilongjiang, 2022, 45（5）: 102−106.
[6]	张鹏, 邵丹, 李江阔, 等. 葡萄冷藏时间对贮后货架期芳香物质的影响[J]. 食品科学,2016,37(2):218−224. [ZHANG P, SHAO D, LI J K, et al. Effect of cold storage time on aromatic substances of grape after storage[J]. Food Science,2016,37(2):218−224.] ZHANG P, SHAO D, LI J K, et al. Effect of cold storage time on aromatic substances of grape after storage[J]. Food Science, 2016, 37（2）: 218−224.
[7]	邵千朔, 黄桂丽, 梁慧敏, 等. 鲜食葡萄保鲜技术研究进展[J]. 江苏农业科学,2024,52(4):17−22. [SHAO Q S, HUANG G L, LIANG H M, et al. Research progress on preservation technology of table grape[J]. Jiangsu Agricultural Sciences,2024,52(4):17−22.] SHAO Q S, HUANG G L, LIANG H M, et al. Research progress on preservation technology of table grape[J]. Jiangsu Agricultural Sciences, 2024, 52（4）: 17−22.
[8]	王曼, 魏佳, 伊丽达娜·迪力夏提, 等. 二氧化硫调控亚硫酸盐代谢对葡萄采后灰霉病的抑制作用[J]. 中国食品学报,2022,22(11):299−308. [WANG M, WEI J, DI LI XIA TI Y L D N, et al. The inhibitory effect of sulfur dioxide on postharvest gray mold of grape by regulating sulfite metabolism[J]. Chinese Journal of Food Science,2022,22(11):299−308.] WANG M, WEI J, DI LI XIA TI Y L D N, et al. The inhibitory effect of sulfur dioxide on postharvest gray mold of grape by regulating sulfite metabolism[J]. Chinese Journal of Food Science, 2022, 22（11）: 299−308.
[9]	王凤超. 二氧化硫处理对鲜食葡萄贮藏病害及生理的影响[D]. 乌鲁木齐:新疆农业大学, 2013. [WANG F C. Effects of sulfur dioxide treatment on storage diseases and physiology of table grapes[D]. Urumqi:University of Xinjiang Agricultural, 2013.] WANG F C. Effects of sulfur dioxide treatment on storage diseases and physiology of table grapes[D]. Urumqi: University of Xinjiang Agricultural, 2013.
[10]	李梓童, 彭丽, 熊思国, 等. 1-MCP和SO₂处理对阳光玫瑰葡萄贮藏品质及风味的影响[J]. 包装工程,2023,44(11):64−77. [LI Z T, PENG L, XIONG S G, et al. Effects of 1-MCP and SO₂ treatments on storage quality and flavor of Sunny Rose grape[J]. Packaging Engineering,2023,44(11):64−77.] LI Z T, PENG L, XIONG S G, et al. Effects of 1-MCP and SO₂ treatments on storage quality and flavor of Sunny Rose grape[J]. Packaging Engineering, 2023, 44（11）: 64−77.
[11]	秦丹, 石雪晖, 林亲录, 等. 葡萄采后病害生物防治研究进展[J]. 食品与机械,2007,122(6):142−144. [QIN D, SHI X H, LIN Q L, et al. Research progress on biological control of postharvest diseases of grape[J]. Food and Machinery,2007,122(6):142−144.] QIN D, SHI X H, LIN Q L, et al. Research progress on biological control of postharvest diseases of grape[J]. Food and Machinery, 2007, 122（6）: 142−144.
[12]	JIANG C M, SHI J L, LIU Y L, et al. Inhibition of Aspergillus carbonarius and fungal contamination in table grapes using Bacillus subtilis[J]. Food Control,2014,35(1):41−48. doi: 10.1016/j.foodcont.2013.06.054
[13]	刘海波, 田世平, 秦国政, 等. 罗伦隐球酵母对葡萄采后病害的拮抗效果[J]. 中国农业科学,2002,43(7):831−835. [LIU H B, TIAN S P, QIN G Z, et al. Antagonistic effect of Cryptococcus lorenensis on postharvest diseases of grape[J]. Agricultural Science in China,2002,43(7):831−835.] doi: 10.3321/j.issn:0578-1752.2002.07.017 LIU H B, TIAN S P, QIN G Z, et al. Antagonistic effect of Cryptococcus lorenensis on postharvest diseases of grape[J]. Agricultural Science in China, 2002, 43（7）: 831−835. doi: 10.3321/j.issn:0578-1752.2002.07.017
[14]	李丽梅, 刘霞, 李喜宏, 等. 常温下黑曲霉对刺伤红提葡萄的致病规律研究及拮抗菌筛选[J]. 食品研究与开发,2020,41(15):35−39. [LI L M, LIU X, LI X H et al. Study on pathogenic rule of Aspergillus niger on punctured red grapes at room temperature and screening of antagonistic bacteria[J]. Food Research and Development,2020,41(15):35−39.] doi: 10.12161/j.issn.1005-6521.2020.15.007 LI L M, LIU X, LI X H et al. Study on pathogenic rule of Aspergillus niger on punctured red grapes at room temperature and screening of antagonistic bacteria[J]. Food Research and Development, 2020, 41（15）: 35−39. doi: 10.12161/j.issn.1005-6521.2020.15.007
[15]	高宇洁, 詹发强, 陈澄, 等. 黑曲霉拮抗菌Xenorhabdus bovienii 445筛选、鉴定及发酵优化[J]. 新疆农业科学,2022,59(5):1203−1215. [GAO Y J, ZHAN F Q, CHEN C, et al. Screening, identification and fermentation optimization of antagonistic Aspergillus niger bacterium Xenorhabdus bovienii 445[J]. Xinjiang Agricultural Sciences,2022,59(5):1203−1215.] GAO Y J, ZHAN F Q, CHEN C, et al. Screening, identification and fermentation optimization of antagonistic Aspergillus niger bacterium Xenorhabdus bovienii 445[J]. Xinjiang Agricultural Sciences, 2022, 59（5）: 1203−1215.
[16]	LI P Q, FENG B Z, YAO Z, et al. Antifungal activity of endophytic bacillus K1 against Botrytis cinerea[J]. Front Microbiol,2022,22(13):935675.
[17]	HAIDAR R, ROUDET J, BONNARD O, et al. Screening and modes of action of antagonistic bacteria to control the fungal pathogen Phaeomoniella chlamydospora involved in grapevine trunk diseases[J]. Microbiol Res,2016,10(192):172−184.
[18]	东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京:科学出版社, 2001:166−171. [DONG X Z, CAI M Y. Common bacteri-al system identification manual[M]. Beijing:Science Press, 2001:166−171.] DONG X Z, CAI M Y. Common bacteri-al system identification manual[M]. Beijing: Science Press, 2001: 166−171.
[19]	MULET M, GOMILA M, RAMÍREZ A, et al. Pseudomonas nosocomialis sp. nov. isolated from clinical specimens[J]. International Journal of Systematic and Evolutionary Microbiology,2019,69(11):3392−3398. doi: 10.1099/ijsem.0.003628
[20]	潘超, 高庆超, 梁颖, 等. 葡萄酒酿造乳酸菌的分离鉴定及糖苷酶活分析[J]. 中国食品学报,2023,23(4):345−353. [PAN C, GAO Q C, LIANG Y, et al. Isolation and identification of lactic acid bacteria from wine production and analysis of glycosidase activity[J]. Chinese Journal of Food Science,2023,23(4):345−353.] PAN C, GAO Q C, LIANG Y, et al. Isolation and identification of lactic acid bacteria from wine production and analysis of glycosidase activity[J]. Chinese Journal of Food Science, 2023, 23（4）: 345−353.
[21]	张丽娟, 黄伟, 王宁, 等. 耐辐射芽胞杆菌JK23的鉴定及其生防活性研究[J]. 中国生物防治学报,2021,37(5):997−1006. [ZHANG L J, HUANG W, WANG N, et al. Identification and biocontrol activity of radiation-tolerant Bacillus JK23[J]. Chinese Journal of Biological Control,2021,37(5):997−1006.] ZHANG L J, HUANG W, WANG N, et al. Identification and biocontrol activity of radiation-tolerant Bacillus JK23[J]. Chinese Journal of Biological Control, 2021, 37（5）: 997−1006.
[22]	全国信息与文献标准化技术委员会第6分委员会. GB 15193.3-2014 食品安全国家标准急性经口毒性试验[S]. 北京:中国标准出版社, 2014:1−6. [The sixth sub-committee of the National Technical Committee for Standardization of Information and Documentation. GB 15193.3-2014 National standard acute oral toxicity test for food safety[S]. Beijing:Standards Press of China, 2014:1−6.] The sixth sub-committee of the National Technical Committee for Standardization of Information and Documentation. GB 15193.3-2014 National standard acute oral toxicity test for food safety[S]. Beijing: Standards Press of China, 2014: 1−6.
[23]	王宁, 黄伟, 鲁致远, 等. 苹果树腐烂病生防链霉菌A144的鉴定及其代谢产物的抑菌活性[J]. 西北农业学报,2023,32(3):440−449. [WANG N, HUANG W, LU Z Y, et al. Identification of Streptomyces A144 in apple tree rot and antibacterial activity of its metabolites[J]. Northwest Agricultural Journal,2023,32(3):440−449.] WANG N, HUANG W, LU Z Y, et al. Identification of Streptomyces A144 in apple tree rot and antibacterial activity of its metabolites[J]. Northwest Agricultural Journal, 2023, 32（3）: 440−449.
[24]	WU S M, LIU G, ZHOU S N, et al. Characterization of Antifungal lipopeptide Biosurfactants produced by marine Bacterium Bacillus sp. CS30[J]. Mar Drugs,2019,17(4):199. doi: 10.3390/md17040199
[25]	JIA Q L, FAN Y Y, DUAN S S, et al. Effects of Bacillus amyloliquefaciens XJ-BV2007 on growth of Alternaria alternata and production of Tenuazonic Acid[J]. Toxins,2023,15(1):53. doi: 10.3390/toxins15010053
[26]	赖呈纯, 赖恭梯, 陈冰星, 等. 拮抗酵母菌的筛选鉴定、生物学特性及其对葡萄灰霉病菌的抑菌活性研究[J]. 中国生物防治学报,2023,39(2):1−14. [LAI C C, LAI G T, CHEN B X, et al. Screening, identification, biological characteristics and antibacterial activity of antagonistic yeast against Botrytis botrytis[J]. Chinese Journal of Biological Control,2023,39(2):1−14.] LAI C C, LAI G T, CHEN B X, et al. Screening, identification, biological characteristics and antibacterial activity of antagonistic yeast against Botrytis botrytis[J]. Chinese Journal of Biological Control, 2023, 39（2）: 1−14.
[27]	刘佳欣. 海洋枯草芽孢杆菌产抗菌脂肽类化合物的分离纯化及鉴定分析[D]. 大连:大连工业大学, 2018. [LIU J X. Isolation, purification and identification of antibacterial liptide compounds produced by Bacillus subtilis marinum[D]. Dalian:Dalian Polytechnic University, 2018.] LIU J X. Isolation, purification and identification of antibacterial liptide compounds produced by Bacillus subtilis marinum[D]. Dalian: Dalian Polytechnic University, 2018.
[28]	魏玉洁, 邹弯, 马文瑞, 等. 应用高通量测序技术研究新疆产区葡萄果实、叶片及果园土壤微生物多样性[J]. 食品科学,2018,39(6):162−170. [WEI Y J, ZHOU W, MA W R et al. High-throughput sequencing technology was used to study the microbial diversity of grape fruits, leaves and orchards in Xinjiang[J]. Food Science,2018,39(6):162−170.] doi: 10.7506/spkx1002-6630-201806026 WEI Y J, ZHOU W, MA W R et al. High-throughput sequencing technology was used to study the microbial diversity of grape fruits, leaves and orchards in Xinjiang[J]. Food Science, 2018, 39（6）: 162−170. doi: 10.7506/spkx1002-6630-201806026
[29]	张琦, 刘应敏, 杨东燕, 等. 贝莱斯芽孢杆菌SM2对番茄灰霉病的生防效果[J]. 中国瓜菜,2024,37(2):66−73. [ZHANG Q, LIU Y M, YANG D Y, et al. Biocontrol effect of Bacillus velezensis SM2 on gray mold of tomato[J]. Chinese Melon,2024,37(2):66−73.] ZHANG Q, LIU Y M, YANG D Y, et al. Biocontrol effect of Bacillus velezensis SM2 on gray mold of tomato[J]. Chinese Melon, 2024, 37（2）: 66−73.
[30]	YIN X T, LI T G, JIANG X L, et al. Suppression of grape white rot caused by Coniella vitis using the potential biocontrol agent Bacillus velezensis GSBZ09[J]. Pathogens,2022,11(2):248. doi: 10.3390/pathogens11020248
[31]	GIKAS E, BAZOTI F N, KATSIMARDOU, M, et al. Determination of colistin A and colistin B inhuman plasma by UPLC–ESI high resolution tandem MS:Application to a pharmacokinetic study[J]. Pharm Biomed Anal,2013,83(5):228−236.
[32]	RAVI A, NANDAYIPURATH V V T, RAJAN S, et al. Effect of zinc oxidenanoparticle supplementation on the enhanced production of surfactin and iturin lipopeptides of endophytic Bacillus sp. Fcl1 and its ameliorated antifungal activity[J]. Pest Manag Sci,2020,77(20):1035−1041.
[33]	孙静, 刘玉娟, 罗晓娇, 等. 杆菌霉素D和壳聚糖复合抑菌作用及其对砂糖橘保鲜效果[J/OL]. 食品工业科技:1−14[2024-04-30]. [SUN J, LIU Y J, LUO X J, et al. Bacteriostatic effect of bacitomycin D and chitosan on preservation of sugar orange[J/OL]. Food Industry Science and Technology:1−14[2024-04-30].] SUN J, LIU Y J, LUO X J, et al. Bacteriostatic effect of bacitomycin D and chitosan on preservation of sugar orange[J/OL]. Food Industry Science and Technology: 1−14[2024-04-30].
[34]	XIONG Z R, COBO M, WHITTAL R M, et al. Purification and characterization of antifungal lipopeptide produced by Bacillus velezensis isolated from raw honey[J]. PLoS One,2022,17(4):e0266470. doi: 10.1371/journal.pone.0266470

Supplements (0)

Cited By

Cited by

Periodical cited type(4)

1.	徐祥，谷贵章，尚佳宇，张进杰，林邦楚，谢红丰，徐大伦. 低盐雪菜中呈味核苷酸关联物的检测及贮藏过程中的变化. 食品科学. 2024(04): 315-322 .
2.	解双瑜，孙波，李智，柳凯，赵子伟. KCl替代NaCl对东北酸菜细菌菌群多样性的影响. 中国酿造. 2023(02): 58-62 .
3.	朱瑜，郑志，朱绍辉，候栋，陈璇，權英珠，朴丙熙，姜林娟. 干酪乳杆菌-山药复合发酵剂对溃疡性结肠炎小鼠肠道菌群的调节及其抗炎作用. 新乡医学院学报. 2023(05): 401-410 .
4.	卢志乐. 桂林家庭酸坛子腌制品食用安全探讨分析. 中国食品工业. 2022(22): 86-90 .