Evaluation of Control Efficacy of Enterococcus mundtii ZC-05 against Bacterial Canker of Kiwifruit and Preliminary Study on Its Biocontrol Mechanism

doi:10.16409/j.cnki.2095-039x.2026.02.014

Abstract

Abstract: Kiwifruit bacterial canker, often termed the “cancer” of kiwifruit, currently lacks effective control methods. In this study, a potential biocontrol strain Enterococcus mundtii ZC-05, previously isolated from stem of Kiwifruit, was labeled with green fluorescent protein(GFP) to investigate its colonization characteristics. Results indicated that protoplast fusion was an effective method for transforming strain ZC-05, successfully yielding the GFP-labeled transformant ZC-05-GFP. Confocal microscopy observation revealed that ZC-05-GFP colonized kiwifruit stems two weeks after trunk injection and reach the leaf via the sap flow. Field trials demonstrated that injection of ZC-05 suspension significantly reduced the incidence of kiwifruit bacterial canker, achieving a control efficacy of 50.96%, which was substantially higher than that of the copper-based bactericide Thiodiazole-Copper(13.46%)(P<0.05). Whole-genome sequencing of ZC-05 identified seven genes encoding type VI secretion system(T6 SS) effector proteins. Furthermore, quantitative real-time PCR analysis indicated that the gene Gm-1574, encoding an antibacterial toxic protein, was significantly up-regulated after co-culture with Pseudomonas syringae pv. actinidiae(Psa-Y), the pathogen of Kiwifruit bacterial canker, suggesting its potential important role in the biocontrol activity of ZC-05. This study clarifies the colonization characteristics of ZC-05, confirms its field efficacy against kiwifruit bacterial canker, and preliminarily elucidates its biocontrol mechanisms. These findings provide a basis for the development and practical application of trunk-injected biocontrol agents in the future.

Key words: kiwifruit bacterial canker, Enterococcus mundtii, GFP labeling, biocontrol mechanism

CLC Number:

S476

GU Jiaying, YIN Zhicheng, LI Nanyi, CAI Xiaojian, ZHANG Xin. Evaluation of Control Efficacy of Enterococcus mundtii ZC-05 against Bacterial Canker of Kiwifruit and Preliminary Study on Its Biocontrol Mechanism[J]. Chinese Journal of Biological Control, 2026, 42(3): 611-620.

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References

[1] Asif M, Liang S, Renjian H, et al. A stitch in time:Sustainable and eco-friendly solutions for kiwifruit bacterial canker[J]. Physiological and Molecular Plant Pathology, 2024, 136(1):102506.
[2] Gan K, Sawa T, Ayumi I, et al. Evaluation of various cultivars of Actinidia species and breeding source Actinidia rufa for resistance to Pseudomonas syringae pv. actinidiae biovar 3[J]. Journal of General Plant Pathology, 2018, 84(6):399-406.
[3] 钟曼茜,翟舒嘉,刘伟,等.我国即食猕猴桃产业发展现状、问题与对策[J].中国果树, 2023, 65(2):122-127.
[4] Pereira C, Costa P, Pinheiro L, et al. Kiwifruit bacterial canker:an integrative view focused on biocontrol strategies[J]. Planta, 2021, 253(2):49.
[5] Zhang Y, Chen Y, Ma J, et al. Controlling citrus Huanglongbing based on soil remediation and biocontrol[J]. European Journal of Plant Pathology, 2024,169(2):379-393.
[6] 田涛,亓雪晨,王琦,等.芽孢杆菌绿色荧光蛋白标记及其在小麦体表定殖的初探[J].植物病理学报, 2004, 34(4):346-351.
[7] Stuurman N, Bras C P, Schlaman H R M, et al. Use of green fluorescent protein color variants expressed on stable broad host range vectors to visualize rhizobia interacting with plants[J]. The American Phytopathological Society, 2000, 13(11):1163-1169.
[8] Prieto P, Mercado-blanco J. Endophytic colonization of olive roots by the biocontrol strain Pseudomonas fluorescens PICF7[J]. FEMS Microbiology Ecology, 2008, 64(2):297-306.
[9] 杨潇湘,黄小琴,张蕾,等.绿色荧光蛋白标记解淀粉芽孢杆菌Bam22在油菜体内的定殖[J].中国农学通报, 2022, 38(1):125-130.
[10] Tian R, Tian Y, Mi Q, et al. Histocytological analysis reveals the biocontrol activity of a rhizospheric bacterium Pseudomonas rhizophila Z98 against kiwifruit bacterial canker[J]. Pesticide Biochemistry and Physiology, 2025, 208(1):106251.
[11] Wang H., Wang N., Tan Y, et al. Paenibacillus polymyxa YLC1:a promising antagonistic strain for biocontrol of Pseudomonas syringae pv. actinidiae,causing kiwifruit bacterial canker[J]. Pest Management Science, 2023, 79(11):4357-4366.
[12] Li W, Lee S Y, Cho Y J, et al. Mediation of induced systemic resistance by the plant growth-promoting rhizobacteria Bacillus pumilus S2-3-2[J].Molecular Biology Reports, 2020, 47(11):8429-8438.
[13] Shao X, Wu Q, Li L, et al. Adapting the inoculation methods of kiwifruit canker disease to identify efficient biocontrol bacteria from branch microbiome[J]. Molecular Plant Pathology, 2024, 25(1):e13399.
[14] 尹志诚,顾佳颖,李南羿,等.猕猴桃溃疡病生防菌群的构建及其生防效应[J].林业科学, 2024, 60(12):101-110.
[15] Peter A M, Lara B, Ben C, et al. Re-programming of Pseudomonas syringae pv. actinidiae gene expression during early stages of infection of kiwifruit[J].BioMed Central Genomics, 2018, 19(1):822.
[16] 李春玲,宋赵依,宗兆锋.非致病性尖镰孢FO47的原生质体融合改良研究[J].西北农林科技大学学报(自然科学版), 2010, 38(10):126-130.
[17] Teh B S, Apel J, Shao Y, et al. Colonization of the intestinal tract of the polyphagous pest Spodoptera littoralis with the GFP-tagged indigenous gut bacterium Enterococcus mundtii[J]. Frontiers in Microbiology, 2016(1), 7:928.
[18] 匡美美,高建有,罗庆,等.湘西州猕猴桃溃疡病发病情况及其影响因素分析[J].湖南农业科学, 2022, 52(1):56-59.
[19] 杨玉文,刘德华,王成梁,等.瓜类细菌性果斑病拮抗菌ZY1的鉴定及其生物学特性[J].中国生物防治学报, 2024, 40(6):1430-1438.
[20] Rio D C, Ares M J R, Hannon G J, et al. Purification of RNA using TRIzol(TRI reagent)[J]. Cold Spring Harbor Protocols, 2010, 2010(6):5439.
[21] Han C, Gu H, Wang J, et al. Regulation of L-Threonine dehydrogenase in somatic cell reprogramming[J]. Stem Cells, 2013, 31(5):953-65.
[22] Chen Q, Yu J J, He J, et al. Isobenzofuranones and isocoumarins from kiwi endophytic fungus Paraphaeosphaeria sporulosa and their antibacterial activity against Pseudomonas syringae pv. actinidiae[J]. Phytochemistry, 2022, 195(1):113050.
[23] De Jong H, Reglinski T, Elmer P A G, et al. Integrated use of Aureobasidium pullulans strain CG163 and acibenzolar-S-methyl for management of bacterial canker in kiwifruit[J]. Plants, 2019, 8(8):287.
[24] Rheinländer P A, Sutherland P W, Elmer P A G. Visualisation of the mode of action of a biological control agent, Aureobasidium pullulans(strain YBCA5)against Pseudomonas syringae pv. actinidiae biovar 3 on the kiwifruit phylloplane[J]. Australasian Plant Pathology, 2021, 50(4):379-388.
[25] Bai J, Liu Y, Liu M, et al. Application of phage therapy against red-fleshed kiwifruit canker[J]. Biological Control, 2022, 169:104893.
[26] 朱海云,马瑜,柯杨,等.猕猴桃细菌性溃疡病生防菌的筛选、鉴定及其防效初探[J].微生物学杂志, 2016, 36(5):90-96.
[27] Biondi E, Schiavi D, Perez F S, et al. Bacillus amyloliquefaciens subsp. plantarum strain D747 to control the kiwifruit bacterial canker disease[J].Proceedings of the X International Symposium on Kiwifruit, 2022, 1332(13):95-102.
[28] Ali M A, Luo J, Ahmed T, et al. Pseudomonas bijieensis strain XL17 within the P. corrugata subgroup producing 2, 4-diacetylphloroglucinol and lipopeptides controls bacterial canker and gray mold pathogens of kiwifruit[J]. Microorganisms, 2022, 10(2):425.
[29] 杜贞娜,晏子英,候忠余,等.猕猴桃溃疡病病原菌的鉴定及生防菌的筛选[J].西南农业学报, 2021, 34(4):755-761.
[30] Daranas N, RosellóG, Cabrefiga J, et al. Biological control of bacterial plant diseases with Lactobacillus plantarum strains selected for their broad-spectrum activity[J].The Annals of applied biology, 2019, 174(1), 92-105.
[31] 殷幼平,袁训娥,李强,等.生防菌枯草芽孢杆菌CQBS03的绿色荧光蛋自基因标记及其在柑橘叶片上的定殖[J].中国农业科学, 2010, 43(17):3555-3563.
[32] Pukatzki S, Ma A T, Sturtevant D, et al. Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system[J]. Proceedings of The National Academy of Sciences, 2006, 103(5):1528-1533.
[33] Bernal P, Allsopp L P, Filloux A, et al. The Pseudomonas putida T6SS is a plant warden against phytopathogens[J]. The International Society for Microbial Ecology Journal, 2017, 11(4):972-987