Identification of new pathogen of walnut black spot disease in Shaanxi Province and screening of laboratory agents

doi:10.14088/j.cnki.issn0439-8114.2024.07.010

Abstract

Abstract: The pathogenic bacteria of black spot disease on the diseased leaves of Juglans regia ‘Robert Livermore' were isolated, and the pathogenic bacteria were identified by Koch's test, and the status of the pathogenic bacteria in taxonomy was determined by pathogen biochemical tests and molecular biology techniques. The results showed that 23 bacterial strains with the consistent colony morphology were isolated from the diseased leaves of the red kernel walnut in the field, and Koch's tieback test was performed on randomly selected strains to cause the disease of red kernel walnut leaves. Through physiological and biochemical phenotype tests, combined with multi-site sequence analysis based on 16S rDNA, gyrB and rpoD gene sequences, the pathogen was identified as Pseudomonas amygdali. Then 12 kinds of agents were tested for indoor toxicity, and 6 agents having an obvious inhibitory effect on the pathogen were screened out. Among them, tetramycin had the best bacteriostatic effect, with an EC₅₀ of 50.353 4 μg/mL, and the bacteriostatic effect of other agents was zhongshengmycin > kasugamycin > thiodiazole-copper > thiazole zinc > copper hydroxide.

Key words: Juglans regia ‘Robert Livermore', walnut black spot disease, Pseudomonas amygdali, new pathogens, screening of laboratory agents, Shaanxi Province

CLC Number:

S436.64

LEI Qiong, LIN Xin, WANG Wen-jun, QU Jia-nan, ZHANG Zhi-you. Identification of new pathogen of walnut black spot disease in Shaanxi Province and screening of laboratory agents[J]. HUBEI AGRICULTURAL SCIENCES, 2024, 63(7): 67-71.

References

[1] 李敏,刘媛,孙翠,等.核桃营养价值研究进展[J].中国粮油学报,2009,24(6):166-170.
[2] 李忠新,杨莉玲,阿布力孜·巴斯提,等.中国核桃产业发展研究[J].中国农机化学报,2013,34(4):23-28.
[3] 张毅,刘斌,陈新乐,等.商洛核桃主要病虫害及防治对策[J].陕西林业科技,2021,49(2):76-81.
[4] 董兆斌,王根宪,王英宏.洛南县美国红仁核桃引种初报[J].陕西林业科技,2017(1):25-28.
[5] BELISARIO A,ZOINA A,PEZZA L,et al.Susceptibility of species of Juglans to pathovars of Xanthomonas campestris[J].European journal of forest pathology,1999,29(1):75-80.
[6] SCOTTON M,BORTOLIN E,FIORIN A,et al.Environmental and pathogenic factors inducing brown apical necrosis on fruit of English (Persian) walnut[J].Phytopathology,2015,105(11):1427-1436.
[7] SCORTICHINI M,MARCHESI U,DI PROSPERO P.Genetic diversity of Xanthomonas arboricola pv. juglandis (synonyms: X. campestris pv. juglandis;X. juglandis pv. juglandis) strains from different geographical areas shown by repetitive polymerase chain reaction genomic fingerprinting[J].Journal of phytopathology,2001,149(6):325-332.
[8] 宫永红. 核桃细菌性黑斑病研究进展[J].北方果树,2012(6):1-4.
[9] FRUTOS D.Bacterial diseases of walnut and hazelnut and genetic resources[J].Journal of plant pathology, 2010,92(1):79-85.
[10] 陈善义,陶万强,王合,等.北京地区核桃黑斑病病原菌的分离、致病性测定和16S rDNA序列分析[J].果树学报,2011,28(3):469-473,549.
[11] 曲文文. 山东省核桃(Juglans regia)主要病害病原鉴定[D].山东泰安:山东农业大学,2011.
[12] 王琳莹. 石棉县核桃黑斑病与炭疽病病原鉴定及其防治技术研究[D].四川雅安:四川农业大学,2016.
[13] 王瀚,王让军,田凤鸣,等.陇南核桃致病性成团泛菌的分离鉴定及其致病性研究[J].福建农业学报,2016,31(10):1086-1090.
[14] 瞿佳,门欣,孙晓宇,等.陕西核桃黑斑病病原菌鉴定及药剂防治研究[J].西北农业学报,2021,30(3):452-461.
[15] 李亚,张珍荫,肖波,等.核桃细菌性黑斑病病原分子鉴定及室内药剂筛选[J].西南林业大学学报(自然科学),2022, 42(5):104-110.
[16] GARRITY G,BELL J,LILBURN T.Bergey's manual of systematic bacteriology[M].Berlin,Germany:Springer verlag,2004.
[17] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.370-384.
[18] CLADERA A,BENNASAR A,BARCELÓ M,et al.Comparative genetic diversity of Pseudomonas stutzeri genomovars, clonal structure, and phylogeny of the species[J].Journal of bacteriol,2004,186(16):5239-5248.
[19] MULET M,BENNASAR A,LALUCAT J,et al.An rpoD-based PCR procedure for the identification of Pseudomonas species and for their detection in environmental samples[J].Molecular and cellular probes,2009,23(3-4):140-147.
[20] KUMAR S,STECHER G,TAMURA K.MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Molecular biology and evolution,2016,33(7),1870-1874.
[21] MALEKI-ZADEH H,FALAHI C,KHODAYGAN P,et al.Bacterial leaf spot and die-back of hazelnut caused by a new pathovar of Pseudomonas amygdali[J].European journal of plant pathology,2022,163:293-303.
[22] LACOBELLIS N S,EVIDENTE A,SURICO G,et al.Production of phytohormones by Pseudomonas amygdali and their role in the hyperplastic bacterial canker of almond[J].Journal of pathology,1990,129(3):177-186.
[23] LAMICHHANE J, MESSÉAN A, MORRIS C. Insights into epidemiology and control of diseases of annual plants caused by the Pseudomonas syringae species complex[J].Journal of general plant pathology,2015,81:331-350.
[24] SŁOMNICKA R,OLCZAK-WOLTMAN H,OSKIERA M,et al. Genome analysis of Pseudomonas syringae pv. lachrymans strain 814/98 indicates diversity within the pathovar[J].European journal of plant pathology,2018,151:663-676.
[25] KASHIHARA S,NISHIMURA T,NOUTOSHI Y,et al.HopAZ1, a type III effector of Pseudomonas amygdali pv. tabaci, induces a hypersensitive response in tobacco wildfire-resistant Nicotiana tabacum ‘N509'[J].Molecular plant pathology,2022,23(6):885-894.