新疆农业科学 ›› 2022, Vol. 59 ›› Issue (7): 1726-1733.DOI: 10.6048/j.issn.1001-4330.2022.07.019
收稿日期:
2021-09-18
出版日期:
2022-07-20
发布日期:
2022-08-04
通信作者:
郭庆元
作者简介:
张卢慧(1995-),女,新疆博乐人,硕士研究生,研究方向为作物病害及防治,(E-mail) 1421023812@qq.com
基金资助:
ZHANG Luhui(), ZHAO Zhiqiang, GUO Qingyuan()
Received:
2021-09-18
Online:
2022-07-20
Published:
2022-08-04
Correspondence author:
GUO Qingyuan
Supported by:
摘要:
【目的】研究鉴定2018年在新疆巴音郭楞蒙古自治州和硕县辣椒种植区内新发现一种辣椒细菌性果实条斑病病原,为该病田间病害规律研究和病害防治提供依据。【方法】采用组织分离法、柯赫氏法则回接证病、形态学观察、生理生化特性测定、16S rDNA分子鉴定等方法,鉴定该病病原菌,运用针刺接种法对辣椒果面进行致病性回接测定。【结果】从田间采集的发病辣椒果面上分离、纯化共获得30个单菌落;获得了4个致病性强的菌株,4个菌株形态特征及生理生化测定结果一致。4个致病菌株均与已报道的阿根廷假单胞菌Pseudomonas argentinensis聚在一个进化发育分支中,并且与模式菌株P. argentinensis CH01T (登录号:AY691188)的相似性达到99.2%。【结论】新疆和硕县辣椒细菌性果实条斑病的病原菌鉴定为P. argentinensis。
中图分类号:
张卢慧, 赵志强, 郭庆元. 辣椒细菌性果实条斑病病原鉴定[J]. 新疆农业科学, 2022, 59(7): 1726-1733.
ZHANG Luhui, ZHAO Zhiqiang, GUO Qingyuan. Identification of Pathogen Causing Fruit Bacterial Streak on Pepper[J]. Xinjiang Agricultural Sciences, 2022, 59(7): 1726-1733.
图1 辣椒细菌性果实条斑病田间发病症状、回接症状及烟草过敏性反应结果 注:a、b:田间自然侵染条件下辣椒表面发病症状;c:回接症状;d:CK;e:菌株烟草过敏性反应
Fig.1 Symptoms of Bacterial Leaf Spot of Pepper in the Field and Back to symptoms and tobacco allergic reaction results Note: a, b: Symptoms of surface disease on pepper under natural infestation in the field; c:Symptoms of tieback; d: control; e: Strain tobacco allergic reaction
图2 病原菌代表菌株 KHL1的形态特征 注:a:菌落在NA培养基上的形态;b:菌体形态;c:菌体大小
Fig.2 Morphological characteristics of the representative isolate KHL1 straina Note: a: The morphology of the colony on NA medium; b: Gram stain results;C: Size of bacteria
项目 Items | 菌株 Strains | 项目 Items | 菌株 Strains | ||||
---|---|---|---|---|---|---|---|
1-4 | 5 | 6 | 1-4 | 5 | 6 | ||
革兰氏染色Gram stain | - | - | - | 海藻糖Trehalose | + | + | + |
氧化酶Oxidase | + | + | + | 半乳糖Galactose | + | + | - |
接触酶Hydrogen peroxidase | + | + | + | 麦芽糖Maltose | + | + | - |
硝酸盐还原Nitratreduction | + | + | - | 阿拉伯糖Arabinose | + | + | + |
甲基红测定Methylred test | - | ND | - | 纤维二糖Cellobiose | - | - | - |
V-P 测定VogesProskauer | - | ND | - | 菊糖Inulin | - | - | - |
吲哚Indole | - | - | - | 肌醇Inositol | - | - | + |
明胶液化Gelatinliquefaction | - | - | - | 木糖xylose | - | - | - |
过氧化氢酶Catalase | + | + | + | 蔗糖Sucrose | + | - | + |
4度生长Under 4℃ growth | - | - | - | 山梨醇Inositol | - | - | - |
D-果糖 D-fructose | + | + | + | 赤藓糖醇Meso-erythritol | - | - | - |
葡萄糖Glucose | + | + | + | 甘露醇Mannitol | + | + | - |
表1 供试4个菌株生理生化特性测定结果
Table 1Phys iological and biochemical characteristics of 4 strains tested
项目 Items | 菌株 Strains | 项目 Items | 菌株 Strains | ||||
---|---|---|---|---|---|---|---|
1-4 | 5 | 6 | 1-4 | 5 | 6 | ||
革兰氏染色Gram stain | - | - | - | 海藻糖Trehalose | + | + | + |
氧化酶Oxidase | + | + | + | 半乳糖Galactose | + | + | - |
接触酶Hydrogen peroxidase | + | + | + | 麦芽糖Maltose | + | + | - |
硝酸盐还原Nitratreduction | + | + | - | 阿拉伯糖Arabinose | + | + | + |
甲基红测定Methylred test | - | ND | - | 纤维二糖Cellobiose | - | - | - |
V-P 测定VogesProskauer | - | ND | - | 菊糖Inulin | - | - | - |
吲哚Indole | - | - | - | 肌醇Inositol | - | - | + |
明胶液化Gelatinliquefaction | - | - | - | 木糖xylose | - | - | - |
过氧化氢酶Catalase | + | + | + | 蔗糖Sucrose | + | - | + |
4度生长Under 4℃ growth | - | - | - | 山梨醇Inositol | - | - | - |
D-果糖 D-fructose | + | + | + | 赤藓糖醇Meso-erythritol | - | - | - |
葡萄糖Glucose | + | + | + | 甘露醇Mannitol | + | + | - |
图3 菌株KHL1、KHL4、KHL6、KHL9 16SrDNA扩增 注:M:2000 D Ladder marker;1-4:菌株KHL1、KHL4、KHL6、KHL9 16SrDNA扩增结果;CK:阴性对照
Fig.3 The results of 16SrDNA amplification of strains KHL1、KHL4、KHL6、KHL9 Note: M: 2000 D Ladder marker; 1-4:16SrDNA amplification results of strains KHL1、 KHL4、KHL6、 KHL9; CK: negative control
图4 供试菌株KHL1、KHL4、KHL6、KHL9基于16SrDNA基因序列的系统发育树
Fig.4 The phylogenetic tree of the tested strains KHL1, KHL4, KHL6, KHL9 based on the 16S rDNA gene sequence
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