马铃薯疮痂病病原菌的分离鉴定及其生长特性分析

doi:10.6048/j.issn.1001-4330.2022.08.021

新疆农业科学 ›› 2022, Vol. 59 ›› Issue (8): 1999-2006.DOI: 10.6048/j.issn.1001-4330.2022.08.021

• 植物保护·农产品分析检测·农业装备工程与机械化 • 上一篇下一篇

马铃薯疮痂病病原菌的分离鉴定及其生长特性分析

宋素琴¹(), 高海峰², 吕卓¹^,³, 唐琦勇¹, 顾美英¹, 张志东¹, 楚敏¹, 朱静¹, 王玮¹

1.新疆农业科学院微生物应用研究所/新疆特殊环境微生物重点实验室,乌鲁木齐 830091
2.新疆农业科学院植物保护研究所/农业部西北荒漠绿洲作物有害生物综合治理重点实验室,乌鲁木齐 830091
3.新疆农业大学食品科学与药学学院,乌鲁木齐 830052

收稿日期:2021-09-28 出版日期:2022-08-20 发布日期:2022-10-01
通信作者: 宋素琴
作者简介:宋素琴（1976-）,女,副研究员,研究方向为微生物活性产物及作物病害,（E-mail） suqin song@163.com
基金资助:
农业部西北荒漠绿洲作物有害生物综合治理重点实验室开放基金资助课题(KFJJ202006);自治区公益性科研院所基本科研业务经费(KY2018012);自治区公益性科研院所基本科研业务经费(KY2019020)

Isolation and Identification of Pathogen of Potato Scab and Its Growth Characteristics from Zepu Area in Xinjiang

SONG Suqin¹(), GAO Haifeng², LV Zhuo¹^,³, TANG Qiyong¹, GU Meiying¹, ZHANG Zhidong¹, CHU Min¹, ZHU Jing¹, WANG Wei¹

1. Xinjiang Key Laboratory of Special Environmental Microorganisms/ Research Institute of Applied Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi, 830091, China
2. Key Laboratory of Intergraded Management of Harmful Crop Vermin of China Northwestern Oasis, MOARA, Institute of Plant Protection,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China
3. College of Food and Pharmaceutical Sciences, Xinjiang Agricultural University, Urumqi, 830052, China

Received:2021-09-28 Online:2022-08-20 Published:2022-10-01
Correspondence author: SONG Suqin
Supported by:
Open Fund for Key Laboratory of Integrated Pest Management on Crop in Northwestern Oasis, Ministry of Agriculture and Rural Affairs, China(KFJJ202006);Basic Scientific R &D Program of Public Welfare Research Institutions of Xinjiang Uygur Autonomous Region of China(KY2018012);Basic Scientific R &D Program of Public Welfare Research Institutions of Xinjiang Uygur Autonomous Region of China(KY2019020)

摘要/Abstract

摘要：

【目的】研究新疆喀什泽普县阿依库勒乡马铃薯疮痂病病原菌的种类及其生长特性。【方法】采用组织分离法结合柯赫氏法则、显微形态和16S rDNA等方法,分离鉴定马铃薯疮痂病病原菌;分析菌株对不同碳源、氮源的利用能力,以及不同pH的生长能力。【结果】新疆泽普马铃薯疮痂病的病原菌为疮痂链霉菌（Streptomyces acidiscabies）,产白色光滑呈自由弯曲状的孢子,能产生黑色素和可溶性色素,可利用葡萄糖等8种碳源和甲硫氨酸（Met）和组氨酸（His）2种氮源,接种健康马铃薯块茎上能产生网纹状甚至凹陷裂口状病斑。【结论】该地区马铃薯疮痂病病原菌最适生长温度为30℃,pH为7,最适碳氮源为葡萄糖和甲硫氨酸,全黑暗利于菌株的生长。

关键词: 马铃薯疮痂病; 分离鉴定; 杀菌剂; 拮抗菌

Abstract:

【Objective】 The purpose of this study is to determine the species and growth characteristics of potato scab pathogen in Aikule Township, Kashgar Prefecture, Xinjiang, for the potato scab disease affected the yield and quality of potato seriously, and caused great losses to potato industry, especially to virus-free seed potato in Xinjiang. 【Method】 The pathogen was isolated and identified by tissue separation method combined with Koch's rule, microscopic morphology and 16 S rDNA. Then different carbon sources and nitrogen sources were selected to test its growth characteristics. 【Result】 Based on its morphological characteristics and 16S rDNA sequence, the pathogen of this disease was identified as Streptomyces acidiscabies from this area, which produced white-smooth curved spores with melanin and soluble pigment. Eight kinds of the carbon sources and two kinds of nitrogen sources were tested its growth period, such as glucose, methionine（Met） and histidine（His） etc. It could produce reticulate netlike scab after inoculatedon healthy potato tubers by smearing the tuber skin with spores suspension. The optimum growth temperature was 30℃, the pH was 7, and the optimum carbon and nitrogen sources were glucose and methionine separately. 【Conclusion】 The pathogen of the potato scab are isolated and identified from this area, and its growth characteristics were proved in this study, which would be the beneficial research basis for controlling this disease in Xinjiang.

Key words: potato common scab; isolation and identification; germicide; antagonists

中图分类号:

S436.22

宋素琴, 高海峰, 吕卓, 唐琦勇, 顾美英, 张志东, 楚敏, 朱静, 王玮. 马铃薯疮痂病病原菌的分离鉴定及其生长特性分析[J]. 新疆农业科学, 2022, 59(8): 1999-2006.

SONG Suqin, GAO Haifeng, LV Zhuo, TANG Qiyong, GU Meiying, ZHANG Zhidong, CHU Min, ZHU Jing, WANG Wei. Isolation and Identification of Pathogen of Potato Scab and Its Growth Characteristics from Zepu Area in Xinjiang[J]. Xinjiang Agricultural Sciences, 2022, 59(8): 1999-2006.

图/表 9

表1 常规PCR反应体系的扩增

Table 1 16s rDNA sequence Amplification of conventional PCR reaction system

Form	Volume（µL）
2XMix	25
F(10 μM)	1
R(10 μM)	1
Bacterial templates	1
ddH₂O	23

图 1 菌株ZPCJ-1的形态特征注： A：菌落正面 B：菌落背面 C：菌株显微（100×） D：菌株显微（40×）

Fig. 1 Morphological characteristics of ZPCJ-1 Note： A：positive colony map of strain; B：reverse colony map of strain; C：micromorphology of strain（100×）;D：micromorphology of strain（40×）

图 2 菌株ZPCJ-1的致病性注：A：接种菌株的马铃薯表面图;B：打孔器法;C、F：菌株对马铃薯表面的致病性;D：针刺法;E：在离体叶片上的实验结果（120 d）

Fig. 2 Determination of pathogenicity of pathogenic bacteria ZPCJ-1 Note：A：Surface map of potato inoculated with bacteria;B：the punch method;C、F：the pathogenicity of the pathogen to the potato skin;D：the acupuncture method;E：experiment on isolated leaves（120 d）

表 2 菌株ZPCJ-1的生物学特性

Table 2 Biological characteristics of ZPCJ-1

Cultural characteristic	Result	Cultural characteristic	Result
Spore morphology	Smooth, free to bend	Inose	++
Spore color	white	Utilization of nitrogen sources
Melanin	+++	Met	+++
Soluble pigment	+++	His	+
Utilization of carbon sources		Sensitivity experiment
Amylaceum	+++	Streptomycin	+++
Saccharose	+++	Penicillin	+++
Fructopyranose	++	Phenol	+++
Wood sugar	++	Crystal violet	+++
Mannitol	++	Starch utilization	+++
Arabinose	++	pH	4.0
Rhamnose	++	/	/

图 3 不同温度下菌株ZPCJ-1生长变化注：图中小写字母表示0.05水平差异显著性,下同

Fig. 3 Effect of temperature on ZPCJ-1 growth of strain Note：Differentlowercaselettersindicatesignificant differenceat P <0.05level,the same as below.

图 4 不同光照下菌株ZPCJ-1生长变化

Fig. 4 Effect of light condition on ZPCJ-1 growth of strain

图 5 不同pH下菌株ZPCJ-1生长变化

Fig. 5 Effect of pH on ZPCJ-1 growth of strain

图 6 不同碳氮源下菌株ZPCJ-1生长变化

Fig. 6 Effect of carbon and nitrogen sources on ZPCJ-1 growth of strain

图 7 用于ZPCJ-1鉴定的PCR扩增电泳图谱及系统发育树构建

Fig. 7 PCR amplification electrophoresis map and construction of phylogenetic tree for identification of ZPCJ-1

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马铃薯疮痂病病原菌的分离鉴定及其生长特性分析

Isolation and Identification of Pathogen of Potato Scab and Its Growth Characteristics from Zepu Area in Xinjiang

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