阿克苏地区枣黑斑病菌 PCR 检测方法的建立与应用

doi:10.6048/j.issn.1001-4330.2018.09.013

新疆农业科学 ›› 2018, Vol. 55 ›› Issue (9): 1674-1681.DOI: 10.6048/j.issn.1001-4330.2018.09.013

• 植物保护·土壤肥料·农业生态环境 • 上一篇下一篇

阿克苏地区枣黑斑病菌 PCR 检测方法的建立与应用

白剑宇^1,2, 毕司进², 宋峰惠¹, 史彦江¹

1.新疆林业科学院经济林研究所,乌鲁木齐 830063;
2.新疆农业大学农学院,乌鲁木齐 830052

收稿日期:2018-08-29 出版日期:2018-09-20 发布日期:2018-12-07
通信作者: 史彦江(1961-),男,山西人,研究员,研究方向为林木引种育种,(E-mail)syj504@126.com
作者简介:白剑宇(1977-),河北承德人,博士后,研究方向为林业有害生物防控,(E-mail)bjyhao2010@sina.com
基金资助:
中国博士后科学基金面上项目“新疆红枣黑斑病菌的田间追踪检测技术研究”(2014M552565XD);新疆维吾尔自治区林业发展补助资金项目“南疆红枣主产区红枣主要病害普查鉴定及田间追踪监测技术研究”(XJLKY2016056)

Development and Application of PCR for Detection of Alternaria alternata Causing Jujube Black Spot in Aksu

BAI Jian-yu^1,2, BI Si-jin², SONG Feng-hui¹, SHI Yan- jiang¹

1.Research Institute of Economic Forestry,Xinjiang Academy of Forestry Sciences,Urumqi 830063,China;
2. College of , Xinjiang Agricultural University, Urumqi 830052, China

Received:2018-08-29 Online:2018-09-20 Published:2018-12-07
Correspondence author: SHI Yan-jiang, (1961-), male, native place: Shanxi. Professor, research field: Forestry pest control, (E-mail)syj504@126.com
Supported by:
China Postdoctoral Science Foundation Project" The Tracking and Detection Technology of Pathogen of the Black Spot of Jujube in Fields in Xinjiang" (2014M552565XD) and Forestry Development Grants Fund Project of Xinjiang Uyghur Autonomous Region "The Main Jujube Diseased Census Identification and Tracking Detection Technology in Fields in Main Producing Areas in Southern Xinjiang" (XJLKY2016056)

摘要/Abstract

摘要： 【目的】建立枣黑斑病菌的分子检测方法,研究枣黑斑病菌田间侵染动态,分析病害防治的最佳时间,为枣黑斑病的高效防治奠定基础。【方法】利用文献公布的枣黑斑病菌(Alternaria alternata) HSP70序列特异性引物,通过PCR技术进行引物特异性验证与灵敏度检测,优化反应体系,建立病原菌分子检测技术,并应用该技术确定枣黑斑病菌侵染动态监测和越冬场所。【结果】该检测技术对枣黑斑病菌的检测的最低浓度为4.886 pg/μL,可在病害症状未显症之前检测到病原菌,对病原菌的越冬场所进行检测与验证病果、病叶及冠下表层土壤是枣黑斑病菌越冬的主要场所。【结论】建立的枣黑斑病菌检测技术能够快速准确的检测和鉴定病原菌,可应用于枣黑斑病菌田间侵染动态和越冬场所的监测与验证,为阿克苏地区枣黑斑病的流行监测和早期防治提供了技术支持。

关键词: 枣黑斑病; HPS70; 分子检测; 侵染动态

Abstract: 【Objective】 The purpose of this project is to develop a molecular detection method of jujube black spot in order to carry out the detection and analysis of field infection dynamic of jujube black spot, so that we can find the optimal time for disease prevention and lay the foundation for the efficient control of the jujube black spot.【Method】The molecular detection technique was established by using unconcealed specific primers of black spot pathogen (Alternaria alternata) HSP70 gene after the sensitivity test by PCR technology and optimized reaction system. The infection dynamic of jujube black spot and the overwinter sites were detected and determined using the molecular detection technique. 【Result】The test results showed that the lowest detection concentration of the jujube black spot pathogen was 4.886 pg/μL and the pathogen could be detected before the symptoms appearance. Meanwhile, the overwintering sites of pathogen could be detected and validated by using this molecular detection technique. Thus, it was proved that the soil surface, diseased leaves, and diseased fruit are the main overwintering sites of jujube black spot.【Conclusion】The established technology can quickly and accurately detect and identify the pathogen, and can be applied to the field infection dynamics of jujube black spot pathogen and the monitoring and verification of the overwintering site. It has provided technical support for epidemic monitoring and early prevention and control of jujube black spot in Aksu area.

Key words: jujube black spot; HPS70; molecular detection; infection dynamics

白剑宇, 毕司进, 宋峰惠, 史彦江. 阿克苏地区枣黑斑病菌 PCR 检测方法的建立与应用[J]. 新疆农业科学, 2018, 55(9): 1674-1681.

BAI Jian-yu, BI Si-jin, SONG Feng-hui, SHI Yan- jiang. Development and Application of PCR for Detection of Alternaria alternata Causing Jujube Black Spot in Aksu[J]. Xinjiang Agricultural Sciences, 2018, 55(9): 1674-1681.

参考文献

[1] 白剑宇,刘艳祥,刘正兴,等. 阿克苏地区枣黑斑病病原鉴定及室内药剂初选[J]. 新疆农业科学,2016,53(9):1 674-1 682.
   BAI Jian-yu, LIU Yan-xiang, LIU Zheng-xing, et al. (2016). Pathogens identification and Fungicides Screening of black spot of jujube indoors in Aksu [J]. Xinjiang Agricultural Sciences, 53(9):1,674-1,682. (in Chinese)
[2] 林忠敏,赵晓军,赵可俊,等. 红枣果实黑斑病的病原分离和鉴定[J].山西农业科学, 2001,29(1) :74-77.
   LIN Zhong-min, ZHAO Xiao-jun, ZHAO Ke-jun, et al. (2001). Red dates fruit black spot The pathogen isolation and identification of fruits black spot of jujube [J]. Journal of Shanxi Agricultural Sciences, 29(1): 74-77. (in Chinese)
[3] 吴玉柱, 黄亚丽谢晨星, 等.冬枣黑斑病病原菌的鉴定[J].中国森林病虫, 2005,(2):1-3.
   WU Yu-zhu, HUANG Ya-li, XIE Chen-xing, et al. (2005). Pathogens identification of the black spot of winter jujube [J]. China Forest Pest and Disease, (2): 1-3. (in Chinese)
[4] 宗淑萍, 杨文香刘大群, 等.冬枣黑点病病原鉴定[J].华北农学报, 2006,(5):105-107.
   ZONG Shu-ping, YANG Wen-lei, LIU Da-qun, et al. (2006). Pathogens identification of black spot of winter jujube[J]. Acta Agriculturae Boreali-Sinica, (5): 105-107. (in Chinese)
[5] 李晓军, 张勇,曲健禄, 等. 杀菌剂对冬枣黑斑病病原细菌的室内抑菌效果[J]. 落叶果树, 2004,(2):50-51.
   LI Xiao-jun, AHANG Yong, QU Jian-lu, et al. (2004). Fungicides indoor antibacterial effect on the pathogens of the black spot of jujube [J]. Deciduous Fruits, (2): 50-51. (in Chinese)
[6] 宗淑萍, 杨文香, 刘大群, 等.冬枣黑点病病原鉴定[J].华北农学报, 2006,(5):105-107.
   ZONG Shu-ping, YANG Wen-lei, LIU Da-qun, et al. (2006). Pathogens identification of black spot of winter jujube [J]. Acta Agriculturae Boreali-Sinica, (5): 105-107. (in Chinese)
[7] 王军,辛玉成. 冬枣黑斑病病原的分离与鉴定[J].青岛农业大学学报, 2007,(1):21-23.
   WANG Jun, XIN Yu-cheng. (2007). Isolation and identification of pathogens of winter jujube black spot [J]. Journal of Qingdao Agricultural University, (1): 21-23. (in Chinese)
[8] 李夏鸣, 郭黄萍胡增丽, 等.枣黑斑病研究[J].山西农业科学, 2009,(11):37-40.
   LI Xia-ming, GUO Huang-ping, HU Zhen-li, et al. (2009). Research on Jujube Black Spot [J]. Journal of Shanxi Agricultural Sciences, (11): 37-40. (in Chinese)
[9] 靳增军, 刘春琴, 甄文超, 等.冬枣黑疔病病原菌鉴定[J].华北农学报, 2006,21(增刊):162-165.
   JIN Zeng-jun, LIU Chun-qin, ZHEN Wen-caho, et al. (2006). Pathogen identification of winter jujube black spot [J]. Acta Agriculturae Boreali-Sinica, 21(Sup.): 162-165. (in Chinese)
[10] 赵文华,李一泉.枣树黑斑病在云南省红河州发生的主要症状、病原分析、发生规律及综合防治策略[J]. 石河子大学学报, 2004,(22): 152-155.
   ZHAO Wen-hua, LI Yi-quan. (2004). The main symptoms, pathogenic analysis, occurence law and comprehensive prevention and control Strategy of jujube black spot in Honghe Prefecture, Yunnan Province [J]. Journal of Shihezi University, (22): 152-155. (in Chinese)
[11] 刘晓琳, 刘玉,马荣,等.新疆枣果黑斑病病原菌鉴定及生物学特性[J].西北林学院学报,2015,3(3):132-138.
   LIU Xiao-lin, LIU Yu, MA Rong, et al. (2015). Identification and biological characteristics of the pathogen causing jujube black spot in Xinjiang [J]. Journal of Northwest Forestry University, 30(3): 132-138. (in Chinese)
[12] 向征, 钟聪慧,胡军,等.新疆枣果黑斑病病原鉴定[J].新疆农业科学,2013,50(5):845-850.
   XIANG Zheng, ZHONG Chong-hui, HU Jun, et al.(2013). Identification of jujube black spot pathogens in Xinjiang, China [J]. Xinjiang Agricultural Sciences, 50(5): 845-850. (in Chinese)
[13] 宋博, 徐兵强,朱晓峰,等.南疆红枣黑斑病病原鉴定及其保守基因序列分析[J].植物病理学报,2017,47(3):411-415.
   SONG Bo, XU Bing-qiang, ZHU Xiao-feng, et al. (2017). Identification of jujube black spot pathogens in Southern Xinjiang and its sequence analysis of conserved genes [J]. Acta Phytopathologica Sinica, 47(3): 411-415. (in Chinese)
[14] Margaret T. Mmbaga, Ainong Shi and Mee-Sook Kim. (2011). Identification of Alternaria alternata as a Causal Agent for Leaf Blight in Syringa Species [J]. Plant Pathol. J. 27(2):120-127.

阿克苏地区枣黑斑病菌 PCR 检测方法的建立与应用

Development and Application of PCR for Detection of Alternaria alternata Causing Jujube Black Spot in Aksu

PDF下载

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 12

编辑推荐

Metrics

[1]	杨茹薇, 刘易, 李江涛, 古丽米拉·热合木土拉, 罗正乾, 徐琳黎, 沈洪飞. 马铃薯病毒病病原RT-PCR分子鉴定[J]. 新疆农业科学, 2023, 60(12): 3032-3040.
[2]	白剑宇, 罗达, 刘正兴, 李宏. 枣树花叶病毒病的分子检测[J]. 新疆农业科学, 2022, 59(3): 657-662.
[3]	路子峰, 时晓磊, 李玉姗, 崔燕华, 仙鹤, 严勇亮, 张金波, 高海峰, 丛花. 153份新疆冬小麦品种3个条锈病抗性基因的分子检测[J]. 新疆农业科学, 2022, 59(12): 2924-2932.
[4]	王冬梅, 潘洪生, 李海强, 丁瑞丰, 阿克旦・吾外士, 刘建, 李号宾. DNA分子检测技术定量评估捕食性天敌对棉蚜的控害功能[J]. 新疆农业科学, 2022, 59(11): 2661-2667.
[5]	孙晓军, 周婷婷, 玉山江·麦麦提, 何伟, 罗文芳, 许建军, 阿布都赛买提·吐尔送. 设施番茄病毒病病原鉴定[J]. 新疆农业科学, 2021, 58(1): 99-106.
[6]	许泰百, 萨吉达木·艾则孜, 吴琼, 李玉华, 李进, 郭庆元. 基于PCR检测的4种土传病害病原菌侵染棉花的动态分析[J]. 新疆农业科学, 2019, 56(5): 910-918.
[7]	王宁, 侯敏, 房世杰, 艾尼江·尔斯满, 侯新强, 杨蓉, 包慧芳. 红枣黑斑病拮抗细菌JK1产芽孢培养基的优化[J]. 新疆农业科学, 2018, 55(6): 1098-1106.
[8]	李伟华, 刘太国, 高海峰, 刘博, 吴伟, 李宁静, 高利, 陈万权. 伊犁河谷小麦条锈病菌源地调查研究[J]. 新疆农业科学, 2017, 54(9): 1679-1687.
[9]	周登攀;郝小军;向本春. 新疆石河子地区设施樱桃病毒病害调查及病原检测[J]. , 2017, 54(4): 715-724.
[10]	包慧芳;王宁;侯敏. 新疆红枣黑斑病病原菌鉴定及拮抗细菌筛选[J]. , 2016, 53(9): 1667-1673.
[11]	董宁;冯宏祖;王兰;纪显洪;于丝丝. 防治红枣黑斑病杀菌剂的室内筛选[J]. , 2015, 52(9): 1700-1706.
[12]	高静涛;牛建新;王雪莲;刘娜;叶春秀;张飞;朱天丁. 枣疯病植原体的分子检测与同源性鉴定[J]. , 2011, 48(4): 662-667.