运用RAPD技术分析检测枣疯病植原体基因

doi:10.7668/hbnxb.2007.05.041

华北农学报 ›› 2007, Vol. 22 ›› Issue (5): 172-175. doi: 10.7668/hbnxb.2007.05.041

所属专题： 植物保护；生物技术；果树专题；

运用RAPD技术分析检测枣疯病植原体基因

樊新萍¹, 田建保¹, A. Bertaccini², 王娟¹, 杨晓宇¹

1. 山西省农业科学院果树研究所, 山西太谷 030815;
2. 意大利博洛尼亚大学农学院植物环境科学技术系, 意大利博洛尼亚 40127

收稿日期:2007-03-12 出版日期:2007-10-28
作者简介:樊新萍(1973-)，女，山西永济人，助理研究员，在读博士，主要从事果树分子生物学方面的研究。
基金资助:
山西省国际科技合作项目(2006081004)

Use of Random Amplified Polymorphic DNA(RAPD)Analysis to Detect Jujube Witches’ Broom Phytoplasmas

FAN Xin-ping¹, TIAN Jian-bao¹, A.Bertaccini², WANG Juan¹, YANG Xiao-yu¹

1. Pomology Institute, Shanxi Academy of Agriculture Sciences, Taigu 030815, China;
2. DiSTA, Patologia Vegetale, Alma Mater Studiorum, Università di Bologna, Bologna 40127, Italy

Received:2007-03-12 Published:2007-10-28

摘要/Abstract

摘要： 枣疯病是中国枣区最重要的病害之一，从表现症状的和未表现症状的3个枣品种(酸枣、辣椒枣和郎枣)的叶脉中提取核酸DNA。用13个引物(12个10 bp碱基和1个11 bp碱基)进行RAPD分析。只有引物AM-14从所收集的表现症状枣品种中扩增出400 bp的特异条带，而其他12个引物未有基因多样性；引物AL-07可以区分郎枣和其他2个品种，所有的样品重复2次，结果一致，与使用植原体特异性引物的常规PCR相比，对于检测不同枣品种枣疯病的枣植原体，RAPD技术是一个快速有效的方法。

关键词: 枣疯病, 植原体, RAPD, 检测

Abstract: Jujube witches' broom is a devastating disease of Ziziphus jujube that occurs in various jujube regions of China. Nucleic acid extracted from midribs of samples collected from three jujube varieties (Suanzao, Lajiaozao, Langzao) from symptomatic and asymptomatic shoots were tested by random amplified polymorphic DNA analyses. Using 13 different 10 and 11 bp random primers the amplification of jujube DNA was achieved from all the samples; AM-14 primer provided amplification of specific DNA fragments of about 400 bp, only from samples collected from symptomatic plants. No genetic variations in these varieties were identified using the other 11 arbitrary primers; only with primer AL-07 it was possible to differentiate Lang-zao from the other two varieties tested. All the experiments were repeated 2 times and the results were consistent. Compared with PCR analyses with phytoplasma-specific primers, RAPD techniques resulted to be an alternative rapid and sensitively method for detecting jujube phytoplasmas presence in different jujube varieties.

Key words: Jujube witches’broom, Phytoplasmas, Random Amplified Polymorphic DNA, Detection

中图分类号:

S436.6

樊新萍, 田建保, A. Bertaccini, 王娟, 杨晓宇. 运用RAPD技术分析检测枣疯病植原体基因[J]. 华北农学报, 2007, 22(5): 172-175. doi: 10.7668/hbnxb.2007.05.041.

FAN Xin-ping, TIAN Jian-bao, A.Bertaccini, WANG Juan, YANG Xiao-yu. Use of Random Amplified Polymorphic DNA(RAPD)Analysis to Detect Jujube Witches’ Broom Phytoplasmas[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(5): 172-175. doi: 10.7668/hbnxb.2007.05.041.

http://www.hbnxb.net/CN/Y2007/V22/I5/172

参考文献

[1] Lee I M, Davis R E. Molecular Biology and Pathogenesis[M]. Washington. DC: American Society for Microbiology, 1992: 379-390.
[2] Lee I M, Hammond R W, Davis R E, et al. Universal amplification and analysis of pathogen 16S rDNA for classification and identification of mycoplasmalike organisms[J]. Phytopathology, 1993, 83: 834-842.
[3] Schneider B, Seemeller E, Smart C D, et al. Molecular and diagnostic procedures in mycoplasmology[M]. San Diego: Academic Press, 1995: 369-380.
[4] Smart C D, Schneider B, Blomquist C L, et al. Phytoplasmaspecie PCR primers based on sequences of the 16S-23S rRNA spacer region[J]. Appl Environ Microbiol, 1996, 62: 2988-2993.
[5] Namba S, Kato S, Iwanam I S, et al. Detection and differentiation of plant-pathogenic mycoplasmalike organisms using polymerase chain reaction[J]. Phytopathology, 1993, 83: 786-91.
[6] Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures[J]. Physiol Plant, 1962, 15: 473-497.
[7] Tian J B, Guo H P, Bertaccini A, et al. Molecular detection of jujube witches'broom phytoplasmas in micropropagated jujube shoots[J]. Hortic Sci, 2000, 35: 1274-1275.
[8] Prince J P, Davis R E, Wolf T K, et al. Molecular detection of diverse myeoplasmalike organisms(MLOs)associated with grapevine yellows and their classification with aster yellows, X-disease, and elm yellows MLOs[J]. Phxtopath, 1993, 83: 1130-1137.
[9] 李登科, 黄丛林, 田建保, 等. 高质量枣树基因组DNA提取方法的研究[J]. 分子植物育种, 2005, 3: 579-583.
[10] Deng S, Hiruki C. Amplification of 16S rRNA genes from culturable and nonculturable mollicutes[J]. Microbiol Meth, 1991. 14: 53-61.
[11] Scheider B, Ahrens U, Kirkpatrick B C, et al. Classification of plant-pathogenic mycoplasma-like organisms using restriction-site analysis of PCR-amplified 16S rDNA[J]. Gen Microbiol, 1993, 139: 519-527.
[12] Zhu S F, Hadidi A, Lee I M, et al. Characterization of the phytoplasmas associated with cherry lethal yellows and jujube witches'-broom disease in China[J]. Acta Hort, 1998, 472: 701-707.
[13] Jung H Y, Sawayanagi T, Kakizawa H, et al. "Candidatus phytoplasma Ziziphi", a novel phytoplasma taxon associated with jujube witches'-broom disease[J]. Int J Syst Evol Microbiol, 2003, 53: 1037-1041.
[14] Fan X P, Paltrinieri S, Pastore M, et al. Molecular detection of Candidatus Phytoplasma Ziziphae'in different jujube varieties[C]//XXVII International Horticultural Congress IHC2006-Seoul, Korea. Acta Horticulturae IHC2006 proceedings. Belgium, 2007, S04-P-15.

[1]	华明艳, 宋兰芳, 崔少杰, 孙海波, 金凤媚. 番茄褐色皱纹果病毒分子检测与基因组部分序列分析[J]. 华北农学报, 2024, 39(3): 179-186.
[2]	李义, 康占海, 董素芬, 王羽, 李艳艳, 赵欣雨, 李星. 75份国内外小麦品种抗叶锈性鉴定及基因分析[J]. 华北农学报, 2023, 38(S1): 362-370.
[3]	刘树森, 郭宁, 孙华, 马红霞, 张海剑, 石洁. 玉米根腐病菌麦根腐平脐蠕孢的LAMP检测方法[J]. 华北农学报, 2023, 38(1): 162-167.
[4]	贾晓君, 董雅凤, 张尊平, 范旭东, 任芳, 张宝东, 胡国君. 苹果4个器官不同部位RNA提取及病毒检测效果比较[J]. 华北农学报, 2022, 37(S1): 283-290.
[5]	贲海燕, 郝永娟, 霍建飞, 姚玉荣, 高苇, 王万立. 草莓白粉病菌LAMP可视化快速检测方法的建立及初步应用[J]. 华北农学报, 2022, 37(6): 182-190.
[6]	王佳荣, 董瑞, 张梦宇, 高璞, 张培培, 李在峰, 刘大群. 40份CIMMYT小麦品系苗期及成株抗叶锈病基因鉴定[J]. 华北农学报, 2022, 37(2): 201-210.
[7]	金凤媚, 薛俊, 孙海波, 郝志愚. 基于小RNA技术的天津地区番茄花叶病毒分子检测与基因组部分序列分析[J]. 华北农学报, 2021, 36(5): 176-183.
[8]	孟雅宁, 严立斌, 张红肖, 赵海涛, 范妍芹. 辣椒抗疫病基因分子标记的筛选及检测[J]. 华北农学报, 2020, 35(S1): 338-341.
[9]	程胜群, 吕文河, 高艳玲, 白艳菊, 范国权, 张威, 张抒, 邱彩玲, 申宇, 董学志, 白雅梅. 马铃薯S病毒RT-qPCR通用检测体系的建立与应用[J]. 华北农学报, 2020, 35(6): 187-194.
[10]	陈利达, 石延霞, 谢学文, 曹金强, 柴阿丽, 李宝聚. 我国不同地区番茄主要病毒病种类的分子检测与分析[J]. 华北农学报, 2020, 35(1): 185-193.
[11]	金凤媚, 宋建, 薛俊, 陈东亮, 王姝, 张越, 孙海波. 北京地区侵染菊花的番茄斑萎病毒分子检测与基因组部分序列分析[J]. 华北农学报, 2020, 35(1): 175-184.
[12]	李欣, 崔秀明, 刘迪秋, 孙恬, 郭爱玲, 陈军. 三七根腐病菌尖孢镰刀菌的Real-time PCR检测方法[J]. 华北农学报, 2019, 34(S1): 324-330.
[13]	张敬敬, 张海英, 潘秀清, 许勇, 任毅, 高秀瑞, 李冰, 史宇凡, 党继革, 杨明珠, 武彦荣. 河北省130份西瓜品种与种质资源抗病基因KASP检测分析[J]. 华北农学报, 2019, 34(2): 110-116.
[14]	刘栋, 郭娜, 马建富, 李爱荣. 外源野生胡麻总DNA遗传转化栽培胡麻及RAPD分子验证[J]. 华北农学报, 2018, 33(S1): 29-32.
[15]	杨蕊, 韩雪纯, 李晴, 陈招荣. 天津地区梨病毒病检测方法筛选及带毒率检测[J]. 华北农学报, 2018, 33(5): 125-129.

选择文件类型/文献管理软件名称

选择包含的内容

运用RAPD技术分析检测枣疯病植原体基因

Use of Random Amplified Polymorphic DNA(RAPD)Analysis to Detect Jujube Witches’ Broom Phytoplasmas

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

运用RAPD技术分析检测枣疯病植原体基因

Use of Random Amplified Polymorphic DNA(RAPD)Analysis to Detect Jujube Witches’ Broom Phytoplasmas

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价