赞皇大枣枣疯病植原体分子分类

doi:10.3864/j.issn.0578-1752.2011.21.011

Abstract

Abstract: 【Objective】 Jujube Witches’ Broom (JWB) is a serious disease caused by phytoplasma. In this paper, the taxonomic status of JWB-associated phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’，which is the only natural triploid cultivar in Chinese jujube, was identified.【Method】16S rDNA gene was amplified by using specific conservative primers and the nested-PCR primers, then the PCR product was cloned and sequenced. Homology analysis, dendrogram and virtual RFLP were also obtained by using related software. 【Result】 One set of the conserved primers was designed to amplify a 1 850 bp fragment (GenBank GU184180) which included 16S rDNA (1 529 bp), adjacent spacer sequence (264 bp) and partial sequence of 23S rDNA (57 bp) by PCR from total DNA extracted from diseased Ziziphus jujuba Mill. ‘Zanhuangdazao’. Nucleotide sequencing of the amplified fragment indicated that JWB-associated phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’ shared nucleotide similarity about 87% to 98% with other phytoplasmas from data of GenBank, and a maximum similarity at 98% with Elm yellows（EY）phytoplasma. A phylogenetic tree based on 16S ribosomal DNA sequences was constructed. The result of virtual RFLP showed a significant difference among JWB-associated phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’ and 4 subgroups. The blast result of tuf also showed that JWB-associated phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’ (JN001985) shared nucleotide similarity about 52.3% to 76.7% with 4 subgroups. 【Conclusion】 JWB-associated phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’ was clustered into the elm yellows group (16Sr V) phytoplasma, and it shared the highest similarity with the B subgroups. Its taxonomic status is consisting with those diploid cultivars.

Key words: Ziziphus jujuba Mill.‘Zanhuangdazao’, JWB-associated phytoplasma, 16S rDNA, molecular classification

YANG Hai-Xu, WANG Yang, ZHAO Yan-Lin, ZHAO Jin, LIU Meng-Jun. Molecular Classification of Jujube Witches’ Broom (JWB) Associated Phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’[J].Scientia Agricultura Sinica, 2011, 44(21): 4429-4437.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2011.21.011

https://www.chinaagrisci.com/EN/Y2011/V44/I21/4429

References

[1]朱天生, 潘一展, 崔延涛, 高瑞, 李向东, 朱水芳. 榆树黄化病植原体的分子监测与鉴定. 植物病理学报, 2008, 38(4): 401-406.

Zhu T S, Pan Y Z, Cui Y T, Gao R, Li X D, Zhu S F. Molecular detection and identification of the phytoplasma associated with Elm yellows in China. Acta Phytopathologica Sinica, 2008, 38(4): 401-406. (in Chinese)

[2]车海彦, 罗大全, 符瑞益, 叶莎冰, 吴云锋. 海南长春花黄化病植原体的16S rDNA 序列分析研究. 植物病理学报, 2009, 39(2): 212-216.

Che H Y, Luo D Q, Fu R Y, Ye S B, Wu Y F. Sequence analysis of 16S ribosomal DNA of phytoplasma associated with periwinkle yellows disease in Hainan. Acta Phytopathologica Sinica, 2009, 39(2): 212-216. (in Chinese)

[3]卢全有. 安徽桑黄化型萎缩病植原体16S rDNA序列分析及分子检测. 植物病理学报, 2010, 40(6): 642-646.

Lu Q Y. Molecular detection and sequence analysis is on 16S ribosomal DNA of phytoplasma associated with mulberry yellow dwarf disease occurring in Anhui province. Acta Phytopathologica Sinica, 2010, 40(6): 642-646. (in Chinese)

[4]李永, 田国忠, 徐启聪, 朴春根, 汪来发, 郭民伟. 臭矢菜丛枝病植原体的分子鉴定研究. 植物病理学报, 2009, 39(4): 377-384.

Li Y, Tian G Z, Xu Q C, Piao C G, Wang L F, Guo M W. Molecular identification of cleome witches′broom phytoplasma. Acta Phytopathologica Sinica, 2009, 39(4): 377-384. (in Chinese)

[5]蔡红, 张华明, 陈海如, 秦洋. 竹丛枝植原体16SrDNA片段克隆与序列分析. 植物保护, 2005, 31(2): 38-40.

Cai H, Zhang H M, Chen H R, Qin Y. Cloning and sequence analysis of 16S ribosomal DNA of phytoplasma associated with witches broom of bamboo. Plant Protection, 2005, 31(2): 38-40. (in Chinese)

[6]张荣, 孙广宇, 张雅梅, 康振生. 小麦蓝矮病植原体16S rDNA序列分析研究. 植物病理学报, 2005, 35(5): 397-402.

Zhang R, Sun G Y, Zhang Y M, Kang Z S. Sequence analysis of 16S ribosomal DNA of phytoplasm a associated with wheat blue dwarf. Acta Phytopathologica Sinica, 2005, 35(5): 397-402. (in Chinese)

[7]车海彦, 罗大全, 符瑞益, 吴云锋. 海南省木豆丛枝病植原体的分子检测及鉴定. 植物病理学报, 2010, 40(2): 113-121.

Che H Y, Luo D Q, Fu R Y,Wu Y F. Molecular detection and identification of a phytoplasma associated with pigeon. Acta Phytopathologica Sinica, 2010, 40(2): 113-121.(in Chinese)

[8]安凤秋, 吴云锋, 孙秀芹, 顾沛雯, 杨英. 小麦蓝矮病植原体延伸因子(EF-Tu) tuf 基因序列的同源性分析. 中国农业科学, 2006, 39(1): 74-80.

An Q F, Wu Y F, Sun X Q, Gu P W, Yang Y. Homologic analysis of tuf gene for eongation factor Tu of phytoplasma from wheat blue dwarf. Scientia Agricultura Sinica, 2006, 39(1): 74-80.(in Chinese)

[9]史英姿, 吴云锋, 顾沛雯, 安凤秋, 杨艳. 泡桐丛枝植原体16S rDNA和延伸因子基因序列分析. 微生物学通报, 2007, 34(2): 291-295.

Shi Y Z, Wu Y F, Gu P W, An F Q, Yang Y. Homologic analysis of 16S rDNA and elongation factor gene of paulownia witches broom phytoplasma. Microbiology, 2007, 34(2): 291-295. (in Chinese)

[10]卢全有. 桑树黄化型萎缩病植原体延伸因子基因的克隆及序列分析. 植物保护, 2010, 36(5): 43-46.

Lu Q Y. Cloning and sequence analysis of tuf gene for elongation factor Tu of phytoplasma associated with mulberry yellow dwarf disease. Plant Protection, 2010, 36(5): 43-46. (in Chinese)

[11]黄云, 王靖, 赵艳琴, 何苗. 桃黄化病病原植原体的鉴定及16S rDNA序列分析. 园艺学报, 2009, 36(3): 341-346.

Huang Y, Wang Y, Zhao Y Q, He M. The identification and sequence analysis of 16s ribosomal DNA of the phytoplasma sp. associated with peach yellow disease. Acta Horticulturae Sinica, 2009, 36(3): 341-346.(in Chinese)

[12]赖帆, 李永, 徐启聪, 田国忠. 植原体的最新分类研究动态. 微生物学通报, 2008, 35(2): 291-295.

Lai F, Li Y, Xu Q C, Tian G Z. The present status on classification of phytoplasmas. Microbiology, 2008, 35(2): 291-295. (in Chinese)

[13]Smart C D, Schneider B, Blomouist C L. Phytoplasma-specific PCR primers based on sequences of the 16-23S rRNA spacer region. Applied and Environmental Microbiology, 1996, 62: 2988-2993.

[14]魏婷, 吴云锋, 侯伟, 武科科, 李毅然, 张珏, 孙润红. 柳树黄化病植原体的分子分类. 微生物学报, 2009, 49(3): 389-394.

Wei T, Wu Y F, Hou W, Wu K K, Li Y R, Zhang Y, Sun R H. Molecular classification of a phytoplasma associated with willow yellow disease. Acta Microbiologica Sinica, 2009,49(3): 389-394. (in Chinese)

[15]张松柏, 张德咏, 罗香文, 张曙光, 李华平. 水稻橙叶病植原体16S rDNA基因的序列分析. 华南农业大学学报, 2009, 30(2): 37-39.

Zhang S B, Zhang D Y, Luo X W, Zhang S G, Li H P. Sequence analysis of 16S ribosomal DNA of phytoplasma associated with rice orange leaf. Journal of South China Agricultural University, 2009，30(2): 37-39. (in Chinese)

[16]徐启聪, 田国忠, 王振亮, 孔繁华, 李永, 王合. 中国各地不同枣树品种上枣疯病植原体的PCR检测及分子变异分析. 微生物学报, 2009, 49(11): 1510-1519.

Xu Q C, Tian G Z, Wang Z L, Kong F H, Li Y, Wang H. Molecular detection and variability of jujube witches' broom phytoplasmas from different cultivars in various regions of China. Acta Microbiologica Sinica, 2009,49(11): 1510-1519. (in Chinese)

[17]王海妮, 吴云锋, 安凤秋, 顾沛文, 张昭亮. 枣疯病和酸枣丛枝病植原体16S rDNA和tuf基因的序列同源性分析. 中国农业科学, 2007, 40(10): 2200-2205.

Wang H N, Wu Y F, An F Q, Gu P W, Zhang Z L. The homology analysis for sequences of 16S rDNA and tuf gene of phytoplasma from jujube witches′ broom and wild jujube witches′ broom. Scientia Agricultura Sinica, 2007, 40(10): 2200-2205. (in Chinese)

[18]Lee S, Han S, Cha B. Mixed in fection of 16S rDNA V and I groups of phytoplasma in a single jujube. The Plant Pathology Journal, 2009, 25(1): 1-4.

[19]牟海青, 赵文军, 邓东亮, 朱水芳. 植原体16S rDNA RFLP 指纹图谱分析软件研究. 植物病理学报, 2010, 40(5): 522-529.

Mou H Q, Zhao W J, Deng D L, Zhu S F. One software of 16S rDNA RFLP fingerprint pattern analys is for phytoplasma identification. Acta Phytopathologica Sinica, 2010, 40(5): 522-529.(in Chinese)

[20]顾沛文, 吴云锋, 安凤秋. 小麦蓝矮植原体寄主范围的鉴定及RFLP分析. 植物病理学报, 2007, 37(4): 390-397.

Gu P W, Wu Y F, An F Q. Host range testing and RFLP analysis of wheat blue dwarf phytoplasma. Acta Phytopathologica Sinica, 2007, 37(4): 390-397.(in Chinese)

[21]薛渝峰. 植原体诱导下抗枣疯病相关基因的差异表达研究[D]. 河北保定: 河北农业大学, 2008.

Xue Y F. The Differential expression of genes related to the resistance of jujube witches broom induced by phytoplasma[D]. Baoding, Hebei: Agricultural University of Hebei, 2008. (in Chinese)

[22] 刘清神, 许东林, 林盘芳, 周国辉. 广州桑树植原体分子检测及多样性初探. 蚕业科学, 2006, 32(1): 1-5.

Liu Q S, Xu D L, Lin P F, Zhou G H. Molecular detection of mulberry phytoplasma in guangzhou and primary study of its 16S rDNA diversity. Science of Sericulture, 2006, 32(1): 1-5. (in Chinese)

[23]Wei W, Davis R E, Lee I M, Zhao Y. Computer-simulated RFLP analysis of 16S rRNA genes : identification of ten new phytoplasma groups. International Journal of Systematic and Evolutionary Microbiology, 2007(57): 1855-1867.

[24]李春香, 杨群. PCR产物直接测序还是克隆测序?——密叶杉属rDNAITS序列的测定方法. 植物学通报, 2002, 19(6): 698-704.

Li C X,Yang Q. Direct sequencing of PCR products or sequencing by cloning PCR products-method of determining internal transcribed spacer sequences of nuclear ribosomal DNA in Athrotaxis. Chinese Bulletin of Botany, 2002, 19(6): 698-704. (in Chinese)

[25]Brenner D, Krieg N, Staley J. Bergey' Manual of Systematic Bacteriology. Second edition. Volume 2. New York: Springer Science and Business Media Inc, 2005.

[26]解焱, 李典谟, John Mackinnon. 中国生物地理区划研究. 生态学报, 2002(10): 1599-1615.

Xie Y, Li D M, Mackinnon J. Preliminary researches on bio-geographical divisions of China. Actaecologica Sinica, 2002(10): 1599-1615. (in Chinese)

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Molecular Classification of Jujube Witches’ Broom (JWB) Associated Phytoplasma from Ziziphus jujuba Mill. ‘Zanhuangdazao’

PDF

Abstract

Cite this article

share this article

References

Related Articles 9

Metrics

Comments

Recommended 0

[1]	SUN Jian-Guang, HU Hai-Yan, LIU Jun, CHEN Qian, GAO Miao, XU Jing, ZHOU Yi-Qing. Growth Promotion Potential and Distribution Features of Nitrogen-Fixing Bacteria in Field Environments [J]. Scientia Agricultura Sinica, 2012, 45(8): 1532-1544.
[2]	WENG Pei-Fang, CHEN Xi, SHEN Xi-Quan, WU Zu-Fang, REN Jing. Microbial Community Diversity Analysis During the Pickled Processing of Mustard Tuber with Low-Salinity [J]. Scientia Agricultura Sinica, 2012, 45(2): 338-345.
[3]	HUANG Xian-qing,WANG Yu-fen,ZHAO Gai-ming,XIE Hua,GAO Xiao-ping,SUN Ling-xia,LI Miao-yun1,ZHANG Qiu-hui1,ZHAO Guang-hui . Identification of A Microorganism Strain Isolated from the Corn Sausage by 16S rDNA Sequence Analysis and Optimization of Preservative Mixture [J]. Scientia Agricultura Sinica, 2010, 43(24): 5112-5120 .
[4]	WANG Ai-hua,YIN You-ping,XIONG Hong-li,LI Yan-fang,LI Jia,XIAN Jia-xu,WANG Zhong-kang . Endophytic Bacterial Diversity Analysis of Huanglongbing Pathogen-Infected Citrus Phloem Tissue in Guangxi [J]. Scientia Agricultura Sinica, 2010, 43(23): 4823-4833 .
[5]	. Genetic Diversity of Potato Scab Pathogens in China Based on 16S rDNA Sequences [J]. Scientia Agricultura Sinica, 2009, 42(2): 499-504 .
[6]	,. Phenotypic and Genetic Diversity of Rhizobia Associated with Glycyrrhiza spp. Grown in Northern Regions of China [J]. Scientia Agricultura Sinica, 2006, 39(7): 1321-1327 .
[7]	,,,,,. Characterization and Identification on the Pathogen of Potato Scab in China [J]. Scientia Agricultura Sinica, 2006, 39(02): 313-318 .
[8]	,,,,. Effct of Continuous Cropping of Different Vegetables on DNA Polymorphorism of Soil Bacterial [J]. Scientia Agricultura Sinica, 2005, 38(10): 2076-2083 .
[9]	,,. Analysis of Rumen Bacterial Diversity of Goat by Denaturing Gradient Gel Electrophoresis and 16S rDNA Sequencing [J]. Scientia Agricultura Sinica, 2004, 37(09): 1374-1378 .