药用植物凤丹(Paeonia suffruticosa)根际土壤细菌群落16S rRNA基因的ARDRA分析

Abstract

PDF HTML阅读 XML下载 导出引用 引用提醒 药用植物凤丹(Paeonia suffruticosa)根际土壤细菌群落16S rRNA基因的ARDRA分析 DOI: 10.5846/stxb201503030405 作者: 作者单位: 安徽师范大学,安徽师范大学,安徽师范大学,安徽师范大学,安徽师范大学,安徽师范大学 作者简介: 通讯作者: 中图分类号: 基金项目: 国家自然科学基金（81173491）；安徽省自然科学基金资助项目（1308085MC42） Bacterial diversity in rhizosphere soil of the medicinal tree peony (Paeonia suffruticosa) revealed by amplified ribosomal DNA restriction analysis Author: Affiliation: Anhui normal university,Anhui normal university,Anhui normal university,,Anhui normal university,Anhui normal university Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:凤丹（Paeonia suffruticosa Andr.）为芍药科多年生植物，是一种重要的传统中药资源。在其生长发育的周期中与土壤微生物尤其是根际土壤微生物有密切的关系。通过构建16S rRNA基因克隆文库及文库的限制性片段长度多态性分析（ARDRA），对中国药用植物凤丹5大主要分布区域的根际土壤细菌群落多样性进行了研究。采用限制性内切酶HinfI和Csp6I对克隆文库中随机挑选的1000个白色克隆子进行了酶切分型，根据酶切图谱的不同，将其分为324个OTUs，并对38个优势OTUs进行了测序和系统发育分析。16S rRNA基因序列分析结果表明，凤丹根际土壤细菌种群主要包括：变形菌门（包括alpha、beta、gamma、detla亚门）、酸杆菌门、放线菌门、拟杆菌门及厚壁菌门等11类细菌，此外还包含了3个未归类的细菌。变形菌门和酸杆菌门为文库中的主要菌群，分别占克隆总数的47.34%和14.36%，其中Pseudomonas sp.、Burkholderia sp.和Arthrobacter sp.为优势菌属。研究结果表明，我国药用植物凤丹5大主要分布区域的根际土壤细菌种群不仅具有丰富的多样性，还存在丰富的潜在新菌种。 Abstract:The medicinal tree peony (Paeonia suffruticosa) is an important resource in traditional Chinese medicine. Soil bacteria, especially rhizosphere bacteria, have strong effects on plant health and growth. In this study, the bacterial community structure and diversity in P. suffruticosa rhizosphere soil in five major distribution areas in China were investigated by amplified ribosomal DNA restriction analysis (ARDRA). The ARDRA patterns of the amplified 16S rRNA gene produced an average of 2-6 bands that ranged in size from 100 to 1302 bp, which indicated that the fragments of the 16S rRNA gene were efficiently double-digested with the restriction endonucleases HinfI and Csp6I. ARDRA-based cluster analysis showed that 702 positive clones were clustered into 74, 69, 76, 73, and 72 operational taxonomic units (OTUs) in the FH, YS, BZ, HZ, and LY libraries, respectively. Most of the OTUs contained 1-3 clones, which implied high diversity in the peony rhizosphere soil; twenty-six OTUs contained 4-6 clones; and five OTUs contained 7-12 clones. Rarefaction analysis showed that the positive clones in each library covered the diversity of bacterial taxa, which was further confirmed by clone library coverage (range:71.32%-81.40%). Diversity analysis indicated high bacterial diversity in the soils (Shannon diversity index 3.949-4.184; Chao1 index range 89.52-107.25). Sequence analysis revealed diverse bacterial phyla in the 16S rRNA gene library, consisting of α, β, γ, and δ subclasses of Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Verrucomicrobia, TM7, Gemmatimonadetes, and three unclassified bacteria. The dominant phyla were Proteobacteria and Acidobacteria (47.34% and 14.36% of the total clones, respectively), and the dominant genera in the clone libraries were Pseudomonas, Burkholderia, and Arthrobacter. In addition, a portion of the clones was only distantly related to sequences in the GenBank database, suggesting that bacteria in rhizosphere soil of the medicinal tree peony were unique and diverse. Furthermore, the relative abundance of Firmicutes (mainly Bacillus spp.) was positively correlated with soil Cu content. The rhizosphere bacteria belonged the same dominant phyla (Proteobacteria, Acidobacteria, and Actinobacteria) and showed high diversity between geo-authentic and non-authentic areas, implying that P. suffruticosa tended to form a similar microenvironment and select similar bacterial communities in the rhizosphere soil. 参考文献 相似文献 引证文献