西藏青稞根际土壤可培养放线菌的遗传多样性及其促生功能分析

doi:10.16036/j.issn.1000-2650.2019.06.005

摘要/Abstract

摘要： 【目的】认识藏区青稞根际土壤中可培养放线菌的遗传多样性及应用价值。【方法】采集藏区青稞根际土壤,对根际样品中的放线菌进行分离,测定分离菌株的溶磷能力、抗病促生（IAA产量）能力,并运用反转录重复因子扩增（BOXA1R-PCR）技术、16S rRNA基因测序和系统发育分析系统分析可培养放线菌的系统发育地位。【结果】通过7种培养基共分离得129株放线菌菌株,其中有51株菌株（占总分离菌株的39.5%）产IAA,8株产量较高（>57 mg/L）,77株（占总分离菌株的60.0%）表现出溶磷活性。基于BOXA1R-PCR图谱,采用平均连锁聚类法（UPGMA）进行聚类分析,129株菌被分成19种不同的遗传图谱类型。基于分离菌株的菌落特征和遗传聚类图谱选取32株代表菌株进行16S rRNA基因序列测定。通过 Mothur 软件对分离菌株进行 OTUs （operational taxonomic units）聚类分析,共得到 5个OTU分类单元,其中以OTU1为主。系统发育显示代表菌株分属于链霉菌属（Streptomyces）,拟诺卡氏菌属（Nocardiopsis）,类诺卡氏菌属（Nocardioides）,小单孢菌属（Micromonospora）,放线菌属（Actinomyces）等 5个属,其中链霉菌属（Streptomyces）为优势种属。【结论】西藏青稞根际土中放线菌多样性明显,这些放线菌中蕴藏着大量的有开发应用前景的菌株。

关键词: 青稞根际, 放线菌, 多样性, 功能分析

Abstract: 【Objective】 To study the genetic diversity and application prospect of the culturable actinomycetes inhabited in the rhizosphere of Highland barley. 【Method】 The rhizosphere samples were collected from five different cultivation areas of highland barley in Tibetan, rhizosphere culturable Actinomycetes strains were isolated by pure cultivation method. The phosphorus solubilizing activities, plant pathogen disease resistance and indole-3-acetic acid (IAA) production of the isolates were determined, finally the genetic diversity and phylogeny characteristics of the isolates were analyzed by transcription factor amplification (BOXA1R-PCR) and 16S rRNA gene sequencing, respectively. 【Result】 Totally, 129 Actinomycetes strains were isolated by using 7 different culture medium. Among the total isolates, 39.5% (51/129) could produce IAA and eight of them have had a higher yield (>57 mg/L), 60.0% (77/129) had phosphate solubilizing activities. Based on the BOXA1R-PCR pattern, function of unweighted pair-group method with arithmetic means(UPGMA) were used to perform clustering analysis for the 129 isolates and resulted into 19 representative genetic patterns. Based on the colony morphology and the clustering results, 32 typical strains were selected for further 16S rRNA gene sequencing. Moreover, the operational taxonomic units(OTUs) analysis was performed using Mothur software and 5 different OTUs were obtained, among which, OTU1 was the dominant group. Phylogeny analysis showed that the representative strains were phylogenetically affiliated to genus of Streptomyces, Nocardiopsis, Nocardioides, Micromon-ospora， and Actinomyces. Streptomyces was the dominant species. 【Conclusion】 Various of Actinomycetes in habited in the Highland barley rhizosphere. These strains are vital resources for screening high efficiency functional strains that could be used in Highland barley cultivation.

Key words: highland barley rhizosphere, Actinomycetes, diversity, functional analysis

中图分类号:

Q938

高雪, 辜运富, 尼玛扎西, 刘国一, 刘婷, 刘怡, 普布贵吉. 西藏青稞根际土壤可培养放线菌的遗传多样性及其促生功能分析[J]. 四川农业大学学报, 2019, 37(06): 777-784.

GAO Xue, GU Yunfu, NYIMA Tashi, LIU Guoyi, LIU Ting, LIU Yi, PUBU Guiji. Analysis of the Genetic Diversity and Promoting Functions of the Culturable Actinomycetes in the Rhizosphere of Highland Barley in Tibet[J]. Journal of Sichuan Agricultural University, 2019, 37(06): 777-784.

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