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浙江大学学报(农业与生命科学版)  2017, Vol. 43 Issue (4): 431-440    DOI: 10.3785/j.issn.1008-9209.2016.12.221
生物科学与技术     
藏东南地区毛壳属真菌多样性及系统发育分析
岳海梅1*,庄华2,潘朝晖3,巩文峰1
1.西藏农牧学院植物科学学院,西藏 林芝 860000;2.西北农林科技大学植物保护学院/旱区作物逆境生物学国家重点实验室,陕西 杨凌 712100;3.西藏高原生态研究所,西藏 林芝 860000
Diversity and phylogenetic analysis of Chaetomium fungus from southeastern Tibet
YUE Haimei1*, ZHUANG Hua2, PAN Zhaohui3, GONG Wenfeng1
(1. Department of Plant Science, Tibet Agricultural and Animal Husbandry College, Linzhi 860000, Xizang, China; 2. College of Plant Protection, Northwest A & F University/State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling 712100, Shaanxi, China; 3. Tibet Plateau Institute of Ecology, Linzhi 860000, Xizang, China)
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摘要:

为分析藏东南地区毛壳属(Chaetomium Kunze)真菌多样性及系统发育情况,从藏东南5个市、县采集土壤、植物残体和动物粪便等标本共355份,采用组织分离法和稀释分离法得到36株毛壳菌,平均分离几率为10.14%。将分离得到的菌株进行多样性指数特征分析,并按形态特征初步分类;运用聚合酶链式反应对菌株的rDNA-ITS和β-tubulin基因序列进行扩增与序列测定;利用MEGA 5.0软件构建rDNA-ITS和β-tubulin基因系统发育进化树。结果显示:林芝县的毛壳属真菌多样性指数最高,为1.178 5,物种均匀度指数也最高,为0.605 6;昌都市的毛壳属真菌丰富度指数最高,达到0.861 4。不同地区毛壳属真菌的优势种也存在差异:工布江达县的优势种为球毛壳(C. globosum);墨脱县和昌都市的优势种为粪生毛壳(C. funicola);察隅县的优势种为旋丝毛壳(C. bostrychodes);林芝县的优势种为反卷毛壳(C. convolutum)。根据形态特征可将36株菌归属为8个种。分析扩增后的rDNA-ITS和β-tubulin基因序列发现,β-tubulin基因的变异位点、单倍型总数、单倍型多样度、核苷酸多样性指数和核苷酸平均差异数与rDNA-ITS序列的这些参数存在显著差异,体现出更大的碱基变异性。系统发育进化树表明,供试的36个菌株被分为7个组,rDNA-ITS和β-tubulin这2个基因片段不仅可用来区分毛壳菌属中形态差异较大的种,如印度毛壳(C. indicum)、大果毛壳(C. megalocarpum)、球毛壳(C. globosum)、粪生毛壳(C. funicola),还可区分形态特征非常相似的印度毛壳(C. indicum)和直立毛壳(C. erectum),同时也可以对一些形态上难以鉴定的种进行初步鉴定,但不能较好地区分反卷毛壳(C. convolutum)、旋丝毛壳(C. bostrychodes)和黑毛壳(C. nigricolor),需要结合更多的基因片段进行区分。综上所述,本文对藏东南地区毛壳属真菌多样性与系统发育情况进行了研究,为丰富西藏毛壳属真菌资源和开发毛壳菌代谢产物奠定了理论基础。

Abstract:

Chaetomium Kunze is mainly distributed in cellulose-containing substrates in the nature, including soil, plant residue, excrement of birds, omnivorous animals and rodents. Persistent organic compounds can be degraded by cellulase produced by Chaetomium Kunze, which also has antagonistic effects on certain microorganisms in the soil. However, littlehas been known about Chaetomium in Tibet. Therefore, a systematic research on resource distribution of Chaetomium in Tibet is necessary and urgent.
In this study, 355 samples of soil, plant residue and animal manure were collected from five regions of southeastern Tibet, in which plant residue and animal manure were separated by tissue separation, and soil samples were separated by dilution separation method. SPSS 13.5 software was used to analyze the diversity index of Chaetomium spp. from different regions, and 36 strains were preliminarily classified according to Arx system. The nucleotide sequence polymorphisms of rDNA-ITS and β-tubulin gene were analyzed by DnaSP version 5.0. The rDNA-ITS and β-tubulin gene sequences were used to analyze the diversity of Chaetomium species in southeastern Tibet.
The results showed that a total of 36 strains were successfully isolated from the 355 samples, and the average isolation rate was 10.14%. The diversity index data indicated that both the diversity index and species evenness index of Chaetomium spp. in Nyingchi was the highest, with the value of 1.178 5 and 0.605 6 respectively. But the richness index of Chaetomium was the highest in Chamdo, with the value of 0.861 4. Differences of dominant populations of Chaetomium were observed among the five regions: the dominant population in Gongbogyamda County, Medog County and Chamdo City, Zayü County, and Nyingchi County was C. globosum, C. funicola, C. bostrychodes, and C. convolutum, respectively. According to the morphological characteristics, the 36 strains were assigned to eight species. Moreover, rDNA-ITS sequences and β-tubulin gene were applied to conduct the diversity analysis, and the result showed that the variation sites, haplotype numbers, haplotype diversity, nucleotide diversity index and nucleotide difference of β-tubulin gene were significantly different from rDNA-ITS sequences, while β-tubulin gene showed greater base variability. The phylogenetic tree of rDNA-ITS and β-tubulin genes indicated that the 36 isolates were divided into seven groups. The two gene fragments can not only distinguish the species of Chaetomium with different morphologies (such as C. indicum, C. megalocarpum, C. globosum and C. funicola), but also those with very similar morphological characteristics (such as C. indicum and C. erectum) and some species hard to distinguish. However, several species such as C. convolutum, C. bostrychodes and C. nigricolor can’t be distinguished by rDNA-ITS and β-tubulin genes, which still need combining more gene fragments to differentiate.
In conclusion, the results confirm the rich resources of Chaetomium fungus in southeastern region of Tibet, providing data for abundant resources of Chaetomium in Tibet, and laying a foundation for the exploitation of the metabolites of Chaetomium.

收稿日期: 2016-12-22 出版日期: 2017-05-09
CLC:  Q 949.32  
基金资助: 国家自然科学基金(31360006);旱区作物逆境生物学国家重点实验室开放基金课题(CSBAA2016006)
通讯作者: 岳海梅(http://orcid.org/0000-0001-9282-2295)     E-mail: yuehm1980@163.com
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引用本文:

岳海梅,庄华,潘朝晖,巩文峰. 藏东南地区毛壳属真菌多样性及系统发育分析[J]. 浙江大学学报(农业与生命科学版), 2017, 43(4): 431-440.

YUE Haimei, ZHUANG Hua, PAN Zhaohui, GONG Wenfeng. Diversity and phylogenetic analysis of Chaetomium fungus from southeastern Tibet. Journal of Zhejiang University (Agriculture and Life Sciences), 2017, 43(4): 431-440.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2016.12.221        http://www.zjujournals.com/agr/CN/Y2017/V43/I4/431

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