毒沟褶菌（Trogia venenata）群体遗传学及鸡油菌复合种分子系统学与生物地理学研究

发布时间：2018-01-14 12:35

  本文关键词：毒沟褶菌（Trogia venenata）群体遗传学及鸡油菌复合种分子系统学与生物地理学研究　出处：《云南大学》2016年博士论文　论文类型：学位论文

  更多相关文章： 毒沟褶菌 鸡油菌复合种 物种多样性 群体遗传 地区特有

【摘要】：毒沟褶菌(Trogia venenata)是一个具有争议的、能导致不明原因死亡(SUD)的致命蘑菇,主要分布在中国的西南地区。为探索其致命机制,研究人员们已经分析了该真菌可能导致人类死亡的相关因素：两个非蛋白氨基酸、重金属钡含量及其他因子。本研究采用多基因联合分子遗传学的方法探讨了该物种的基因型多样性、群体结构、自然条件下的生殖模式以及群体间的遗传分化水平对其致命因子的影响。鸡油菌属真菌具有重要的经济、生态和科研价值,代表类群是鸡油菌复合种Cantharellus cibarius species complex (CCSC)。云南地区是现代物种多样性中心之一,鸡油菌物种丰富。但是,由于缺乏系统学和生物地理学研究,目前该属真菌的系统学研究主要局限于欧美、非洲等地区。本研究利用多基因片段作为分子标记,结合GenBank中已有的数据,对鸡油菌复合种的物种多样性、分子系统学和生物地理学等进行了探讨,主要结果如下：1.毒沟褶菌(T.venenata)群体遗传学研究本研究于2010-2011年进行了多次实地调查和样品采集,选择了七个地区进行了采样,采样点主要集中于楚雄大姚县、大理市祥云县、鹤庆县和宾川县以及腾冲县等地区,其中五个地区有不明原因死亡实例,共采集了232份样品。采用了ITS、β-tub、rpb2和tef1-α四个基因片段,运用多基因联合分析的方法对毒沟褶菌(T. venenata)进行群体遗传学研究。本研究中ITS片段用于对样本进行分子鉴定,其他三个蛋白编码基因用于分子遗传学分析。结果表明,β-tub、rpb2和tef1-α三个基因片段能够识别丰富的遗传变异位点,可以准确反映各个地理群体间的遗传分化水平。单倍型多样性分析、分子遗传差异(FST)和群体结构分析揭示T. venenata群体内的遗传多样性较高,分子变异主要分布于地方群体内部,群体之间存在明显的结构特征—7个地理群体属于4个遗传群体,群体间有较低水平的基因交流,孢子传播及有性生殖过程对保持群体遗传多样性和稳定性起到重要作用；哈迪-温伯格平衡检测和基于单核苷酸多态性位点(SNPs)的连锁平衡分析证明T. venenata群体间遵循随机交配模式,近亲繁殖方式也是该菌生殖模式的重要组成部分。Mantel检测结果显示T. venenata群体之间的遗传距离与地理距离之间具有微弱的正相关关系,其遗传分化与钡含量呈负相关关系。2.鸡油菌复合种(CCSC)分子系统学与生物地理学研究本研究选取了云南省16个地区和5个欧洲地区共414份鸡油菌样品,采用了tef1-α, rpb1，chs1和ATP6四个基因,对其进行了系统学研究和系统发育种的识别。首先,采用309份样品的tef1-α序列进行Blast比对,结果显示我们的样品与GenBank数据库中的6个鸡油菌属真菌种的序列相似度最高。将符合要求的序列进行比对和保守区的选择,通过GenAlEx分析后,共识别出47种基因型。其次,使用基因型序列(47种)和参考序列(47个)组成的数据集进行系统发育分析共识别出7个系统发育分支(A-G),其中末端分支A、E、F、G仅含有云南地区的样品,分支B、C、D为来自欧洲的样本。通过多基因谱系一致性物种识别(GCPSR)方法鉴定出4个已知种,即B分支属于C. tenuithrix、D分支属于C. amethysteus、E分支属于伤锈鸡油菌(C. ferruginascens)和F分支属于C. lewisii; 1个新的系统发育种,即分支G；2个分支(A和C)可继续拆分为多个物种,即样品中的疣孢鸡油菌(C. tuberculosporus)和鸡油菌(C. cibarius)存在多个隐存种,揭示鸡油菌在欧洲和云南地区具有很高的物种多样性,自然界中仍然存在着大量还未被描述的种。此外,依据子囊菌和担子菌的分化时间(约582Mya)为基点,基于rpb1和tef1-a联合基因的分子钟分析显示鸡油菌的初始分化时间为209±32 Mya (121-335Mya,95% HPD)。该类真菌广泛分布于北温带地区,基于最大似然法分析和Bayesian Binary MCMC analysis两种方法进行的历史分布区重建结果支持该真菌主要起源于非洲南部以及美国南部地区。随后,该类真菌进而向非洲北部、欧亚大陆扩散和传播,且有明显的地区特有现象,推测可能与冰川进退导致的多次传播与隔离事件紧密相关,但也不排除人为引种其共生树种所致。
[Abstract]:Poison ditch (Trogia venenata) pleat bacterium is a controversial, can cause unexplained death (SUD) of the deadly mushrooms, mainly distributed in the Chinese southwest. To explore the lethal mechanism, researchers have analyzed the fungi may cause human death related factors: two non protein amino acids, the content of heavy metals in barium and other factors. This research adopts multi gene combined with molecular genetics of genotype of the species diversity, community structure, natural conditions, reproductive mode and genetic differentiation among populations on the fatal factors. Cantharelloid fungi of the genus has important economic, ecological and scientific value. Representative groups are Cantharellus cibarius species complex cantharelloid composite (CCSC). The Yunnan area is one of the center of modern species diversity, species richness of Cantharellus. However, due to lack of system science and Study on the current system of biogeography, fungi research mainly confined to Europe, Africa and other regions. This study uses the multi gene as a molecular marker, combined with the existing GenBank data of Cantharellus species complex species diversity, molecular systematics and biogeography are discussed, the main results are as follows: 1. poison ditch pleat bacterium (T.venenata) genetic studies in this study, 2010-2011 conducted a number of field investigation and sample collection, selected seven regions were sampled, the sampling points are mainly concentrated in the Chuxiong Dayao County, Dali City, Heqing county and the area of auspicious clouds County, Binchuan county and Tengchong County, one of the five areas the unexplained death case, 232 samples were collected. Using ITS, beta -tub, rpb2 and tef1- alpha four gene fragments of the virus, using the method of channel pleat bacterium combination gene analysis (T. venenata) group. Pass the study. In this study, ITS fragments for molecular identification of samples, molecular genetic analysis for the other three protein encoding genes. The results show that the beta -tub, rpb2 and tef1- alpha three gene fragments to identify abundant genetic variation, can accurately reflect the different level of genetic differentiation among populations and haplotype analysis. Diversity, molecular genetic differences (FST) genetic analysis reveals the T. venenata group in high diversity and population structure of molecular variation is mainly distributed in local groups within groups between structural features evident in 7 geographical populations belong to 4 genetic groups, between groups have lower levels of gene flow, and spore propagation the process of sexual reproduction to an important role in maintaining the genetic diversity and stability; Hardy Weinberg equilibrium test and single nucleotide polymorphism (SNPs) based on chain balance Venenata analysis showed that the T. groups follow the random mating mode, an important part of the results of.Mantel detection methods is the inbreeding coefficient of reproductive patterns of bacteria showed weak positive correlation between genetic distance and geographic distance between T. and venenata groups, the genetic differentiation and barium content was negatively related to.2. (CCSC) and cantharelloid complex biogeography this study selected 16 areas of Yunnan province and 5 in Europe a total of 414 samples of cantharelloid molecular phylogeny, using rpb1, CHS1 and tef1- alpha, ATP6 four gene, has carried on the system research and development system for recognition. Firstly, compare Blast with 309 samples tef1- alpha sequences, the results show that 6 of our sample chanterelle and GenBank database in a sequence of fungi of the genus. The highest similarity of alignment and conserved regions to meet the requirements of sequence selection Choose, through GenAlEx analysis, identified 47 genotypes. Secondly, using gene sequence (47 species) and the reference sequence (47) phylogenetic analysis identified 7 phylogenetic branches consisting of data sets (A-G), the end of E, F, A branch, G contains only Yunnan area branch of B, C, samples, D samples from Europe. By multi gene pedigree consistency species identification (GCPSR) method identified 4 known species, which belongs to the C. tenuithrix D B branch, C. branch belonging to amethysteus, E belong to the branch injury (C. ferruginascens) and rust cantharelloid F branch belonging to C. lewisii; 1 new system development, namely branch G; 2 branches (A and C) may continue to be split into multiple species, namely in the sample (C. tuberculosporus) cantharelloid wart spore (C. cibarius) and cantharelloid there are many cryptic species, revealing the cantharelloid has very high species diversity in Europe and the Yunnan region In nature, there are still a large number of undescribed species. In addition, on the basis of ascomycetes and basidiomycetes differentiation time (about 582Mya) as the starting point, rpb1 and tef1-a combined with molecular clock gene analysis showed that the initial differentiation time is 209 + 32 cantharelloid based on Mya (121-335Mya, 95% HPD). The fungi are widely distributed in the north temperate zone, the maximum likelihood method and Bayesian Binary MCMC analysis two methods based on the reconstruction of historical distribution results support the fungus mainly originated in southern Africa and the southern United States. Subsequently, the fungi and then to North Africa, Eurasia, spread, and endemic phenomenon is obvious. May lead to multiple and glacier spread and isolation events are closely related, but does not rule out the introduction of human symbiotic species.

【学位授予单位】：云南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：Q949.32
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本文编号：1423622