本文關(guān)鍵詞：毒溝褶菌（Trogia venenata）群體遺傳學(xué)及雞油菌復(fù)合種分子系統(tǒng)學(xué)與生物地理學(xué)研究　出處：《云南大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 毒溝褶菌 雞油菌復(fù)合種 物種多樣性 群體遺傳 地區(qū)特有

【摘要】：毒溝褶菌(Trogia venenata)是一個(gè)具有爭(zhēng)議的、能導(dǎo)致不明原因死亡(SUD)的致命蘑菇,主要分布在中國的西南地區(qū)。為探索其致命機(jī)制,研究人員們已經(jīng)分析了該真菌可能導(dǎo)致人類死亡的相關(guān)因素：兩個(gè)非蛋白氨基酸、重金屬鋇含量及其他因子。本研究采用多基因聯(lián)合分子遺傳學(xué)的方法探討了該物種的基因型多樣性、群體結(jié)構(gòu)、自然條件下的生殖模式以及群體間的遺傳分化水平對(duì)其致命因子的影響。雞油菌屬真菌具有重要的經(jīng)濟(jì)、生態(tài)和科研價(jià)值,代表類群是雞油菌復(fù)合種Cantharellus cibarius species complex (CCSC)。云南地區(qū)是現(xiàn)代物種多樣性中心之一,雞油菌物種豐富。但是,由于缺乏系統(tǒng)學(xué)和生物地理學(xué)研究,目前該屬真菌的系統(tǒng)學(xué)研究主要局限于歐美、非洲等地區(qū)。本研究利用多基因片段作為分子標(biāo)記,結(jié)合GenBank中已有的數(shù)據(jù),對(duì)雞油菌復(fù)合種的物種多樣性、分子系統(tǒng)學(xué)和生物地理學(xué)等進(jìn)行了探討,主要結(jié)果如下：1.毒溝褶菌(T.venenata)群體遺傳學(xué)研究本研究于2010-2011年進(jìn)行了多次實(shí)地調(diào)查和樣品采集,選擇了七個(gè)地區(qū)進(jìn)行了采樣,采樣點(diǎn)主要集中于楚雄大姚縣、大理市祥云縣、鶴慶縣和賓川縣以及騰沖縣等地區(qū),其中五個(gè)地區(qū)有不明原因死亡實(shí)例,共采集了232份樣品。采用了ITS、β-tub、rpb2和tef1-α四個(gè)基因片段,運(yùn)用多基因聯(lián)合分析的方法對(duì)毒溝褶菌(T. venenata)進(jìn)行群體遺傳學(xué)研究。本研究中ITS片段用于對(duì)樣本進(jìn)行分子鑒定,其他三個(gè)蛋白編碼基因用于分子遺傳學(xué)分析。結(jié)果表明,β-tub、rpb2和tef1-α三個(gè)基因片段能夠識(shí)別豐富的遺傳變異位點(diǎn),可以準(zhǔn)確反映各個(gè)地理群體間的遺傳分化水平。單倍型多樣性分析、分子遺傳差異(FST)和群體結(jié)構(gòu)分析揭示T. venenata群體內(nèi)的遺傳多樣性較高,分子變異主要分布于地方群體內(nèi)部,群體之間存在明顯的結(jié)構(gòu)特征—7個(gè)地理群體屬于4個(gè)遺傳群體,群體間有較低水平的基因交流,孢子傳播及有性生殖過程對(duì)保持群體遺傳多樣性和穩(wěn)定性起到重要作用；哈迪-溫伯格平衡檢測(cè)和基于單核苷酸多態(tài)性位點(diǎn)(SNPs)的連鎖平衡分析證明T. venenata群體間遵循隨機(jī)交配模式,近親繁殖方式也是該菌生殖模式的重要組成部分。Mantel檢測(cè)結(jié)果顯示T. venenata群體之間的遺傳距離與地理距離之間具有微弱的正相關(guān)關(guān)系,其遺傳分化與鋇含量呈負(fù)相關(guān)關(guān)系。2.雞油菌復(fù)合種(CCSC)分子系統(tǒng)學(xué)與生物地理學(xué)研究本研究選取了云南省16個(gè)地區(qū)和5個(gè)歐洲地區(qū)共414份雞油菌樣品,采用了tef1-α, rpb1，chs1和ATP6四個(gè)基因,對(duì)其進(jìn)行了系統(tǒng)學(xué)研究和系統(tǒng)發(fā)育種的識(shí)別。首先,采用309份樣品的tef1-α序列進(jìn)行Blast比對(duì),結(jié)果顯示我們的樣品與GenBank數(shù)據(jù)庫中的6個(gè)雞油菌屬真菌種的序列相似度最高。將符合要求的序列進(jìn)行比對(duì)和保守區(qū)的選擇,通過GenAlEx分析后,共識(shí)別出47種基因型。其次,使用基因型序列(47種)和參考序列(47個(gè))組成的數(shù)據(jù)集進(jìn)行系統(tǒng)發(fā)育分析共識(shí)別出7個(gè)系統(tǒng)發(fā)育分支(A-G),其中末端分支A、E、F、G僅含有云南地區(qū)的樣品,分支B、C、D為來自歐洲的樣本。通過多基因譜系一致性物種識(shí)別(GCPSR)方法鑒定出4個(gè)已知種,即B分支屬于C. tenuithrix、D分支屬于C. amethysteus、E分支屬于傷銹雞油菌(C. ferruginascens)和F分支屬于C. lewisii; 1個(gè)新的系統(tǒng)發(fā)育種,即分支G；2個(gè)分支(A和C)可繼續(xù)拆分為多個(gè)物種,即樣品中的疣孢雞油菌(C. tuberculosporus)和雞油菌(C. cibarius)存在多個(gè)隱存種,揭示雞油菌在歐洲和云南地區(qū)具有很高的物種多樣性,自然界中仍然存在著大量還未被描述的種。此外,依據(jù)子囊菌和擔(dān)子菌的分化時(shí)間(約582Mya)為基點(diǎn),基于rpb1和tef1-a聯(lián)合基因的分子鐘分析顯示雞油菌的初始分化時(shí)間為209±32 Mya (121-335Mya,95% HPD)。該類真菌廣泛分布于北溫帶地區(qū),基于最大似然法分析和Bayesian Binary MCMC analysis兩種方法進(jìn)行的歷史分布區(qū)重建結(jié)果支持該真菌主要起源于非洲南部以及美國南部地區(qū)。隨后,該類真菌進(jìn)而向非洲北部、歐亞大陸擴(kuò)散和傳播,且有明顯的地區(qū)特有現(xiàn)象,推測(cè)可能與冰川進(jìn)退導(dǎo)致的多次傳播與隔離事件緊密相關(guān),但也不排除人為引種其共生樹種所致。
[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.

【學(xué)位授予單位】：云南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q949.32
，

本文編號(hào)：1423622