【摘要】：獼猴桃屬植物存在普遍的雜交漸滲現(xiàn)象,其屬下分類(lèi)一直以來(lái)都存在爭(zhēng)議,因此,該屬植物的系統(tǒng)發(fā)育關(guān)系有待于進(jìn)一步揭示。隨著測(cè)序技術(shù)的快速發(fā)展,葉綠體基因組被廣泛用于植物物種的系統(tǒng)發(fā)育關(guān)系研究。相較于編碼區(qū)而言,葉綠體基因組非編碼區(qū)序列更有利于解析低分類(lèi)階元植物物種系統(tǒng)發(fā)育關(guān)系。為獲得更多獼猴桃葉綠體基因組相關(guān)數(shù)據(jù),本研究對(duì)毛花獼猴桃(Actinidia eriantha)、軟棗獼猴桃(Actinidiaarguta)和狗棗獼猴桃(Actinidiakolomikta)進(jìn)行了葉綠體全基因組的測(cè)定與注釋。分析與比較了已經(jīng)測(cè)得的九個(gè)獼猴桃屬植物葉綠體基因組結(jié)構(gòu),同時(shí)通過(guò)比對(duì)尋找葉綠體基因間隔區(qū)變異位點(diǎn),開(kāi)發(fā)了葉綠體基因間隔區(qū)內(nèi)具有多態(tài)性位點(diǎn)的分子標(biāo)記,對(duì)獼猴桃屬植物屬內(nèi)系統(tǒng)發(fā)育關(guān)系進(jìn)行重建。主要研究結(jié)果如下:(1)毛花獼猴桃、軟棗獼猴桃和狗棗獼猴桃葉綠體基因組均為環(huán)狀雙鏈DNA,總長(zhǎng)度在156,484-157,425bp之間,有典型的4個(gè)區(qū)域:一對(duì)反向重復(fù)區(qū)長(zhǎng)度在23,892-24,226bp之間,大的單拷貝區(qū)域長(zhǎng)度在88,093-88,639bp之間,小的單拷貝區(qū)域長(zhǎng)度在20,475-20,579bp之間。三個(gè)基因組均編碼113個(gè)不同的基因,包括79個(gè)蛋白編碼基因,30個(gè)tRNA基因和4個(gè)rRNA基因,其中有16個(gè)基因在重復(fù)區(qū)有一個(gè)拷貝,一個(gè)基因(trnfM-CAU)在大的單拷貝區(qū)域有一個(gè)拷貝。毛花獼猴桃、軟棗獼猴桃和狗棗獼猴桃三個(gè)葉綠體基因組中包含的重復(fù)序列總數(shù)目依次為39、25、33;軟棗獼猴桃葉綠體基因組的重復(fù)序列中不存在60bp以下的重復(fù)序列。使用位點(diǎn)模型檢測(cè)到y(tǒng)cf2,accD和一個(gè)遺傳系統(tǒng)基因rpl20在獼猴桃的進(jìn)化過(guò)程中經(jīng)歷了正選擇作用。(2)利用mVISTA對(duì)葉綠體基因組序列一致性差異進(jìn)行比較分析,共檢測(cè)到24個(gè)基因間隔區(qū)序列存在較大差異。選擇其中4個(gè)基因間隔區(qū)區(qū)間:rps16-trnQ(UUG)、ndhC-trnV(UAC)、ndhF-rpl32 和 trnE(UUC)-trnT(GGU)進(jìn)行引物設(shè)計(jì),用于獼猴桃屬植物的系統(tǒng)發(fā)育關(guān)系重建。研究結(jié)果顯示四個(gè)基因間隔區(qū)的序列更好地揭示了獼猴桃系統(tǒng)發(fā)育關(guān)系;除了支持凈果組作為一個(gè)獨(dú)立進(jìn)化分支外,其他3個(gè)組(斑果組、星毛組和糙毛組)都不成單系;現(xiàn)有的獼猴桃分類(lèi)系統(tǒng)有待于進(jìn)一步完善。
[Abstract]:There is a general phenomenon of hybrid infiltration in Actinidia, and the classification of Actinidia has always been controversial. Therefore, the phylogenetic relationship of Actinidia needs to be further revealed. With the rapid development of sequencing, chloroplast genome is widely used to study phylogenetic relationship of plant species. Compared with the coding region, the sequence of chloroplast genome noncoding region is more favorable to the phylogenetic relationship of plant species with low taxonomic order. In order to obtain more relevant data of chloroplast genome of Actinidia chinensis, the whole chloroplast genome of Actinidia (Actinidia eriantha), soft date kiwifruit (Actinidiaarguta) and dog jujube kiwifruit (Actinidiakolomikta) were determined and annotated in this study. The chloroplast genome structure of nine kiwifruit plants was analyzed and compared. At the same time, a molecular marker with polymorphic loci in chloroplast spacer region was developed by comparing and searching for variation sites in chloroplast spacer region. The phylogenetic relationship of genus Actinidia was reconstructed. The main results were as follows: (1) the chloroplast genomes of Actinidia chinensis, Actinidia chinensis and Actinidia angustifolia were all circular double-stranded DNA, with total length of 156484-157425bp. There are four typical regions: the length of a pair of reverse repeat regions is between 23892-24226bp, the length of large single copy region is between 88093-88639bp, and the length of small single copy region is between 20475-20579bp. The three genomes all encode 113 different genes, including 79 protein coding genes, 30 tRNA genes and 4 rRNA genes, 16 of which have a copy in the repeat region. A gene (trnfM-CAU) has a copy in a large single copy region. The total number of repeats contained in the chloroplast genomes of Actinidia deliciosa, Actinidia chinensis and Actinidia angustifolia was 390.2533, and the repeat sequence of Actinidia chinensis chloroplast genome did not contain the repeat sequence below 60bp. The site model was used to detect the positive selection of ycf2,accD and a genetic system gene rpl20 in the evolution of kiwifruit. (2) the difference of chloroplast genome sequence consistency was analyzed by mVISTA. A total of 24 gene spacer sequences were detected. Rps16-trnQ (UUG), ndhC-trnV (UAC), ndhF-rpl32 and trnE (UUC)-trnT (GGU) were selected for primer design to reconstruct phylogenetic relationship of Actinidia. The results showed that the sequence of the four spacers better revealed the phylogenetic relationship of kiwifruit, except for supporting the net fruit group as an independent evolutionary branch, none of the other three groups (fruiting group, stellate group and strigose group) were single. The existing kiwifruit classification system needs to be further improved.
【學(xué)位授予單位】：中國(guó)科學(xué)院武漢植物園
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：Q943.2

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