【摘要】：銀杏是珍貴的經(jīng)濟(jì)樹種,果、葉、材等用途廣泛,有著較高的經(jīng)濟(jì)價(jià)值,銀杏是裸子植物中古老的孑遺植物,有極其重要的理論價(jià)值。我國(guó)銀杏古樹資源較為豐富,但受環(huán)境變化等諸多因素影響,正在逐漸減少,因而對(duì)銀杏古樹進(jìn)行保護(hù)與利用,顯得尤為緊迫。目前,國(guó)內(nèi)部分地區(qū)已經(jīng)開展了對(duì)其本地銀杏半同胞家系的研究,且多為家系形態(tài)指標(biāo)研究,對(duì)全國(guó)范圍內(nèi)收集銀杏種實(shí)并進(jìn)行半同胞家系研究未見報(bào)道。本研究在全國(guó)范圍內(nèi)收集98個(gè)縣市130株銀杏古樹種實(shí),播種育苗后構(gòu)建銀杏半同胞家系種質(zhì)資源圃,隨后對(duì)銀杏半同胞家系進(jìn)行生長(zhǎng)指標(biāo)測(cè)定、核型分析,AFLP分子標(biāo)記研究。本研究分別從生長(zhǎng)量、細(xì)胞水平、分子水平對(duì)銀杏半同胞家系進(jìn)行變異規(guī)律分析,并初步評(píng)價(jià)出苗期生長(zhǎng)優(yōu)良的半同胞家系;根據(jù)銀杏古樹種實(shí)形態(tài)特性,分析銀杏古樹種子分布規(guī)律,為進(jìn)一步的銀杏良種評(píng)價(jià)提供參考。此外,本研究利用BSR技術(shù)對(duì)銀杏半同胞家系進(jìn)行高通量測(cè)序,發(fā)掘與株高生長(zhǎng)性狀相關(guān)的基因,為后續(xù)開展功能基因研究和分子育種實(shí)踐提供有價(jià)值信息,以及為遺傳連鎖圖譜構(gòu)建,生長(zhǎng)性狀主效QTL的精確定位等研究提供有力工具。本研究的主要結(jié)果如下:(1)根據(jù)銀杏半同胞家系連續(xù)三年的生長(zhǎng)量,初步選擇出72 (廣西興安)等優(yōu)良銀杏半同胞家系8個(gè)。根據(jù)銀杏半同胞家系遺傳多樣性結(jié)果可知,銀杏半同胞家系北部區(qū)、西部和南區(qū)遺傳多樣性高于中部區(qū)和東部區(qū)家系。此外,28個(gè)銀杏半同胞家系核型分析表明,銀杏從貴州、四川地區(qū)向西南及東北方向進(jìn)化。因此,根據(jù)銀杏的進(jìn)化趨勢(shì)及遺傳多樣性分析,最終選出北部區(qū)、東部區(qū)、西部區(qū)和南部區(qū)優(yōu)良銀杏半同胞家系各2-3個(gè)。(2)本研究采用的8對(duì)AFLP引物對(duì)297份銀杏半同胞家系進(jìn)行擴(kuò)增,檢測(cè)到譜帶1727條,均為多態(tài)帶;8對(duì)引物中僅有4對(duì)引物有單態(tài)帶,由此可見,銀杏半同胞家系表現(xiàn)了較高的多態(tài)性水平。由Gst估算的銀杏半同胞家系基因流值Nm為1.4085,表明銀杏半同胞家系間存在一定程度的基因交流,但家系間的基因流明顯處于較低水平。(3)根據(jù)銀杏種實(shí)形態(tài)的差異,本研究將古樹種子分布地點(diǎn)劃分為5個(gè)區(qū)域:東部區(qū)、中部區(qū)、北部區(qū)、南部區(qū)和西部區(qū)。根據(jù)種子營(yíng)養(yǎng)成分分析可知,不同古樹種子營(yíng)養(yǎng)成分與地理區(qū)域關(guān)系不顯著。東部區(qū)種子主要分布于山東、江蘇、浙江三省,該區(qū)古樹種實(shí)相對(duì)較大,可重點(diǎn)進(jìn)行核用銀杏良種選育。(4)本研究用BSR技術(shù)對(duì)貴州盤縣銀杏半同胞家系高生長(zhǎng)株和矮生長(zhǎng)株兩個(gè)混池進(jìn)行轉(zhuǎn)錄組測(cè)序,識(shí)別差異表達(dá)基因601個(gè),其中有513個(gè)基因得到功能注釋。對(duì)高生長(zhǎng)株和低生長(zhǎng)株兩個(gè)樣本的reads進(jìn)行SNP分析,BSR關(guān)聯(lián)到119個(gè)基因,其中58個(gè)基因得到功能注釋。
[Abstract]:Ginkgo biloba is a precious economic tree species, fruit, leaf, wood and other widely used, has a higher economic value, ginkgo biloba is an ancient relict plant in gymnosperms, has extremely important theoretical value. The ancient ginkgo tree resources are abundant in China, but due to the environmental change and many other factors, they are gradually decreasing, so it is particularly urgent to protect and utilize the ancient ginkgo trees. At present, some local Ginkgo biloba half-sibling families have been studied in some parts of our country, and most of them are family morphological indexes. No report has been reported on the collection of ginkgo seeds and half-sib families in the whole country. In this study, 130 ancient ginkgo trees were collected from 98 counties and cities in China. After seeding and seedling raising, the germplasm resource nursery of Ginkgo biloba half-sib family was constructed, and then the growth index, karyotype analysis and AFLP molecular marker study were carried out on Ginkgo Half-sib families. In this study, the variation rules of Ginkgo biloba half-sib families were analyzed in terms of growth, cell level and molecular level, and preliminary evaluation of half-sib families with good growth at seedling stage was carried out. According to the seed morphological characteristics of ancient ginkgo trees, the seed distribution of ancient ginkgo trees is analyzed, which provides a reference for further evaluation of improved ginkgo varieties. In addition, the high throughput sequencing of Ginkgo biloba semi-sib families was carried out by using BSR technique, and the genes related to plant height growth traits were discovered, which provided valuable information for further functional gene research and molecular breeding practice. It also provides powerful tools for the construction of genetic linkage map and the precise mapping of QTL for growth traits. The main results of this study are as follows: (1) according to the growth of Ginkgo Half-sib families for three consecutive years, 8 excellent Ginkgo Half-sib families 72 (Xingan, Guangxi) were preliminarily selected. According to the genetic diversity of Ginkgo biloba half-sib families, the genetic diversity of Ginkgo biloba half-sib families in the northern, western and southern regions was higher than that in the central and eastern regions. In addition, the karyotype analysis of 28 semi-sib families of Ginkgo biloba showed that Ginkgo biloba evolved from Guizhou and Sichuan to southwest and northeast. Therefore, according to the evolutionary trend and genetic diversity analysis of Ginkgo biloba, the northern and eastern regions were selected. There were 2-3 excellent Ginkgo biloba half-sib families in the western and southern regions. (2) eight pairs of AFLP primers were used to amplify 297Ginkgo biloba half-sib families and 1727 bands were detected, all of which were polymorphic. There were only 4 pairs of primers with monomorphic bands in 8 pairs of primers. It can be seen that Ginkgo biloba semi-sib families showed a high level of polymorphism. The Nm estimated by Gst was 1.4085, which indicated that there was a certain degree of gene exchange among the half-sib families of Ginkgo biloba, but the gene flow among families was obviously at a lower level. (3) according to the differences of seed morphology of Ginkgo biloba. In this study, the seed distribution sites of ancient trees were divided into five regions: eastern region, central region, northern region, southern region and western region. According to the analysis of seed nutrient composition, there was no significant relationship between the nutritional composition and geographical region of different ancient trees. Seeds in the eastern region are mainly distributed in Shandong, Jiangsu and Zhejiang provinces, where ancient tree species are relatively large. (4) in this study, BSR technique was used to sequence the two mixed pools of high and low growth plants of Ginkgo biloba Half-sib families in Panxian, Guizhou Province, and 601 differentially expressed genes were identified. Of these, 513 genes were annotated. The reads of two samples of high and low growth plants was analyzed by SNP. The BSR was associated with 119 genes, of which 58 genes were annotated.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S792.95

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本文編號(hào)：2299052