本文關(guān)鍵詞： 花生 SSR 熒光檢測(cè)法 遺傳多樣性 指紋圖譜　出處：《曲阜師范大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：花生是重要的油料作物和經(jīng)濟(jì)作物。中國(guó)擁有較為豐富的種質(zhì)資源，但是隨著針對(duì)性育種工作的廣泛進(jìn)行，花生栽培種遺傳背景越來越狹窄。遺傳多樣性研究是進(jìn)行花生栽培種育種改良的基礎(chǔ)，而品種的指紋圖譜鑒定可為育種工作的知識(shí)產(chǎn)權(quán)保護(hù)提供科學(xué)依據(jù)。本文以篩選出的100對(duì)SSR引物分析144份花生栽培種的遺傳多樣性，并對(duì)銀染和熒光檢測(cè)技術(shù)兩種SSR PCR擴(kuò)增產(chǎn)物的檢測(cè)方法進(jìn)行對(duì)比，同時(shí)選取23對(duì)引物的159個(gè)標(biāo)記構(gòu)建群體的指紋圖譜，進(jìn)而為育種提供分子依據(jù)、為生產(chǎn)提供服務(wù)。主要研究結(jié)果如下： 1.144份花生栽培種的遺傳多樣性分析：應(yīng)用100對(duì)SSR引物對(duì)144個(gè)花生栽培種進(jìn)行PCR擴(kuò)增，共檢測(cè)到409個(gè)等位變異，每對(duì)SSR引物平均能夠檢測(cè)到4.09個(gè)等位變異；聚類分析結(jié)果表明，100對(duì)SSR引物能夠?qū)⒐┰嚨乃衅贩N區(qū)分開，聚類結(jié)果與栽培花生類型、產(chǎn)地及親緣關(guān)系呈正相關(guān)；品種遺傳相似系數(shù)為0.68-0.97。可見，總體上栽培花生遺傳基礎(chǔ)較為狹窄，特別是國(guó)內(nèi)現(xiàn)有的育成品種。多樣性分析顯示，地方品種和國(guó)外品種與其他品種間遺傳距離較大，作為親本應(yīng)用于育種可拓寬選育品種的遺傳背景。 2.銀染和熒光檢測(cè)技術(shù)的對(duì)比：選取9對(duì)多態(tài)性及特異性較好的引物，分別用銀染法和熒光檢測(cè)法掃描了85個(gè)栽培品種的SSR標(biāo)記多態(tài)性。與銀染法相比，，熒光檢測(cè)法具有靈敏度高、準(zhǔn)確性強(qiáng)、效率高、在遺傳多樣性分析方面更精確可靠等優(yōu)點(diǎn)。此外，通過高通量PCR、高通量上樣檢測(cè)及大片段檢測(cè)優(yōu)化等方法可以提高熒光檢測(cè)法檢測(cè)效率，有效降低實(shí)驗(yàn)成本。 3.144份花生栽培種SSR指紋圖譜的構(gòu)建：應(yīng)用23對(duì)等位變異數(shù)目大于5的SSR引物構(gòu)建了144個(gè)供試材料的指紋圖譜，以0、1形式統(tǒng)計(jì)，并將0、1字符串轉(zhuǎn)換為十六進(jìn)制，結(jié)果表明每一個(gè)品種的指紋圖譜均為唯一代碼。下一步將通過F1群體或親緣關(guān)系較近的品種對(duì)構(gòu)建的指紋圖譜進(jìn)行驗(yàn)證，以期為花生育種提供科學(xué)準(zhǔn)確的鑒定手段。
[Abstract]:Peanut is an important oil crop and cash crop. China has abundant germplasm resources, but with the extensive work of targeted breeding. The genetic diversity of peanut cultivars is becoming more and more narrow, which is the basis of peanut breeding. The identification of fingerprinting can provide a scientific basis for the protection of intellectual property rights in breeding. The genetic diversity of 144 peanut cultivars was analyzed with 100 pairs of SSR primers selected in this paper. Silver staining and fluorescence detection were compared between two SSR PCR amplification methods, and 159 markers of 23 pairs of primers were selected to construct the fingerprint of the population. The main results are as follows: Analysis of genetic diversity of 1.144 peanut cultivars: #number0# pairs of SSR primers were used to amplify 144 peanut cultivars and 409 alleles were detected. An average of 4.09 alleles per pair of SSR primers could be detected. The results of cluster analysis showed that all cultivars could be distinguished by SSR primer, and the cluster results were positively correlated with the type, origin and relatives of cultivated peanut. The genetic similarity coefficient of cultivars was 0.68-0.97.The genetic basis of cultivated peanut was relatively narrow in general, especially the existing varieties in China. The diversity analysis showed. The genetic distance between local varieties and foreign varieties and other varieties is great. As parents, the genetic background of selected varieties can be widened. 2. Comparison of silver staining and fluorescence detection: 9 pairs of primers with good polymorphism and specificity were selected. The polymorphism of SSR markers in 85 cultivars were scanned by silver staining method and fluorescence detection method respectively. Compared with silver staining method, fluorescent detection method has higher sensitivity, higher accuracy and higher efficiency. In addition, the efficiency of fluorescence detection can be improved by high-throughput PCR, high-throughput sample detection and optimization of large fragment detection. Effectively reduce the cost of the experiment. Construction of SSR fingerprinting of 3. 144 peanut cultivars: using 23 pairs of SSR primers with allelic variation of more than 5, the fingerprints of 144 tested materials were constructed, and the results were analyzed in the form of 0 ~ 1. The result shows that the fingerprint of each variety is the unique code. The next step is to verify the fingerprint map by F1 population or the closely related variety. In order to provide a scientific and accurate identification means for peanut breeding.
【學(xué)位授予單位】：曲阜師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：S565.2

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