【摘要】：小麥(Triticum aestivum)是世界上種植最廣泛的谷物,它提供了大約20%的人類所消耗的熱量。預(yù)計(jì)到2050年對小麥的需求可能會增加60%。因此,提高小麥產(chǎn)量尤為迫切。千粒重、單位面積穗數(shù)、穗粒數(shù)作為產(chǎn)量三要素是提高小麥產(chǎn)量的重要途徑。除此之外,種質(zhì)資源也在提高小麥產(chǎn)量上起重要作用。目前,小麥育種普遍存在種質(zhì)資源匱乏等問題,迫切需要新種質(zhì)的發(fā)現(xiàn)和創(chuàng)造。人工誘導(dǎo)產(chǎn)生突變體、構(gòu)建突變體庫,可以為小麥功能基因研究和小麥遺傳改良提供基礎(chǔ)材料。本研究以煙農(nóng)15和經(jīng)EMS誘變煙農(nóng)15獲得的農(nóng)藝性狀差異較大的110個(gè)株系為試驗(yàn)材料,以小麥12個(gè)主要產(chǎn)量性狀為研究對象,利用RNA-Seq技術(shù)開發(fā)高通量單核苷酸多態(tài)性(SNP)和插入缺失(InDel)標(biāo)記用于關(guān)聯(lián)分析,為小麥遺傳育種提供候選關(guān)聯(lián)標(biāo)記及基因。主要結(jié)果如下:1.本研究對煙農(nóng)15開花后9天(DPA9)、18天(DPA18)、27天(DPA27)三個(gè)時(shí)期樣本進(jìn)行RNA-Seq技術(shù)測序,采用參考基因組拼接的方法對小麥轉(zhuǎn)錄組進(jìn)行組裝,得到較完整的轉(zhuǎn)錄組。共獲得195601條轉(zhuǎn)錄本,平均長度1988bp。2.本研究對110個(gè)突變體株系開花后9天(DPA9)、18天(DPA18)、27天(DPA27)三個(gè)時(shí)期RNA等量等濃度混合后進(jìn)行RNA-Seq測序,在已拼接好的轉(zhuǎn)錄組基礎(chǔ)上,共在110個(gè)突變體株系中開發(fā)了126,980個(gè)標(biāo)記,其中包括101,876個(gè)SNP標(biāo)記,25,104個(gè)Indel標(biāo)記。3.對突變體群體進(jìn)行表型分析和相關(guān)性分析,結(jié)果表明突變體株系的12個(gè)產(chǎn)量性狀均有較大的變異,變異系數(shù)在4.12~111.90%之間,變異最小的性狀為粒長、變異最大性狀為頂部不育小穗數(shù)。多數(shù)性狀之間相關(guān)性顯著。4.利用126,980個(gè)標(biāo)記對產(chǎn)量相關(guān)性狀進(jìn)行關(guān)聯(lián)分析,共檢測到84個(gè)標(biāo)記與9個(gè)產(chǎn)量性狀在P7.87E-6水平存在顯著關(guān)聯(lián),其中,與株高、小穗數(shù)、頂部不育小穗數(shù)、不育小穗數(shù)、可育小穗數(shù)、穗數(shù)、千粒重、粒長、粒寬顯著關(guān)聯(lián)的標(biāo)記數(shù)分別為2、1、38、5、3、1、6、25、3個(gè);檢測到16個(gè)標(biāo)記與穗粒數(shù)在P7.87E-8水平存在顯著關(guān)聯(lián);檢測到25個(gè)標(biāo)記與穗長在P1E-6水平存在顯著關(guān)聯(lián)。單個(gè)標(biāo)記位點(diǎn)的變異解釋率范圍在18.209%-52.993%。
[Abstract]:Wheat (Triticum aestivum) is the most widely grown grain in the world, providing about 20 percent of the calories consumed by humans. Demand for wheat is expected to increase by 60 percent by 2050. Therefore, increasing wheat yield is particularly urgent. 1000-grain weight, panicle number per unit area and grain number per panicle are three important factors to improve wheat yield. In addition, germplasm resources also play an important role in improving wheat yield. At present, wheat breeding is generally lack of germplasm resources, so the discovery and creation of new germplasm are urgently needed. Artificial induction of mutants and construction of mutants library can provide basic materials for wheat functional gene research and wheat genetic improvement. In this study, 110 lines obtained from Yannong 15 and Yannong 15 mutated by EMS were used as experimental materials, and 12 main yield characters of wheat were studied. High throughput single nucleotide polymorphisms (SNP) and insertion deletion (InDel) (InDel) markers were developed by RNA-Seq for association analysis to provide candidate association markers and genes for wheat genetics and breeding. The main results are as follows: 1. In this study, three samples of Yannong 15 were sequenced by RNA-Seq technique in three periods (DPA9, DPA18, 27 days (DPA27). The transcriptome of wheat was assembled by reference genome splicing method, and the complete transcriptome was obtained. A total of 195601 transcripts were obtained, with an average length of 1988bp.2. In this study, RNA-Seq sequencing was carried out on 110 mutants 9 days after anthesis (DPA9), 18 days after anthesis (DPA18) and 27 days after anthesis (DPA27). On the basis of spliced transcriptome, 126980 markers were developed in 110 mutants. These include 101876 SNP tags and 25104 Indel tags. Phenotypic analysis and correlation analysis of the mutant population showed that the 12 yield traits of the mutants had great variation, the coefficient of variation was between 4.12 and 111.90%, and the least variation was grain length. The maximum variation was the number of sterile spikelets at the top. The correlation between most traits was significant. 4. A total of 84 markers and 9 yield traits were found to be significantly correlated at P7.87E-6 level with 126980 markers, including plant height, spikelet number, number of sterile spikelets at the top, number of fertile spikelets, and number of fertile spikelets. The number of markers significantly correlated with panicle number, 1000-grain weight, grain length and grain width were 2 ~ (1) 1 ~ (38) ~ (35) ~ (35) ~ (3) ~ (1) ~ (1) ~ (1) ~ 625, 3, respectively, and 16 markers were significantly correlated with grain number per spike at P7.87E-8 level, and 25 markers were found to be significantly correlated with ear length at P1E-6 level. The interpretation rate of single marker loci ranged from 18.209 to 52.993.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S512.1

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