本文關(guān)鍵詞：番茄控制早開花的主效QTL的精細(xì)定位　出處：《中國農(nóng)業(yè)科學(xué)院》2015年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 番茄 早開花 QTL-Sequence mi RNA Ycf2

【摘要】：開花標(biāo)志著植物從營養(yǎng)生長階段進(jìn)入生殖生長階段,該過程受遺傳和環(huán)境共同影響。在相同環(huán)境條件下,早開花意味著植物能夠利用較少的能量進(jìn)入生殖生長,這對于植物規(guī)避惡劣環(huán)境(如低溫)完成生命周期有重要意義。番茄上,開花時間是一個由多基因控制的數(shù)量性狀,開花早晚與熟性有較高的相關(guān)性。同時,早熟意味著較高的產(chǎn)值。因此,闡明開花時間的遺傳基礎(chǔ)具有重要意義。本研究以071-440(晚開花,P2)與Bone MM(早開花,P1)兩個栽培番茄構(gòu)建的F2群體為研究對象,利用混合分組(BSA)QTL-Seq技術(shù)定位主效QTLs,結(jié)合基因表達(dá)分析篩選候選基因,并對兩個已報道的mi RNA進(jìn)行了表達(dá)模式分析,獲得的主要結(jié)果如下:1.基于QTL-Seq定位了7個與番茄早開花性狀相關(guān)的主效QTL。以構(gòu)建的1200個F2單株為研究對象,選取極端早熟和極端晚熟的各40個單株的DNA等量混合,組成兩個子代池,與親本池共同測序。采用QTL-Seq分析將番茄的早開花性狀定位于1號染色體上,其物理區(qū)域為1.6Mb-71.8Mb。根據(jù)SNP-index的結(jié)果,在此區(qū)域發(fā)現(xiàn)了7個主效QTL位點(diǎn),分別位于23.5Mb-24Mb、24Mb-28Mb、35Mb-40Mb、52.5Mb-54Mb、59Mb-62Mb、69Mb-72Mb和70Mb-74Mb之間。根據(jù)番茄基因組注釋的結(jié)果,在上述區(qū)域預(yù)測得到了與早開花性狀相關(guān)的7個基因,分別為EF1(Solyc01g017060),EF2(Solyc01g018000),EF3(Solyc01g034220),EF4(Solyc01g057370),EF5(Solyc01g058640),EF6(Solyc01g073700)和EF7(Solyc01g081360)。2.結(jié)合傳統(tǒng)QTL分析,篩選獲得了1個與番茄早開花相關(guān)的QTL。根據(jù)QTL-Seq分析的結(jié)果,在上述7個區(qū)域設(shè)計了143個Indel標(biāo)記,隨機(jī)挑選136個F2單株進(jìn)行QTL分析。QTL分析結(jié)果表明:番茄早開花的QTL位于標(biāo)記Fl-Indel 2和Fl-Indel 8之間,對應(yīng)的物理位置為23.5Mb-25.3Mb。該區(qū)域與測序分析預(yù)測的23.5Mb-24Mb區(qū)間存在較高的一致性,即EF1所在的區(qū)域。3.對測序分析預(yù)測的7個基因,設(shè)計引物,利用實時熒光定量q PCR對7個基因在雙親間的表達(dá)進(jìn)行了研究。結(jié)果表明7個基因中EF1在雙親中相對表達(dá)量最高,相對表達(dá)水平分別為219.34(P1)和73.46(P2),達(dá)到顯著差異水平(P?0.05)。因此EF1可能參與了番茄開花時間的調(diào)控。經(jīng)BLAST分析,該基因與番茄的Ycf2基因相似度較高。4.研究了mi R156和mi R172在番茄不同葉位(從下部向上,依次為第1,3,4,6,9,12片葉)的表達(dá)模式。結(jié)果顯示,mi R156和mi R172在P1(早開花)中的表達(dá)隨葉位增加表達(dá)量增加,均在第12片急劇升高,達(dá)到最高值,分別為1.12倍和4.88倍,而P1的始花節(jié)位為第6片葉。這兩個mi RNA在P2(第12片葉始花)中的表達(dá)存在差異,mi R156在第3片葉時相對表達(dá)量最高(2.18倍),mi R172在第9片葉時相對表達(dá)量最高(62.12倍),第12片葉時表達(dá)量下降。因此,這兩個mi RNA的表達(dá)與早開花沒有必然的聯(lián)系。
[Abstract]:Marks the flowering plant growth stage from nutrients into the reproductive growth stage, the process is affected by both genetic and environmental. Under the same condition, early flowering means that plants can use less energy to reproductive growth, the plant to avoid bad environment (such as temperature) has important significance to complete the life cycle. Tomato, flowering time is a quantitative character controlled by multi genes, flowering and maturity have higher correlation. At the same time, early means higher value. Therefore, has important significance in elucidating the genetic basis of the flowering time. In this study, 071-440 (late flowering, P2) and Bone MM (early flowering, two cultivated P1) construction of tomato F2 group as the research object, using the hybrid group (BSA) QTL-Seq technology to locate the main effect QTLs, combined with gene expression analysis of candidate gene screening, and two reported mi RNA expression pattern analysis, The main results are as follows: 1. QTL-Seq and 7 tomato early flowering related traits of major QTL. to construct 1200 F2 individuals based on as the research object, selects the 40 individual extreme prematurity and extreme late mixture of DNA, composed of two sub generation pool, together with their parents by QTL-Seq sequencing pool. Analysis of Tomato Early Flowering Traits on chromosome 1, the physical area is 1.6Mb-71.8Mb. according to the results of SNP-index, this region revealed 7 major QTL sites located in 23.5Mb-24Mb, 24Mb-28Mb, 35Mb-40Mb, 52.5Mb-54Mb, 59Mb-62Mb, 69Mb-72Mb and 70Mb-74Mb between the tomato genome annotation. According to the results, in the regional forecast with Early Flowering Traits of 7 genes, respectively. EF1 (Solyc01g017060), EF2 (Solyc01g018000), EF3 (Solyc01g034220), EF4 (Solyc01g057370), EF5 (Solyc01g058640), EF6 (Solyc01g07 3700) and EF7 (Solyc01g081360).2. combined with traditional QTL analysis, and obtained 1 tomato early flowering related QTL. according to the result of QTL-Seq screening, in the above 7 areas to design 143 Indel markers, 136 randomly selected F2 plants were analyzed by QTL.QTL analysis results showed that the tomato early flowering QTL in marker Fl-Indel 2 and Fl-Indel 8, corresponding to the physical location of the 23.5Mb-25.3Mb. region and sequencing analysis of 23.5Mb-24Mb interval prediction is consistent, namely 7 genes,.3. EF1's regional prediction and analysis of sequencing primers designed on the expression of 7 genes between the parents were studied using real-time fluorescence quantitative Q PCR. The results showed that 7 genes in the EF1 parent relative expression is highest, the relative expression levels were 219.34 (P1) and 73.46 (P2), reached the level of significant difference (P? 0.05). Therefore, EF1 may be involved in the flowering of Tomato Time control. By BLAST analysis,.4. Ycf2 gene similarity of the gene and study of the MI R156 and tomato mi R172 in tomato leaves in different positions (from lower to upper, followed by the 1,3,4,6,9,12 leaf) expression patterns. The results showed that MI R156 and MI R172 in P1 (early flowering) expression in with the increase of leaf expression increase, increased sharply in twelfth, reached the highest value, respectively 1.12 times and 4.88 times, while the P1 node of the first flower for sixth leaves. The two mi RNA in P2 (twelfth leaf Shihua) there are differences in the expression of MI R156 in the third leaf the relative expression was the highest (2.18 times), MI R172 in the ninth leaf relative expression was the highest (62.12 times), Twelfth piece of leaves decreased. Therefore, the expression of the two mi RNA is not necessarily associated with early flowering.

【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S641.2

【參考文獻(xiàn)】

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本文編號：1400428