番茄控制早开花的主效QTL的精细定位

发布时间：2018-01-09 07:05

本文关键词：番茄控制早开花的主效QTL的精细定位　出处：《中国农业科学院》2015年博士论文　论文类型：学位论文

更多相关文章：番茄 早开花 QTL-Sequence mi RNA Ycf2

【摘要】：开花标志着植物从营养生长阶段进入生殖生长阶段,该过程受遗传和环境共同影响。在相同环境条件下,早开花意味着植物能够利用较少的能量进入生殖生长,这对于植物规避恶劣环境(如低温)完成生命周期有重要意义。番茄上,开花时间是一个由多基因控制的数量性状,开花早晚与熟性有较高的相关性。同时,早熟意味着较高的产值。因此,阐明开花时间的遗传基础具有重要意义。本研究以071-440(晚开花,P2)与Bone MM(早开花,P1)两个栽培番茄构建的F2群体为研究对象,利用混合分组(BSA)QTL-Seq技术定位主效QTLs,结合基因表达分析筛选候选基因,并对两个已报道的mi RNA进行了表达模式分析,获得的主要结果如下:1.基于QTL-Seq定位了7个与番茄早开花性状相关的主效QTL。以构建的1200个F2单株为研究对象,选取极端早熟和极端晚熟的各40个单株的DNA等量混合,组成两个子代池,与亲本池共同测序。采用QTL-Seq分析将番茄的早开花性状定位于1号染色体上,其物理区域为1.6Mb-71.8Mb。根据SNP-index的结果,在此区域发现了7个主效QTL位点,分别位于23.5Mb-24Mb、24Mb-28Mb、35Mb-40Mb、52.5Mb-54Mb、59Mb-62Mb、69Mb-72Mb和70Mb-74Mb之间。根据番茄基因组注释的结果,在上述区域预测得到了与早开花性状相关的7个基因,分别为EF1(Solyc01g017060),EF2(Solyc01g018000),EF3(Solyc01g034220),EF4(Solyc01g057370),EF5(Solyc01g058640),EF6(Solyc01g073700)和EF7(Solyc01g081360)。2.结合传统QTL分析,筛选获得了1个与番茄早开花相关的QTL。根据QTL-Seq分析的结果,在上述7个区域设计了143个Indel标记,随机挑选136个F2单株进行QTL分析。QTL分析结果表明:番茄早开花的QTL位于标记Fl-Indel 2和Fl-Indel 8之间,对应的物理位置为23.5Mb-25.3Mb。该区域与测序分析预测的23.5Mb-24Mb区间存在较高的一致性,即EF1所在的区域。3.对测序分析预测的7个基因,设计引物,利用实时荧光定量q PCR对7个基因在双亲间的表达进行了研究。结果表明7个基因中EF1在双亲中相对表达量最高,相对表达水平分别为219.34(P1)和73.46(P2),达到显著差异水平(P?0.05)。因此EF1可能参与了番茄开花时间的调控。经BLAST分析,该基因与番茄的Ycf2基因相似度较高。4.研究了mi R156和mi R172在番茄不同叶位(从下部向上,依次为第1,3,4,6,9,12片叶)的表达模式。结果显示,mi R156和mi R172在P1(早开花)中的表达随叶位增加表达量增加,均在第12片急剧升高,达到最高值,分别为1.12倍和4.88倍,而P1的始花节位为第6片叶。这两个mi RNA在P2(第12片叶始花)中的表达存在差异,mi R156在第3片叶时相对表达量最高(2.18倍),mi R172在第9片叶时相对表达量最高(62.12倍),第12片叶时表达量下降。因此,这两个mi RNA的表达与早开花没有必然的联系。
[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.

【学位授予单位】：中国农业科学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：S641.2

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