本文選題：擬南芥 + START結(jié)構(gòu)域　； 參考：《中國(guó)農(nóng)業(yè)科學(xué)》2017年17期

【摘要】：【目的】分析擬南芥中僅含START結(jié)構(gòu)域(the lipid/sterol-binding St AR-related lipid transfer protein domains)亞家族成員的啟動(dòng)子元件及表達(dá)特性,闡明啟動(dòng)子元件與表達(dá)特性的相互關(guān)系,為明確僅含START結(jié)構(gòu)域亞家族的生物學(xué)功能,解析植物生長(zhǎng)發(fā)育機(jī)理、更好地促控植物生長(zhǎng)提供理論依據(jù)�！痉椒ā坷蒙镄畔W(xué)方法分析僅含START結(jié)構(gòu)域亞家族6個(gè)成員的親緣關(guān)系及各自啟動(dòng)子區(qū)所包含的所有元件,對(duì)啟動(dòng)子元件的功能進(jìn)行分析和分類(lèi);利用real-time PCR方法研究亞家族成員的表達(dá)特性;用突變體分析確認(rèn)基因的功能;分析通過(guò)啟動(dòng)子元件分析預(yù)測(cè)的基因功能與通過(guò)突變體分析確認(rèn)的基因功能間的相關(guān)性,為基因的功能和作用機(jī)制分析提供科研思路和方法。【結(jié)果】生物信息學(xué)分析表明,擬南芥中僅含START結(jié)構(gòu)域亞家族成員共6個(gè),分為3個(gè)分支,每個(gè)分支一對(duì)基因,每一對(duì)基因中均有一個(gè)基因的啟動(dòng)子中含有高轉(zhuǎn)錄水平元件5′UTR Py-rich stretch,6個(gè)亞家族成員的啟動(dòng)子區(qū)均含有多個(gè)光、激素和逆境響應(yīng)元件,也存在各自特異的響應(yīng)元件;用real-time PCR分析基因的表達(dá)量,結(jié)果與生物信息學(xué)預(yù)測(cè)結(jié)果一致:At3g13062的表達(dá)量明顯高于同分支的At1g55960,At3g23080的表達(dá)量明顯高于同分支的At4g14500,At1g64720的表達(dá)量明顯高于同分支的At5g54170。組織定位分析表明亞家族成員有特異的時(shí)空表達(dá)特性:At3g13062和At1g55960在幼苗期就有明顯表達(dá),但At3g13062主要在子葉、下胚軸和根中表達(dá),而At1g55960主要在根中表達(dá)量較多;在成苗期擬南芥中,At3g13062主要在葉片中表達(dá),在根中表達(dá)量較少;而At1g55960在成苗期擬南芥的葉片和根系的表達(dá)量均較多;At3g23080和At4g14500在幼苗期表達(dá)量較少,而在成苗期表達(dá)量增加,At3g23080主要分布于蓮座葉中,At4g14500主要分布于蓮座葉葉柄部。上述結(jié)果表明僅含START結(jié)構(gòu)域亞家族成員可能在擬南芥不同時(shí)期和不同的部位分別行使著功能。利用生物信息學(xué)對(duì)啟動(dòng)子元件分析發(fā)現(xiàn),各個(gè)基因均具有胚乳特意表達(dá)的Skn-1 motif元件,暗示著該家族基因可能參與調(diào)控胚乳的發(fā)育,進(jìn)而影響種子的活力和萌發(fā);通過(guò)對(duì)At3g23080和At4g14500的T-DNA插入突變體種子的分析表明,At3g23080和At4g14500表達(dá)量下降均會(huì)導(dǎo)致種子活力和萌發(fā)率的下降,這與生物信息學(xué)預(yù)測(cè)結(jié)果一致。【結(jié)論】擬南芥中僅含START結(jié)構(gòu)域的亞家族有6個(gè)成員;6個(gè)亞家族成員可能在光、激素和逆境調(diào)控的發(fā)育過(guò)程中具有重要的作用;含START結(jié)構(gòu)域的亞家族6個(gè)成員雖然同源關(guān)系較近,但具有各自的時(shí)空表達(dá)特性,在擬南芥的不同組織和不同發(fā)育時(shí)期執(zhí)行著各自的功能。基因啟動(dòng)子元件與基因功能有直接關(guān)系,啟動(dòng)子區(qū)具有胚乳特意表達(dá)Skn-1 motif元件的At3g23080和At4g14500表達(dá)量下降均會(huì)導(dǎo)致種子活力和萌發(fā)率下降。
[Abstract]:[objective] to analyze the promoter elements and expression characteristics of the members of the lipid/sterol-binding St AR-related lipid transfer protein domains) subfamily containing only the START domain in Arabidopsis thaliana, and to elucidate the relationship between the promoter elements and the expression characteristics. In order to clarify the biological function of the subfamily containing only START domain, the mechanism of plant growth and development was analyzed. [methods] bioinformatics was used to analyze the relationship among the six members of the START domain subfamily and all the elements contained in their respective promoter regions. The function of promoter element was analyzed and classified, the expression characteristics of subfamily members were studied by real-time PCR method, the function of gene was confirmed by mutants analysis. The correlation between gene function predicted by promoter element analysis and gene function confirmed by mutant analysis was analyzed, which provided scientific research ideas and methods for gene function and mechanism analysis. [results] Bioinformatics analysis showed that, In Arabidopsis thaliana, there are only 6 members of START domain subfamily, which are divided into three branches, each branching a pair of genes. In each pair of genes, the promoter of one gene contains a high transcription level element 5'UTR Py-rich retch, and the promoter regions of the six subfamily members contain multiple light, hormone and stress response elements, as well as their own specific response elements. Real-time PCR was used to analyze the expression of the gene. The results were consistent with the bioinformatics prediction. The expression of 1: At3g13062 was significantly higher than that of At1g55960U / At3g23080 with the same branch, which was significantly higher than that of At4g14500 / At1g64720 with the same branch. Tissue localization analysis showed that the members of the subfamily had special temporal and spatial expression characteristics: At3g13062 and At1g55960 were expressed obviously at seedling stage, but At3g13062 was mainly expressed in cotyledons, hypocotyls and roots, while At1g55960 was mainly expressed in roots. At3g13062 was expressed mainly in leaves and less in roots in Arabidopsis thaliana at seedling stage, while At1g55960 expression in leaves and roots of Arabidopsis thaliana was more than that in leaves and roots of Arabidopsis thaliana at seedling stage, and the expression of At3g23080 and At4g14500 in seedling stage was less than that in seedling stage. At the seedling stage, At3g23080 was mainly distributed in the leaves of rosette, and At4g14500 was mainly distributed in the petiole of rosette leaves. These results suggest that only members of the START domain subfamily may function at different stages and parts of Arabidopsis thaliana. By bioinformatics analysis of promoter elements, it was found that all genes had Skn-1 motif elements specifically expressed in endosperm, suggesting that the family genes might be involved in regulating the development of endosperm, thus affecting the vigor and germination of seeds. The analysis of At3g23080 and At4g14500 T-DNA inserted mutant seeds showed that the decreased expression of At3g23080 and At4g14500 resulted in the decrease of seed vigor and germination rate. [conclusion] in Arabidopsis thaliana, there are 6 members of the subfamily with only START domain, 6 members may play an important role in the development of light, hormone and stress regulation. Although the six members of the subfamily with START domain share close homology, they have their own spatio-temporal expression characteristics and perform their own functions in different tissues and developmental stages of Arabidopsis thaliana. The gene promoter element is directly related to gene function. The decrease of At3g23080 and At4g14500 expression in the promoter with endosperm specially expressing Skn-1 motif elements will lead to the decrease of seed vigor and germination rate.
【作者單位】： 河北農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院/河北省植物生理和分子病理學(xué)重點(diǎn)實(shí)驗(yàn)室;
【基金】：河北省自然科學(xué)基金(C2017204060)
【分類(lèi)號(hào)】：Q943.2

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1 趙士洞;從START常務(wù)委員會(huì)第八次會(huì)議看START建設(shè)的新進(jìn)展[J];地球科學(xué)進(jìn)展;1995年01期

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