【摘要】：果樹果實發(fā)育和成熟調(diào)控機理研究是果樹學科的核心問題之一。然而,多年來有關(guān)果樹果實發(fā)育和成熟機理的研究基本上都限定在果實生理、生化代謝層面的探討上,而對操縱果實生理、生化代謝的更為深刻的信號轉(zhuǎn)導機制幾乎沒有實質(zhì)性了解。MRLK家族(Malectin-like domain-containing receptor-like kinases)是一類含有Malectin結(jié)構(gòu)域的類受體蛋白激酶家族。因Malectin結(jié)構(gòu)域具有寡聚糖結(jié)合特性,因此推測MRLKs可能和寡糖信號轉(zhuǎn)導有著密切的關(guān)系。果樹果實發(fā)育和成熟過程中不可避免地發(fā)生細胞壁代謝和寡聚糖信號轉(zhuǎn)導,因此推測MRLKs在果樹果實發(fā)育和成熟調(diào)控中可能起著重要的作用。有鑒于此,本研究以非呼吸躍變型果實草莓為材料,對MRLKs在果樹果實發(fā)育和成熟調(diào)控中的作用進行了研究。主要結(jié)果如下:1、首先基于生物信息學分析,首次在草莓基因組中鑒定出62個MRLK家族成員,按照其在染色體上的分布位置命名為FvMRLK1-FvMRLK62。根據(jù)FvMRLK蛋白家族胞外的Malectin結(jié)構(gòu)域特征,該家族可分成6個亞家族。FvMRLK家族成員的內(nèi)含子、外顯子和Malectin保守結(jié)構(gòu)域等基因結(jié)構(gòu)和生物遺傳進化分析表明,遺傳進化相同的家族具有相似的結(jié)構(gòu)特征,相似的結(jié)構(gòu)決定具有相近的功能。2、基因表達譜分析表明,FvMRLK家族成員在根、莖、葉和果等各種組織中均有表達,但是,根中的表達量相對高于其他組織。其中,FvMRLK家族成員中有33個成員在果實中表達。在這33個成員中,60%以上成員的基因表達特征和果實發(fā)育與成熟進程有著密切的關(guān)系。具體來說,在果實發(fā)育的早期,表達量很高,隨著果實成熟表達水平急劇下降。進一步分析表明,FvMRLK家族大部分成員都可對操縱草莓果實發(fā)育和成熟的內(nèi)、外因子(如激素和環(huán)境因素)產(chǎn)生靈敏的應(yīng)答反應(yīng),這些結(jié)果表明FvMRLKs在草莓果實發(fā)育和成熟調(diào)控中可能起著重要的作用。3、為了進一步證明草莓MRLK家族在草莓果實發(fā)育和成熟調(diào)控中的作用,通過草莓果實瞬時基因操縱技術(shù),研究了草莓MRLK家族其中的一個成員,FaMRLK47,在草莓果實發(fā)育和成熟的作用。結(jié)果表明,FaMRLK47的過量表達(over-expression)可顯著抑制草莓果實的成熟,相反,FaMRKL47-RNAi沉默可促進草莓果實成熟。與此相一致,FaMRLK47-OE可抑制一系列果實成熟標志基因的表達;相反,FaMRLK47-RNAi可促進草莓果實成熟標志基因的表達。該研究結(jié)果為FaMRLK家族參與草莓果實發(fā)育和成熟提供了直接證據(jù)。4、鑒于FaMRLK家族在草莓果實發(fā)育和成熟過程中的重要作用,進一步探討了寡聚糖信號在草莓果實發(fā)育和成熟調(diào)控中的作用。成功建立了草莓果實中寡糖分離和分析體系。初步證明,寡聚糖可以作為信號在草莓果實發(fā)育和成熟調(diào)控中起著重要作用。為更加深入地探討FaMRKL家族的分子識別,研究建立了 FaMRLK47的表達和純化體系。等溫滴定微量熱技術(shù)分析初步證實FaMRLK47和寡聚糖信號具有識別特征。該研究結(jié)果為深入解析FaMRLKs信號轉(zhuǎn)導機理奠定了重要的基礎(chǔ)。綜上所述,本研究首次發(fā)現(xiàn)草莓MRLKs在果實發(fā)育和成熟過程中有重要的調(diào)控作用,MRLKs調(diào)控草莓果實發(fā)育和成熟可能和寡糖信號識別有著密切關(guān)系。該研究不僅為深入解析草莓果實發(fā)育和成熟機理提供了重要信息,同時對今后草莓分子育種或品質(zhì)改良也有重要意義。
[Abstract]:The research of fruit development and mature control mechanism of fruit tree is one of the core problems of fruit tree. However, the research on the mechanism of fruit development and maturation of fruit trees for many years is basically limited to the research of the physiological and biochemical metabolic level of the fruit, but the more profound signal transduction mechanism of the physiological and biochemical metabolism of the fruit is hardly known. The MRLK family (Malectin-like domain-containing receptor-like kinastes) is a family of class-like receptor protein kinases that contain the Malectin domain. Since the Malectin domain has an oligosaccharide binding property, it is presumed that the MRLKs may have a close relationship with the oligosaccharide signal transduction. In the process of fruit development and maturation of fruit tree, cell wall metabolism and oligosaccharide signal transduction are inevitable, so it is suggested that MRLKs may play an important role in fruit development and maturation regulation of fruit trees. In the light of the above, the study on the role of MRLKs in fruit development and maturation of fruit trees was studied in this paper. The main results are as follows: 1. First, based on the bioinformatics analysis, 62 MRLK family members were identified for the first time in the strawberry genome, and named FvMRLK1-FvMRLK62 according to their distribution position on the chromosome. According to the Malectin domain characteristic of the FvMRLK protein family, the family can be divided into 6 subfamilies. The genetic structure and the biogenetic evolution analysis of the intron, exon, and Malectin conserved domain of the FvMRLK family show that the same family with the same genetic evolution has similar structural characteristics and similar structural decisions have similar functions. The stem, leaf and fruit were expressed in various tissues, but the amount of expression in the root was higher than that of other tissues. Of these, 33 of the members of the FvMRLK family are expressed in the fruit. Among the 33 members, more than 60% of the members had a close relationship to the expression of the gene and the development of the fruit. In particular, in the early stage of fruit development, the expression level is high, and as the mature expression level of the fruit drops sharply. Further analysis shows that most members of the FvMRLK family can respond to the development and maturation of strawberry fruits, such as hormones and environmental factors, and these results show that FvMRLKs may play an important role in the development and maturation of strawberry fruit. In order to further demonstrate the role of the strawberry MRLK family in the development and maturation of strawberry fruit, a member of the strawberry MRLK family and the FMRLK47, a member of the strawberry MRLK family, were studied by the instant gene manipulation of strawberry fruit, and the effect of the strawberry MRLK family on the development and maturation of strawberry fruit was studied. The results showed that over-expression of FMRLK47 could significantly inhibit the ripening of strawberry fruit. In contrast, the FMRLK47-OE can inhibit the expression of a series of fruit mature marker genes; instead, the FMRLK47-RNAi can promote the expression of the mature marker gene of the strawberry fruit. The results showed that the FMRLK family had direct evidence for the development and maturation of strawberry fruit. 4. In view of the important role of the FMRLK family in the development and maturation of strawberry fruit, the role of the oligosaccharide signal in the development and maturation of strawberry fruit was further discussed. The oligosaccharide separation and analysis system in strawberry fruit was successfully established. It is proved that the oligosaccharide can play an important role in the development and maturation of strawberry fruit. In order to further study the molecular recognition of the FMRKL family, the expression and purification system of FMRLK47 was established. The analysis of the micro-thermal technology of isothermal titration showed that the FMRLK47 and the oligosaccharide signal had the characteristics of identification. The results of this study provide an important basis for the in-depth analysis of the signal transduction mechanism of the FMRLKs. In conclusion, for the first time in this study, it is found that the strawberry MRLKs plays an important role in the development and maturation of the fruit, and the MRLKrs is closely related to the development and maturation of the strawberry and the identification of the oligosaccharide signal. The research not only provides important information for deeply analyzing the development and maturation mechanism of strawberry fruit, but also has great significance to the future of strawberry molecular breeding or quality improvement.
【學位授予單位】：中國農(nóng)業(yè)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：S668.4

【參考文獻】

相關(guān)期刊論文 前10條

1 Hong-Ze Liao;Meng-Meng Zhu;Hong-Hui Cui;Xin-Yu Du;Yu Tang;Li-Qun Chen;De Ye;Xue-Qin Zhang;;MARIS plays important roles in Arabidopsis pollen tube and root hair growth[J];Journal of Integrative Plant Biology;2016年11期

2 張卿;邢宇;曹慶芹;秦嶺;;熒光定量PCR技術(shù)優(yōu)化及在草莓上的應(yīng)用[J];北京農(nóng)學院學報;2016年04期

3 張卿;劉帥;邢宇;曹慶芹;秦嶺;;草莓果實總RNA提取方法的比較[J];中國農(nóng)學通報;2015年31期

4 Wenwei Lin;Xiyu Ma;Libo Shan;Ping He;;Big Roles of Small Kinases:The Complex Functions of Receptor-Like Cytoplasmic Kinases in Plant Immunity and Development[J];Journal of Integrative Plant Biology;2013年12期

5 盧航;趙小明;白雪芳;杜昱光;;寡聚半乳糖醛酸誘導煙草抗煙草花葉病毒研究初探[J];植物保護;2008年03期

6 陳麗;陳曉藝;李沖偉;張亞楠;李憲臻;;寡糖農(nóng)藥作用機理及生物來源研究進展[J];安徽農(nóng)業(yè)科學;2008年13期

7 劉元召;趙小明;姜華;杜昱光;;寡聚半乳糖醛酸對大豆生長及植保素產(chǎn)生的影響[J];山西農(nóng)業(yè)科學;2008年03期

8 瞿明仁;寡糖研究進展[J];中國飼料;2004年01期

9 倪紅,楊艷燕,閻達中;寡糖的開發(fā)現(xiàn)狀及其應(yīng)用研究進展[J];湖北大學學報(自然科學版);2003年02期

10 香紅星,董仲華,劉亞力;功能性寡糖的研究應(yīng)用進展(上)[J];飼料研究;2001年07期

相關(guān)博士學位論文 前1條

1 張運紅;高活性寡糖篩選及其促進植物生長的生理機制研究[D];華中農(nóng)業(yè)大學;2011年

相關(guān)碩士學位論文 前1條

1 孫麗娜;寡聚半乳糖醛酸的酶解制備及其銅螯合物對小鼠生長特性的影響[D];中國農(nóng)業(yè)科學院;2007年

，

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