茉莉酸调控的二倍体马铃薯离体块茎发育差异蛋白质组学分析

发布时间：2018-03-10 05:24

本文选题：茉莉酸　切入点：马铃薯　出处：《甘肃农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：马铃薯(Solanum tuberosum L.)作为第四大主要作物,在农业生产中占据着重要的地位。马铃薯的块茎即可食用又可用于无性繁殖,因此研究块茎的的形成和发育对于提高马铃薯的产量和品质具有重要的意义。马铃薯块茎的发育是一个复杂的过程,其受许多因素的影响,包括光、温度、营养物质和激素等。茉莉酸在诱导马铃薯块茎形成和发育方面具有重要的调节作用。为探明茉莉酸对马铃薯块茎发育的影响,本研究以二倍体马铃薯(DM)试管苗为实验材料,通过用不同浓度(0、0.5、5、50μΜ)的茉莉酸处理马铃薯的茎段,对诱导形成的试管薯进行形态学和生理学分析,同时研究茉莉酸调控的试管薯差异蛋白质组变化,阐明茉莉酸调控的马铃薯块茎发育生理和分子机制。在本研究中,获得如下结果:1、茉莉酸对块茎在形态和生理方面的影响。茉莉酸浓度为0.5和5μΜ时,马铃薯块茎体积增大,同时还有利于单薯鲜干重的积累,但是茉莉酸浓度为50μΜ时抑制块茎体积的增大以及单薯鲜干重的积累。当茉莉素浓度为0.5、5和50μΜ时,马铃薯块茎的结薯率分别为对照的2.6、4.7和5.1倍,块茎的长宽比较对照分别下降了19.8%、21.9%和27.3%。茉莉酸促进块茎结薯期提前,在50μΜ茉莉酸处理下,第5天块茎开始形成。马铃薯块茎中淀粉含量在0.5和5μΜ的茉莉酸处理下较对照分别下降了21.1%和5.4%,而在50μΜ茉莉酸处理下较对照上升了28.3%。经0.5、5和50μΜ的茉莉酸处理后,马铃薯块茎的可溶性糖的含量分别是对照的1.9、4.1和2.9倍。马铃薯块茎还原糖的含量在0.5和5μΜ的茉莉酸处理下分别低于对照16.9%和14.2%,而在50μΜ的茉莉酸处理下高于对照6.5%。经0.5、5和50μΜ的茉莉酸处理后,马铃薯块茎脂氧合酶活性较对照分别降低了43.0%、78.4%和87.8%。2、茉莉酸调控的马铃薯块茎离体发育差异蛋白质组学分析。在0、0.5、5和50μΜ的茉莉酸处理下分别检测到620±13.1、611±19.9、608±3.1和616±10.1个蛋白质点,并且不同处理下检测到的蛋白质点差异不显著。在4个处理下总共鉴定出41个差异表达蛋白质点,其中施加外源茉莉酸后有14个诱导产生的新的蛋白质。采用MALDI-TOF/TOF质谱对所鉴定的差异表达的蛋白质点进行分析后并功能分类,可将其分为代谢相关蛋白、储藏相关蛋白、蛋白翻译和降解相关蛋白、细胞防御相关蛋白、分子伴侣、通道蛋白和细胞壁相关蛋白。3、研究发现,经5μΜ的茉莉酸处理后块茎中NAD依赖性苹果酸脱氢酶上调表达,可能通过催化苹果酸生成草酰乙酸促进三羧酸循环为块茎的发育提供能量。茉莉酸处理后果糖激酶先上调表达再下调表达,说明在离体发育过程中,茉莉酸可能通过影响果糖激酶的表达来影响马铃薯块茎淀粉含量。茉莉酸处理后果糖二磷酸醛缩酶上调表达,说明茉莉酸可能通过参与糖酵解的过程加速糖类物质代谢。在块茎的发育过程中,茉莉酸处理后马铃薯块茎中Kunitz型蛋白酶抑制剂上调表达,表明在块茎发育过程中茉莉酸有助于马铃薯块茎储藏类蛋白的积累。茉莉酸处理后肌动蛋白-7下调表达,可能会通过抑制细胞壁的合成和细胞形状的维持来影响薯形的变化。
[Abstract]:Potato (Solanum tuberosum L.) as the fourth major crops, occupies an important position in agricultural production. The potato tuber can be eaten and can be used for asexual reproduction, the formation and development is of great significance to improve the yield and quality of potato tubers. So the study of potato tuber development is a complex process. It is affected by many factors, including light, temperature, nutrients and hormones. Jasmonic acid has an important regulatory role in the induction of tuber formation and development. In order to investigate the jasmonic acid effect on potato tuber development, based on the diploid potato (DM) seedlings as experimental material, through the use of different concentrations (0,0.5,5,50 UM) stem segments of jasmonic acid to induce the formation of potato, microtuber of morphological and physiological analysis, microtuber differences and jasmonic acid regulation research Proteome, clarify the regulation of jasmonic acid in potato tuber growth physiology and molecular mechanism. In this study, the results obtained are as follows: 1. The effect of jasmonic acid on the morphology and physiology of the tubers. Jasmonic acid concentration of 0.5 and 5 um, potato tuber size, but also conducive to tuber fresh dry weight but the accumulation of jasmonic acid concentration increases suppresses the tuber size 50 um and the accumulation of tuber fresh and dry weight. When JA concentration is 0.5,5 and 50 um, Tuberization of potato tuber rate respectively 2.6,4.7 and 5.1 times, the length and width of tuber ratio were decreased by 19.8% 21.9%, 27.3%. and jasmonic acid promote tuber tuber in the period ahead, 50 um jasmonic acid treatment, fifth days begin to form. Tubers of potato starch content in jasmonic acid treatment of 0.5 and 5 um under were decreased by 21.1% and 5.4%, and in 50 um jasmonic acid Treatment increased by 28.3%. by Ja 0.5,5 and 50 m, the soluble sugar content of potato tubers were 1.9,4.1 and 2.9 times. The control of potato tuber reducing sugar content in jasmonic acid treatment of 0.5 and 5 um respectively under 16.9% and 14.2% lower than that of the control, but higher than in jasmonic acid 50 um under the control of 6.5%. by Ja 0.5,5 and 50 um after potato tuber lipoxygenase activity than that of the control were reduced by 43%, 78.4% and 87.8%.2, jasmonic acid regulation of potato tuber in vitro development of proteomic analysis. In jasmonic acid 0,0.5,5 and 50 um. Were detected in 620 + 13.1611 + 19.9608 + 3.1 and 616 + 10.1 protein spots, and different treatments of protein spots detected no significant difference in the 4 treatments. We identified 41 differentially expressed protein spots, including exogenous jasmonic acid There are 14 induced new proteins. By using MALDI-TOF/TOF mass spectrometry to the differential expression of the identified protein spots were analyzed and the functions of classification, can be divided into metabolism related proteins, storage related protein, protein translation and degradation of proteins, cell defense related proteins, molecular chaperones, protein and cell wall related protein.3, the study found that by Ja 5 um after tuber NAD dependent malate dehydrogenase expression may promote energy three acid cycle is tuber growth by catalytic malate oxaloacetate. After jasmonic acid treatment to upregulate the expression of fructokinase and down-regulation of expression in vitro development process jasmonic acid, may influence the starch content of potato tuber by influencing the expression of fructokinase. Expression of fructose two phosphate aldolase up-regulated by jasmonic acid, jasmonic acid may explain the mo Through the process of glycolysis accelerated carbohydrate metabolism. In the development process of tuber, the expression of Kunitz type potato protease inhibitors increase after jasmonic acid treatment, showed that the tuber acid contributes to potato tuber storage protein accumulation during the development of Mo Li jasmine. After acid treatment could down regulate the expression of actin -7. To change the shape by maintaining the inhibition of cell wall synthesis and cell shape.

【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S532

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