马铃薯块茎离体发育过程茉莉酸调控的差异蛋白质组分析

发布时间：2018-03-25 20:26

本文选题：马铃薯　切入点：块茎　出处：《甘肃农业大学》2017年硕士论文

【摘要】：马铃薯块茎是一种营养储藏器官,其发育过程及其复杂,受多种环境因素和植物激素的调控。本研究以离体培养的马铃薯匍匐茎为试验材料,探究马铃薯块茎经过茉莉酸(Jasmonate acid,JA)及茉莉酸抑制剂水杨苷羟肟酸(Salicylhydroxamic acid,SHAM)处理后在形态学,生理学和蛋白质组学等方面的变化,这为进一步阐明马铃薯离体块茎发育过程受JA的调控奠定基础。本研究的结果如下:1.JA对马铃薯块茎在形态和生理方面的影响。JA处理马铃薯匍匐茎后,马铃薯块茎的鲜重、干重、直径和结署率与对照相比均显著增加,LOX(Lipoxygenase,LOX)的活性反而降低,这揭示了在块茎发育中JA有助于贮藏物质的积累。以下数据说明,JA促进了块茎中单糖转变成淀粉,有利于块茎中淀粉的积累,JA处理下,随着JA浓度的增加,淀粉含量先升高再降低,且在0.5μM JA处理下含量最高,为15.6 mg/g,块茎中先降低再升高的还原糖含量和可溶性糖含量,他们在JA(0.5μM)处理下含量均最低,分别是0.5 mg/g和2.5mg/g;与对照相比,在0.5μM JA和不同浓度SHAM共同处理下,淀粉含量先升高后降低,其中0.5μM JA和300μM SHAM共同处理下,淀粉含量最高,为22.8 mg/g,0.5μM JA和700μM SHAM共同处理下淀粉含量最低,为5.5 mg/g;0.5μM JA和SHAM共同处理下,SHAM浓度不断的增加,块茎的还原糖含量显著增加,在5μM JA和SHAM共同处理时,淀粉含量先升高再降低,其中在5μM JA和1μM SHAM共同处理下淀粉含量最高,为25.5 mg/g;与5μM JA相比,块茎中先降低再升高的还原糖含量,在5μM JA和1μM SHAM共同处理下含量最低,为0.4 mg/g,5μM JA和700μM SHAM共同处理下还原糖的含量最高,为5.9 mg/g;与5μM JA相比,5μM JA和不同浓度(1、300、700μM)的SHAM共同处理下可溶性糖含量逐渐升高。2.分析不同浓度(0、0.5、5μM)的JA或(0.5、5μM)JA和SHAM共同处理的马铃薯离体块茎中表达的差异蛋白质,结果表明不同浓度(0、0.5、5μM)的JA处理下,鉴定到44个差异蛋白质点,这些差异蛋白涉及能量和代谢、储藏和防御、氧化还原、转录和翻译、物质转运与细胞结构等功能范畴。JA(0.5μM)和SHAM共同处理下,共鉴定了37个差异蛋白质,这些蛋白涉及能量和代谢、储藏和防御、氧化还原、转录和翻译以及物质转运等功能范畴。JA(5μM)和SHAM共同处理下块茎蛋白质,鉴定到了31个差异蛋白质,这些蛋白质涉及能量和代谢、储藏和防御、氧化还原、转录和翻译以及物质转运等功能范畴。3.研究发现,ADP-葡萄糖焦磷酸酶被JA诱导表达,有助于块茎发育中淀粉含量的增加。能量和物质代谢中的糖酵解途径、三羧酸循环途径和磷酸戊糖途径被JA调控来提供生长发育所需的能量,但是JA降低了块茎发育过程中物质合成的能量消耗。JA通过清除植物体产生的过剩的甲醇、甲醛和甲酸等次生代谢物质以及自由基活性氧等物质引起块茎生长发育的适应性应答,提高了块茎发育的耐受力和抵抗性。
[Abstract]:Potato tuber is a kind of nutrient storage organ, its development process is complex, and it is regulated by many environmental factors and plant hormones. In this study, potato stolons cultured in vitro were used as experimental materials. To investigate the morphological, physiological and proteomic changes of potato tubers treated with Jasmonate acididine (JAA) and salicylhydroxamic acid (Salicylhydroxamic acidic acid SHAM), a jasmonic acid inhibitor. The results are as follows: 1. The effect of JA on the morphology and physiology of potato tuber. The fresh and dry weight of potato tuber after treated with JA. Compared with the control, both the diameter and the rate of formation increased the activity of LOXLX Lipoxygenase (LOX) decreased, which indicated that JA was beneficial to the accumulation of storage materials during tuber development. The following data indicated that JA promoted the conversion of monosaccharide into starch in tuber. The starch content increased and then decreased with the increase of JA concentration in tuber, and the highest content was 15.6 mg / g under 0.5 渭 M JA treatment. The content of reducing sugar and soluble sugar in tuber decreased first and then increased with the increase of JA concentration. Under JA(0.5 渭 M treatment, the content of starch was the lowest, 0.5 mg/g and 2.5 mg / g, respectively. Compared with the control, the starch content increased first and then decreased under the combination of 0.5 渭 M JA and different concentration of SHAM, and the starch content was the highest under the combined treatment of 0.5 渭 M JA and 300 渭 M SHAM. The starch content was the lowest under the treatment of 22.8 mg / g 0.5 渭 M JA and 700 渭 M SHAM, which was 5.5 mg / g 0.5 渭 M JA and SHAM, and the reducing sugar content of tuber increased significantly. When treated with 5 渭 M JA and SHAM, the starch content increased first and then decreased. The starch content was the highest at 5 渭 M JA and 1 渭 M SHAM, 25.5 mg / g. Compared with 5 渭 M JA, the content of reducing sugar in tubers decreased first and then increased, and the lowest in 5 渭 M JA and 1 渭 M SHAM. The content of reducing sugar was the highest under the co-treatment of 0.4 mg / g 5 渭 M JA and 700 渭 M SHAM. Compared with 5 渭 M JA and 1 300 700 渭 M of different concentrations of SHAM, the soluble sugar content increased gradually. 2. The differential protein expression in potato tubers treated with 5 渭 M JA or 0.5 渭 M)JA and SHAM was analyzed. The results showed that 44 different protein spots were identified under different concentrations of JA (0.5 渭 M). These differential proteins were involved in energy and metabolism, storage and defense, redox, transcription and translation. A total of 37 differentially expressed proteins were identified by co-treatment of substance transport and cell structure with SHAM. These proteins are involved in energy and metabolism, storage and defense, and redox. In combination with SHAM, 31 differential proteins were identified, which were involved in energy and metabolism, storage and defense, and redox. The expression of ADP- glucose pyrophosphatase induced by JA is beneficial to the increase of starch content in tuber development and the glycolysis pathway in energy and substance metabolism. The tricarboxylic acid cycle pathway and the pentose phosphate pathway are regulated by JA to provide energy for growth and development, but JA reduces the energy consumption of substance synthesis during tuber development. JA removes excess methanol from plants. Secondary metabolites such as formaldehyde and formic acid as well as free radical reactive oxygen species caused adaptive responses to tuber growth and development and increased tolerance and resistance to tuber development.
【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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