【摘要】 茭白(Zizania latifolia)是我国南方重要的水生蔬菜,温度对其田间生产具有较大的影响。以浙江省主栽双季茭白品种浙茭911为对照,采用双向电泳技术(two-dimensional electrophoresis,2-DE)对茭白耐冷栽培品种龙茭2号叶片的差异表达蛋白进行比较鉴定分析,获得蛋白点数分布在822~955,差异蛋白点筛选表明,低温及恢复处理中,龙茭2号差异表达蛋白上调数量均高于浙茭911,差异表达蛋白下调数量均低于浙茭911;质谱成功鉴定了6个耐冷相关差异表达蛋白:IAA(indole acetic acid)生长素水解酶(IAA auxin amido hydrolase,IAA-AH)、甲酸四氢叶酸连接酶(putative formate-tetra-hydrofolate ligase,FSH)、苹果酸脱氢酶malate dehydrogenase,MDH)、6-磷酸甘露糖还原酶(mannose-6-phosphate reductase,M6PR)、ATP合成酶(ATP synthase CF1 alpha chain,ATPase)和分子伴侣cpn60-1(chaperonin cpn60-1,cpn60-1),分别与植物激素代谢、核酸合成、能量代谢、光合作用及蛋白折叠等功能相关。IAA-AH、MDH和分子伴侣cpn60-1在两个茭白品种中均表现出低温处理后表达量上调,恢复处理后表达量下调,其他3个蛋白在两个品种间表达变化存在差异;5℃低温处理时,龙茭2号中差异蛋白表达较为稳定,并能较好地恢复至对照水平。基于茭白转录组测序信息查询,克隆获得6个差异蛋白点相关基因的编码区序列并进行了荧光定量PCR验证分析。本研究从蛋白和分子水平阐明了双季茭白龙茭2号耐冷相关的部分蛋白及其表达变化,有助于揭示龙茭2号孕茭特性及其耐冷机制,为田间选育不同采收期的茭白耐冷新品种提供理论依据。更多还原

【Abstract】 As a main aquatic vegetable in southern China, Zizania latifolia could be affected by temperature in the field. The two double-cropping cultivars of Z. latifolia were comparatively analyzed by the method of two-dimensional electrophoresis(2-DE), including the chilling tolerance cultivar of Longjiao 2 and the main cultivar of Zhejiao 911. Among the 822 to 955 protein spots of leaves, differential expression protein spots were selected and analyzed. It was found that the cultivar of Longjiao 2 contained more up-regulated spots and fewer down- regulated spots than that of Zhejiao 911 under the treatments of low temperate and recovery,respectively. Six differential expression chilling tolerance protein spots were successfully identified by matrixassisted laser desorption/ionization time-of- flight/time- of- flight mass spectrometry(MALDI- TOF- TOF MS),including IAA auxin amido hydrolase(IAA- AH), putative formate- tetra- hydrofolate ligase(FSH), malate dehydrogenase(MDH), mannose-6-phosphate reductase(M6PR), ATP synthase CF1 alpha chain(ATPase) and the molecular chaperon cpn60-1, which were related to the plant hormone metabolism, nucleic acid synthesis,energy metabolism, photosynthesis and protein folding, respectively. The expression trend of IAA-AH, MDH and the molecular chaperon of cpn60-1 were similar in the two cultivators of Longjiao 2 and Zhejiao 911 with increasing expression in the treatments of low temperature and decreasing expression in the following treatments of recovery, while the other three were expressed differently between the two cultivators. Especially in the treatments with the temperate of 5 degrees, there were more steady changes of expression and better recovery in the cultivator of Longjiao 2 than that in Zhejiao 911. Based on transcriptome sequencing of Z.latifolia, parts of the coding sequences of 6 differential expression proteins were cloned and the changes of expression were analyzed by Real- time PCR. In this research, the proteins related to chilling tolerant of Longjiao 2 were identified and discussed, which could be contributed to the illumination of the special features of Longjiao 2 and chilling tolerance. The results would provide theoretical basis to the breeding of Z. latifolia cultivars which could be harvested in different times.更多还原