笃斯越橘响应低温和短光周期的蛋白质组分析

以3年生笃斯越橘苗为材料,将苗木分别置于对照(23℃,日照长度14 h),低温短日照(4℃,日照长度10 h)和低温长日照(4℃,日照长度14 h)的人工气候室中。处理21 d后,采用同位素标记相对和绝对定量(iTRAQ)蛋白质组学技术测定枝条蛋白质表达变化情况。结果显示:通过质谱鉴定出5 972个蛋白质,600个差异表达蛋白,其中在低温短日照与对照比对组中,丰度显著上调蛋白有140个,丰度显著下调蛋白有114个;在低温长日照与对照比对组中,丰度显著上调蛋白有255个,丰度显著下调蛋白有122个;在低温短日照与低温长日照比对组中,丰度显著上调蛋白有39个,丰度显著下调蛋白有187个。这些蛋白主要参与(1)RNA代谢,(2)蛋白质翻译后修饰,(3)碳水化合物转运和代谢,(4)能量代谢和转换,(5)脂质代谢,(6)次生代谢产物合成,(7)抗氧化与胁迫防御,(8)光合作用,(9)无机离子转运和代谢。结果表明与抗冻性有关的蛋白差异表达主要受低温诱导,少数蛋白表达受低温和光周期共同影响,低温短光周期更有利于抗冻性形成。低温锻炼可显著提高RNA代谢、蛋白质翻译后修饰、抗氧化和防御反应、次生代谢产物合成以及脂质代谢过程中许多蛋白质的丰度,提示这些代谢途径和相关蛋白可能在笃斯越橘抗冻机制中起重要作用。

Proteome Analysis of Response of Vaccinium uligiuosum Seedlings to Low-temperature and Short Photoperiod

The three-year-old seedlings of Vaccinium uligiuosum L. were selected as testing materials. The seedlings were exposed to the control（23 ℃，daylength 14 h），low-temperature and short day（4 ℃，daylength 10 h） and low-temperature and long day（4 ℃，daylength 14 h）under controlled environment chamber for 21 d respectively. The changes in the protein expression of shoot tissues after treatment were recorded. iTRAQ-based quantitative proteomics techniques were used in this study. The results showed that of 5 972 proteins and 600 differential expression proteins identified by a mass spectrometric analysis，140 proteins were significantly up-regulated and 114 were significantly down-regulated in low temperature and short-day vs control；255 proteins were significantly up-regulated and 122 were significantly down-regulated in low temperature and long-day vs control；39 proteins were significantly up-regulated and 187 were significantly down-regulated in low temperature and short-day vs low temperature and long-day. They included proteins were mainly involved in（1）RNA metabolism；（2）protein posttranslational modification；（3）carbohydrate metabolic；（4）energy production and conversion；（5）lipid metabolism；（6）secondary metabolites biosynthesis；（7）antioxidant and stress defense；（8）photosynthesis；（9）inorganic ion transport and metabolism. The differential expression proteins related to freezing tolerance were mainly induced by low temperature. In addition，expression of a few proteins is affected by both low temperature and photoperiod，and low temperature and short photoperiod are more favorable to freezing resistance acquisition. Cold acclimation increased significantly abundance of protein involved in RNA metabolism，protein posttranslational modification，antioxidant and defense responses，secondary metabolites biosynthesis，and lipid metabolism，revealed these metabolic pathways and related proteins may play an important role in freezing resistance mechanism in Vaccinium uligiuosum L.