笃斯越橘响应低温和短光周期的蛋白质组分析
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摘要
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。
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.
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.
引文
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Banados M P,Strik B.2006.Manipulation of the annual growth cycle of blueberry using photoperiod.Acta Horticulturae,715(715):65-71.
Chen M,Markham J E,Cahoon E B.2012.SphingolipidΔ8 unsaturation is important for glucosylceramide biosynthesis and low-temperature performance in Arabidopsis.Plant Journal for Cell&Molecular Biology,69(5):769-781.
Chen Qi,Yuan Jin-hai,Liu Zi-gang,Liu Wan-cang,Fang Yan,Zhao Xin-wang,Ma Li,Pu Yuan-yuan,Zhao Yan-ning,Zeng Xiu-cun.2017.Differentially expressed proteins in response to low temperature in Brassica campestris‘Longyou 7’seedlings.Chinese Journal of Eco-Agriculture,25(3):381-390.(in Chinese)陈奇,袁金海,刘自刚,孙万仓,方彦,赵新旺,马骊,蒲媛媛,赵艳宁,曾秀存.2017.白菜型冬油菜“陇油7号”叶片响应低温胁迫的差异蛋白鉴定与分析.中国生态农业学报,25(3):381-390.
Cheng Xi-yong,Hu Xiao-xian,Li Hai-xia,Dong Zhong-dong,Ren Yan,Chen Shu-lin,Zhan Ke-hui,Xu Hai-xia.2016.Identification and analysis of differentially expressed proteins in wheat youngs pike under cold stress.Journal of Triticeae Crops,36(10):1342-1349.(in Chinese)程西永,呼晓贤,李海霞,董中东,任妍,陈树林,詹克慧,许海霞.2016.低温胁迫下小麦幼穗中差异表达蛋白质鉴定.麦类作物学报,36(10):1342-1349.
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Di Bao,Meng Yu,Zhang Gang,Guo Ming.2013.Effects of photoperiod and temperature on electrical impedance spectroscopy of Catalpa bungei.Journal of Agricultural University of Hebei,36(5):54-59.(in Chinese)邸葆,孟昱,张钢,郭明.2013.不同光周期和温度对楸树电阻抗参数的影响.河北农业大学学报,36(5):54-59.
Die J V,Arora R,Rowland L J.2016.Global patterns of protein abundance during the development of cold hardiness in blueberry.Environmental&Experimental Botany,124:11-21.
Fraser T C M,Qia B,Elhussein S,Chatrattanakunchai S,Stobart A K,Lazarus C M.2004.Expression of the isochrysis C18Δ9 polyunsaturated fatty acid specific elongase component alters Arabidopsis glycerolipid profiles.Plant Physiol,135(2):859-866.
Gao F,Zhou Y,Zhu W,Li X F,Fan L M,Zhang G F.2009.Proteomic analysis of cold stress-responsive proteins in Thellungiella rosette leaves.Planta,230(5):1033-1046.
Guan Q,Wu J,Zhang Y,Jiang C,Liu R,Chai C,Zhu J.2013.A DEAD box RNA helicase is critical for pre-mRNA splicing cold-responsive gene regulation,and cold tolerance in Arabidopsis.Plant Cell,25(1):342-356.
Heidarvand L,Amiri R M.2010.What happens in plant molecular responses to cold stress?Acta Physiologiae Plantarum,32(3):419-431.
Hooks M A.2010.Modelling the peroxisomal carbon leak during lipid mobilization in Arabidopsis.Biochemical Society Transactions,38(5):1230-1233.
Huang C K,Shen Y L,Huang L F,Wu S J,Yeh C H,Lu C A.2015.The DEAD-box RNA helicase AtRH7/PRH75 participates in pre-rRNAprocessing,plant development and cold tolerance in Arabidopsis.Plant Cell Physiology,57(1):174-191.
Ifuku K.2014.The PsbP and PsbQ family proteins in the photosynthetic machinery of chloroplasts.Plant Physiology and Biochemistry,81:108-114.
Jackson S D.2008.Plant responses to photoperiod.New Phytologist,181(3):517-531.
Konishi T,Shinohara K,Yamada K,Sasaki Y.1996.Acetyl-Co A carboxylase in higher plants:most plants other than Gramineae have both the prokaryotic and the eukaryotic forms of this enzyme.Plant&Cell Physiol,37(2):117-122.
Kraujalyte V,Venskutonis P R,Pukalskas A,?esonien?L,Daubaras R.2015.Antioxidant properties,phenolic composition and potentiometric sensor array evaluation of commercial and new blueberry(Vaccinium corymbosum)and bog blueberry(Vaccinium uliginosum)genotypes.Food Chemistry,188:583-590.
Lee C M,Thomashow M F.2012.Photoperiodic regulation of the C-repeat binding factor(CBF)cold acclimation pathway and freezing tolerance in Arabidopsis thaliana.Proceedings of the National Academy of Sciences,USA,109:15054-15059.
Li C,Puhakainen T,Welling A,Viher?-Aarnio A,Ernstsen A,Junttilad O,Heinoa P,Palvaa E T.2002.Cold acclimation in silver birch(Betula pendula)development of freezing tolerance in different tissues and climatic ecotypes.Physiologia Plantarum,116(4):478-488.
Lütz C.2010.Cell physiology of plants growing in cold environments.Protoplasma,244:53-73.
Neilson K A,Mariani M,Haynes P A.2011.Quantitative proteomic analysis of cold-responsive proteins in rice proteomics.Proteomics,11(9):1696-1706.
Oquist G,Huner N P.2003.Photosynthesis of overwintering evergreen plants.Annual Review of Plant Biology,54(1):329-355.
Ortega-GarcíaF,Peragón J.2009.The response of phenylalanine ammonia-lyase,polyphenol oxidase and phenols to cold stress in the olive tree(Olea europaea,L.cv.Picual).Journal of the Science of Food&Agriculture,89(9):1565-1573.
Palonen P.2006.Vegetative growth,cold acclimation,and dormancy as affected by temperature and photoperiod in six red raspberry(Rubus idaeus L.)cultivars.European Journal of Horticultural Science,71(1):1-6.
Panadero J,Pallotti C,Rodriguez-Vargas S,Randez-Gil F,Prieto J A.2006.A downshift in temperature activates the high osmolarity glycerol(HOG)pathway,which determines freeze tolerance in Saccharomyces cerevisiae.The Journal of Biological Chemistry,281(2):4638-4645.
Qi Jun-feng.2010.Banana leaves under low temperature stress SSH library construction and differential protein on study[M.D.Dissertation].Haikou:Hainan University.(in Chinese)祁君凤.2010.低温胁迫香蕉叶片SSH文库的构建及其差异蛋白的初步研究[硕士论文].海口:海南大学.
Rawsthorne S.2002.Carbon flux and fatty acid synthesis in plants.Progress in Lipid Research,41(2):182-196.
Renaut J,Hausman J F,Bassett C,Artlip T,Cauchie H M,Witters E,Wisniewski M.2008.Quantitative proteomic analysis of short photoperiod and low-temperature responses in bark tissues of peach(Prunus persica L.Batsch).Tree Genetics&Genomes,4(4):589-600.
Renaut J,Lutts S,Hoffmann L,Hausman J F.2004.Responses of poplar to chilling temperatures:proteomic and physiological aspects.Plant Biology,6(1):81-90.
Rowland L J,Panta G R,Mehra S,Parmentier-Line C.2004.Molecular genetic and physiological analysis of the cold-responsive dehydrins of blueberry.Journal of Crop Improvement,10(1):53-76.
Schmitz R J,Hong L,Michaels S,Amasino R M.2005.FRIGIDA-ESSEN TIAL1 interacts genetically with FRIGIDA and FRIGIDA LIKE1 to promote the winter annual habit of Arabidopsis thaliana.Development,132(24):5471-5478.
Schottler M A.2004.The role of plastocyanin in the adjustment of the photosynthetic electron transport to the carbon metabolism in tobacco.Plant Physiology,136(4):4265-4274.
Shockey J M,Fulda M S,Browse J.2003.Arabidopsis contains a large superfamily of acyl-activating enzymes.Phylogenetic and biochemical analysis reveals a new class of acyl-coenzyme a synthetases.Plant Physiology,132(2):1065-1076.
Singh R,Dubey A K,Sanyal I.2019.Optimisation of adventitious shoot regeneration and Agrobacterium-mediated transformation in Canna×generalis(Canna Lily).Horticultural Plant Journal,5(1):39-46.
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