基于蛋白质组学研究光质对番茄果实挥发性物质的影响机理
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摘要
以‘Micro-Tom’番茄(Solanum lycopersicum L.)为试材,种子发芽后30d移入人工气候室,分别进行红/蓝光组合(1:1、3:1、5:1、7:1)处理,以白光处理为对照。在番茄果实绿熟期、转色期、成熟期取样,测定果实挥发性物质含量及蛋白组数据。结果表明,在转色期和成熟期,对番茄风味有积极作用的己醛、反式–2–己烯醛、β–紫罗兰酮、牻牛儿丙酮、6–甲基–5–庚烯–2–酮、2–苯乙醇、愈伤木酚的含量在红/蓝光3︰1组合处理中最高,而对风味有消极作用的水杨酸甲酯的含量较低,并且在成熟期为0。进一步分析了红/蓝光3︰1组合处理与对照相比的差异蛋白,在成熟过程中番茄果实共鉴定到12个与挥发性物质产生有关的差异蛋白,其中8个上调表达,4个下调表达。通过对12个差异蛋白进行mRNA水平的验证发现,只有转色期的苯丙酮酸互变异构酶(MIF)在蛋白质和mRNA水平表达不一致,其他11个蛋白在两个水平表达均一致。
We used tomato(Solanumly copersicum L.)‘Micro-Tom'as the test material,after 30 days of seed germination. The seedlings were transferred to the artificial climate chamber and exposed to the following light conditions:red/blue combined light(1:1),red/blue combined light(3:1),red/blue combined light(5:1),red/blue combined light(7:1),and white light as the control. Fruits were harvested at the mature green,breaker,and ripe stages,then we determined the fruit volatile compounds content and proteomic. The results showed that in breaker and ripe stages of tomato fruits,hexanal,trans-2-hexenal,β-ionone,geranylacetone,6-methyl-5-hepten-2-one,2-phenylethanolx,callus lignan,which have positive effects on tomato flavor were the highest in the red/blue combined light(3:1)treatment. The content of methyl salicylate,which has a negative effect on tomato flavor was low,and no methyl salicylate was detected at the ripe stage. To further elucidate the effects of light quality on volatile compounds content of tomato fruit,total protein was extracted from the control and red/blue combined light(3:1). A subsequent proteomic analysis revealed that twelve proteins related to volatile compounds were differentially abundant between the control and red/blue combined light(3:1)during ripening. Among the twelve differentially abundant proteins related to volatile compounds,eight were up-regulated and four were down-regulated. The differential abundances of the twelve proteins were validated by determining the expression levels of the corresponding genes by qRT-PCR. The expression levels of eleven of the genes were consistent with the abundances of the corresponding proteins. There was a lack of consistency in the gene expression levels and protein abundances for phenylpyruvate tautomerase(MIF).
We used tomato(Solanumly copersicum L.)‘Micro-Tom'as the test material,after 30 days of seed germination. The seedlings were transferred to the artificial climate chamber and exposed to the following light conditions:red/blue combined light(1:1),red/blue combined light(3:1),red/blue combined light(5:1),red/blue combined light(7:1),and white light as the control. Fruits were harvested at the mature green,breaker,and ripe stages,then we determined the fruit volatile compounds content and proteomic. The results showed that in breaker and ripe stages of tomato fruits,hexanal,trans-2-hexenal,β-ionone,geranylacetone,6-methyl-5-hepten-2-one,2-phenylethanolx,callus lignan,which have positive effects on tomato flavor were the highest in the red/blue combined light(3:1)treatment. The content of methyl salicylate,which has a negative effect on tomato flavor was low,and no methyl salicylate was detected at the ripe stage. To further elucidate the effects of light quality on volatile compounds content of tomato fruit,total protein was extracted from the control and red/blue combined light(3:1). A subsequent proteomic analysis revealed that twelve proteins related to volatile compounds were differentially abundant between the control and red/blue combined light(3:1)during ripening. Among the twelve differentially abundant proteins related to volatile compounds,eight were up-regulated and four were down-regulated. The differential abundances of the twelve proteins were validated by determining the expression levels of the corresponding genes by qRT-PCR. The expression levels of eleven of the genes were consistent with the abundances of the corresponding proteins. There was a lack of consistency in the gene expression levels and protein abundances for phenylpyruvate tautomerase(MIF).
引文
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Chang Pei-pei.2015.Identification and analysis of volatile components and quality traits in tomato fruits[M.D.Dissertation].Yangling:Northwest A&F University.(in Chinese)常培培.2015.番茄果实芳香物质和品质性状的鉴定与分析[硕士论文].杨凌:西北农林科技大学.
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Coleman H D,Park J Y,Nair R,Chapple C,Mansfield S D.2008.RNAi-mediated suppression of p-coumaroyl-CoA 3'-hydroxylase in hybrid poplar impacts lignin deposition and soluble secondary metabolism.Proceedings of the National Academy of Sciences of the United States of America,105(11):4501-4506.
Dignum M J W,Kerler J,Verpoorte R.2001.β-Glucosidase and peroxidase stability in crude enzyme extracts from green beans of Vanilla planifolia andrews.Phytochemical Analysis,12:174-179.
Dong Fei,Wang Chuanzeng,Sun Xiudong,Zhang Xianzheng,Ren Yuqian,Zhou Lijun,Wang Dan,Liu Shiqi.2018.Studies on the sugar metabolism mechanism of tomato fruit under red light revealed by proteomic analysis.Acta Horticulturae Sinica,45(10):1941-1951.(in Chinese)董飞,王传增,孙秀东,张现征,任煜倩,周丽君,王丹,刘世琦.2018.基于蛋白质组学研究红光对番茄果实糖代谢的影响.园艺学报,45(10):1941-1951.
Dong Fei,Wang Chuan-zeng,Zhang Xian-zheng,Qin Yu,Liu Shi-qi.2017.Effects of different light qualities on physiological and photosynthetic characteristics of cherry tomato seedlings.Plant Physiology Journal,53(7):1208-1214.(in Chinese)董飞,王传增,张现征,秦瑜,刘世琦.2017.不同光质对樱桃番茄幼苗生理与光合特性的影响.植物生理学报,53(7):1208-1214.
Du Tian-hao,Zhou Xiao-ting,Zhu Lan-ying,Zhang Jing,Zou Zhi-rong.2016.Effect of melatonin treatment on tomato fruit quality and volatile compounds under salt stress.Food Science,37(15):69-76.(in Chinese)杜天浩,周小婷,朱兰英,张静,邹志荣.2016.褪黑素处理对盐胁迫下番茄果实品质及挥发性物质的影响.食品科学,37(15):69-76.
Ge Qian,Kuang Jing,Wu Yu-cui,Zhang Yuan,Xiao Ya-ping,Wang Zhe-zhi.2013.Cloning and expression analysis of cinnamyl alcohol dehydrogenase(SmCAD)gene in Salvia miltiorrhiza Bunge.Plant Science Journal,31(3):261-268.(in Chinese)葛茜,邝静,武玉翠,张媛,肖娅萍,王喆之.丹参肉桂醇脱氢酶基因(SmCAD)的克隆及表达分析.植物科学学报,31(3):261-268.
Goff S A,Klee H J.2006.Plant volatile compounds:sensory cues for health and nutritional value?Science,311:815-819.
Guo Xiao-hong.2009.Sequence and function analysis of peroxidase in Tamarix hispida[Ph.D.Dissertation].Harbin:Northeast Forestry University.郭晓红.2009.柽柳过氧化物酶基因的序列分析及功能验证[博士论文].哈尔滨:东北林业大学.
Jin Ya-zhong.2014.Identification,classification and expression regulation of alcohol dehydrogenases gene family members in melon(Cucumis melo var.makuwa Makino)[Ph.D.Dissertation].Shenyang:Shenyang Agricultural University.(in Chinese)靳亚忠.2014.甜瓜醇脱氢酶基因家族成员鉴定、分类及表达调控[博士论文].沈阳:沈阳农业大学.
Jose L R,Yury M T,Antonio J M,Arnaud G B,Antonio G.2014.The expanded tomato fruit volatile landscape.Journal of Experimental Botany,65:4613-4623.
Kader A A.2008.Flavor quality of fruits and vegetables.Journal of the Science of Food and Agriculture,88:1863-1868.
Kanisawa T.1993.Flavor development in vanilla beans.Kouryou,180:113-123.
Lee E J,Facchini P J.2011.Tyrosine aminotransferase contributes to benzylisoquinoline alkaloid biosynthesis in opium poppy.Plant Physiology,157:1067-1078.
Lee J H J,Jayaprakasha G K,Rush C M,Crosby K M,Patil B S.2018.Production system influences volatile biomarkers in tomato.Metabolomlcs,14:99-111.
Leong G,Archavlis A,Derbesy M.1989.Research on the glucoside fraction of the vanilla bean.Journal of Essential Oil Research,1:33-41.
Li Y,Xin G F,Wei M,Shi Q H,Yang F J,Wang X F.2017.Carbohydrate accumulation and sucrose metabolism responses in tomato seedling leaves when subjected to different light qualities.Scientia Horticulturae,225:490-497.
Li Yan,Wang Li-wei,Wen Lian-lian,Wei Min,Shi Qing-hua,Yang Feng-juan,Wang Xiu-feng.2017.Effects of red and blue light qualities on main fruit quality of tomato during color-turning period.Acta Horticulturae Sinica,44(12):2372-2382.(in Chinese)李岩,王丽伟,文莲莲,魏珉,史庆华,杨凤娟,王秀峰.2017.红蓝光质对转色期间番茄果实主要品质的影响.园艺学报,44(12):2372-2382.
Liang Xin-yuan.2017.Role of lipoxygenase and ethylene in tomato fruit ripening and aroma synthesis[M.D.Dissertation].Shenyang:Shenyang Agricultural University.(in Chinese)梁馨元.2017.脂氧合酶和乙烯在番茄果实成熟及香气合成中的作用[硕士论文].沈阳:沈阳农业大学.
Luo C H,Lin Q M,Lin S Y,Meng C,Wang H.2017.Cosynthesis of L-homophenylalanine and 2-phenylethanol by recombinant Saccharomyces cerevisiae expressing aspartate aminotransferase from Escherichia coli BL21(DE3).Journal of Bioscience and Bioengineering,123:1-7.
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