基于代谢组学与蛋白组学的广东烟区沙泥田烤烟上部叶挂灰成因分析

Analysis of the causes of ash hanging in flue-cured tobacco upper leaves in sandy mud field in Guangdong tobacco-growing areas based on metabolomics and proteomics

  • 摘要: 【目的】 基于代谢组学和蛋白组学联合分析广东烟区沙泥田与紫色土烤烟上部叶物质代谢和蛋白表达差异,为减少沙泥田烟叶挂灰提供参考依据。【方法】 以烤后不易挂灰的紫色土烟叶(对照)和烤后易挂灰的沙泥田烟叶为研究对象,选择2种土壤类型烟叶上部叶欠熟、尚熟、成熟和过熟4个时期的鲜烟叶样品,测定多酚氧化酶(PPO)活性;再以成熟时期烟叶进行代谢组学和蛋白组学联合分析,鉴定沙泥田上部成熟烟叶中的差异代谢物和差异表达蛋白,并分析筛选影响沙泥田上部烟叶褐变的关键通路。【结果】 上部叶成熟过程中,紫色土烟叶PPO活性极显著低于沙泥田烟叶(P<0.01),且二者在成熟时期差值最大。从2种土壤类型成熟时期烟叶样品中共筛选出差异代谢物129个,差异表达蛋白1314个,差异代谢物、差异表达蛋白均以上调为主;差异代谢物的聚类和KEGG代谢通路分析主要富集到酪氨酸代谢、苯丙氨酸—酪氨酸—色氨酸生物合成、苯丙素生物合成等通路上。差异表达蛋白的GO功能注释和KEGG信号通路富集分析主要富集到苯丙素生物合成、酪氨酸代谢、异喹啉生物碱生物合成和植物激素信号传导等过程。多酚类物质生物合成过程中,沙泥田烟叶差异表达蛋白苯丙氨酸解氨酶、4-香豆酸辅酶A连接酶、羟基肉桂酰辅酶A莽草酸酯/奎宁酸羟基肉桂酰转移酶下调;差异代谢物阿魏酸、松柏醇、芥子醇下调。脱落酸生物合成和代谢过程中,沙泥田烟叶差异表达蛋白八氢番茄红素脱氢酶、玉米黄质环氧化酶、9-顺式环氧类胡萝卜素双加氧酶、黄氧素脱氢酶下调;差异代谢物红花菜豆酸、二氢红花菜豆酸下调。【结论】 与紫色土烤烟上部叶相比,沙泥田烤烟上部叶生长过程中更易受环境胁迫,导致烟叶素质下降,脱落酸合成增加,激素信号激活,酚类代谢增强,PPO活性上升,在烘烤过程中易导致酶促褐变更加剧烈,从而造成烤后烟叶易挂灰。

     

    Abstract: 【Objective】 Based on the combined analysis of metabolomics and proteomics,the differences in material metabolism and protein expression of flue-cured tobacco upper leaves in sandy mud field and pruple soil of Guangdong tobacco-growing areas were analyzed to provide reference for reducing ash deposition of tobacco leaves in sandy mud field. 【Method】 Purple soil flue-cured tobacco leaves that were less prone to ash hanging after curing(control)and sandy mud field flue-cured tobacco leaves that were prone to ash hanging after curing were used as the research objects. The fresh tobacco leaves of the upper leaves of the 2 soil types at 4 stages of before maturity,just mature,mature and overmature were selected to determine the activity of polyphenol oxidase(PPO). Subsequently,a combined metabolomic and proteomic analysis was conducted on mature tobacco leaves to identify differential metabolites and differentially expressed proteins present in mature upper leaves from sandy mud fields. Additionally,this study aimed to analyze and screen for key pathways influencing browning in these upper tobacco leaves grown in sandy mud field. 【Result】 During the maturation of the upper leaves,the PPO activity of the purple soil tobacco leaves was extremely significantly lower than that of the sandy mud field tobacco leaves(P<0.01),and the difference between the two was the largest during the maturation period. A total of 129 differential metabolites and 1314 differentially expressed proteins were screened from the mature tobacco samples of the 2 soil types,and the differential metabolites and differentially expressed proteins were mainly up-regulated. The clustering of differential metabolites and KEGG metabolic pathway analysis were mainly enriched in tyrosine metabolism,phenylalanine-tyrosine-tryptophan biosynthesis,phenylpropanoid biosynthesis. GO functional annotation and KEGG signal pathway enrichment analysis of differentially expressed proteins were mainly enriched in phenylpropanoid biosynthesis,tyrosine metabolism,isoquinoline alkaloids biosynthesis and plant hormone signal transduction. During the biosynthesis of polyphenols,the differentially expressed proteins of phenylalanine ammonia lyase,4-coumaric acid coenzyme A ligase and hydroxycinnamoyl coenzyme A shikimate/quinic acid hydroxycinnamoyl transferase were down-regulated in sandy mud field tobacco leaves. The differentially expressed proteins of phytoene desaturase,zeaxanthin epoxidase,9-cis-epoxycarotenoid dioxygenase and xanthooxin dehydrogenase were down-regulated in sandy mud field tobacco leaves. Differential metabolites phaseic acid and dihydrophaseic acid were down-regulated. 【Conclusion】 Compared with the flue-cured tobacco upper leaves in purple soil,the flue-cured tobacco upper leaves in sandy mud field are more susceptible to environmental stress during the growth process,resulting in a decrease in the quality of tobacco leaves,an increase in abscisic acid synthesis,activation of hormone signals,enhancement of phenolic metabolism,and an increase in PPO activity. During the curing process,it is easy to lead to more severe enzymatic browning,resulting in easy ash hanging of cured tobacco leaves.

     

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