硫化氢控制鲜切苹果褐变的可变剪切基因分析

An Analysis of Alternative Splicing Events During Browning Inhibition of Fresh-cut Apples by Hydrogen Sulfide Treatment

利用RNA-seq测序技术分析了H2S处理和对照鲜切苹果两个贮藏时间点(0和6 d)样品的可变剪切,从中挖掘与H2S控制鲜切苹果褐变相关的基因可变剪切。结果表明,4组样品中基因可变剪切均以第1个外显子可变剪切(TSS)和最后1个外显子可变剪切(TTS)为主。与对照组同时期样品相比,H_(2)S处理0和6 d特有的可变剪切基因分别有921和2423个,GO分析结果表明两组样品的特有可变剪切基因主要富集在黄酮合成、细胞质、膜结构、水解酶、氧化还原酶活性等,KEGG富集分析表明上述基因主要富集在植物—病原菌相互作用、植物激素信号转导、碳水化合物代谢、氧化磷酸化、苯丙烷生物合成等代谢通路。此外,H_(2)S处理抑制了多酚氧化酶(PPO)、过氧化物酶(POD)、脂氧合酶(LOX)和磷脂酶D(PLD)等基因的表达,同时提高了苯丙氨酸解氨酶(PAL)和NADH脱氢酶基因的表达。综上,H2S处理可能是通过改变过氧化物酶、苯丙烷代谢、膜结构以及能量代谢相关基因的可变剪切及表达模式,从而控制鲜切苹果褐变反应。

In order to explore the alternative splicing of the browning inhibition of fresh-cut apples by hydrogen sulfide(H_(2)S)treatment,RNA-Seq technology was used to identify the alternative splicing occurring in H_(2)S-treated and H_(2)O-treated fresh-cut apples at two storage time(0 and 6 d).The results showed that the dominate alternative splicing types were alternative transcription start site(TSS)and alternative transcription termination site(TTS)in these four samples.The number of the specific alternative splicing genes of H2S-treated fresh-cut apples at 0 and 6 d were 921 and 2423,respectively.GO annotation showed that the specific alternative splicing genes in H2S-treated fresh-cut apples at 0 and 6 d were mainly enriched in flavonoid biosynthetic process,cytoplasm,membrane,hydrolase and oxidoreductase.KEGG analysis showed that these genes mainly enriched in plant-pathogen interaction,plant hormone signal transduction,carbohydrate metabolism,phenylpropanoid biosynthesis and oxidative phosphorylation.Furthermore,expression of polyphenol oxidase(PPO),peroxidase(POD),lipoxidase(LOX)and phospholipase D(LPD)genes was down-regulated after H_(2)S treatment,whilst expression of phenylalanine ammonia-lyase(PAL)and NADH dehydrogenase was induced.In conclusion,H_(2)S controlled the browning of fresh-cut apples through changing the alternative splicing and expression pattern of peroxide,phenylpropanoid metabolism,membrane and energy metabolism related genes.