荔枝P型ATP酶基因家族鉴定及其在采后贮藏过程中响应拮抗菌N-1的表达模式分析

Identification of P-type ATPase Gene Family and Analysis of Expres-sion Pattern in Response to Antagonistic N-1 During Postharvest Storage in Litchi

P型ATP酶是植物体内一种重要的膜转运蛋白,在果实能量代谢和酸代谢中发挥重要作用,并广泛参与植物离子运输、抗逆、细胞信号转导等各项生命活动,然而在荔枝或其他无患子科植物中尚未对P型ATP酶基因家族进行全面分析,本研究基于‘妃子笑’荔枝转录组数据,通过对这些基因进行生理生化分析、生物信息学分析和表达分析,共鉴定了28个P型ATP酶基因家族成员基因,探讨P型ATP酶在荔枝采后贮藏过程中的潜在功能。结构和系统发育分析表明该基因可以分为5个亚家族,同一亚族的基因在基因结构和motif基序方面具有很大的相似性,其蛋白二级结构主要以α-螺旋为主,亚细胞定位预测分析发现LcPAs多定位于线粒体(21/28)。通过转录组分析,在采后贮藏期间,荔枝P型ATP酶家族基因中上调表达的基因明显高于下调表达基因,暗示其可能在荔枝采后品质劣变过程中发挥着积极的抵御作用。使用拮抗菌N-1处理荔枝可以有效抑制其果实采后褐变和病害的发生,结合RT-qPCR结果,发现拮抗菌N-1处理能诱导LcPA1、LcPA7、LcPA8和LcPA19在整个贮藏过程中的上调表达,并维持了较高的ATP酶活性,进一步说明P型ATP酶家族基因在抑制荔枝病变过程中发挥着重要作用。本研究为了解荔枝P型ATP酶基因家族基因的进化和功能分析提供了参考,为深入了解拮抗菌N-1保鲜机理的分子调控机理提供新的证据。

P-type ATPase is an important membrane transporter in plants.It plays a significant role in plant energy me-tabolism and acid metabolism.In addition,the enzyme is widely involved in plant ion transport,stress resistance,cell signal transduction and other life activities.However,p-type ATPase gene family has not been comprehensively ana-lyzed in litchi or other sapindaceae plants.In this study,based on the transcriptome data of‘Feizixiao’litchi,28 p-type ATPase genes were identified through physiological and biochemical analysis,bioinformatics analysis and expression analysis,to explore the potential function of P-type ATPase during postharvest storage of litchi.The structure and phy-logenetic analysis showed that the gene could be divided into 5 subfamilies,the genes in the same subfamily had similar gene structure and motifs.The secondary structure of protein was mainly consisted ofα-helix,and the N-terminal do-main of LcPAs was more conserved than the C-terminal domain.Subcellular localization prediction analysis showed that LcPAs were mostly located in mitochondria (21/28). According to transcriptome analysis, the up-regulated genes of P type ATPase family genes in litchi were significantly higher than the down-regulated genes during postharst storage, which suggesting that they may play a positive role in the resistance of quality deterioration of litchi after harvest. As a safe and healthy preservation technology, the mechanism of antagonistic biological control is very complex. Results showed that the antagonistic N-1 treatment could effectively inhibit the browning and disease occurrence of litchi fruits after harvest. Combined with RT-qPCR results, it was found that antagonistic N-1 treatment could induce up-regulated expression of LcPA1, LcPA7, LcPA8 and LcPA19 during the whole storage process, which further indicating that P-type ATPase family genes would play an important role in the antagonistic N-1 inhibition of litchi lesions. This study would provide a reference for understanding the evolution and biological function analysis of p-type ATPase gene family in litchi, it also provide new evidence for further understanding the molecular regulation mechanism of antagonistic N-1 preservation mechanism.