西葫芦过氧化物酶POD1基因的分离及表达分析

Isolation and Expression of Peroxidase POD1 Gene fromCucurbita pepo

通过西葫芦转录组测序,筛选得到1条功能注释为过氧化物酶(peroxidase,POD)的基因全长序列(c47620.graph_c0)。分析发现,该序列全长1 245bp,包含1个996bp的开放读码框(open reading frame,ORF),预测编码331个氨基酸,理论分子量(molecular weight,Mw)为35.70kDa,等电点(isoelectric point,pI)为5.40,编码的蛋白质与黄瓜、甜瓜和苦瓜等常见瓜果类蔬菜同源蛋白质的相似性在84%、84%和81%,显示其高度的保守性,基因命名为CpPOD1,GenBank登录号为MF988285。荧光定量PCR分析结果表明,CpPOD1基因具有组织表达特异性,在叶片中表达丰度最高,其次为根,在花中的表达最低。CpPOD1基因在低温弱光条件下,其表达量随时间增加而显著上调。初步推测,CpPOD1基因在西葫芦低温弱光胁迫下存在一定上调表达,猜测可能参与调控下游基因对低温弱光胁迫的响应有关。

A functional annotation as peroxidase(POD)gene(i.e.,c47620.graph_c0)was screened from the transcriptomic sequencing of Cucurbita pepo.It was found that thesequence length was 1 245 bp,including a 996bp open reading frame(ORF)encoded a protein of 331 amino acids.The protein had a predicted molecular weight of 35.70 kDa and a hypothetical isoelectric point of 5.40.The similarity of the proteins to the homologous proteins of cucumber,melon and balsam pear were 84%,84%and 81%,respectively,witha highly conservative nature.It was designated as CpPOD1 gene with a GenBank accession code of MF988285.The RTqPCR revealed the geneto betissue-specific,andits expression in C.pepo highest in the leaves,followed by roots and flowers.Under low temperature and dim light,the CpPOD1 expression was up-regulated over time.It suggested that the gene expression correlatedwith the low temperature/dim light stress as C.pepo regulated the downstream responses.This study could lead to further understanding of the function of CpPOD1 gene inzucchinion cold-and-low-lighttolerance and the regulation mechanism providing evidence and theoretical basis for improved breeding of C.pepo.