核农学报2018，Vol.32Issue(7)：1320-1329,10.DOI:10.11869/j.issn.100-8551.2018.07.1320
石榴PAL基因的克隆与表达分析
Cloning and Expression Analysis of PAL Gene in Pomegranate (Punica granatum L.)
摘要
Abstract
In order to explore the function and expression characteristics of PAL gene,and reveal the fruit coloration and browning mechanism in pomegranate.The major cultivar Taishanhong and Taishansanbaitian were selected as the experimental materials.The full length of PAL gene was cloned by RT-PCR and RACE technology,and its expression was analyzed in different cultivars during development.The result showed that the PAL gene was cloned from the cDNA template of Taishanhong pomegranate (the GenBank number is KY094504).The ORF sequence of PAL gene is 2 205 bp,and encoded 734 amino acids,which containing phenylalanine and histidine ammonia-lyase conserved structure,PLN02457 binding sites and PLN02457 superfamily conserved domains.The predicted molecular weight of PAL is 79 693.87 Da,which is stable protein and hydrophilic protein.There were serine,threonine and tyrosine three phosphorylation sites in PAL protein.The secondary structure of PAL protein was rich in α-helix (47.55％) and random coil (30.52％).qRT-PCR analysis indicated that the expression level of PAL decreased first and then increased in two pomegranate cultivars during fruit development with higher expression level in Taishanhong fruit.The browning degree of two pomegranate cultivars decreased gradually with development,and the browning degree of Taishansanbaitian was significantly higher than Taishanhong the anthocyanin content of Taishanhong increased gradually with development,while had slightly change in the Taishansanbaitian,the anthocyanin content of Taishanhong was higher than Taishansanbaitian.The expression level of PAL gene was positively correlated with the browning degree in two pomegranate cultivars,there were significantly negative correlation between expression level of PAL gene and anthocyanin content in Taishanhong.The study provides theoritical basis for further explorating the colouration and browning mechanism in pomegranate fruit.关键词
石榴/PAL/基因克隆/生物信息学/表达分析Key words
pomegranate/PAL/gene clone/bioinformatics/expression analysis引用本文复制引用
冯立娟,尹燕雷,焦其庆,杨雪梅..石榴PAL基因的克隆与表达分析[J].核农学报,2018,32(7):1320-1329,10.基金项目
山东省自然科学基金(ZR2015YL056),山东省农业重大应用技术创新项目“北方石榴增产高效栽培技术研究” （ZR2015YL056）
