[1]陈桂信,王玉珍,赵利,等.果实CCoAOMT与LAR基因的筛选与表达分析[J].北方园艺,2015,39(09):87-94.[doi:10.11937/bfyy.201509025]
　CHEN Gui-xin,WANG Yu-zhen,ZHAO Li,et al.Screening and Expression of CCoAOMT and LAR Genes from Nai’s Fruit[J].Northern Horticulture,2015,39(09):87-94.[doi:10.11937/bfyy.201509025]
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果实CCoAOMT与LAR基因的筛选与表达分析

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第39卷 期数: 2015年09 页码: 87-94 栏目: 出版日期: 2015-05-15
Title:
Screening and Expression of CCoAOMT and LAR Genes from Nai’s Fruit
文章编号:
1001-0009（2015）09-0087-08
作者:
陈桂信1; 2 ; 王玉珍1; 2 ; 赵利1; 2 ; 姜翠翠3; 吕恃衡1; 潘东明1; 2
(1.福建农林大学 园艺学院，福建 福州 350002；2.福建农林大学 园艺产品贮运保鲜研究所，福建 福州 350002；3.福建省农业科学院 果树研究所，福建 福州 350013)
Author(s):
CHEN Gui-xin1; 2 ; WANG Yu-zhen1; 2 ; ZHAO Li1; 2 ; JIANG Cui-cui3; LYU Shi-heng1; PAN Dong-ming1; 2
（1.College of Horticulture，Fujian Agriculture and Forestry University，Fuzhou,Fujian 350002;2.Institute of Storage Science and Technology of Horticultural Products，Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002;3.Fruit Research Institute，Fujian Academy of Agricultural Sciences，Fuzhou,Fujian 350013）
关键词:
PsCCoAOMT; PsLAR; 稀释池PCR; 实时荧光定量PCR
Keywords:
Nai（Prunus salicina）; PsCCoAOMT; PsLAR; dilute-pool PCR; real-time PCR
分类号:
S 665.9
DOI:
10.11937/bfyy.201509025
文献标志码:
A
摘要:
以果实为研究材料,探讨了PsCCoAOMT和PsLAR基因在果实不同发育时期的表达动态，采用两步法逆转录与抑制PCR技术相结合的方法构建含果实不同发育时期的均一化全长cDNA文库，通过稀释池PCR法筛选出酚类物质代谢途径中的关键酶基因PsCCoAOMT和PsLAR。结果表明：PsCCoAOMT和PsLAR全长cDNA序列分别为 1 195 bp和1 625 bp ，对应的ORF分别为744 bp和1 050 bp，分别编码247和349个氨基酸。PsCCoAOMT相对分子量为27 893.8，等电点为5.42，属于亲水性稳定蛋白；PsLAR相对分子量为38 483.1，等电点为5.40，具有亲水性，属于不稳定蛋白。同源性分析表明，PsCCoAOMT氨基酸序列与白梨相似性达96%，PsLAR氨基酸序列与蔷薇科植物高度同源。荧光定量结果表明，PsCCoAOMT在整个生长发育过程表达量差异都不明显，整体呈下降趋势，花后50 d表达量最低；PsLAR在整个生长发育过程表达量呈明显的下降趋势，前期高后期低，花后125 d表达量最低。
Abstract:
Dynamic changes of expression of PsCCoAOMT and PsLAR genes at different development stages in Nai’s fruits were explored，a normalized full-length cDNA library was constructed with Nai’s fruits at different developmental stages as research materials by combination of two-step reverse transcription and suppressive PCR.Full-length cDNAs of PsCCoAOMT and PsLAR encoding key enzymes in metabolic pathway of phenolics in Nai’s fruit were screened by dilute- poold PCR method.The results showed that the full length cDNAs of PsCCoAOMT and PsLAR genes were 1 195 bp with 744 bp of ORF encoding 247 amino acids and 1 625 bp with 1 050 bp of ORF encodingand 349 amino acids,respectively.The protein encoded by PsCCoAOMT was a stable hydrophilic protein whose relative molecular weight ofmolecular weight was 27 893.8 and pI was 5.42；the protein encoded by PsLAR was an unstable hydrophilic protein whose relative molecular weight ofmolecular weight was 38 483.1 and pI was 5.40.Homologous analysis showed that the homology of PsCCoAOMT compared with that of Pyrus x bretschneideri was the highest by 96%，the PsLARs were highly homologous in Rosaceous plants.The results of Real-time PCR showed that the change of PsCCoAOMT was not obvious and tended to reduce overally in the whole process of development of fruit and reached to minimum after 50 days of anthesis；that of PsLAR gradually reduced in the whole process of development of fruit，that of the gene reduced to zero after 125 days of anthesis.

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备注/Memo

第一作者简介：陈桂信（1967-），男，博士，副教授，硕士生导师，研究方向为园艺植物遗传育种与生物技术。E-mail：guixinchen@126.com.

责任作者:潘东明（1956-），男，本科，教授，博士生导师，研究方向为果树采后生理与分子生物学。E-mail：pdm666@126.com.
基金项目：福建农林大学“园艺植物种质与高优生产技术创新”创新团队基金资助项目（CXTD12013）。

更新日期/Last Update: 2015-08-01