《植物生理学报》 2011, 47(3): 244-248

菠萝果实发育相关RFD1基因的克隆及表达

王尉^{1, 2, 3}, 张秀梅³, 谢江辉^3,*

1安徽农业大学生命科学学院, 合肥230036; 2中山大学生命科学学院植物与微生物互作实验室, 广州510006; 3中国热带农业 科学院南亚热带作物研究所, 广东湛江524091

通信作者：谢江辉；E-mail: xiejhww@163.com；Tel: 020-39332974

摘　要：

Blast菠萝果实不同发育时期的cDNA文库测序结果, 并用信息生物学软件和相关网站进行拼接和分析, 设计特异 引物RT-PCR, 得到一个2 750 bp、编码761个氨基酸的全长基因序列, 命名为RFD1。预测该蛋白是一个亲水性、且具有 多个磷酸化位点的分泌性蛋白, 可能位于叶绿体中。半定量PCR分析发现RFD1基因表达量在果实发育期不断增加, 70 d 达到最高; 而在70 d果实的不同部位并没有显著差异性, 仅果实外部略有增高。

关键词：菠萝; “电子”克隆; 果实发育; 表达

收稿：2010-09-28   修定：2010-12-06

资助：“948”项目[2006-G34(A)]、农业部行业计划资助项目(nyhyzx07-030)和中央级公益性科研院所基本科研业务费专项(200701)。

Cloning and Expression of RFD1 Gene Related to Development from Pineapple [Ananas comosusr (L.) Merr.] Fruit

WANG Wei^1,2,3, ZHANG Xiu-Mei³, XIE Jiang-Hui^3,*

¹Anhui Agricultural University, School of Life Science, Hefei 230036, China; ²Sun Yat-Sen University, School of Life Sciences, Laboratory of Plant and Microbial Interaction, Guangzhou 510006, China; ³China Southern Subtropical Crop Research Institute of CATAS, Zhanjiang, Guangdong 524091, China

Corresponding author: XIE Jiang-Hui; E-mail: xiejhww@163.com; Tel: 020-39332974

Abstract:

In this study, cDNA library of pineapple fruit sequences of different development stages were screened by Blast. And then, sequences were spliced and analyzed by some informatics softwares and related webs. According to the spliced result, specific primers of RT-PCR were designed and performed. We got a 2 750 bp of the full length gene encoding 761 amino acids which was named as RFD1 (related to fruit development). The results indicated that RFD1 protein was a hydrophilic, multiple phosphorylation sites of the secreted protein, and may be located in chloroplasts. Semi-quantitative PCR analysis showed that RFD1 gene expression was gradually increased during fruit development periods, and reached the peak in the 70 d. Though the expression of RFD1 gene in different fruit parts had no significant differences, that in the outer was slightly increased in the 70 d after flowering.

Key words: pineapple (Ananas comosusr); in silico cloning; fruit development; expression