Full Length Research Paper

Cloning and expression of cell wall acid invertase gene fragment from poinsettia (Euphorbia pulcherrima wild.)

Jun Tao^1*, Xiao-Yong Xu¹*, Ju Yu¹, Chun-Yan Cao¹ and Guo-Hua Liang²

¹School of Horticulture and Plant Protection, Yang Zhou University, Yang Zhou 225009, P.R. China.

²School of Agriculture, Yang Zhou University, Yang Zhou 225009, P.R. China.

*Corresponding author. E-mail: taojun@yzu.edu.cn and xyxu@yzu.edu.cn.

Accepted 13 November, 2009

   Abstract

A fragment of invertase gene containing catalytic sites of cysteine was cloned from poinsettia (Euphorbia pulcherrima wild.) by using the polymerase chain reaction (PCR) method. The length of the fragment was 521 bp, encoding 173 amino acids and containing a part of open reading frames, but no intron. It had a high homology to previously cloned cell wall acid invertase genes in other plants by sequence comparison, so it could be speculated that the fragment belongs to putative cell-wall acid invertase gene sequence of poinsettia (termed as EpCWINV, GenBank no. EU274662). Gene expression analyses in different organs including roots, stems, leaves, flowers among others and during different growth periods of leaves by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method and real-time quantitative PCR approach revealed the highest level of expression in roots, the lower in stems and intermediate in leaves and flowers, as well as a continuous rising level in leaves and a continuous declining level in petioles during the developmental stages of the leaves.

Key words: Poinsettia, invertase gene, cloning, gene expression, RT-PCR.