Cloning and Expression Pattern Analysis of Actin from Dendrobium 'Sonia Hiasakul'

XIN Yao;LU Shunjiao;LI Chonghui;YANG Guangsui;YIN Junmei;Institute of Tropical Agriculture and Forestry, Hainan University;Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Crop Gene Resources and Germplam Enhancement in Southern China, Ministry of Agriculture;Hainan Engineering Technology Research Center of Tropical Ornamental Plant Germplasm Innovation and Utilization;

Dendrobium.'Sonia Hiasakul';;Actin;;clone;;expression pattern

A full-length cDNA Actin(DenActin) of Dendrobium 'Sonia Hiasakul' was cloned, and its evolution status and the expression pattern were analysed. A 1 596 bp length complete sequence of DenActin, including an open reading frame of 1 134 bp encoding 377 amino acids was cloned. A 403 bp 3′-terminal of DenActin fragment containing an intron was cloned from gDNA. Homologous alignment analysis showed that it shared over 85% nucleotide sequence similarity to other plants and more than 97% in amino acid level. The phylogenetic tree constructed by maximum likelihood method suggested that the relationship of Den. 'Sonia Hiasakul' with Den. moniliforme was most intimate and it was classified as a major group with other Orchidaceae plants. The semi-quantitative PCR analysis showed that DenActin was stably expressed in various organs of Den. 'Sonia Hiasakul' such as seedling leaves, medium seeding stems tips, roots, leaves and stems, roots, leaves, stalk, flower with a diameter of 1 mm, 3 mm, 5 mm, and 9 mm of mature plant. Cloning and expression pattern analysis of DenActin would provide a candidate reference gene for the expression analysis of functional genes in Den. 'Sonia Hiasakul', and the sequence of 3′-terminal of gDNA containing one intron would provide convenience for primer design to avoid gDNA contamination in quantitative real-time PCR analysis.