(87) Molecular Characterization of Cold Acclimation in Poncirus and Citrus

Abstract

Commercial citrus species, some of the most important fruit crops worldwide, are sensitive to sub-freezing temperatures. Poncirus trifoliata, a species closely related to commercial citrus and tolerant to –30 °C, has been used in breeding programs or as a rootstock to impart greater freeze tolerance. Gene expression of P. trifoliata and C. unshiu (Satsuma mandarin) were investigated and compared under slow and fast cold-acclimation regimes. The mRNA differential display-polymerase chain reaction (DDRT-PCR) and cDNA-AFLP, coupled with quantitative relative RT-PCR or real-time PCR were used. Many unique gene fragments were isolated and found to be up- or down–regulated as a result of exposure to low temperature. The up-regulated fragments in Poncirus show high similarities to genes involved in osmotic regulation (betaine/proline transporter, water channel protein, and nitrate transporter), oxidative stress (aldoketo reductase, early light induced protein), and protein interaction (tetratricopeptide-repeat protein, F-box protein, and ribosomal protein L15). In C. unshiu the up-regulated genes show high similarities to genes involved in transcription (zinc finger and GTP-binding protein-related), signal transduction (14–3–3 protein and extension-like protein), protein synthesis and amino acid translocation (permease and ribosomal proteins), chromosome folding (chromosome condensation, structural maintenance of chromosomes-like protein), and carbohydrate metabolism (glycosyl transferase). Several genes involved in photosynthesis, defense and cell wall metabolism were down regulated. Characterization of cold responsive genes will be discussed.