Cloning and Expression Analysis of DcPSY2 Gene Under Abiotic Stress in Carrot (Daucus carota)
DING Xu, WANG Ya-Hui, LI Tong, ZHANG Rong-Rong, XU Zhi-Sheng, XIONG Ai-Sheng*
College of Horticulture, Nanjing Agricultural University / State Key Laboratory of Crop Genetics and Germplasm Enhancement / Ministry of Agriculture and Rural Affairs Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing 210095, China
Abstract：Carotenoids, as an important nutrient, are very common in carrot (Daucus carota). Lycopene, a kind of carotenoids, plays important roles in growth, development, and response to abiotic stress in higher plant. Phytoene synthase (PSY) is one of the key enzymes of lycopene biosynthesis. In this study, DcPSY2 (GenBank No. NM_001329167.1) was cloned from carrot cv. 'Junchuanhong' by RT-PCR (reverse transcription PCR). It contained an ORF with the length of 1 317 bp and encoded 438 amino acids. The amino acid sequences of DcPSY2 and the PSY proteins of 17 other plants were compared, the results showed that the overall similarity of PSY proteins was 82.22 %, which indicated that PSY proteins were conserved. Evolutionary relationship exhibited that DcPSY2 protein had the closest evolutionary relationship with the PSY protein in mahogany (Bixa orellana). DcPSY2 protein was a hydrophilic and non-secretory protein without signal peptide. It belonged to isoprenoid biosyn C1 superfamily and was distributed in various organelles. Secondary structure prediction demonstrated that DcPSY2 contained 53.65 % α-helix, 5.48 % β-fold, 12.79 % extended main chain and 28.08 % random curl. The results of qRT-PCR showed that DcPSY2 gene expression changed significantly in response to 4 kinds of abiotic stresses, including high temperature (38 ℃), low temperature (4 ℃), drought (200 g/L PEG6000) and high salt (200 mmol/L NaCl). Under high temperature or high salt treatments, the relative expression level of DcPSY2 reached the peak after 2 h. Whereas under low temperature or drought treatments, the expression level of DcPSY2 increased significantly at 1 h (P<0.05). The results provide a basic material for the functional study of DcPSY2 gene in carrot growth and its application study in stress resistance breeding.
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