Abstract: 【Objective】 The Omega -3 fatty acid desaturase (ω-3 FAD) is a key enzyme in the fatty acid biosynthesis pathway of plant. Through analysis of omega-3 fatty acid desaturase gene structure and its expression in different tissues from Paeonia ostii, this study will lay a foundation for the research of regulating effects of FAD3 gene on fatty acid biosynthesis and provide a theoretical basis for the formation in the process of regulation. 【Method】 The FAD gene from Paeonia ostii (FAD3) was cloned using RT-PCR and RACE-PCR strategies. Nucleotide sequence was analyzed using Vector NTI Advance 11 software. Homology was analyzed using BLAST. A phylogenetic tree was constructed using neighbor-joining of MEGA 7.0. The secondary structure and three-dimensional model of FAD3 were predicted using ExPASy and Phyre2, respectively. Expression profiles of FAD3 at different developmental stages and in different tissues of the P. ostii were assayed using real-time quantitative PCR. 【Result】 The FAD gene in P. ostii was cloned and named as FAD3 (GenBank accession number: KX906966). The full length of FAD3 cDNA is 1 723 bp, contains a 1 308 bp open reading frame (ORF) encoding a putative protein of 435 amino acids with a molecular mass of 49.9 kD and an isoelectric point (pI) of 7.42. The 3′-untranslated region of 287 bp and 5′-untranslated region of 99 bp were obtained from the FAD3 gene of P. ostii. N end without signal peptide, fat coefficient is 83.08, instability index 35.67, the grand average of hydrophobicity value of -0.222. By FAD3 protein secondary structure prediction, FAD3 mainly in the alpha helix and random coil, followed by less extended strand, beta turn content; multiple sequence alignment results showed that it contains two conserved domains of FAD3 gene. Phylogenetic tree analysis showed that the P. ostii has the closest evolutionary relationship with P. lactiflora. Subcellular localization analysis of TMHMM and Target P indicated that it might be targeted to the endoplasmic reticulum with three transmembrane regions. FAD3 gene was expressed in the root, stem, leaf, petal, pistil, stamen and seed of P. ostii by the tissue-specificity expression. The expressive content of FAD3 gene in different tissues of P. ostii was different: The highest was leaf, the next was pistil, and the lowest level was in stamen; In different periods of seeds, the highest expression was seeds of 10d, the next was 20d, in 60d expression was the lowest. 【Conclusion】 The full length cDNA sequence of FAD3 gene was successfully cloned from P. ostii, It shows a variety of expression patterns in different tissues, which laid a foundation for the further study on the function and expression regulation mechanism of FAD3 gene in the process of unsaturated fatty acid biosynthesis.

Key words: Paeonia suffruticosa Andr., fatty acid desaturase, cloning, real-time PCR