Cloning and expression analysis of peony PoLPAT2 gene

doi:10.3969/j.issn.1004-1524.20240504

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (2): 321-328.DOI: 10.3969/j.issn.1004-1524.20240504

• Horticultural Science • Previous Articles Next Articles

Cloning and expression analysis of peony PoLPAT2 gene

ZHANG Meiying¹^,²(), MO Qian¹^,³, QI Xiushuang³, TONG Ningning², KONG Fan², LIU Zheng’an², LYU Changping⁴^,⁵^,^*(), PENG Liping²^,³^,^*()

1. College of Horticulture, Hunan Agricultural University, Changsha 410128, China
2. Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, China National Botanical Garden, Beijing 100093, China
3. Tianxiangyuan (Liaoning) Biotechnology Co., Ltd., Huludao 125300, Liaoning, China
4. College of Landscape Architecture and Horticultural Design, Hunan Agricultural University, Changsha 410128, China
5. Hunan Mid-Subtropical Quality Plant Breeding and Utilization Engineering Technology Research Center, Hunan Agricultural University, Changsha 410128, China

Received:2024-06-09 Online:2025-02-25 Published:2025-03-20
Contact: LYU Changping,PENG Liping

Abstract

Abstract:

Lysophosphatidic acid acyltransferase (LPAT) catalyzes the acylation of lysophosphatidic acid (LPA) at the sn-2 position to form phosphatidic acid (PA), which is one of the key rate-limiting enzymes in oil synthesis. Peony (Paeonia suffruticosa) is an emerging woody oil plant. Studying the evolution and expression patterns of the peony LPAT gene family is of great significance for further exploring the mechanisms of oil synthesis in peony seeds. In this study, we cloned the PoLPAT2 gene from peony seeds and analyzed its physicochemical properties, phylogenetic relationships, gene structure, and expression patterns using bioinformatics methods. The results showed that the coding sequence length of PoLPAT2 was 1 194 bp, encoding 397 amino acids, consisting of 20 different amino acids, and located on the plasma membrane. The relative molecular weight of the protein is 44.82 ku, with a molecular formula of C₂₀₆₅H₃₂₃₅N₅₄₁O₅₄₄S₁₅, a total of 6 400 atoms, a theoretical isoelectric point of 9.67. It contains 37 phosphorylation sites and exhibits high aliphatic properties. Classified as a hydrophilic protein, it possesses 3 transmembrane regions, lacks signal peptides, and has an instability index of 43.68, indicating it is an unstable protein. Protein structure prediction indicates that PoLPAT2 protein consists of four secondary structures, namely irregular coil, alpha helix, beta helix, and elongation chain. Phylogenetic analysis show that PoLPAT2 in peony is most closely related to homologs in grapes and walnuts. qRT-PCR results showed that the expression level of this gene was peaked at 105 d post-late seed development, and the overall trend showed a decrease followed by an increase, indicating that PoLPAT2 may be related to the stage of seed development. This findings provide reference for the functional verification of the PoLPAT2 gene and the improvement of oil content in peony seeds.

Key words: peony, lysophosphatidyl acyltransferase (LPAT), gene cloning, bioinformatics analysis

CLC Number:

S685.11

ZHANG Meiying, MO Qian, QI Xiushuang, TONG Ningning, KONG Fan, LIU Zheng’an, LYU Changping, PENG Liping. Cloning and expression analysis of peony PoLPAT2 gene[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 321-328.

Figures/Tables 9

Fig.1 Agarose gel electrophoresis of PoLPAT2

Fig.2 Sequence analysis of amino acids PoLPAT2 and other LPAT2s

Fig.3 Phylogenetic analysis of PoLPAT2 with its homologues from other species

Fig.4 Hydrophilic/hydrophobic distribution curve of PoLPAT2 protein

Fig.5 Signal peptide prediction of PoLPAT2 protein

Fig.6 Transmembrane domain prediction of PoLPAT2 protein

Fig.7 Secondary structure prediction of PoLPAT2 protein Blue, α-helix;Green, β-turn:Red, Extended strand;Purple, Random coli.

Fig.8 Prediction of tertiary structure of PoLPAT2 protein

Fig.9 Relative expression levels of PoLPAT2 in seeds at different developmental stages

References 15

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Cloning and expression analysis of peony PoLPAT2 gene

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