Cloning and Expression Characteristic Analysis of PlSPL1 Gene in Paeonia lactiflora Pall.

doi:10.7668/hbnxb.20194773

Abstract

Abstract:

To investigate the nature and function of PlSPL1 (SPL)gene of Paeonia lactiflora,further elucidate the commonalities and characteristic differences of the PlSPL1 gene in different species,and explore the role of PlSPL1 in the degree of stem straightening in Paeonia lactiflora. The full-length sequence of the PlSPL1 gene was obtained by RACE technology using the stalk of herbaceous peony Hongfeng as the research material,and the structural,physicochemical properties,and phylogenetic relationships of PlSPL1 were analyzed and predicted using bioinformatics software,then,the expression level of PlSPL1 in different developmental stages of herbaceous peony stems was analyzed using qRT-PCR technology,and subcellular localization analysis of the protein was conducted using laser confocal microscopy technology.The results showed that the open reading frame of the PlSPL1 gene was 3 000 bp,encoding 999 amino acids.The protein had a molecular formula of C₄₈₆₉H₇₆₈₂N₁₄₀₆O₁₄₉₇S₄₃,a molecular weight of 111.25 ku,a theoretical isoelectric point of 6.26,encoding a hydrophilic unstable acidic protein,phosphorylation modification predominantly on serine,no signal peptide,with transmembrane structure,and had a secondary structure consisting mainly of random coil.Phylogenetic tree analysis revealed that the PlSPL1 protein was most closely related to peony,followed by a closer relationship with grape;protein sequence comparison analysis revealed that the PlSPL1 protein had a conserved SBP domain which was unique to SPL transcription factor family.The relative expression analysis found that PlSPL1 gradually showed a decreasing trend with stem development,indicating that PlSPL1 was negatively regulating the stem development of herbaceous peony and it was hypothesised that PlSPL1 played important roles in degree of stem straightness;and the subcellular localisation showed that the PlSPL1 protein was localised in the nucleus.The above results indicate that PlSPL1 is participated in the stalk development of herbaceous peony.

Key words: Paeonia lactiflora Pall., SPL, Gene cloning, Expression pattern, Subcellular location

XU Huajie, LU Lili, TANG Yuhan, ZHAO Daqiu, MENG Jiasong, TAO Jun. Cloning and Expression Characteristic Analysis of PlSPL1 Gene in Paeonia lactiflora Pall.[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(2): 55-61. doi: 10.7668/hbnxb.20194773.

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Figures/Tables 10

Fig.1 Detection of PCR amplification product of PlSPL1 gene in herbaceous peony DNA Marker 5000;1.PCR product.

Fig.2 Phylogenetic tree constructed based on amino acid sequences of PlSPL1 and Arabidopsis thaliana AtSPL1 encoded protein

Fig.3 Analysis of PlSPL1 protein phosphorylation sites

Fig.4 Signal peptide prediction of PlSPL1 protein in herbaceous peony

Fig.5 Transmembrane structure prediction of PlSPL1 protein in herbaceous peony

Fig.6 Prediction of secondary structure of PlSPL1 protein

Fig.7 Phylogenetic tree analysis of SPL proteins in 12 plants including herbaceous peony

Fig.8 Analysis of conserved sequence of SPL protein in 12 plants including herbaceous peony

Fig.9 Expression pattern of PlSPL1 at different developmental stages of the stem Different lowercase letters indicate significant differences(P<0.05).

Fig.10 Subcellular localization of PlSPL1 protein in tobacco epidermis

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