Volume 55 Issue 6
Jun.  2024
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CHOU Li-ping, CHAO Rui-qiang, LIANG Jin-jun, WEN Peng-fei, ZHANG Peng-fei. 2024: Expression and vector construction of ZjPG gene in Ziziphus jujuba Mill.. Journal of Southern Agriculture, 55(6): 1682-1691. DOI: 10.3969/j.issn.2095-1191.2024.06.013
Citation: CHOU Li-ping, CHAO Rui-qiang, LIANG Jin-jun, WEN Peng-fei, ZHANG Peng-fei. 2024: Expression and vector construction of ZjPG gene in Ziziphus jujuba Mill.. Journal of Southern Agriculture, 55(6): 1682-1691. DOI: 10.3969/j.issn.2095-1191.2024.06.013
shu

Expression and vector construction of ZjPG gene in Ziziphus jujuba Mill.

  • 1 College of Horticulture, Shanxi Agricultural University, Jinzhong, Shanxi 030801, China;

  • 2 Shanxi Shilou Jujube Science and Technology Courtyard, Lüliang, Shanxi 032500, China

Funds: 

The Open Bidding Project of Science and Technology Major Special Plan of Shanxi (20220114 0601027)

Shanxi Key Research and Development Plan Project (201903D221072)

Position Expert Project of Fruit Tree Industry System in Shanxi (2023CYJSTX07-13)

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  • Received Date: January 23, 2024
    Graphical Abstract
    • Abstract
  • 【Objective】 This study aimed to clone the polygalacturonase（PG） gene from Chinese jujube（Ziziphus jujuba Mill.）, analyze the expression pattern of ZjPG gene and construct an overexpression vector, providing a theoretical reference for exploring the molecular mechanism of ZjPG gene in regulating fruit cracking and breeding crack-resistant jujube varieties. 【Method】 Using Z. jujuba Mill. cv. Huping as experimental material, the ZjPG gene（ID: LOC107426373） was cloned. The ZjPG protein was analyzed bioinformatically by online tools such as ExPASy, SOPMA and SWISSMODEL. A phylogenetic tree based on PG amino acid sequence was constructed by using MEGA 7.0 software, and the cis-acting elements of the ZjPG promoter were analyzed. The relative expression levels of the ZjPG gene were detected by quantitative real-time PCR（qRT-PCR） in different tissues of Z. jujuba Mill. cv. Huping（young leaves, mature leaves, flowers, fruits at expansion period, white mature period, crisp ripening period and full-ripening periods） as well as normal and cracked fruits at the crisp ripening period, which had been all treated with different concentrations of methyl jasmonate（MeJA, 0.2 and 0.4 mol/L） and abscisic acid（ABA, 0.2 mol/L）. The ZjPG gene overexpression vector was constructed. 【Result】 The total length of coding region（CDS） sequence of ZjPG gene in Z. jujuba Mill. cv. Huping was 1353 bp, with 99.70% CDS sequence similarity and 99.33% amino acid sequence similarity to the ZjPG gene in NCBI database, indicating the successful cloning of the ZjPG gene. The molecular weight of ZjPG protein was 49145.28 Da, encoding 450amino acids residues, with a theoretical isoelectric point of 6.27. It was a stable hydrophilic protein without transmembrane structural domains, containing a conserved sequence of Pgu1. Its secondary structure consisted of 12.67% α-helix, 32.22% extended strand, 8.00% β-sheet and 47.11% random coil. Phylogenetic evolutionary tree showed that ZjPG was closely related to AtPG from Arabidopsis thaliana and VvPG from Vitis vinifera. ZjPG and PG protein motifs of other nine species were highly conserved. The ZjPG gene promoter contained multiple hormone-responsive cis-acting elements. The qRT-PCR analysis showed that ABA and MeJA could induce the expression of ZjPG gene. The relative expression level of the ZjPG gene was significantly higher in cracked fruits than in normal fruits（P＜0.05）. The ZjPG gene was expressed in leaves, flowers and fruits of Z. jujuba Mill. cv. Huping, with the highest relative expression level in full-ripening period fruits. The pCAMBIA-1300-ZjPG expression vector was successfully constructed. 【Conclution】 The CDS sequence of the Zj PG gene from Z. jujuba Mill. cv. Huping is successfully cloned and its overexpression vector is successfully constructed.The PG protein sequences are highly conserved among different species. The expression of the ZjPG gene is regulated by ABA and MeJA signals, and highly expressed in cracked fruits. The ZjPG gene expression shows tissue and spatiotemporal specificity.
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