The function of CchABCG5 gene in Camellia chekiangoleosa Hu

doi:10.3969/j.issn.1004-1524.20240746

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (7): 1407-1416.DOI: 10.3969/j.issn.1004-1524.20240746

• Crop Science • Previous Articles Next Articles

The function of CchABCG5 gene in Camellia chekiangoleosa Hu

MIAO Bailing^a(), CHEN Juanjuan^a, LI Liangjie^a, CHU Zongli^b, DONG Xiangxiang^a^,^*()

College of Horticulture; b. College of Agriculture, Xinyang Agriculture and Forestry University, Xinyang 464000, Henan, China

Received:2024-08-19 Online:2025-07-25 Published:2025-08-20

Abstract

Abstract:

ABCG5 is a transporter involved in plant growth and development. To investigate the function of the ABCG5 gene, Camellia chekiangoleosa Hu was used as experimental material. The coding sequence(CDS) of the CchABCG5 gene was obtained by RT-PCR amplification, and the gene function was preliminarily analyzed by bioinformatics analysis, subcellular localization and genetic transformation of Arabidopsis thaliana. The results showed that the CDS length of CchABCG5 gene was 1 935 bp, encoding 644 amino acids, with one nucleotide binding domain and one transmembrane domain, and it was a WBC-type half-sized transporter. Subcellular localization analysis showed that CchABCG5 protein was localized in the cell membrane, and phylogenetic analysis indicated that the CchABCG5 protein shares the closest evolutionary relationship with that of tea plant (Camellia sinensis). CchABCG5-overexpression Arabidopsis thaliana exhibited a reduced number of rosette leaves, an early flowering phenotype, and significantly elevated expression levels of flowering-related genes, as well as increased lipid content in seeds and leaves. Therefore, it was speculated that CchABCG5 gene played a role in flowering regulation and lipid transport in Camellia chekiangoleosa Hu.

Key words: Camellia chekiangoleosa Hu, ABCG5, transporter, gene cloning, subcellular localization, genetic transformation

CLC Number:

S794.4

MIAO Bailing, CHEN Juanjuan, LI Liangjie, CHU Zongli, DONG Xiangxiang. The function of CchABCG5 gene in Camellia chekiangoleosa Hu[J]. Acta Agriculturae Zhejiangensis, 2025, 37(7): 1407-1416.

Figures/Tables 10

Table 1 Primer information

基因Gene	正向引物Forward primer(5’-3’)	反向引物Reverse primer(5’-3’)	用途Usage
CchABCG5	GGTACCATGGAGACACAAGGTTGTGAGATTG	GTCGACAAGAAAAGCACCCTTAAGGCCC	基因克隆Gene cloning
CchABCG5	ACAGCAATCACTGCACCAAG	TGCTTCTACACTCACCTTCATCA	相对表达量分析
			Relative expression analysis
AtACT2	AAGCTGGGGTTTTATGAATGG	TTGTCACACACAAGTGCATCAT	相对表达量分析
			Relative expression analysis
AtCO	ATTCTGCAAACCCACTTGCT	CCTCCTTGGCATCCTTATCA	相对表达量分析
			Relative expression analysis
AtFT	CTGGAACAACCTTTGGCAAT	AGCCACTCTCCCTCTGACAA	相对表达量分析
			Relative expression analysis
AtFUL	GGAGAAGAAAACGGGTCAGC	ACTCGTTCGTAGTGGTAGGAC	相对表达量分析
			Relative expression analysis

Fig.1 Cloning of the CchABCG5 gene and restriction enzyme digestion of its overexpression vector M, DL 5 000 marker; 1, Target gene fragment; 2, Restriction enzyme digestion bands of pRI 101-AN-CchABCG5 plasmid.

Fig.2 Coding sequence and amino acid sequence of CchABCG5 protein *, Terminator.

Fig.3 Phylogenetic tree of ABCG5 proteins from different plant species

Fig.4 Structural domain and multiple sequence alignment of CchABCG5 protein

Fig.5 Kanamycin screening (A) and PCR verification (B) of CchABCG5-overexpression Arabidopsis thaliana M, DL 5 000 marker; 1-3, Overexpression lines; -, Negative control, +, Positive control.

Fig.6 Subcellular localization of CchABCG5 protein in Arabidopsis thaliana protoplasts GFP, Green fluorescence; mCherry, Cell membrane marker; Bright, Bright field; Merged, Superposition field.

Fig.7 Phenotype of Arabidopsis thaliana A, The flowering phenotype of Arabidopsis thaliana; B, Bolting time of Arabidopsis thaliana; C, The number of lotus leaves of Arabidopsis thaliana. Col-0, Wild-type Arabidopsis thaliana; OE1-OE3, three CchABCG5-overexpression Arabidopsis thaliana lines. Duncan’s multiple range test was used to compare the significance of differences, the bars marked without the same lowercase letters indicated significant difference between different lines (P<0.05). The same as below.

Fig.8 Relative expression levels of flowering-related genes in Arabidopsis thaliana t-test was used to analyze the statistical significance of differences, ** indicated P<0.01.

Fig.9 The lipid content in seeds (A) and leaves (B) of Arabidopsis thaliana

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The function of CchABCG5 gene in Camellia chekiangoleosa Hu

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