Cloning of light regulator LjCOP1 in Lonicera japonica and its expression characteristics

doi:10.3969/j.issn.1004-1524.20230896

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (6): 1290-1299.DOI: 10.3969/j.issn.1004-1524.20230896

• Horticultural Science • Previous Articles Next Articles

Cloning of light regulator LjCOP1 in Lonicera japonica and its expression characteristics

WAN Yan¹^,²(), ZHOU Xiaochun³, FANG Hailing¹^,²^,^*(), LIN Yi¹, QI Xiwu², YU Xu², CHEN Zequn², LIANG Chengyuan²^,^*()

1. Nanjing University of Chinese Medicine, Nanjing 210023, China
2. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China
3. Yancheng Dafeng District Forest Farm, Yancheng 224100, Jiangsu, China

Received:2023-07-19 Online:2024-06-25 Published:2024-07-02

Abstract

Abstract:

Light is one of the most important environmental factors to regulate growth, development and metabolism of plants, and COP1 (Constitutively photomorphogenic1) is a molecular switch for light signal transduction. In this study, LjCOP1 gene was cloned from honeysuckle (Lonicera japonica Thunb.) based on the honeysuckle transcriptome database, and analyzed for its bioinformatics and expression pattern. The results showed that LjCOP1 contains 13 exons and 12 introns, with a coding sequence (CDS) length of 2 034 bp encoding 677 amino acid residues. Multiple sequence comparison showed that LjCOP1 is highly conserved with other plant COP1 sequences, and all of them contain special structural domains such as ring zinc finger binding domain, coiled-coil domain and WD40 repeats. Protein structure prediction based on SWISS-MODEL showed that LjCOP1 was highly conserved with AtCOP1 of Arabidopsis thaliana at the level of protein tertiary structure. The analysis of promoter cis-acting progenitor showed that pLjCOP1 contained a large number of hormone-responsive, light-responsive, and stress-responsive elements. Phylogenetic analysis showed that LjCOP1 and LsCOP1 in lettuce genes were closely related. Furthermore, expression pattern analysis showed that LjCOP1 was expressed in different parts of honeysuckle and during the period of flower development, and was highly expressed in leaves. Compared with the dark condition, the highest gene abundance of LjCOP1 was found in white light, and responded more strongly to dark-to-blue light.

Key words: Lonicera japonica, LjCOP1 gene, expression characteristic

CLC Number:

S567.9

WAN Yan, ZHOU Xiaochun, FANG Hailing, LIN Yi, QI Xiwu, YU Xu, CHEN Zequn, LIANG Chengyuan. Cloning of light regulator LjCOP1 in Lonicera japonica and its expression characteristics[J]. Acta Agriculturae Zhejiangensis, 2024, 36(6): 1290-1299.

Figures/Tables 8

Table 1 Primers used for cloning and expression of LjCOP1 gene

引物名称 Primer name	序列 Primer sequence(5'-3')
LjCOP1-F	ATGGCTGCTGATAGTTCAGTTGGG
LjCOP1-R	TCATCTTTTCGTTGCGACAAAAAAG
LjCOP1pro-F	TATAAAGCCTTCAATCGCTAAG
LjCOP1pro-R	ATCCCTGTCCAACTCATCC
qLjCOP1-F	GAGAAACATAAGCCAACC
qLjCOP1-R	TATTCGCCATTTACCG
qLjActin-F	TGCTGGATTCTGGTGATGGT
qLjActin-R	ATTTCCCGCTCTGCTGTG

Fig.1 Electrophoresis results (A) of LjCOP1 gene PCR products and gene structure (B), and comparison of COP1 sequences in different plants (C) Rectangles in Figure B are exons, lines are introns; The horizontal lines in Figure C represent conserved RING, coiled coil and WD40 domains.

Fig.2 Prediction of the secondary structure of LjCOP1 protein of L. japonica Orange helix is α-helix, purple arrow is extended strand, and pink line is random curl.

Fig.3 Prediction of tertiary structure of LjCOP1 protein in L. japonica(A) and comparison between the tertiary structure of LjCOP1 and AtCOP1 (B) The orange fragment is the protein structure of Arabidopsis COP1, and the green fragment is the SWISS-MODEL prediction result of LjCOP1 protein of L. japonica.

Fig.4 Analysis of promoter and regulatory elements of LjCOP1 gene in L. japonica The red font represents the start codon and the upstream position is negative.

Fig.5 Phylogenetic analysis of the LjCOP1 in L. japonica

Fig.6 Temporal and spatial expression analysis of LjCOP1 gene in L. japonica A, Expression analysis of LjCOP1 gene in different tissue parts of L. japonica;B, Expression analysis of LjCOP1 gene in different flowering stages of L. japonica,in Figure B, YA is the young alabastrum, GA is the green alabastrum, WAI is the white alabastrum I, WAII is the white alabastrum Ⅱ, SF is the silvery flower, and GF is the golden flower. The bars with different lowercase letters indicate the significant difference (P<0.05). The same as below.

Fig.7 Analysis of LjCOP1 gene expression in L. japonica under different light conditions A, Dark; B, White light; C, Red light; D, Blue light.

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Cloning of light regulator LjCOP1 in Lonicera japonica and its expression characteristics

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