乳白石蒜<em>LaDFR1</em>基因及启动子克隆与功能研究

doi:10.3969/j.issn.1674-7968.2023.07.006

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Abstract The new variety Lycoris albiflora 'Astro Girl' has gradually changing flower colors, which is a special flower color of Lycoris. Previous studies have found that the dihydroflavonol 4-reductase (DFR) genes are the key enzyme genes for the anthocyanins biosynthesis in the petals of Lycoris. In order to further study the DFR genes function, LaDFR1 and its promoter sequence were cloned by RT-PCR and Genome walking technology from petals in Lycoris chinensis×radiata 'Astro Girl' in this study. Bioinformatics and expression pattern analysis of LaDFR1 were carried out. The function of the LaDFR1 promoter was further investigated by overexpressing it in tobacco (Nicotiana tabacum) leaves. The results showed that, (1) LaDFR1 cDNA with 1 161 bp ORF was cloned, encoding 386 amino acids. The conserved domain of amino acid sequence of LaDFR1 had NADPH and substrate binding sites, which belonged to NADB Rossmann super gene family. Sequence alignment and phylogenetic analysis showed that the consistency of DFR amino acid sequence between L. chinensis×radiata 'Astro Girl' and Nelumbo nucifera was 80.43%. (2) LaDFR1 was mainly expressed in petals, followed by filaments, and the lowest expression in scape. The expression of LaDFR1 gene was the lowest in the first flowering day, and the expression increased gradually with the extension of flowering time. (3) LaDFR1 promoter sequence with 1 570 bp was obtained, which contained multiple light, abscisic acid (ABA) induction, MYB and NAC binding cis acting elements. The LaDFR1pro: PBI121 integration vector was constructed to transiently transform tobacco leaves. The GUS activity induced by light was higher than dark, which indicated that the LaDFR1 promoter had photoinduced activity. In summary, the LaDFR1 gene and its promoter sequence were successfully obtained in this study, which may be involved in the synthesis and accumulation of anthocyanins in the petals of L. chinensis×radiata 'Astro Girl'. This study can provide target genes for further study on the regulation of color changes in Lycoris albiflora and color improvement using genetic engineering.

Key words： Lycoris albiflora Dihydroflavonol 4-reductase (DFR) Gene clone Expression analysis

Received: 05 May 2022

Corresponding Authors: *gaoyanhui408@126.com

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	ZHU Yi-Chen
	XUE Hui-Min
	ZHOU Qi
	ZONG Jie
	GAO Yan-Hui

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ZHU Yi-Chen,XUE Hui-Min,ZHOU Qi, et al. Cloning and Function Study of LaDFR1 Gene and Promoter from Lycoris albiflora[J]. 农业生物技术学报, 2023, 31(7): 1396-1404.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2023.07.006 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2023/V31/I7/1396

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