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为探索草莓光形态建成抑制子FaCOP1的功能及其表达特异性,采用同源克隆法获得‘丰香’草莓FaCOP1的完整开放阅读框序列,对该序列及其编码的氨基酸序列进行相关生物信息学分析,同时采用qRT-PCR分析该基因的时空表达以及在不同光质处理下的表达模式。结果表明,FaCOP1开放阅读框全长1 989 bp,Gen Bank登录号为KX583676,编码662个氨基酸,蛋白质分子量为74.7187 kD,理论等电点为6.54,具有环形锌指域,卷曲螺旋域和WD40重复序列等3个保守结构域。序列比对以及系统进化树分析发现,COP1进化过程中具有高度保守性以及物种间的差异性。qRT-PCR结果表明,FaCOP1在草莓的根、茎、叶、花及成熟果实中均有表达,花中的表达量最高,其次是叶和根,而茎和成熟果实中最少。随着草莓果实的发育(从小绿期到全红期)FaCOP1的转录水平总体呈现递减的规律,与果实花青素积累模式相反。草莓叶片和果实的FaCOP1表达均能够被白、红、蓝及红蓝光(1︰1)诱导。FaCOP1在转录水平上没有阻遏FaHY5的表达,其关系需在蛋白质水平上进一步验证。
In order to explore the function and expression specificity of FaCOP1, an inhibitor of strawberry light morphology, a complete open reading frame (ORF) sequence of FaCOP1 was obtained by homologous cloning. The bioinformatics analysis was carried out on this sequence and its encoded amino acid sequence Meanwhile, the temporal and spatial expression of the gene was analyzed by qRT-PCR and the expression pattern under different light quality treatments. The results showed that the open reading frame of FaCOP1 was 1 989 bp in length and GenBank accession number KX583676 encoded 662 amino acids with a molecular weight of 74.7187 kD and a theoretical isoelectric point of 6.54 with a circular zinc finger domain, coiled-coil domain and WD40 repeats Sequence and other three conserved domains. Sequence alignment and phylogenetic tree analysis showed that COP1 was highly conserved and inter-species divergent. The results of qRT-PCR showed that FaCOP1 was expressed in roots, stems, leaves, flowers and mature fruits of strawberry. The expression level of FaCOP1 was the highest in flowers, followed by leaves and roots, while it was the lowest in stem and mature fruits. With the development of strawberry fruit (from small green to red), the transcriptional level of FaCOP1 showed a decreasing pattern in general, contrary to the pattern of anthocyanin accumulation in fruit. FaCOP1 expression in strawberry leaves and fruits was induced by white, red, blue and red-blue light (1︰1). FaCOP1 does not repress FaHY5 expression at the transcriptional level, and its relationship needs to be further verified at the protein level.