枸杞LbSPL6基因的克隆及表达分析
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引用本文:张红雨,梁新华,石 晶.枸杞LbSPL6基因的克隆及表达分析[J].西北植物学报,2021,41(3):377~385
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作者单位
张红雨,梁新华,石 晶* (宁夏大学 生命科学学院银川 750021) 
基金项目:国家自然科学基金(31401444,41661108);
中文摘要:从实验室前期对枸杞(Lycium barbarum L.)花发育过程转录组测序结果推测,枸杞Squamosa启动子结合蛋白(Squamosa promoter binding protein like,SPL)转录因子可能在枸杞花发育过程中发挥重要功能。该研究以宁夏特色植物资源枸杞为材料,采用RACE方法克隆LbSPL6基因,通过生物信息学及基因表达分析对该基因进行初步研究。结果表明:(1)成功克隆获得LbSPL6基因,其开放阅读框全长1 524 bp,编码507个氨基酸,分子量为55.34 kD;序列分析表明LbSPL6蛋白中包含3个保守基序,且氨基酸序列与茄科植物同源蛋白的氨基酸序列高度相似。(2)qRT PCR分析证实,LbSPL6基因在枸杞花器官中表达,并且在花药发育的四分体时期及单核花粉时期表达量较高;亚细胞定位实验证明,LbSPL6蛋白定位于细胞核中。该研究结果为进一步研究枸杞LbSPL6转录因子在花发育过程中的功能和作用机制奠定了基础。
中文关键词:枸杞  LbSPL6基因  基因克隆  表达分析
 
Cloning and Expression Analysis of LbSPL6 in Lycium barbarum
Abstract:Based on the transcriptome sequencing of flower development in Lycium barbarum, it is suggested that the Squamosa promoter binding protein like (SPL) transcription factor may play an important role in flower development of L. barbarum. In this study, we used RACE method to clone the LbSPL6 gene from a characteristic plant resource in Ningxia——L. barbarum. Then we studied the gene function through bioinformatics and gene expression analysis. The results showed that: (1) the open reading frame of LbSPL6 gene was 1 524 bp in length, which encoded 507 amino acids, and a molecular weight of 55.34 kD; Sequence analysis showed that the LbSPL6 protein contained three conserved motifs; The amino acid sequence of LbSPL6 protein was similar to that of Solanaceae homologous protein. (2) qRT PCR analysis results showed that the LbSPL6 gene was expressed in the floral organs of L. barbarum. Moreover, the expression of LbSPL6 was higher in the tetrad stage and single nucleus pollen stage of anther development. Subcellular localization confirmed the nuclear localization of LbSPL6 protein. These results further elucidated the function of LbSPL6 transcription factor in the development of L. barbarum flowers.
keywords:Lycium barbarum L.  LbSPL6 gene  gene cloning  expression analysis
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