Cloning and Analysis of GmCOL4 Gene in Glycine max

CONSTANS (CO) is a key gene in photoperiodic flowering pathway and locates between the circadian clock gene and the meristem identity gene. For elucidating the function of CO in flowering behavior of soybean, a CO-like gene, designated GmCOL4, was cloned from Glycine max cv. Kennong 18 using reverse transcription-polymerase chain reaction (RT-PCR) and bioinformatics methods. This gene encodes a protein embedded 2 conserved domains, B-box and CCT, which belong to subgroup III. Phylogenetic analysis based on critical amino acid sequences indicated that GmCOL4 is close to COL9 with similarity of 64.3%. The expression profiles of GmCOL4 revealed by quantitative real-time RT-PCR (qRT-PCR) showed a similar pattern to that of COL9. GmCOL4 was largely regulated by biological circadian, while light had weak effect on it. GmCOL4 was expressed mainly in leaves with the highest expression at anthesis stage. Therefore, GmCOL4 is one of the important genes for regulation of flowering in soybean.

摘要CONSTANS(CO)是植物光周期开花途径中的关键基因之一。通过RT-PCR和生物信息学的方法, 克隆了大豆GmCOL4基因并分析其结构特征, 用实时荧光定量PCR (quantitative real-time RT-PCR, qRT-PCR)研究了其转录特点。结果表明, GmCOL4的4个外显子编码一个具有B-box和CCT保守结构域的CO-like蛋白, 在序列上与拟南芥(Arabidopsis thaliana) COL9相似性最高, 为64.3%。分析其转录特征发现, GmCOL4表达主要受生物节律的影响, 受光的调节作用较弱。器官特异性表达分析发现, GmCOL4主要在大豆叶片中表达, 表达模式与COL9相似。