Abstract：
CONSTANS-LIKE (COL ) genes are a key signalling molecule that regulates plant growth and development during the photoperiod. Our preliminary experiments showed that the photoperiod greatly influence the formation of Tetrastigma hemsleyanum root tubers. In this study, we examined the oscillation patterns and expression characteristics of COL genes in leaves of T. hemsleyanum under different photoperiod conditions. Six genes were selected as candidate reference genes for further analyses: (1) 18S ribosomal RNA (18S rRNA ); (2) α-tubulin (TUBA ); (3) 30S ribosomal RNA (30S rRNA ); (4) TATA binding protein (TBP ); (5) elongation factor 1α (EF-1α ); and (6) RNA polymerase II (RPII ). The geNorm, NormFinder, and BestKeeper software programs were used to evaluate expression stability. Two ThCOL genes were screened in the T. hemsleyanum transcriptome library, and their expression patterns under different photoperiod conditions were analysed using quantitative reverse transcription PCR. The genes EF-1α , TUBA , and 18S rRNA were used to analyse the expression profiles of CONSTANS genes (ThCOL4 and ThCOL5 ) under different photoperiods. The expression peaks of ThCOL4 and ThCOL5 appeared at different times, demonstrating that their oscillation patterns were influenced by the photoperiod. We speculate that these two ThCOL genes may be involved in different biological processes.
摘要：
康斯坦斯样（COL）基因是在光周期期间调节植物生长和发育的关键信号分子。我们的初步实验表明，光周期极大地影响了四根hemsleyanum根块茎的形成。在这项研究中,我们研究了在不同光周期条件下Hemsleyanum叶片中COL基因的振荡模式和表达特征。选择六个基因作为候选参考基因进行进一步分析：（1）18S核糖体RNA（18SrRNA）；（2）α-微管蛋白（TUBA）；（3）30S核糖体RNA（30SrRNA）；（4）TATA结合蛋白（TBP）；（5）延伸因子1α（EF-1α）；（6）RNA聚合酶II（RPII）。geNorm,NormFinder,和BestKeeper软件程序用于评估表达稳定性。在T.hemsleyanum转录组文库中筛选了两个ThCOL基因，并使用定量逆转录PCR分析了它们在不同光周期条件下的表达模式。基因EF-1α，TUBA,和18SrRNA用于分析CONSTANS基因（ThCOL4和ThCOL5）在不同光周期下的表达谱。ThCOL4和ThCOL5的表达峰出现在不同的时间,证明它们的振荡模式受到光周期的影响。我们推测这两个ThCOL基因可能参与不同的生物学过程。
