Mizuno Nobuyuki, Nitta Miyuki, Sato Kazuhiro, Nasuda Shuhei
Laboratory of Plant Genetics, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Genes Genet Syst. 2012;87(6):357-67. doi: 10.1266/ggs.87.357.
An X-ray mutant showing an early flowering phenotype has been identified in einkorn wheat (Triticum monococcum L.), for which a major QTL for heading time was previously mapped in the telomeric region on the long arm of chromosome 3A. Recent advances in Triticeae genomics revealed that the gene order in this region is highly conserved between wheat and barley. Thus, we adopted a hypothetical gene order in barley, the so-called GenomeZipper, to develop DNA markers for fine mapping the target gene in wheat. We identified three genes tightly linked to the early heading phenotype. PCR analysis revealed that early-flowering is associated with the deletion of two genes in the mutant. Of the two deleted genes, one is an ortholog of the LUX ARRHYTHMO (LUX)/PHYTOCLOCK 1 (PCL1) gene found in Arabidopsis, which regulates the circadian clock and flowering time. We found distorted expression patterns of two clock genes (TOC1 and LHY) in the einkorn pcl1 deletion mutant as was reported for the Arabidopsis lux mutant. Transcript accumulation levels of photoperiod-response related genes, a photoperiod sensitivity gene (Ppd-1) and two wheat CONSTANS-like genes (WCO1 and TaHd1), were significantly higher in the einkorn wheat mutant. In addition, transcripts of the wheat florigen gene (WFT) accumulated temporally under short-day conditions in the einkorn wheat mutant. These results suggest that deletion of WPCL1 leads to abnormally higher expression of Ppd-1, resulting in the accumulation of WFT transcripts that triggers flowering even under short-day conditions. Our observations from gene mapping, gene deletions, and expression levels of flowering related genes strongly suggest that WPCL1 is the most likely candidate gene for controlling the early flowering phenotype in the einkorn wheat mutant.
在一粒小麦(Triticum monococcum L.)中鉴定出一个表现出早花表型的X射线突变体,此前已在3A染色体长臂的端粒区域定位到一个控制抽穗期的主要数量性状位点(QTL)。小麦族基因组学的最新进展表明,该区域的基因顺序在小麦和大麦之间高度保守。因此,我们采用了大麦中的一种假设基因顺序,即所谓的基因组拉链,来开发DNA标记,以便在小麦中精细定位目标基因。我们鉴定出三个与早抽穗表型紧密连锁的基因。PCR分析表明,早花与突变体中两个基因的缺失有关。在这两个缺失的基因中,一个是拟南芥中发现的LUX ARRHYTHMO(LUX)/PHYTOCLOCK 1(PCL1)基因的直系同源基因,该基因调节昼夜节律和开花时间。我们发现,一粒小麦pcl1缺失突变体中两个生物钟基因(TOC1和LHY)的表达模式出现紊乱,这与拟南芥lux突变体的报道一致。在一粒小麦突变体中,光周期反应相关基因、一个光周期敏感性基因(Ppd-1)和两个小麦CONSTANS样基因(WCO1和TaHd1)的转录本积累水平显著更高。此外,在短日条件下,一粒小麦突变体中开花素基因(WFT)的转录本在特定时间积累。这些结果表明,WPCL1的缺失导致Ppd-1异常高表达,从而导致WFT转录本积累,即使在短日条件下也能触发开花。我们从基因定位、基因缺失以及开花相关基因表达水平方面的观察结果强烈表明,WPCL1是控制一粒小麦突变体早花表型最有可能的候选基因。