Shinozuka H, Hisano H, Ponting R C, Cogan N O I, Jones E S, Forster J W, Yamada T
Field Science Center for Northern Biosphere, Hokkaido University, Kita 11, Nishi 10, Kita, Sapporo 060-0811, Japan.
Theor Appl Genet. 2005 Dec;112(1):167-77. doi: 10.1007/s00122-005-0119-8. Epub 2005 Oct 21.
The alpha-subunit of the casein protein kinase CK2 has been implicated in both light-regulated and circadian rhythm-controlled plant gene expression, including control of the flowering time. Two putative CK2alpha genes of perennial ryegrass (Lolium perenne L.) have been obtained from a cDNA library constructed with mRNA isolated from cold-acclimated crown tissue. The genomic organisation of the two genes was determined by Southern hybridisation analysis. Primer designs to the Lpck2a-1 and Lpck2a-2 cDNA sequences permitted the amplification of genomic products containing large intron sequences. Amplicon sequence analysis detected single nucleotide polymorphisms (SNPs) within the p150/112 reference mapping population. Validated SNPs, within diagnostic restriction enzyme sites, were used to design cleaved amplified polymorphic sequence (CAPS) assays. The Lpck2a-1 CAPS marker was assigned to perennial ryegrass linkage group (LG) 4 and the Lpck2a-2 CAPS marker was assigned to LG2. The location of the Lpck2a-1 gene locus supports the previous conclusion of conserved synteny between perennial ryegrass LG4, the Triticeae homoeologous group 5L chromosomes and the corresponding segment of rice chromosome 3. Allelic variation at the Lpck2a-1 and Lpck2a-2 gene loci was correlated with phenotypic variation for heading date and winter survival, respectively. SNP polymorphism may be used for the further study of the role of CK2alpha genes in the initiation of reproductive development and winter hardiness in grasses.
酪蛋白激酶CK2的α亚基与光调节和昼夜节律控制的植物基因表达有关，包括对开花时间的控制。从用冷驯化的冠组织中分离的mRNA构建的cDNA文库中获得了多年生黑麦草（Lolium perenne L.）的两个假定的CK2α基因。通过Southern杂交分析确定了这两个基因的基因组结构。针对Lpck2a - 1和Lpck2a - 2 cDNA序列设计的引物允许扩增包含大内含子序列的基因组产物。扩增子序列分析在p150/112参考作图群体中检测到单核苷酸多态性（SNP）。在诊断性限制性酶切位点内经过验证的SNP被用于设计酶切扩增多态性序列（CAPS）分析。Lpck2a - 1 CAPS标记被定位到多年生黑麦草连锁群（LG）4，Lpck2a - 2 CAPS标记被定位到LG2。Lpck2a - 1基因座的位置支持了之前关于多年生黑麦草LG4、小麦族同源群5L染色体和水稻第3号染色体相应区段之间保守同线性的结论。Lpck2a - 1和Lpck2a - 2基因座的等位变异分别与抽穗期和冬季存活率的表型变异相关。SNP多态性可用于进一步研究CK2α基因在禾本科植物生殖发育起始和抗寒性中的作用。
