In addition to Circadian clock associated 1 (CCA1) and Late elongated hypocotyl (LHY), Reveille/LHY-CCA1-LIKE (RVE/LCL) genes are known as clock regulators in plants. So far, the association of RVE expression with anthocyanin accumulation has been reported only in Arabidopsis and pear. In this study, Brassica oleracea RVE5 (BoRVE5) was selected as a candidate gene to develop a PCR marker to predict anthocyanin accumulation under low temperature (LT) condition, based on the observation that the lengths of AT simple sequence repeats in 3′ untranslated region of BoRVE5 tend to inversely correlates with anthocyanin amount. Among BoRVE5 genomic DNA sequences of purple cabbages and green cabbages—both high anthocyanin accumulators (HAAs) and low anthocyanin accumulators (LAAs) of green cabbages under LT condition, sequence polymorphisms changing its amino acid sequence were identified between LAAs and HAAs/purple cabbages. We named the LAA-type gene as BoRVE5a and HAA-type as BoRVE5b, and found their expression levels are associated with anthocyanin accumulation phenotypes. Using these polymorphisms, a molecular marker discriminating LAAs from HAAs and purple cabbages was developed and validated using cabbage lines with diverse phenotypes, including 57 green and 59 purple cabbages. This marker is expected to be even more valuable if used in combination with other markers for anthocyanin phenotypes, such as those developed from BoMYBL2-1 or BoDFR1.

除了与生物钟相关的 1 ( CCA1 ) 和晚期伸长的下胚轴( LHY ) 之外，Reveille / LHY-CCA1-LIKE ( RVE / LCL ) 基因被称为植物中的时钟调节器。到目前为止，仅在拟南芥和梨中报道了RVE表达与花青素积累的关联。本研究中，甘蓝RVE5（BoRVE5) 被选为候选基因以开发 PCR 标记以预测低温 (LT) 条件下的花青素积累，基于观察到BoRVE5 3' 非翻译区中 AT 简单序列重复的长度倾向于与花青素量成反比. 在紫色卷心菜和绿色卷心菜的BoRVE5基因组DNA序列中，LT条件下绿色卷心菜的高花青素蓄积物（HAA）和低花青素蓄积物（LAA），在LAAs和HAAs/紫色卷心菜之间鉴定了改变其氨基酸序列的序列多态性。我们将 LAA 型基因命名为BoRVE5a，将 HAA型基因命名为BoRVE5b，并发现它们的表达水平与花青素积累表型相关。使用这些多态性，开发了一种分子标记，可将 LAA 与 HAA 和紫色卷心菜区分开来，并使用具有不同表型的卷心菜品系（包括 57 种绿色和 59 种紫色卷心菜）进行验证。如果与其他花青素表型标记（例如从BoMYBL2-1或BoDFR1开发的标记）结合使用，则预计该标记会更有价值。