College of Forestry, Shenyang Agricultural University, Shenyang, China.
Key Laboratory of Forest Tree Genetics, Breeding and Cultivation of Liaoning Province, Shenyang, China.
Theor Appl Genet. 2021 Jan;134(1):159-169. doi: 10.1007/s00122-020-03688-9. Epub 2020 Oct 4.
Overexpression and virus-induced gene silencing verified BoDFR1 conferred the anthocyanin accumulation in pink-leaved ornamental kale. Leaf color is an essential trait in the important horticultural biennial plant ornamental kale (Brassica oleracea var. acephala). The identity of the gene conferring this striking trait and its mode of inheritance are topics of debate. Based on an analysis of F, F, BCP, and BCP ornamental kale populations derived from a cross between a pink-leaved P28 and white-leaved D10 line, we determined that the pink leaf trait is controlled by a semi-dominant gene. We cloned two genes potentially involved in anthocyanin biosynthesis in ornamental kale: Bo9g058630 and Bo6g100940. Based on their variation in sequence, we speculated that Bo9g058630, encoding the kale dihydroflavonol-4 reductase (BoDFR1) enzyme, plays a critical role in the development of the pink leaf trait. Indeed, an InDel marker specific for BoDFR1 completely co-segregated with the pink leaf trait in our F population. We then generated the 35Spro: DFR-GUS overexpression vector, which we transformed into D10. Overexpression of BoDFR1 indeed restored some anthocyanin accumulation in this white-leaved parental line. In addition, we targeted BoDFR1 in P28 using virus-induced gene silencing. Again, silencing of BoDFR1 resulted in a substantial decrease in anthocyanin accumulation. This work lays the foundation for further exploration of the mechanism underlying anthocyanin accumulation in pink-leaved ornamental kale.
过量表达和病毒诱导的基因沉默证实 BoDFR1 导致了观赏羽衣甘蓝中花色苷的积累。叶色是二年生重要园艺植物观赏羽衣甘蓝（Brassica oleracea var. acephala）的一个重要特征。赋予这种显著特征的基因及其遗传模式是一个争论的话题。基于对来自粉色叶 P28 和白色叶 D10 系杂交的 F、F、BCP 和 BCP 观赏羽衣甘蓝群体的分析，我们确定粉色叶性状受一个半显性基因控制。我们克隆了两个可能参与观赏羽衣甘蓝花色苷生物合成的基因：Bo9g058630 和 Bo6g100940。基于它们序列的差异，我们推测 Bo9g058630 编码羽衣甘蓝二氢黄酮醇 4-还原酶（BoDFR1）酶，在粉色叶性状的发育中起关键作用。事实上，一个针对 BoDFR1 的 InDel 标记在我们的 F 群体中与粉色叶性状完全共分离。然后，我们生成了 35Spro:DFR-GUS 过表达载体，并将其转化到 D10 中。BoDFR1 的过表达确实恢复了该白色叶亲本系中一些花色苷的积累。此外，我们使用病毒诱导的基因沉默靶向 P28 中的 BoDFR1。再次，BoDFR1 的沉默导致花色苷积累大量减少。这项工作为进一步探索粉色叶观赏羽衣甘蓝中花色苷积累的机制奠定了基础。
