Yuan Kaiwen, Zhao Xinyu, Sun Wenru, Yang Limei, Zhang Yangyong, Wang Yong, Ji Jialei, Han Fengqing, Fang Zhiyuan, Lv Honghao
State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Hortic Res. 2023 Jun 29;10(8):uhad133. doi: 10.1093/hr/uhad133. eCollection 2023 Aug.
comprises several important vegetable and ornamental crops, including curly kale, ornamental kale, cabbage, broccoli, and others. The accumulation of anthocyanins, important secondary metabolites valuable to human health, in these plants varies widely and is responsible for their pink to dark purple colors. Some curly kale varieties lack anthocyanins, making these plants completely green. The genetic basis of this trait is still unknown. We crossed the curly kale inbred line BK2019 (without anthocyanins) with the cabbage inbred line YL1 (with anthocyanins) and the Chinese kale inbred line TO1000 (with anthocyanins) to generate segregating populations. The no-anthocyanin trait was genetically controlled by a recessive gene, . We generated a linkage map and mapped to a 256-kb interval on C09. We identified one candidate gene, , in the target genomic region; this gene is homologous to , which encodes a dihydroflavonol-4-reductase-like (DFR-like) protein in . In BK2019, a 1-bp insertion was observed in the second exon of and directly produced a stop codon. To verify the candidate gene function, CRISPR/Cas9 gene editing technology was applied to knock out . We generated three mutants, two of which were completely green with no anthocyanins, confirming that corresponds to Different insertion/deletion mutations in exons were found in all six of the other no-anthocyanin kale varieties examined, supporting that independent disruption of resulted in no-anthocyanin varieties of . This study improves the understanding of the regulation mechanism of anthocyanin accumulation in subspecies.
包括几种重要的蔬菜和观赏作物，如羽衣甘蓝、观赏羽衣甘蓝、卷心菜、西兰花等。这些植物中花青素（对人体健康有重要价值的次生代谢产物）的积累差异很大，这也是它们呈现从粉色到深紫色的原因。一些羽衣甘蓝品种缺乏花青素，使得这些植物完全呈绿色。该性状的遗传基础仍不清楚。我们将羽衣甘蓝自交系BK2019（无花青素）与卷心菜自交系YL1（有花青素）和芥蓝自交系TO1000（有花青素）杂交，以产生分离群体。无花青素性状受一个隐性基因遗传控制。我们构建了一个连锁图谱，并将该基因定位到C09染色体上一个256 kb的区间。我们在目标基因组区域鉴定出一个候选基因；该基因与在[具体物种]中编码二氢黄酮醇-4-还原酶样（DFR样）蛋白的[基因名称]同源。在BK2019中，在该基因的第二个外显子中观察到一个1 bp的插入，直接产生了一个终止密码子。为了验证候选基因的功能，应用CRISPR/Cas9基因编辑技术敲除该基因。我们获得了三个该基因的突变体，其中两个完全呈绿色且无花青素，证实了该基因对应于[基因名称]。在所有检测的其他六个无花青素羽衣甘蓝品种中均发现了该基因外显子的不同插入/缺失突变，支持该基因的独立破坏导致了无花青素羽衣甘蓝品种的产生。本研究增进了对羽衣甘蓝亚种花青素积累调控机制的理解。
