Abstract：
CONCLUSIONS: BraANS.A3 was the key gene controlling purple leaf color in pak choi, and two short fragments of promoter region in green pak choi might be interfering its normal expression. Pak choi (B. rapa L. ssp. chinensis) is an influential and important vegetable with green, yellow, or purple leaves that is cultivated worldwide. The purple leaves are rich in anthocyanins, but the underlying genetics and evolution have yet to be extensively studied. Free-hand sections of the purple leaves indicated that anthocyanins mainly accumulate throughout the adaxial and abaxial epidermal leaf cells. Segregation analyses of an F2 population of a B. rapa ssp. chinensis L. purple leaf mutant ZBC indicated that the purple trait is controlled by an incompletely dominant nuclear gene. Bulked segregant analysis (BSA) showed that the key genes controlling the trait were between 24.25 and 38.10 Mb on chromosome A03 of B. rapa. From the annotated genes, only BraA03g050560.3C, homologous to Arabidopsis AtANS, was related to the anthocyanin synthesis pathway. Genome annotation results and transcriptional sequencing analyses revealed that the BraANS.A3 gene was involved in the purple leaf trait. qRT-PCR analyses showed that BraANS.A3 was highly upregulated in ZBC but hardly expressed in the leaves of an inbred homozygous line of B. campestris ssp. chinensis L. green leaf mutant WTC, indicating that BraANS.A3 played a key role catalyzing anthocyanin synthesis in ZBC. Full-length sequence alignment of BraANS.A3 in WTC and ZBC showed that it was highly conserved in the gene region, with significant variation in the promoter region. In particular, the insertion of two short fragments of the promoter region in WTC may interfere with its normal expression. The promoter regions of ANS in six Brassica species all had multiple cis-elements involved in responses to abscisic acid, light, and stress, suggesting that ANS may be involved in multiple metabolic pathways or biological processes. Protein-protein interactions predicted that BraANS.A3 interacts with virtually all catalytic proteins in the anthocyanin synthesis pathway and has a strong relationship with Transparent Testa 8 (TT8). These results suggest that BraANS.A3 promotes anthocyanin accumulation in purple pak choi and provide new insights into the functional analysis of anthocyanin-related genes in Chinese cabbage and transcriptional regulatory networks.
摘要：
结论：BraANS。A3是控制小白菜紫叶颜色的关键基因，绿白菜启动子区的两个短片段可能会干扰其正常表达。白菜(B.rapaL.ssp.中国）是一种有影响力的重要绿色蔬菜，黄色,或全世界种植的紫色叶子。紫色的叶子富含花青素，但是潜在的遗传学和进化还没有得到广泛的研究。紫色叶子的徒手切片表明，花色苷主要在整个正面和背面表皮叶细胞中积累。B.rapassp的F2种群的分离分析。紫叶突变体ZBC表明紫色性状受不完全显性核基因控制。批量分离分析（BSA）表明，控制该性状的关键基因在B.rapa的A03染色体上的24.25至38.10Mb之间。从注释的基因中，只有BraA03g050560.3C，同源拟南芥AtANS，与花青素合成途径有关。基因组注释结果和转录测序分析显示，BraANS。A3基因参与紫叶性状。qRT-PCR分析显示BraANS.A3在ZBC中高度上调，但在油菜纯合品系的叶片中几乎不表达。中国绿叶突变体WTC，表明Braans。A3在ZBC中催化花色苷合成中起关键作用。BraANS的全长序列比对。在WTC和ZBC中的A3显示它在基因区域高度保守，在启动子区域有显著的变异。特别是,WTC启动子区的两个短片段的插入可能会干扰其正常表达。6种芸苔属植物ANS的启动子区都有多个顺式元件参与对脱落酸的反应，光,和压力，提示ANS可能参与多种代谢途径或生物过程。蛋白质-蛋白质相互作用预测BraANS。A3与花青素苷合成途径中的几乎所有催化蛋白相互作用，并与透明Testa8（TT8）有很强的关系。这些结果表明BraANS。A3促进紫小白菜花色苷的积累，为大白菜花色苷相关基因的功能分析和转录调控网络提供新的见解。
