Academy of Agricultural and Forestry Sciences, Qinghai University, Xining, Qinghai, China.
Qinghai Key Laboratory of Hulless Barley Genetics and Breeding, Xining, Qinghai, China.
BMC Genomics. 2023 Apr 4;24(1):166. doi: 10.1186/s12864-023-09240-5.
WD40 transcription factors, a large gene family in eukaryotes, are involved in a variety of growth regulation and development pathways. WD40 plays an important role in the formation of MYB-bHLH-WD (MBW) complexes associated with anthocyanin synthesis, but studies of Qingke barley are lacking.
In this study, 164 barley HvWD40 genes were identified in the barley genome and were analyzed to determine their relevant bioinformatics. The 164 HvWD40 were classified into 11 clusters and 14 subfamilies based on their structural and phylogenetic protein profiles. Co-lineage analysis revealed that there were 43 pairs between barley and rice, and 56 pairs between barley and maize. Gene ontology (GO) enrichment analysis revealed that the molecular function, biological process, and cell composition were enriched. The Kyoto Encyclopedia of Genes and Genomes (KEGG) results showed that the RNA transport pathway was mainly enriched. Based on the identification and analysis of the barley WD40 family and the transcriptome sequencing (RNA-seq) results, we found that HvWD40-140 (WD40 family; Gene ID: r1G058730), HvANT1 (MYB family; Gene ID: HORVU7Hr1G034630), and HvANT2 (bHLH family; Gene ID: HORVU2Hr1G096810) were important components of the MBW complex related to anthocyanin biosynthesis in Qingke, which was verified via quantitative real-time fluorescence polymerase chain reaction (qRT-PCR), subcellular location, yeast two-hybrid (Y2H), and bimolecular fluorescent complimentary (BiFC) and dual-luciferase assay analyses.
In this study, we identified 164 HvWD40 genes in barley and found that HvnANT1, HvnANT2, and HvWD40-140 can form an MBW complex and regulate the transcriptional activation of the anthocyanin synthesis related structural gene HvDFR. The results of this study provide a theoretical basis for further study of the mechanism of HvWD40-140 in the MBW complex related to anthocyanin synthesis in Qingke.
WD40 转录因子是真核生物中一个大型基因家族，参与多种生长调控和发育途径。WD40 在与花青素合成相关的 MYB-bHLH-WD（MBW）复合物的形成中起着重要作用，但青稞中 WD40 的研究还很缺乏。
本研究在大麦基因组中鉴定了 164 个大麦 HvWD40 基因，并对其相关的生物信息学进行了分析。根据结构和系统发育蛋白图谱，164 个 HvWD40 被分为 11 个簇和 14 个亚家族。共线性分析表明，大麦和水稻之间有 43 对，大麦和玉米之间有 56 对。基因本体（GO）富集分析表明，分子功能、生物过程和细胞组成富集。京都基因与基因组百科全书（KEGG）结果表明，RNA 转运途径主要富集。基于大麦 WD40 家族的鉴定和分析以及转录组测序（RNA-seq）结果，我们发现 HvWD40-140（WD40 家族；基因 ID：r1G058730）、HvANT1（MYB 家族；基因 ID：HORVU7Hr1G034630）和 HvANT2（bHLH 家族；基因 ID：HORVU2Hr1G096810）是与青稞中花青素生物合成相关的 MBW 复合物的重要组成部分，这通过定量实时荧光聚合酶链反应（qRT-PCR）、亚细胞定位、酵母双杂交（Y2H）和双分子荧光互补（BiFC）和双荧光素酶测定分析得到了验证。
本研究在大麦中鉴定了 164 个 HvWD40 基因，发现 HvnANT1、HvnANT2 和 HvWD40-140 可以形成 MBW 复合物，并调节花青素合成相关结构基因 HvDFR 的转录激活。本研究结果为进一步研究 HvWD40-140 在与青稞中花青素合成相关的 MBW 复合物中的作用机制提供了理论依据。
