Key Laboratory of Southwest China Wildlife Resources Conservation (Ministry of Education), College of Life Science, China West Normal University, Nanchong, Sichuan, People's Republic of China.
School of Agricultural Science, Xichang University, Xichang, Sichuan, People's Republic of China.
PeerJ. 2022 Aug 24;10:e13902. doi: 10.7717/peerj.13902. eCollection 2022.
Crop male sterility has great value in theoretical research and breeding application. HTS-1, whose stamens transformed into pistils or pistil-like structures, is an important male sterility material selecting from Chinese Spring three-pistil (CSTP) wheat. However the molecular mechanism of pistillody development in HTS-1 remains a mystery. RNA-seq data of 11 wheat tissues were obtained from the National Center for Biotechnology Information (NCBI), including the stamens of CSTP and the pistils and pistillodic stamen of HTS-1. The Salmon program was utilized to quantify the gene expression levels of the 11 wheat tissues; and gene quantification results were normalized by transcripts per million (TPM). In total, 58,576 genes were used to construct block-wise network by co-expression networks analysis (WGCNA) R package. We obtained all of modules significantly associated with the 11 wheat tissues. AgriGO V2.0 was used to do Gene Ontology (GO) enrichment analysis; and genes and transcription factors (TFs) in these significant modules about wheat pistillody development were identified from GO enrichment results. Basic local alignment search tool (BLAST) was used to align HTS-1 proteins with the published pistillody-related proteins and TFs. Genes about wheat pistillody development were analyzed and validated by qRT-PCR. The MEturquoise, MEsaddlebrown, MEplum, MEcoral1, MElightsteelblue1, and MEdarkslateblue modules were significantly corelated to pistillodic stamen (correlation < 0.05). Moreover, 206 genes related to carpel development (GO:0048440) or gynoecium development (GO:0048467) were identified only in the MEturquoise module by Gene Ontology (GO) analysis, and 42 of 206 genes were hub genes in MEturquoise module. qRT-PCR results showed that 38 of the 42 hub genes had highly expressed in pistils and pistillodic stamens than in stamens. A total of 15 pistillody development-related proteins were validated by BLAST. Transcription factors (TFs) were also analyzed in the MEturquoise module, and 618 TFs were identified. In total, 56 TFs from 11 families were considered to regulate the development of pistillodic stamen. The co-expression network showed that six of HB and three of BES1 genes were identified in 42 hub genes. This indicated that TFs played important roles in wheat pistillody development. In addition, there were 11 of ethylene-related genes connected with TFs or hub genes, suggesting the important roles of ethylene-related genes in pistillody development. These results provide important insights into the molecular interactions underlying pistillody development.
作物雄性不育在理论研究和育种应用中具有重要价值。HTS-1 的雄蕊转变为雌蕊或雌蕊状结构，是从小麦三雌蕊中国春（CSTP）中选择的重要雄性不育材料。然而，HTS-1 中雌蕊发育的分子机制仍然是一个谜。从国家生物技术信息中心（NCBI）获得了 11 种小麦组织的 RNA-seq 数据，包括 CSTP 的雄蕊和 HTS-1 的雌蕊和雌蕊状雄蕊。利用 Salmon 程序对 11 种小麦组织的基因表达水平进行定量；并通过每百万转录本（TPM）对基因定量结果进行标准化。总共使用共表达网络分析（WGCNA）R 包构建了 58576 个基因的块状网络。我们获得了与 11 种小麦组织显著相关的所有模块。使用 Agrigo V2.0 进行基因本体论（GO）富集分析；并从 GO 富集结果中鉴定出这些与小麦雌蕊发育显著模块相关的基因和转录因子（TFs）。使用基本局部比对搜索工具（BLAST）将 HTS-1 蛋白与已发表的雌蕊发育相关蛋白和 TFs 进行比对。通过 qRT-PCR 对与小麦雌蕊发育相关的基因进行分析和验证。MEturquoise、MEsaddlebrown、MEplum、MEcoral1、MElightsteelblue1 和 MEdarkslateblue 模块与雌蕊状雄蕊显著相关（相关系数 <0.05）。此外，通过基因本体论（GO）分析，仅在 MEturquoise 模块中鉴定出与心皮发育（GO：0048440）或雌蕊发育（GO：0048467）相关的 206 个基因，并且 206 个基因中有 42 个是 MEturquoise 模块中的枢纽基因。qRT-PCR 结果表明，42 个枢纽基因中有 38 个在雌蕊和雌蕊状雄蕊中的表达高于雄蕊。通过 BLAST 验证了总共 15 个与雌蕊发育相关的蛋白质。还分析了 MEturquoise 模块中的转录因子（TFs），并鉴定出 618 个 TFs。总共，来自 11 个家族的 56 个 TF 被认为可以调节雌蕊状雄蕊的发育。共表达网络显示，在 42 个枢纽基因中鉴定出 6 个 HB 和 3 个 BES1 基因。这表明 TFs 在小麦雌蕊发育中发挥重要作用。此外，有 11 个与乙烯相关的基因与 TFs 或枢纽基因相连，这表明乙烯相关基因在雌蕊发育中起着重要作用。这些结果为雌蕊发育的分子相互作用提供了重要的见解。
