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Comprehensive transcriptome-based characterization of differentially expressed genes involved in microsporogenesis of
Yang Xie1, Wei Zhang1, Yan Wang1
1National Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
Abstract:
Microsporogenesis is an indispensable period for investigating microspore development and cytoplasmic male sterility (CMS) occurrence. Radish CMS line plays a critical role in elite F1 hybrid seed production and heterosis utilization. However, the molecular mechanisms of microspore development and CMS occurrence have not been thoroughly uncovered in radish. In this study, a comparative analysis of radish floral buds from a CMS line (NAU-WA) and its maintainer (NAU-WB) was conducted using next generation sequencing (NGS) technology. Digital gene expression (DGE) profiling revealed that 3504 genes were significantly differentially expressed between NAU-WA and NAU-WB library, among which 1910 were upregulated and 1594 were downregulated. Gene ontology (GO) analysis showed that these differentially expressed genes (DEGs) were mainly enriched in extracellular region, catalytic activity, and response to stimulus. KEGG enrichment analysis revealed that the DEGs were predominantly associated with flavonoid biosynthesis, glycolysis, and biosynthesis of secondary metabolites. Real-time quantitative PCR analysis showed that the expression profiles of 13 randomly selected DEGs were in high agreement with results from Illumina sequencing. Several candidate genes encoding ATP synthase, auxin response factor (ARF), transcription factors (TFs), chalcone synthase (CHS), and male sterility (MS) were responsible for microsporogenesis. Furthermore, a schematic diagram for functional interaction of DEGs from NAU-WA vs. NAU-WB library in radish plants was proposed. These results could provide new information on the dissection of the molecular mechanisms underlying microspore development and CMS occurrence in radish.
Insights
This study reveals key genes involved in microspore development and cytoplasmic male sterility (CMS) in radish. Findings offer insights into radish CMS molecular mechanisms for hybrid seed production.
Area of Science:
- Plant Science
- Molecular Biology
- Genetics
Background:
- Microsporogenesis is crucial for understanding microspore development and cytoplasmic male sterility (CMS).
- Radish CMS lines are vital for hybrid seed production, but their molecular mechanisms remain unclear.
- Investigating radish CMS can enhance elite F1 hybrid seed production and heterosis utilization.
Purpose of the Study:
- To uncover the molecular mechanisms of microspore development and CMS occurrence in radish.
- To perform a comparative analysis of gene expression between a radish CMS line and its maintainer.
- To identify candidate genes associated with microsporogenesis and male sterility in radish.
Main Methods:
- Comparative analysis of floral buds from radish CMS (NAU-WA) and maintainer (NAU-WB) lines.
- Next-generation sequencing (NGS) for digital gene expression (DGE) profiling.
- Gene Ontology (GO) and KEGG pathway enrichment analyses.
- Real-time quantitative PCR (RT-qPCR) for validating gene expression.
Main Results:
- 3504 significantly differentially expressed genes (DEGs) identified between CMS and maintainer lines.
- DEGs enriched in extracellular region, catalytic activity, and response to stimulus.
- Key pathways affected include flavonoid biosynthesis, glycolysis, and secondary metabolite biosynthesis.
- Candidate genes for ATP synthase, ARF, TFs, CHS, and MS identified.
Conclusions:
- The study provides novel insights into the molecular basis of microspore development and CMS in radish.
- Identified DEGs and candidate genes offer targets for further research into radish male sterility.
- Findings contribute to understanding and potentially improving hybrid seed production in radish.
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