In this chapter, we summarize the strategies about molecular cloning and functional confirmation of plant genic male-sterility (GMS) genes and their applications for hybrid breeding and seed production via biotechnology-based male-sterility (BMS) systems in crop plants. The main content includes four sections: (1) GMS gene cloning strategies, including forward genetic approaches (e.g., map-based cloning, T-DNA or transposon tagging, and MutMap method) and reverse genetic approaches (e.g., homology-based cloning, anther-specific expression gene screening, and other reverse genetic methods); (2) functional confirmation methods of GMS genes, including transgenic complementation, targeted mutagenesis, allelic mutant test and sequencing, anther spatiotemporal expression analysis, and cytological observation; (3) application value assessment of GMS genes and mutants, such as genetic stability analysis of male sterility controlled by GMS genes under different genetic backgrounds and multiple environments, and genetic effects driven by GMS genes on plant heterosis and analysis of potential linkage with bad traits; (4) development and application of BMS systems based on GMS genes and/or their mutants, including transgenic construct-driven non-transgenic seed systems (e.g., seed production technology (SPT) and multi-control sterility (MCS)), and transgenic male-sterility systems (e.g., roundup hybridization systems (RHS1 and RHS2) and Barnase/Barstar system). Finally, we summarize and provide our perspectives on the studies of GMS genes and development of cost-effective and environment-friendly BMS systems in crop plants.
Part of the book: Synthetic Biology
Gene transcription and transcriptional regulation are crucial biological processes in all cellular life. Through the next-generation sequencing (NGS) technology, transcriptome data from different tissues and developmental stages can be easily obtained, which provides us a powerful tool to reveal the transcriptional landscape of investigated tissue(s) at special developmental stage(s). Anther development is an important process not only for sexual plant reproduction but also for genic male sterility (GMS) used in agriculture production. Plant comparative transcriptomics has been widely used to uncover molecular mechanism of GMS. Here, we focused on researches of anther developmental process and plant GMS genes by using comparative transcriptomics method. In detail, the contents include the following: (1) we described the commonly used flowchart in comparative transcriptomics; (2) we summarized the comparative strategies used to analyze transcriptome data; (3) we presented a case study on a maize GMS gene, ZmMs33; (4) we described the methods and results previously reported on gene co-expression and gene regulatory networks; (5) we presented the workflow of a case study on gene regulatory network reconstruction. The further development of comparative transcriptomics will provide us more powerful theoretical and application tools to investigate molecular mechanism underlying anther development and plant male sterility.
Part of the book: Transcriptome Analysis