Molecular Plant Breeding 2024, Vol.15, No.6, 429-441 http://genbreedpublisher.com/index.php/mpb 436 5.3 Integrative model of environmental regulation of MS genes An integrative model of environmental regulation of MS genes in rice involves the interaction of temperature, light, and water stress. High temperatures have been shown to cause MS by altering gene expression during pollen development, with specific genes being repressed in the anther under high-temperature conditions (Endo et al., 2009). Additionally, the expression of critical genes and the accumulation of metabolites in the metabolism of sugar, lipid, and phenylpropanoid are essential for MS formation, as demonstrated by transcriptomic and metabolomic analyses (Sun et al., 2022). The regulatory mechanisms of MS also involve miRNAs and their target genes, which respond to temperature changes and influence various metabolic pathways (Sun et al., 2021b). Furthermore, the TF OsAL5 links drought stress response and TGMS, highlighting the complex interactions between environmental factors in regulating MS genes (Wen et al., 2021). 5.4 Implications for breeding and crop management Understanding the regulatory mechanisms of temperature, light, and water or humidity on the expression of MS genes in rice has significant implications for breeding and crop management. Breeding programs can focus on developing genetically diverse TGMS lines with critical sterility-inducing temperatures that are not affected by sudden interruptions with lower temperatures, ensuring stable MS under varying environmental conditions (Viraktamath and Virmani, 2001). Additionally, manipulating the expression of key regulatory genes, such as OsAL5, can enhance drought tolerance and improve the resilience of TGMS lines to temperature fluctuations (Wen et al., 2021). Integrating knowledge of miRNA-mediated regulation and metabolic pathways involved in MS can further aid in the development of rice varieties with optimized fertility and stress tolerance (Sun et al., 2021b). Overall, a comprehensive understanding of the cross-talk between environmental factors and MS genes will enable the development of more robust and high-yielding rice cultivars, contributing to food security in the face of climate change. 6 Research Methods for the Regulation of MS Gene Expression 6.1 Techniques for studying gene expression in response to environmental factors To understand the regulatory mechanisms of temperature, light, and water or humidity on the expression of MS genes in rice, various techniques are employed to study gene expression in response to these environmental factors. One such approach is the use of Environmental Gene Regulatory Influence Networks (EGRINs), which integrate multiple genome-scale measurements to uncover how gene expression is coordinated in response to environmental signals. EGRINs have been used to study the response of rice cultivars to high temperatures, water deficit, and agricultural field conditions by integrating time-series transcriptome data, patterns of nucleosome-free chromatin, and the occurrence of known cis-regulatory elements (Wilkins et al., 2016). 6.2 High-throughput sequencing and transcriptomics High-throughput sequencing and transcriptomics are pivotal in identifying and quantifying gene expression changes under different environmental conditions. For instance, transcriptome analysis of anther development stages in maize has identified numerous TF genes that are differentially expressed, providing insights into the regulatory pathways underlying MS (Jiang et al., 2021). Similarly, proteomic analysis using isobaric tags for relative and absolute quantification (iTRAQ) has been employed to study the proteomic changes in TGMS rice lines, revealing differentially expressed proteins involved in various biosynthetic and metabolic processes (Fang et al., 2022). 6.3 CRISPR-Cas9 and other gene editing tools CRISPR-Cas9 and other gene editing tools have revolutionized the study of gene function by enabling precise modifications in the genome. In rice, CRISPR/Cas9 has been used to create novel mutants of the TGMS gene tms5, resulting in lines with specific mutations that exhibit complete MS at high temperatures and restored fertility at lower temperatures (Fang et al., 2022). Additionally, CRISPR interference (CRISPRi) provides a method for sequence-specific control of gene expression, allowing for targeted silencing of transcription and facilitating the study of gene functions and regulatory systems (Larson et al., 2013).
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