Molecular Plant Breeding 2024, Vol.15, No.1, 27-33 http://genbreedpublisher.com/index.php/mpb 29 By combining systems biology with multi-omics datasets, our understanding of the molecular regulatory networks for crop improvement can be enhanced. The concepts of “phenotype to genotype” and “genotype to phenotype” can facilitate crop breeding improvements, especially under environmental stress conditions (Jamil et al., 2020). Undoubtedly, with innovations in technology and methods, Breeding 5.0 will further enhance breeding efficiency, precision, and sustainability. Through genome editing and precise genome design, genetic improvements in plants will become more accurate and controllable. The application of artificial intelligence and big data will expedite the breeding process and optimize breeding strategies. The use of multi-omics integration and systems biology approaches will deepen our understanding of plant traits and genomes, providing more comprehensive support for achieving breeding objectives. 3 Emerging Fields and Applications Breeding 5.0 will actively explore emerging fields and applications to meet the demands of precise agriculture and personalized breeding. It will utilize technologies such as fluorescence-based hybridization and microscopic imaging to deepen our understanding of plant biology. Additionally, there will be a focus on enhancing environmental adaptability and developing breeding strategies to address climate change, thereby promoting sustainable agriculture and food security. The development of these emerging fields and applications will further drive innovation and progress in plant breeding. 3.1 Precise agriculture and personalized breeding Breeding 5.0 will integrate with precise agriculture to achieve more personalized breeding strategies. Precise agriculture utilizes technologies such as sensor technology, remote sensing, and geographic information systems, along with tools like cameras and mobile robots, to analyze the growth status of crops in real farmland settings. This provides crucial information for precise management and breeding experiments (Weyler et al., 2021). By combining the technologies and methods of Breeding 5.0, we can accurately select and cultivate crop varieties tailored to specific environmental conditions and agricultural needs. This personalized breeding approach will make agricultural production more sustainable, improve yields and quality, and reduce resource waste and environmental impact. 3.2 Advancements in fluorescence-based hybridization and microscopic imaging technologies Fluorescence in situ hybridization (FISH) technology and microscopic imaging technologies have made significant progress in their application in plant breeding. These technologies play a vital role not only in gene localization and expression analysis but also in understanding the molecular mechanisms of plant growth and development. In the era of Breeding 5.0, the application of fluorescence-based hybridization and microscopic imaging technologies in breeding will see further development. Fluorescence-based hybridization technology helps researchers visually observe and analyze gene expression and protein interactions in plants, providing in-depth insights into plant biological processes. For instance, by using FISH technology combined with immunofluorescence technique, specific proteins and DNA sequences can be located on the maize synaptonemal complex. This technology is widely applied in the study of plant meiotic chromosome, crucial for understanding the gene recombination process (Stack et al., 2020). FISH and microscopic imaging technologies are employed to precisely measure gene expression at the single-cell level, significantly contributing to the understanding of the impact of spatial or temporal-dependent processes on plant growth and development (Lucic et al., 2021). With advancements in microscopic imaging technology, we can observe the structure and functionality of plant cells and tissues in high resolution and real-time. For example, FISH technology combined with high-throughput and super-resolution microscopy has been used to map and spatially define the contact frequency between different genomic regions. These methods greatly contribute to the understanding of the packaging of the human genome in the cell nucleus (Mao et al., 2020). Newly developed microscopic imaging methods, such as Expansion Microscopy (ExM), make it possible to achieve nanoscale imaging of ribonucleic acid (RNA) on traditional
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