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Apply Immuno-FISH in Zea Maize

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The process of meiosis, a fundamental mechanism in plant sexual reproduction, orchestrates the creation of germ cells with genetically diverse homologous chromosomes, primarily through the intricate event of crossing-over (CO). CO plays a pivotal role in promoting genetic variation by enabling the exchange of genetic material among chromosomes. Despite DNA Double Strand Breaks (DSBs) initiating chromosome recombination, not all DSB events culminate in COs. This study seeks to revolutionize the CO process in plants to expedite plant breeding and produce novel genotypes harboring traits unattainable through traditional methods. In pursuit of this goal, transgenic Zea mays lines have been developed, introducing a bespoke recombinant protein, Cas9:SPO11. This engineered protein is adept at guiding DSBs to precise chromosome regions during meiosis, imparting a degree of control over the recombination landscape. Our hypothesis posits that these targeted regions, under the influence of Cas9:SPO11, experience heightened recombination rates compared to their wild-type counterparts. The innovation lies in the potential to channel recombination events towards desired genomic loci, thereby creating new avenues for directed trait incorporation. The present study is poised to investigate the anticipated increase in CO events at the designated target loci in the transgenic lines when contrasted with the natural recombination pattern of wild-type maize. This investigation hinges on a multimodal approach, combining the power of Immuno-Fluorescent in situ Hybridization (Immuno-FISH) with protein immunolabeling. These techniques afford visualization of chromosome dynamics, elucidating the spatial and temporal aspects of the recombination process. The research aligns with a significant body of literature emphasizing the indispensability of CO in generating genetic diversity and facilitating adaptive evolution. Moreover, the role of DSBs as initiators of recombination events has been underscored by studies in various organisms, including model plant species. The exploration of CRISPR-Cas9 technology in the context of enhancing recombination dynamics is an emerging avenue, with diverse applications ranging from basic research to agricultural innovation. As the scope of plant breeding broadens to address contemporary challenges, such as climate resilience and nutritional enhancement, the ability to strategically modulate recombination events offers an enticing proposition. The outcomes of this study could revolutionize crop improvement strategies, allowing for the accelerated development of plant varieties with desirable traits. Ultimately, this research underscores the potential of leveraging genetic recombination mechanisms to usher in a new era of precision agriculture.

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2023

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dissertation or thesis

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