Date of Award

6-2-2025

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Agricultural and Environmental Sciences

First Advisor

Ahmad Aziz

Abstract

This study delves into the use of the androgenesis pathway of microspores for genetic transformation and double haploid production in Sorghum bicolor. Focusing on the environmental and genetic factors that affect androgenesis, in vitro culture techniques were aimed to be refined for utilizing microspores towards callus induction more effectively. These efforts yielded promising approaches for agronomic traits enhancements. This research enhances the methodology for plant regeneration from microspores, facilitating the generation of homozygous lines with potentially improved traits. Although the current findings do not mark a radical departure from existing knowledge, they lay essential groundwork for future genetic studies in sorghum and possibly other cereal grains encountering similar genetic modification challenges. Microspores were successfully isolated and induced into embryogenic calli, confirming their viability for double haploid production. Cryopreservation protocols enabled long-term storage with post-thaw viability exceeding 50%, ensuring year-round research. Protoplast-based transformation using polyethylene glycol facilitated EGFP gene uptake and expression, demonstrating Agrobacterium-free genetic modification in sweet sorghum. Optimized isolation techniques reduced contamination, minimizing phenolic exudation and microbial interference. By documenting our approach, this work contributes to the broader dialogue on sustainable agriculture and food security, underscoring the value of incremental scientific progress in plant biotechnology. Looking forward, this study highlights the need for continued research to unlock the full potential of sorghum and other crops. The insights gained here aim to inspire further work, advancing our understanding of sorghum biology and serving as a steppingstone toward more resilient, productive crops through biotechnological innovation.

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