Understanding Tomato Regeneration and Maize Transformation Through Bioengineering

Tomato and maize are major crops and genetic models for studying gene function. Different genotypes exhibit varying responses to transformation. In plant bioengineering, some genotypes are not amenable to transformation and regeneration, while others are. Improving the bioengineering of tomato and maize is crucial. In this study, we aim to enhance transformation efficiency in genotypes that show a low response to transformation. We intend to generate overexpression constructs for genes A, B, and C, identified from previous GWAS studies, linked to plant regeneration in tomatoes. In another study, we tested different Ancymidol concentrations to increase maize leaf transformation in genotypes LH287, LH245, and 90DJD28, which showed a low response at lower Ancymidol concentrations. Genes A and C were amplified using cDNA from cotyledons and roots. Gene A was cloned into a construct with the 35S promoter and Nos terminator using Nimble Cloning. Cloning was confirmed by colony PCR, and construct sequencing will confirm cloning. In maize transformation, higher Ancymidol concentrations were tested on six genotypes at 0, 3.4 mg/L (1X), and 6.8 mg/L (2X). GFP expression was observed after two days. Higher GFP expression in 2X Ancymidol may improve transformation, especially in genotypes LH287, 90DJD28, and LH245, showing that 2X Ancymidol enhances transformation frequency in maize leaf tissues. In the future, we aim to generate overexpression constructs and transform low-regenerative genotypes of tomatoes. Additionally, we will further explore maize leaf transformation to determine how increased Ancymidol concentrations can improve the stable transformation of maize plants.

During this REU experience, I have learned the importance of interdisciplinary collaboration. Plant science is often intersecting and complexly interrelated to other STEM fields such as bioinformatics, robotics, physics, and many others. This integrated work taught me the importance of science communication at every education and interest level. Being able to simplify and condense complex research is a crucial skill that I will carry forward into my future science career. The research that I was able to conduct at BTI set me up to have a foundation in bioengineering principles and a deeper understanding of fundamental laboratory skills such as PCR, RNA isolation, and data analysis. BTI’s holistic approach to this internship allowed me to gain a comprehensive understanding of computational strategies, interdisciplinary collaboration, and real-world applications.