Abstract Detail

Crops and Wild Relatives

Gafford, Danielle [1], Pisias, Michael [2], Yang, Bing [3], Pires, Chris [4].

From Cotyledon to Clone; Tissue Culture Optimization for Brassica Transformation.

Due to the effects of climate change decreasing crop efficiency, the need for crop improvement has become more pressing to accommodate increasing demands. Various methods like breeding and gene editing using CRISPR, have taken the forefront for crop improvement. To effectively use CRISPR, a compatible tissue culture method is usually a necessary step for determining efficiency and success in transformation. A variety of known successful tissue culture methods include cotyledon, leaf disk, hypocotyl, protoplast, and microspore culture. While there are pros and cons to each method, many factors influence tissue culture efficiency such as hormone concentration, vitamins, and age of the explant. The genus Brassica has a wide diversity of important crop species and can be genetically engineered through CRISPR to tolerate pressures of climate change. However, while tissue culture methods for specific domesticates of Brassica exist, there is no general protocol that works for the broader genus due to difficulties in addressing the specific needs of each species. My work in testing various BAP concentrations (6-Benzylaminopurine), a shooting hormone, and NAA (1-Naphthaleneacetic acid), a rooting hormone, and their success in producing regenerated plants from across both domestic and wild species provides insight for developing a successful general tissue culture methodology. To establish this method across the genus, cotyledon cuttings of domesticate and wild Brassica are tested with various BAP and NAA concentrations. I have taken B.oleracea, B.barreleri, and B.rapa cotyledon cuttings and have compared regeneration rates amongst various BAP and NAA concentrations. While each species responds differently to varying hormone concentrations and conditions, finding a viable baseline that works across the broader spectrum of Brassica has the potential to streamline and optimize transformation efficiency, resources, and time needed to reach end results for a variety of experiments.

1 - 111 S Rosehill Rd , Lone Jack, MO, 64070, United States
2 - University of Missouri, Division of Biological Sciences, 1201 Rollins St, 311 Bond Life Sciences Center, Columbia, MO, 65211, United States
3 - University of Missouri, College of Agriculture Food and Natural Resource , 340E Bond Life Sciences Center, Columbia, MO, 65201, USA
4 - University Of Missouri, 371 Bond Life Sciences Center, 1201 Rollins Street, Columbia, MO, 65211, United States

Keywords:
Tissue Culture
Brassica oleracea
Brassicaceae
CRISPR
quantitative genetics.

Presentation Type: Poster
Session: P1, Crops and Wild Relatives Posters
Location: Virtual/Virtual
Date: Monday, July 19th, 2021
Time: 5:00 PM(EDT)
Number: P1CW008
Abstract ID:846
Candidate for Awards:None