Abstract Detail

Applications of CRISPR technology across the plant tree of life

Tavernier, Emilie-Katherine [1], Pisias, Michael [2], McDaniel, Stuart [3], Yang, Bing [4], Pires, Chris [5].

Bryophytes to Brassica: Diverse Applications of CRISPR.

Recent advances in targeted genome editing technologies, like CRISPR, have enabled a wide spectrum of applications from evaluating genetic pathways to agricultural improvement. Here, we present new applications of CRISPR to understand sex chromosome evolution in bryophytes, and de novo domestication in the genus Brassica from wild crop relatives. However, each of these models requires unique methods of transformation. The haploid vegetative state of moss is accessible by enzymolysis and direct plasmid uptake, whereas Brassica species are reliant on regeneration of tissues from explant culture and Agrobacterium mediated transformation.
In evolutionary biology, the mechanisms by which new genes are incorporated into regulatory networks have yet to be explained. In species with genetically determined sexes, sex chromosomes may facilitate the evolution of dimorphism. Sex-limited inheritance and chromosomal suppressed recombination allow genes to functionally specialize within males or females. Typically, a newly inherited gene will acquire a sex-specific function for itself. However, in the case of “genetic hijacking”, a chromosomally sex-linked gene will override the developmental program and alter the downstream signaling cascade. Here, we describe experiments using CRISPR to genetically manipulate the dioecious moss, Ceratodon purpureus, and the hermaphroditic moss, Physcomitrella patens to test the influence of the “genetic hijacking model” on the evolution of sex-linked genes.
In our Brassica study, we want to overcome a limitation of CRISPR shared among eudicots, which is that tissue culture methods are often genotype-specific. Extensive work has been done in Brassica to explore tissue culture and transformation techniques in specific model accessions (e.g., B. napus cv. Westar, or B. oleracea doubled haploid line). However, there is no general protocol for Brassica transformation that works within even one species for both domesticated forms and wild relatives. We are optimizing transformation in model and non-model species of Brassica to enable the application of de novo domestication. Here, we present a targeted genome editing approach that uses a major phenotype inducing MADs-box genes in a wild crop relative. Presently, gene editing technologies across model and non-model systems require highly specialized methodologies.
Here we address inherent limitations within genomic editing, transformation, and regeneration efficiency, and their respective solutions. Our shared insights from our work in bryophytes and Brassica demonstrate that CRISPR can be developed across a range of plant lineages. Future applications of our work in Ceratodon and Brassica will expand ease of access to CRISPR across a range of plant lineages.

1 - University of Florida, PO Box 11852, Gainsville, Florida, 32611, United States
2 - University of Missouri, Division of Biological Sciences, 1201 Rollins St, 311 Bond Life Sciences Center, Columbia, MO, 65211, United States
3 - Biology Department, University Of Florida, Gainesville, FL, 32611, United States
4 - University of Missouri, Plant Sciences, 1201 Rollins St , Columbia, MO, 65211, USA
5 - University Of Missouri, 371 Bond Life Sciences Center, 1201 Rollins Street, Columbia, MO, 65211, United States

Keywords:
CRISPR
Gene editing
de novo domestication
Bryophytes
Brassicaceae
sex chromosomes.

Presentation Type: Symposium Presentation
Session: SY6, Applications of CRISPR technology across the plant tree of life
Location: /
Date: Friday, July 23rd, 2021
Time: 11:15 AM(EDT)
Number: SY6004
Abstract ID:805
Candidate for Awards:A. J. Sharp Award,Margaret Menzel Award,Economic Botany Section best student paper