Theses and Dissertations
Issuing Body
Mississippi State University
Advisor
Reichert, Nancy A.
Committee Member
Klink, Vincent
Committee Member
Peng, Zhaohua
Committee Member
Young, Margaret M.
Date of Degree
5-1-2020
Original embargo terms
Worldwide
Document Type
Graduate Thesis - Open Access
Major
Biological Sciences
Degree Name
Master of Science
College
College of Arts and Sciences
Department
Department of Biological Sciences
Abstract
The potential for Hibiscus cannabinus L. (kenaf) improvement via genome editing using the CRISPR/Cas9 system to generate gene knock-outs was explored. Studies included target gene identification, target guide RNA (gRNA) selection, plant tissue (explant) choice and media composition for plant regeneration. A putative kenaf phytoene desaturase gene (pds, GEED01047592.1) was identified in the kenaf transcriptome, and molecularly confirmed. Kenaf seedling tissues were transformed via Agrobacterium tumefaciens containing the cas9 gene (endonuclease required for gene knock-out) and each gRNA separately; putative transgenic calli and adventitious shoots arose on a medium containing 1-naphthaleneacetic acid, thidiazuron and silver nitrate. Tissues appeared chlorotic/albino and shoots remained diminutive/dwarf-like. These unique morphologies had also been noted by researchers who successfully knocked out the pds gene in other plant species. Cas9 DNA was detected in these putative transgenic kenaf tissues, but initial DNA sequencing analysis did not confirm knock-out/mutations in targeted areas of the pds gene.
URI
https://hdl.handle.net/11668/16695
Recommended Citation
Li, Kangqi, "Genome editing in kenaf: Initial studies and target gene characterization" (2020). Theses and Dissertations. 2404.
https://scholarsjunction.msstate.edu/td/2404