Detecting and utilizing genetic variations in okra and grapes to improve food crops

Abstract

Okra (Abelmoschus esculentus (L.) Moench) is one of the most heat and drought tolerant vegetable crops and is cultivated throughout the world for its fibrous pods containing seeds. Okra seeds contain oil, and certain small artisanal producers press okra seeds to make oil that is commercially sold. However, okra seeds do not have a high enough oil content to be a sustainable alternative to other popular seed oils such as palm, canola, and sunflower oil. The goal of this study was to increase the oil content of okra seeds through traditional breeding (self-pollination and open pollination) and selection methods and determine which natural breeding of okra along with which genetic variants allows for the highest seed oil content. To do this, we conducted time-domain nuclear magnetic resonance (TD-NMR) analysis on the seeds to determine which ones have a high seed oil content, extracted DNA from the samples with the highest and lowest oil content, and assessed the genetic diversity in the selected okra samples using SSR markers. We also analyzed previously sequenced grapevine genome data for lines genetically engineered using CRISPR/Cas9 by a collaborator by aligning a reference genome with 14 mutant lines and one wild type (WT) line. This analysis allowed us to understand the position and association of genetic variants across the genome, especially in the phytoene desaturase 1 (pds1) gene, in grapevine leaf phenotypes. Through these analyses on okra and grape, we were able to determine the okra F2 line OO006 had the highest seed oil content progeny and that there is a remarkable genetic difference between okra samples with high and low seed oil content. While we were unable to conclude the variants associated with seed oil content, we were able to learn the methodology required to do so through the grapevine samples.