CRISPR/Cas9-Mediated Mutagenesis of the Granule-Bound Starch Synthase Gene in the Potato Variety Yukon Gold to Obtain Amylose-Free Starch in Tubers
Autor: | Stephany Toinga-Villafuerte, Maria Isabel Vales, Joseph M. Awika, Keerti S. Rathore |
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Rok vydání: | 2022 |
Předmět: |
amylose
amylopectin CRISPR/Cas9 gene knockout GBSS sgRNAs rapid visco analyzer Solanum tuberosum targeted mutagenesis fungi Organic Chemistry Amylopectin food and beverages Starch General Medicine Catalysis Computer Science Applications Inorganic Chemistry Starch Synthase Mutagenesis Yukon Territory Amylose Gold Physical and Theoretical Chemistry CRISPR-Cas Systems Molecular Biology Spectroscopy |
Zdroj: | International Journal of Molecular Sciences; Volume 23; Issue 9; Pages: 4640 |
ISSN: | 1422-0067 |
Popis: | Potato (Solanum tuberosum L.) is the third most important food crop after rice and wheat. Its tubers are a rich source of dietary carbohydrates in the form of starch, which has many industrial applications. Starch is composed of two polysaccharides, amylose and amylopectin, and their ratios determine different properties and functionalities. Potato varieties with higher amylopectin have many food processing and industrial applications. Using Agrobacterium-mediated transformation, we delivered Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR/Cas9) reagents to potato (variety Yukon Gold) cells to disrupt the granule-bound starch synthase (gbssI) gene with the aim of eliminating the amylose component of starch. Lugol-Iodine staining of the tubers showed a reduction or complete elimination of amylose in some of the edited events. These results were further confirmed by the perchloric acid and enzymatic methods. One event (T2-7) showed mutations in all four gbss alleles and total elimination of amylose from the tubers. Viscosity profiles of the tuber starch from six different knockout events were determined using a Rapid Visco Analyzer (RVA), and the values reflected the amylopectin/amylose ratio. Follow-up studies will focus on eliminating the CRISPR components from the events and on evaluating the potential of clones with various amylose/amylopectin ratios for food processing and other industrial applications. |
Databáze: | OpenAIRE |
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