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Multiplex CRISPR-Cas9 Editing of Starch Branching Enzyme II and Vacuolar Invertase Simultaneously Enhances Resistant Starch Content and Cold-Induced Sweetening Resistance in Solanum chacoense.

Created on 13 Jul 2026

Authors

Xiaoqin Li, Cuihua Xin, Jiangbo Guo, Yu Zheng, Fuxin Huang, Xia Li, Haixia Guo, Yang Xu, Wei Shao

Published in

Plant science : an international journal of experimental plant biology. Pages 113319. Jul 12, 2026. Epub Jul 12, 2026.

Abstract

Potato processing suffers from a high glycemic index due to amylopectin-rich starch and from undesirable color and acrylamide formation during frying, mainly caused by cold-induced sweetening (CIS). To address both issues simultaneously, we used CRISPR-Cas9 to knock out two key genes in diploid Solanum chacoense: ScSBE II (starch branching enzyme II), which controls amylopectin biosynthesis, and ScVInv (vacuolar invertase), a central regulator of CIS. Knockout of ScSBEⅡ increased tuber fresh weight-based absolute amylose content by ~5-fold versus wild type. Amylose proportion in total starch elevated from 22% to 54%, while amylopectin abundance declined 1.5-fold, substantially optimizing the amylose/amylopectin mass ratio of tuber starch. These lines also showed a 4-fold reduction in rapidly digestible starch (RDS), 3.5‑fold and 1.2‑fold increases in slowly digestible starch (SDS) and resistant starch (RS), respectively, and markedly improved pasting properties. Enzyme assays confirmed a 2.5‑fold reduction in ScSBE II activity. In wild‑type (WT) tubers, cold storage (4 °C, 7 d) increased ScVInv activity ~5‑fold (to 48μg·min⁻¹·g⁻¹) and reducing sugars 6‑fold (from 11 to 68mg·g⁻¹). Notably, ScVInv single‑knockout and ScSBE II/ScVInv double‑knockout lines produced chips with lighter color and much lower acrylamide than WT or ScSBE II single‑knockout lines. This dual‑gene editing strategy creates novel potato germplasm with enhanced resistant starch (health benefit) and superior processing quality (safer, visually appealing fried products).

PMID:
42437612
Bibliographic data and abstract were imported from PubMed on 13 Jul 2026.

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