Plant Biology and Biotechnology Department, Sirius University of Science and Technology, Olympic Avenue 1, 354340 Sochi, Russia.
Department of Genetics and Biotechnology, Saint-Petersburg State University, Universitetskaya Emb 7/9, 199034 Saint-Petersburg, Russia.
Int J Mol Sci. 2024 Sep 17;25(18):10011. doi: 10.3390/ijms251810011.
Grape is one of the most economically significant berry crops. Owing to the biological characteristics of grapes, such as the long juvenile period (5-8 years), high degree of genome heterozygosity, and the frequent occurrence of inbreeding depression, homozygosity during crossbreeding leads to loss of varietal characteristics and viability. CRISPR/Cas editing has become the tool of choice for improving elite technical grape varieties. This study provides the first evidence of a decrease in the total fraction of phenolic compounds and an increase in the concentration of peroxide compounds in grape callus cells upon the addition of chitosan to the culture medium. These previously unreported metabolic features of the grape response to chitosan have been described and used for the first time to increase the probability of selecting plant cells with knockout characterised by an oxidative burst in response to the presence of a pathogen modulated by chitosan in the high-metabolite black grape variety 'Merlot'. This was achieved by using a CRISPR/Cas9 editing vector construction with the peroxide sensor HyPer as a reporter. This research represents the first CRISPR/Cas9 editing of 'Merlot', one of the most economically important elite technical grape varieties.
葡萄是最具经济意义的浆果作物之一。由于葡萄的生物学特性，如幼年期长（5-8 年）、基因组杂合度高以及自交衰退频繁，杂交过程中的纯合性导致品种特征和活力丧失。CRISPR/Cas 编辑已成为改良优质酿酒葡萄品种的首选工具。本研究首次证明，在培养基中添加壳聚糖可降低葡萄愈伤组织细胞中总酚类化合物的分数，并增加过氧化物化合物的浓度。这些以前未报道的葡萄对壳聚糖反应的代谢特征已被描述，并首次用于增加选择具有氧化爆发的植物细胞的可能性，该爆发是由壳聚糖存在调节的，对病原体的反应以过氧化物传感器 HyPer 为报告载体的 CRISPR/Cas9 编辑载体构建。这是对最具经济意义的优质酿酒葡萄品种之一“梅洛”的首次 CRISPR/Cas9 编辑。
