通过CRISPR/Cas9系统的核糖核蛋白复合物传递破坏花青素途径,改变马铃薯(Solanum tuberosum L.)块茎的颜色

IF 2.3 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Plant Cell, Tissue and Organ Culture Pub Date : 2024-04-16 DOI:10.1007/s11240-024-02743-3
Anders Wulff-Vester, Mariette Andersson, May Bente Brurberg, Per Hofvander, Muath Alsheikh, Wendy Harwood, Trine Hvoslef-Eide
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引用次数: 0

摘要

马铃薯是挪威传统饮食的重要组成部分,该作物面临着病虫害以及日益严峻的气候变化等诸多挑战。基因组编辑可为提高挪威马铃薯栽培品种应对新气候挑战的能力提供工具。作为在挪威栽培品种中使用CRISPR/Cas9的概念验证,我们将两个马铃薯栽培品种 "德西丽 "和 "南森 "的表皮颜色从红色改为黄色。我们的方法包括使用原生质体,并在温室的密闭室内连续培养了三代克隆块茎,以评估编辑后的植株在不同时间和不同温度条件下的稳定性。我们发现,原生质体法非常适合实现 CRISPR/Cas9 的应用。结果表明,黄皮在三代块茎繁殖过程中是一致的。我们在原生质体再生者中发现了一些疑似体细胞变异。我们在第二个生长周期的高温条件下(最高温度接近 40ºC)观察到的一些变异,在第三个培养周期的低温条件下消失了。
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Colour change in potato (Solanum tuberosum L.) tubers by disruption of the anthocyanin pathway via ribonucleoprotein complex delivery of the CRISPR/Cas9 system

Potato is an important part of the traditional Norwegian diet, and the crop faces several challenges with respect to pests and diseases, as well as the increasingly challenging changes in climate. Genome editing may provide tools to improve the resilience of Norwegian potato cultivars to new climate challenges. We have altered the skin colour of two potato cultivars, ‘Desirée’ and ‘Nansen’ from red to yellow, as a proof-of-concept for the use of CRISPR/Cas9 in a Norwegian cultivar. Our method has involved the use of protoplasts and we have grown the regenerants for three successive clonal tuber generations to evaluate the stability of the edited plants over time and under varying temperature conditions in contained rooms in a greenhouse. We found that the protoplast method is well suited to achieving CRISPR/Cas9 applications. The results show that the yellow skin is consistent over the three generations of tuber propagation. We found some suspected somaclonal variation in the protoplast regenerants. Some of the variation which we observed under high temperatures (up to nearly 40ºC) during the second growth cycle, disappeared when cultivated under lower temperatures in the third cultivation cycle.

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来源期刊
Plant Cell, Tissue and Organ Culture
Plant Cell, Tissue and Organ Culture 生物-生物工程与应用微生物
CiteScore
5.40
自引率
13.30%
发文量
203
审稿时长
3.3 months
期刊介绍: This journal highlights the myriad breakthrough technologies and discoveries in plant biology and biotechnology. Plant Cell, Tissue and Organ Culture (PCTOC: Journal of Plant Biotechnology) details high-throughput analysis of gene function and expression, gene silencing and overexpression analyses, RNAi, siRNA, and miRNA studies, and much more. It examines the transcriptional and/or translational events involved in gene regulation as well as those molecular controls involved in morphogenesis of plant cells and tissues. The journal also covers practical and applied plant biotechnology, including regeneration, organogenesis and somatic embryogenesis, gene transfer, gene flow, secondary metabolites, metabolic engineering, and impact of transgene(s) dissemination into managed and unmanaged plant systems.
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