Targeted modification of CmACO1 by CRISPR/Cas9 extends the shelf-life of Cucumis melo var. reticulatus melon.

IF 4.9 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Satoko Nonaka, Maki Ito, Hiroshi Ezura
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引用次数: 1

Abstract

The gaseous plant hormone ethylene is a regulator of fruit shelf-life, one of the essential traits in fruits. Extending fruit shelf-life reduces food loss, thereby expected to contribute to food security. The enzyme 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) is the final step of the ethylene production pathway. Its suppression via antisense technology has been demonstrated to extend the shelf-life of melon, apple, and papaya. Genome editing technology is an innovative technique for plant breeding. Because the genome editing technology would not leave the exogenous genes in the final crop products, the crops via genome editing can be considered non-genetically modified yields; compared to conventional breeding, such as mutation breeding, the breeding term would be expected to be relatively short. These points include the advantage of this technique in utilization for commercial applications. We attempted to extend the shelf-life of the Japanese luxury melon (Cucumis melo var. reticulatus, 'Harukei-3') via modification of the ethylene synthesis pathway with the genome editing technology, CRISPR/Cas9 system. The Melonet-DB (https://melonet-db.dna.affrc.go.jp/ap/top) showed that the melon genome had the five CmACOs and the gene CmACO1 predominantly expressed in harvested fruits. From this information, CmACO1 was expected to be a key gene for shelf-life in melons. Based on this information, the CmACO1 was selected as the target of the CRISPR/Cas9 system and introduced the mutation. The final product of this melon did not have any exogenous genes. The mutation was inherited for at least two generations. In the T2 generation, the fruit phenotypes 14 days after harvest were as follows: ethylene production was reduced to one-tenth that of the wild type, pericarp colour remained green, and higher fruit firmness. Early fermentation of the fresh fruit was observed in the wild-type fruit but not in the mutant. These results show that CmACO1 knockout via CRISPR/Cas9 extended the melon's shelf-life. Moreover, our results suggest that genome editing technology would reduce food loss and contribute to food security.

Abstract Image

Abstract Image

Abstract Image

利用CRISPR/Cas9靶向修饰CmACO1,延长了甜瓜的贮藏期。
气态植物激素乙烯是水果货架期的调节剂,是水果的基本性状之一。延长水果保质期可减少食物损失,从而有望促进粮食安全。酶1-氨基环丙烷-1-羧酸氧化酶(ACO)是乙烯生产途径的最后一步。通过反义技术抑制其可延长甜瓜、苹果和木瓜的保质期。基因组编辑技术是一种创新的植物育种技术。由于基因组编辑技术不会在最终的作物产品中留下外源基因,因此通过基因组编辑的作物可以被认为是非转基因产量;与传统育种(如突变育种)相比,育种周期将相对较短。这些要点包括该技术在商业应用方面的优势。我们试图通过基因组编辑技术CRISPR/Cas9系统修饰乙烯合成途径来延长日本奢侈甜瓜(Cucumis melo var. reticulatus, 'Harukei-3')的保质期。Melonet-DB (https://melonet-db.dna.affrc.go.jp/ap/top)显示,甜瓜基因组包含5个CmACOs, CmACO1基因主要在收获的果实中表达。根据这些信息,CmACO1有望成为决定甜瓜货架期的关键基因。基于这些信息,我们选择CmACO1作为CRISPR/Cas9系统的靶标,并引入突变。这种甜瓜的最终产物没有任何外源基因。这种突变至少遗传了两代人。T2代收获后14天的果实表型为:乙烯产量降至野生型的十分之一,果皮颜色保持绿色,果实硬度更高。在野生型果实中观察到新鲜果实的早期发酵,而在突变体果实中则没有。这些结果表明,通过CRISPR/Cas9敲除CmACO1可以延长甜瓜的保质期。此外,我们的研究结果表明,基因组编辑技术将减少粮食损失,促进粮食安全。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
7.00
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审稿时长
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