ClaPEPCK4: target gene for breeding innovative watermelon germplasm with low malic acid and high sweetness.

IF 4.5 2区农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Gm Crops & Food-Biotechnology in Agriculture and the Food Chain Pub Date : 2025-12-01 Epub Date: 2025-01-14 DOI:10.1080/21645698.2025.2452702

Congji Yang, Jiale Shi, Yuanyuan Qin, ShengQi Hua, Jiancheng Bao, Xueyan Liu, Yuqi Peng, Yige Gu, Wei Dong

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Abstract

Malic acid markedly affects watermelon flavor. Reducing the malic acid content can significantly increase the sweetness of watermelon. An effective solution strategy is to reduce watermelon malic acid content through molecular breeding technology. In this study, we measured the TSS and pH of six watermelon varieties at four growth nodes. The TSS content was very low at 10 DAP and accumulated rapidly at 18, 26, and 34 DAP. Three phosphoenolpyruvate carboxykinase (PEPCK) genes of watermelon were identified and analyzed. The ClaPEPCK4 expression was inversely proportional to malate content variations in fruits. In transgenic watermelon plants, overexpressing the ClaPEPCK4 gene, malic acid content markedly decreased. In the knockout transgenic watermelon plants, two SNP mutations and one base deletion occurred in the ClaPEPCK4 gene, with the malic acid content in the leaves increasing considerably and the PEPCK enzyme activity reduced to half of the wild-type. It is interesting that the ClaPEPCK4 gene triggered the closure of leaf stomata under dark conditions in the knockout transgenic plants, which indicated its involvement in stomatal movement. In conclusion, this study provides a gene target ClaPEPCK4 for creating innovative new high-sweetness watermelon varieties.

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ClaPEPCK4：低苹果酸高甜度西瓜创新种质的靶基因。

苹果酸对西瓜风味有显著影响。降低苹果酸含量可以显著提高西瓜的甜度。通过分子育种技术降低西瓜苹果酸含量是有效的解决策略。本研究测定了6个西瓜品种在4个生育期的TSS和pH值。TSS含量在10 DAP时很低，在18、26和34 DAP时迅速积累。对西瓜磷酸烯醇丙酮酸羧激酶（PEPCK） 3个基因进行了鉴定和分析。ClaPEPCK4的表达量与果实中苹果酸含量的变化成反比。在转基因西瓜植株中，过表达ClaPEPCK4基因，苹果酸含量显著降低。在敲除转基因西瓜植株中，ClaPEPCK4基因发生了2个SNP突变和1个碱基缺失，叶片中苹果酸含量显著增加，PEPCK酶活性降至野生型的一半。有趣的是，ClaPEPCK4基因在敲除转基因植株中触发了黑暗条件下叶片气孔的关闭，表明其参与了气孔运动。综上所述，本研究为高甜度西瓜品种创新提供了ClaPEPCK4基因靶点。

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来源期刊

Gm Crops & Food-Biotechnology in Agriculture and the Food Chain Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

8.10

自引率

10.30%

发文量

期刊介绍： GM Crops & Food - Biotechnology in Agriculture and the Food Chain aims to publish high quality research papers, reviews, and commentaries on a wide range of topics involving genetically modified (GM) crops in agriculture and genetically modified food. The journal provides a platform for research papers addressing fundamental questions in the development, testing, and application of transgenic crops. The journal further covers topics relating to socio-economic issues, commercialization, trade and societal issues. GM Crops & Food aims to provide an international forum on all issues related to GM crops, especially toward meaningful communication between scientists and policy-makers. GM Crops & Food will publish relevant and high-impact original research with a special focus on novelty-driven studies with the potential for application. The journal also publishes authoritative review articles on current research and policy initiatives, and commentary on broad perspectives regarding genetically modified crops. The journal serves a wide readership including scientists, breeders, and policy-makers, as well as a wider community of readers (educators, policy makers, scholars, science writers and students) interested in agriculture, medicine, biotechnology, investment, and technology transfer. Topics covered include, but are not limited to: • Production and analysis of transgenic crops • Gene insertion studies • Gene silencing • Factors affecting gene expression • Post-translational analysis • Molecular farming • Field trial analysis • Commercialization of modified crops • Safety and regulatory affairs BIOLOGICAL SCIENCE AND TECHNOLOGY • Biofuels • Data from field trials • Development of transformation technology • Elimination of pollutants (Bioremediation) • Gene silencing mechanisms • Genome Editing • Herbicide resistance • Molecular farming • Pest resistance • Plant reproduction (e.g., male sterility, hybrid breeding, apomixis) • Plants with altered composition • Tolerance to abiotic stress • Transgenesis in agriculture • Biofortification and nutrients improvement • Genomic, proteomic and bioinformatics methods used for developing GM cops ECONOMIC, POLITICAL AND SOCIAL ISSUES • Commercialization • Consumer attitudes • International bodies • National and local government policies • Public perception, intellectual property, education, (bio)ethical issues • Regulation, environmental impact and containment • Socio-economic impact • Food safety and security • Risk assessments