玉米二酰甘油激酶基因耐低温分析

doi:10.16035/j.issn.1001-7283.2026.04.004

摘要/Abstract

摘要： 二酰甘油激酶（DGK）可催化二酰甘油生成磷脂酸，参与植物对多种非生物胁迫的响应。采用CRISPR/ Cas9技术敲除玉米（Zea mays L.）ZmDGK1基因，获得zmdgk1^CR T₃代纯合株系（遗传转化受体为自交系B104），将3个zmdgk1^CR株系和B104进行低温胁迫处理，测定表型和生理指标。结果表明，3个zmdgk1^CR株系幼苗表现出较轻的叶片失绿、黄化和萎蔫现象；根系生长受到轻微抑制，根系总长较B104增加11.30%~16.17%。在活性氧、细胞膜损伤、抗氧化酶活性、渗透调节物质和光合色素相关指标中，zmdgk1^CR株系与B104均存在显著差异。与B104相比，zmdgk1^CR株系的O₂^-.含量降低24.61%~30.71%，丙二醛含量降低5.19%~18.18%，超氧化物歧化酶活性提高3.31%~19.88%，总叶绿素含量增加8.78%~27.23%。此外，通过对低温处理后zmdgk1^CR株系的叶片和根系进行DAB、NBT、H₂DCF-DA和PI染色，证实ZmDGK1基因缺失可减少活性氧积累、降低细胞膜损伤，同时提高玉米的抗氧化能力、渗透调节能力及光合色素积累，表明ZmDGK1基因对玉米幼苗耐低温性具有负调控作用。

关键词: 玉米, 二酰甘油激酶, 基因编辑（CRISPR/Cas9）, 耐低温性, 活性氧

Abstract:

Diacylglycerol kinase (DGK) catalyzes the conversion of diacylglycerol (DAG) to phosphatidic acid (PA), participating in plant responses to various abiotic stresses. This study employed CRISPR/Cas9 technology to knock out the ZmDGK1 gene in maize (Zea mays L.), generating zmdgk1^CR T₃ homozygous lines (with inbred line B104 as the genetic transformation recipient). Three zmdgk1^CR lines and B104 were subjected to low-temperature stress, and their phenotypic and physiological indicators were measured. Results showed that the zmdgk1^CR seedlings exhibited less severe leaf chlorosis, yellowing, and wilting. Root growth was only slightly inhibited, with total root length increased by 11.30%-16.17% compared to that of B104. Analysis of low- temperature tolerance-related physiological indicators revealed significant differences between the zmdgk1^CR lines and B104 in parameters related to reactive oxygen species, cell membrane damage, antioxidant enzyme activity, osmotic regulators, and photosynthetic pigments. Specifically, the zmdgk1^CR lines exhibited a 24.61%- 30.71% reduction in O₂^-. content and a 5.19%-18.18% reduction in malondialdehyde content, while showing a 3.31%-19.88% increase in superoxide dismutase activity and an 8.78%-27.23% increase in total chlorophyll content. Furthermore, DAB, NBT, H₂DCF-DA, and PI staining of leaves and roots from zmdgk1^CR lines after low-temperature treatment confirmed that ZmDGK1 gene deletion reduced reactive oxygen species accumulation and mitigated cell membrane damage, while enhancing antioxidant capacity of maize, osmotic regulation, and photosynthetic pigment accumulation. These findings collectively indicate that the ZmDGK1 gene negatively regulates low-temperature tolerance in maize seedlings.

Key words: Maize, Diacylglycerol kinase, Gene editing (CRISPR/Cas9), Low-temperature tolerance, Reactive oxygen species

王鑫琦, 车欣洋, 张海洋, 王旭, 李玉涵, 赵硕, 刘思贝, 王雪贺缘, 贺琳, 徐晶宇. 玉米二酰甘油激酶基因耐低温分析[J]. 作物杂志, 2026 (4): 27–35

Wang Xinqi, Che Xinyang, Zhang Haiyang, Wang Xu, Li Yuhan, Zhao Shuo, Liu Sibei, Wang Xueheyuan, He Lin, Xu Jingyu. Analysis of Low-Temperature Tolerance of Diacylglycerol Kinase Gene in Maize[J]. Crops, 2026(4): 27–35

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