玉米低温响应基因ZmNTL1和ZmNTL5的鉴定及功能分析

doi:10.3724/SP.J.1006.2025.53011

摘要/Abstract

摘要：

低温是限制东北地区玉米产量的主要逆境之一。膜NAC转录因子在植物抵御逆境过程中发挥着重要作用，探究其功能将为玉米耐低温育种提供一定的理论依据。本研究通过生物信息技术分析了9条玉米NTL基因的分子特性，利用已发表的玉米低温RNA-seq数据，发现6个成员在根和叶中同时受到低温诱导显著上调表达。选取蛋白同源性达到80.04%的ZmNTL1与ZmNTL5基因，采用基因敲除技术构建了该2条基因的双敲材料zmntl1zmntl5。在低温胁迫条件下，zmntl1zmntl5株系的叶片离子渗透率显著高于B104株系；而其地上部干重和鲜重、地下部干重和鲜重以及叶绿素含量均显著低于B104株系；zmntl1zmntl5株系的O₂^·?和H₂O₂含量显著高于B104对照株系，而GST、SOD、POD和CAT活性均显著低于B104株系。此外，低温胁迫下ZmGST24、ZmNCED3、ZmDREB1.6和ZmDREB2A基因在zmntl1zmntl5株系中的表达量较B104株系显著降低。综上所述，ZmNTL1与ZmNTL5的缺失降低了植物对低温的耐受性，表明二者是玉米低温耐受性的正调控因子。本研究为耐冷玉米品种改良和新品种培育提供了重要的理论基础和宝贵的遗传资源。

关键词: 玉米, 低温, RNA-seq, NTL, 基因编辑

Abstract:

Low temperature is one of the major abiotic stresses limiting maize yield in Northeast China. Membrane-bound NAC transcription factors play important roles in plant stress responses. Investigating their function provides a theoretical basis for breeding maize with enhanced cold tolerance. In this study, the molecular characteristics of nine maize NTL genes were analyzed using bioinformatics approaches. Analysis of publicly available RNA-seq data revealed that six of these genes were significantly upregulated by low temperature stress in both roots and leaves. Among them, ZmNTL1 and ZmNTL5, which share 80.04% sequence homology, were selected for further study. A double knockout mutant (zmntl1zmntl5) was generated using gene-editing techniques. Under low temperature stress, the ion leakage rate in the leaves of zmntl1zmntl5 was significantly higher than in the wild-type B104. The fresh and dry weights of both aboveground and belowground tissues, as well as chlorophyll content, were significantly lower in zmntl1zmntl5 compared to B104. In contrast, zmntl1zmntl5 exhibited significantly higher levels of O₂^? and H₂O₂, while the activities of antioxidant enzymes GST, SOD, POD, and CAT were significantly reduced. Furthermore, the expression levels of ZmGST24, ZmNCED3, ZmDREB1.6, and ZmDREB2A were markedly decreased in zmntl1zmntl5 compared to B104 under cold stress. In summary, the absence of ZmNTL1 and ZmNTL5 compromises cold tolerance in maize, suggesting that both genes function as positive regulators in the cold stress response. This study provides a valuable theoretical foundation and genetic resources for the improvement of cold-tolerant maize varieties and the development of new cultivars.

Key words: maize, low temperature, RNA-seq, NTL, gene editing

高源, 王宇琦, 姜佳宁, 赵健雄, 王雪贺缘, 王浩宇, 张芮嘉, 徐晶宇, 贺琳. 玉米低温响应基因ZmNTL1和ZmNTL5的鉴定及功能分析[J]. 作物学报, 0, (): 0-.

GAO Yuan, WANG Yu-Qi, JIANG Jia-Ning, ZHAO Jian-Xiong, WANG Xue-He-Yuan, WANG Hao-Yu, ZHANG Rui-Jia, XU Jing-Yu, HE Lin. Identification and functional analysis of low temperature responsive genes ZmNTL1 and ZmNTL5 in maize[J]. Acta Agronomica Sinica, 0, (): 0-.

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