哈密瓜脱氢抗坏血酸还原酶基因的鉴定及冷胁迫表达分析

doi:10.7506/spkx1002-6630-20201109-084

食品科学 ›› 2022, Vol. 43 ›› Issue (2): 117-125.doi: 10.7506/spkx1002-6630-20201109-084

哈密瓜脱氢抗坏血酸还原酶基因的鉴定及冷胁迫表达分析

宋文，张琴，宁明，周发科，杨新泉，单春会，唐凤仙

（石河子大学食品学院，新疆特色果蔬贮藏加工教育部工程研究中心，新疆石河子 832000）

出版日期:2022-01-25 发布日期:2022-01-29
基金资助:
国家自然科学基金地区科学基金项目（31360412）；新疆生产建设兵团重点领域创新团队建设计划项目（2017CB012）

Identification and Cold Stress-responsive Expression Analysis of Dehydroascorbate Reductase Gene in Hami Melon (Cucumis melo var. saccharinus)

SONG Wen, ZHANG Qin, NING Ming, ZHOU Fake, YANG Xinquan, SHAN Chunhui, TANG Fengxian

(Engineering Research Center for Storage and Processing of Xinjiang Characteristic Fruits and Vegetables, Ministry of Education, College of Food, Shihezi University, Shihezi 832000, China)

Online:2022-01-25 Published:2022-01-29

摘要/Abstract

摘要： 为阐明哈密瓜脱氢抗坏血酸还原酶（dehydroascorbate reductase，DHAR）基因的基本性质及在响应冷胁迫过程中的表达水平，基于生物信息学对2 个哈密瓜DHAR基因（CmDHAR）的基本信息、多序列比对、系统进化关系、共线性关系、基因结构域特征、蛋白质三级结构、蛋白质-蛋白质相互作用及冷胁迫下的抗坏血酸（ascorbic acid，AsA）含量、DHAR活力、DHAR基因的表达水平及其基因集富集分析（gene set enrichment analysis，GSEA）进行研究。结果表明，2 个CmDHAR编码蛋白氨基酸的长度分别为297 aa与206 aa，等电点分别为8.67与5.99，分子质量为33 082.9 Da与22 891.6 Da。多序列比对结果表明CmDHAR基因具有保守性较高的谷胱甘肽-S-转移酶（glutathione S-transferase，GST）-N端功能结构域。基因结构分析结果表明CmDHAR包含6 个外显子、6 个保守基序、3 类顺式作用元件。共线性分析表明哈密瓜与西葫芦、笋瓜和南瓜在物种进化过程中更为接近。蛋白质三级结构预测CmDHAR2主要包含α-螺旋、无规卷曲、延伸链和β-转角等。蛋白互作分析表明CmDHAR家族蛋白与哈密瓜其他GST家族蛋白之间联系紧密。冷胁迫下耐冷型哈密瓜“伽师瓜-310”中的AsA含量、DHAR活力及CmDHAR基因表达量明显高于不耐冷型的“金皇后-308”。GSEA结果表明CmDHAR基因参与结构分子活性、大分子复合物、细胞膜封闭内腔以及抗氧化活性等生物过程。本研究阐明了CmDHAR的生物信息学特点，证明了DHAR基因在哈密瓜抵御冷胁迫过程中的重要性。

关键词: 哈密瓜；谷胱甘肽-S-转移酶；脱氢抗坏血酸还原酶；冷胁迫

Abstract: To clarify the basic properties of the dehydroascorbate reductase (DHAR) gene in Hami melon and its expression level during cold stress, we investigated the basic information, multiple sequence alignment, phylogenetic relationship, collinearity and domain characteristics of two DHAR genes, protein three-dimensional (3D) models and protein-protein interaction, as well as ascorbic acid (AsA) contents, DHAR activities and DHAR gene expression levels under cold stress, and we performed gene set enrichment analysis (GSEA). The results showed that the lengths of the proteins encoded by the two CmDHAR genes consisted of 297 and 206 amino acids, respectively. Their isoelectric points were 8.67 and 5.99, respectively. Their molecular masses were 33 082.9 and 22 891.6 Da, respectively. Multiple sequence alignment analysis showed that the CmDHAR genes had a highly conserved glutathione S-transferase (GST) N-terminal functional domain. Gene structure analysis indicated that CmDHAR contained six exons, six conserved motifs and three types of cis-acting elements. Collinearity analysis indicated that Hami melon was evolutionarily closer to zucchini, winter squash and pumpkin. The 3D models revealed that the secondary structure of CmDHAR2 mainly included α-helix, random coil, extended strand and β-turn. Protein-protein interaction analysis showed that CmDHAR family members were closely related to other GST family members. The AsA content, DHAR activity and CmDHAR gene expression level in the cold-tolerant cultivar “Jiashi-310” (JS) were significantly higher than those in the cold-sensitive cultivar “Golden Empress-308” (GE) under cold stress. GSEA results revealed that the CmDHAR genes participated in the biological processes of ‘structural molecule activity’, ‘macromolecular complex’, ‘membrane-enclosed lumen’ and ‘antioxidant activity’. This study has proved that the CmDHAR genes play an important role in the resistance of Hami melon to cold stress.

Key words: Hami melon; glutathione S-transferase; dehydroascorbate reductase; cold stress

中图分类号:

TS255.3

宋文，张琴，宁明，周发科，杨新泉，单春会，唐凤仙. 哈密瓜脱氢抗坏血酸还原酶基因的鉴定及冷胁迫表达分析[J]. 食品科学, 2022, 43(2): 117-125.

SONG Wen, ZHANG Qin, NING Ming, ZHOU Fake, YANG Xinquan, SHAN Chunhui, TANG Fengxian. Identification and Cold Stress-responsive Expression Analysis of Dehydroascorbate Reductase Gene in Hami Melon (Cucumis melo var. saccharinus)[J]. FOOD SCIENCE, 2022, 43(2): 117-125.

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哈密瓜脱氢抗坏血酸还原酶基因的鉴定及冷胁迫表达分析

Identification and Cold Stress-responsive Expression Analysis of Dehydroascorbate Reductase Gene in Hami Melon (Cucumis melo var. saccharinus)

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