鸭梨DHN3基因鉴定及在果心褐变中的表达分析

doi:10.7506/spkx1002-6630-20220615-159

食品科学 ›› 2023, Vol. 44 ›› Issue (12): 124-131.doi: 10.7506/spkx1002-6630-20220615-159

鸭梨DHN3基因鉴定及在果心褐变中的表达分析

寇晓敏，李庆秀，李玲，张陈云，闫师杰，梁丽雅，邓冰

（1.天津农学院食品科学与生物工程学院，天津 300392；2.天津农学院农学与资源环境学院，天津 300392；3.山西农业大学食品科学与工程学院，山西晋中 030801）

出版日期:2023-06-25 发布日期:2023-06-30
基金资助:
国家自然科学基金面上项目（32072278）；天津市自然科学基金重点项目（20JCZDJC00420）

Identification and Expression Analysis of the DHN3 Gene during the Core Browning of Late-Harvested Yali Pears (Pyrus bretschneideri Rehd.)

KOU Xiaomin, LI Qingxiu, LI Ling, ZHANG Chenyun, YAN Shijie, LIANG Liya, DENG Bing

(1. College of Food Science and Bioengineering, Tianjin Agricultural University, Tianjin 300392, China; 2. College of Agronomy and Resource Environment, Tianjin Agricultural University, Tianjin 300392, China; 3. College of Food Science and Engineering, Shanxi Agricultural University, Jinzhong 030801, China)

Online:2023-06-25 Published:2023-06-30

摘要/Abstract

摘要： 为探究脱水素（dehydrin，DHN）蛋白与晚采鸭梨（Pyrus bretschneideri Rehd.）果心褐变之间的关系，基于生物信息学方法对鸭梨PbDHN3基因进行鉴定和特性分析，并分析急速降温和缓慢降温条件下晚采鸭梨果心褐变发生过程中PbDHN3基因的表达模式。结果表明，PbDHN3开放阅读框564 bp，编码187 个氨基酸，且具有DHN的Dehydrin superfamily保守结构域，含有2 个Y片段、1 个S片段、3 个K片段，属于Y2SK3型DHN。生物信息学分析表明PbDHN3分子质量19.77 kDa，等电点7.98，为亲水性稳定蛋白；二级结构包含α-螺旋、无规卷曲、延伸链和β-转角，且定位于细胞核，不含信号肽与跨膜结构，为非分泌蛋白。同源性分析结果表明PbDHN3与苹果等5 种植物的DHN相似性达到89.74%以上，系统发育分析显示PbDHN3和P. ussuriensis×P. communis的DHN亲缘关系最近。启动子分析结果表明PbDHN3含有基本启动子元件和其他顺式作用元件，与陆地棉GhDHN1及小麦TaDHN2启动子区预测结果一致，都包含激素响应元件、光响应元件等。实时聚合酶链式反应分析结果表明，急速降温条件下鸭梨果心部位DHN3基因在褐变过程中呈显著上调表达，与果心褐变紧密相关，为进一步探究该基因的功能，明确鸭梨果心褐变发生机制提供理论依据。

关键词: 鸭梨；褐变；脱水素；基因鉴定；生物信息学分析

Abstract: In order to explore the relationship between dehydrin (DHN) and core browning in late-harvested Yali pears, this study identified and characterized the PbDHN3 gene based on bioinformatics methods, and analyzed the expression pattern of this gene during the core browning process under rapid cooling and slow cooling conditions. The results showed that the open reading frame (ORF) of PbDHN3 was 564 bp in length, encoding 187 amino acids, and having a conserved domain of dehydrin superfamily comprising two Y segments, one S segment and three K segments (Y2SK3). Bioinformatics analysis showed that PbDHN3 was a hydrophilic stable protein with a molecular mass of 19.77 kDa and an isoelectric point of 7.98. The secondary structure consists of α-helix, irregular coil, extended chain and β-turn. PbDHN3 was located in the nucleus and contained no signal peptide or transmembrane structure. It was a non-secretory protein. The results of homology analysis showed that PbDHN3 was more than 89.74% similar to the DHN of five other plants such as apples. Phylogenetic analysis showed that PbDHN3 had the closest relationship with the DHN of Pyrus ussuriensis × Pyrus communis. The results of promoter analysis showed that PbDHN3 contained basic promoter elements and other cis-acting elements, which were consistent with the predicted results of the upland cotton GhDHN1 and the wheat TaDHN2 promoter regions, and all of them included hormone response elements and light response elements. The results of real-time fluorescence polymerase chain reaction (PCR) analysis showed that the expression of the DHN3 gene was significantly up-regulated during the core browning process of Yali pears under rapid cooling conditions, which was closely related to the browning of the core. This study provides a theoretical basis for further exploring the function of the PbDHN3 gene and clarifying the mechanism of core browning of Yali pears.

Key words: pears; browning; dehydrin; gene identification; bioinformatics analysis

中图分类号:

TS255.3

寇晓敏，李庆秀，李玲，张陈云，闫师杰，梁丽雅，邓冰. 鸭梨DHN3基因鉴定及在果心褐变中的表达分析[J]. 食品科学, 2023, 44(12): 124-131.

KOU Xiaomin, LI Qingxiu, LI Ling, ZHANG Chenyun, YAN Shijie, LIANG Liya, DENG Bing. Identification and Expression Analysis of the DHN3 Gene during the Core Browning of Late-Harvested Yali Pears (Pyrus bretschneideri Rehd.)[J]. FOOD SCIENCE, 2023, 44(12): 124-131.

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鸭梨DHN3基因鉴定及在果心褐变中的表达分析

Identification and Expression Analysis of the DHN3 Gene during the Core Browning of Late-Harvested Yali Pears (Pyrus bretschneideri Rehd.)

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