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葡萄(Vitis vinifera L.)中乙醛酸酶样基因家族的全基因组分析及其对霜霉病感染的表达谱分析。

Genome-wide analysis of glyoxalase-like gene families in grape (Vitis vinifera L.) and their expression profiling in response to downy mildew infection.

机构信息

College of Horticulture, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.

State Key Laboratory of Crop Stress Biology for Arid Areas, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.

出版信息

BMC Genomics. 2019 May 9;20(1):362. doi: 10.1186/s12864-019-5733-y.

Abstract

BACKGROUND

The glyoxalase system usually comprises two enzymes, glyoxalase I (GLYI) and glyoxalase II (GLYII). This system converts cytotoxic methylglyoxal (MG) into non-toxic D-lactate in the presence of reduced glutathione (GSH) in two enzymatic steps. Recently, a novel type of glyoxalase III (GLYIII) activity has observed in Escherichia coli that can detoxify MG into D-lactate directly, in one step, without a cofactor. Investigation of the glyoxalase enzymes of a number of plant species shows the importance of their roles in response both to abiotic and to biotic stresses. Until now, glyoxalase gene families have been identified in the genomes of four plants, Arabidopsis, Oryza sativa, Glycine max and Medicago truncatula but no similar study has been done with the grapevine Vitis vinifera L.

RESULTS

In this study, four GLYI-like, two GLYII-like and three GLYIII-like genes are identified from the genome database of grape. All these genes were analysed in detail, including their chromosomal locations, phylogenetic relationships, exon-intron distributions, protein domain organisations and the presence of conserved binding sites. Using quantitative real-time PCR analysis (qRT-PCR), the expression profiles of these genes were analysed in different tissues of grape, and also when under infection stress from downy mildew (Plasmopara viticola). The study reveals that most VvGLY-like genes had higher expressions in stem, leaf, tendril and ovule but lower expressions in the flower. In addition, most of the VvGLY-like gene members were P. viticola responsive with high expressions 6-12 h and 96-120 h after inoculation. However, VvGLYI-like1 was highly expressed 48 h after inoculation, similar to VvPR1 and VvNPR1 which are involved in the defence response.

CONCLUSIONS

This study identified the GLYI-like, GLYII-like and GLYIII-like full gene families of the grapevine. Based on a phylogenetic analysis and the presence of conserved binding sites, we speculate that these glyoxalase-like genes in grape encode active glyoxalases. Moreover, our study provides a basis for discussing the roles of VvGLYI-like, VvGLYII-like and VvGLYIII-like genes in grape's response to downy mildew infection. Our results shed light on the selection of candidate genes for downy mildew tolerance in grape and lay the foundation for further functional investigations of these glyoxalase genes.

摘要

背景

糖氧还蛋白系统通常由两种酶组成,即糖氧还酶 I(GLYI)和糖氧还酶 II(GLYII)。该系统在还原型谷胱甘肽(GSH)存在下,通过两步酶促反应将细胞毒性的甲基乙二醛(MG)转化为无毒的 D-乳酸。最近,在大肠杆菌中观察到一种新型的糖氧还酶 III(GLYIII)活性,它可以直接将 MG 解毒为 D-乳酸,无需辅助因子。对多种植物糖氧还酶的研究表明,它们在应对非生物和生物胁迫方面的作用非常重要。到目前为止,已在拟南芥、水稻、大豆和紫花苜蓿的基因组中鉴定出糖氧还酶基因家族,但在葡萄基因组中尚未进行类似的研究。

结果

在这项研究中,从葡萄基因组数据库中鉴定出四个 GLYI 样、两个 GLYII 样和三个 GLYIII 样基因。详细分析了所有这些基因,包括它们的染色体位置、系统发育关系、外显子-内含子分布、蛋白质结构域组织以及保守结合位点的存在。通过定量实时 PCR 分析(qRT-PCR),分析了这些基因在葡萄不同组织中的表达谱,以及在霜霉病(Plasmopara viticola)感染胁迫下的表达谱。研究表明,大多数 VvGLY 样基因在茎、叶、卷须和胚珠中的表达较高,而在花中的表达较低。此外,大多数 VvGLY 样基因成员对 P. viticola 有反应,接种后 6-12 小时和 96-120 小时表达量较高。然而,VvGLYI-like1 在接种后 48 小时表达量较高,与参与防御反应的 VvPR1 和 VvNPR1 相似。

结论

本研究鉴定了葡萄的 GLYI 样、GLYII 样和 GLYIII 样全长基因家族。基于系统发育分析和保守结合位点的存在,我们推测这些糖氧还酶样基因在葡萄中编码具有活性的糖氧还酶。此外,本研究为讨论 VvGLYI-like、VvGLYII-like 和 VvGLYIII-like 基因在葡萄对霜霉病侵染的反应中的作用提供了依据。我们的研究结果揭示了候选基因在葡萄对霜霉病抗性选择中的作用,并为进一步研究这些糖氧还酶基因的功能奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dd61/6509763/9ce0aefe09db/12864_2019_5733_Fig1_HTML.jpg

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