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中华猕猴桃(红心)抗坏血酸过氧化物酶基因家族的鉴定与表达谱分析。

Identification and expression profiling analysis of ascorbate peroxidase gene family in Actinidia chinensis (Hongyang).

机构信息

College of Forestry, Jiangxi Agricultural University/Jiangxi Provincial Key Laboratory of Silviculture, Nanchang, 330045, Jiangxi, People's Republic of China.

College of Agronomy, Jiangxi Agricultural University/Kiwifruit Institute of Jiangxi Agricultural University, Nanchang, 330045, Jiangxi, People's Republic of China.

出版信息

J Plant Res. 2020 Sep;133(5):715-726. doi: 10.1007/s10265-020-01206-y. Epub 2020 Jun 6.

Abstract

Ascorbate peroxidase (APX) is one of the important antioxidant enzymes in the active oxygen metabolism pathway of plants and animals, especially it is the key enzyme to clear HO in chloroplast and the main enzyme of vitamin C metabolism. However, knowledge about APX gene family members and their evolutionary and functional characteristics in kiwifruit is limited. In this study, we identified 13 members of the APX gene family in the kiwifruit (cultivar: Hongyang) genome according the APX proteins conserved domain of Arabidopsis thaliana. Phylogenetic analysis by maximum likelihood split these 13 genes into four groups. The APX gene family members were distributed on nine chromosomes (Nos. 4, 5, 11, 13, 20, 21, 23, 25, 28). Most of the encoded hydrophilic and lipid-soluble enzymes were predicted to be located in the cytoplasm, nucleus and chloroplast. Among them, AcAPX4, AcAPX5, AcAPX8, AcAPX12 were transmembrane proteins, and AcAPX8 and AcAPX12 had the same transmembrane domain. The gene structure analysis showed that AcAPXs were composed of 4-22 introns, except that AcAPX10 was intron-free. Multiple expectation maximization for motif elicitation program (MEME) analyzed 13 APX protein sequences of Actinidia chinensis and identified 10 conserved motifs ranging in length from 15 to 50 amino acid residues. Additionally, the predicted secondary structures of the main motifs consisted of α-helix and random coils. The gene expression of fruits in different growth stages and bagging treatment were determined by qRT-PCR. The results showed that 8 AcAPXs had the highest expression levels during the color turning period and only the gene expression of AcAPX3 was consistent with the ascorbic acid content; five AcAPXs were consistent with the ascorbic acid content after bagging. Our data provided evolutionary and functional information of AcAPX gene family members and revealed the gene expression of different members in different growth stages and bagging treatments These results may be useful for future studies of the structures and functions of AcAPX family members.

摘要

抗坏血酸过氧化物酶 (APX) 是植物和动物活性氧代谢途径中的重要抗氧化酶之一,特别是它是清除叶绿体中 HO 的关键酶,也是维生素 C 代谢的主要酶。然而,关于 APX 基因家族成员及其在猕猴桃中的进化和功能特征的知识有限。在这项研究中,我们根据拟南芥 APX 蛋白保守结构域,从猕猴桃(品种:红阳)基因组中鉴定出 13 个 APX 基因家族成员。最大似然分裂的系统发育分析将这 13 个基因分为 4 组。APX 基因家族成员分布在 9 条染色体上(第 4、5、11、13、20、21、23、25、28 号)。大多数编码的亲水性和脂溶性酶预计位于细胞质、核和叶绿体中。其中,AcAPX4、AcAPX5、AcAPX8、AcAPX12 是跨膜蛋白,AcAPX8 和 AcAPX12 具有相同的跨膜结构域。基因结构分析表明,AcAPXs 由 4-22 个内含子组成,除 AcAPX10 外无内含子。 motif elicitation program (MEME) 对猕猴桃的 13 个 APX 蛋白序列进行了多重预期最大似然分析,鉴定出 10 个长度在 15-50 个氨基酸残基之间的保守基序。此外,主要基序的预测二级结构由α-螺旋和无规卷曲组成。通过 qRT-PCR 测定了不同生长阶段和套袋处理果实的基因表达。结果表明,8 个 AcAPXs 在转色期表达水平最高,只有 AcAPX3 的基因表达与抗坏血酸含量一致;套袋后 5 个 AcAPXs 与抗坏血酸含量一致。我们的数据提供了 AcAPX 基因家族成员的进化和功能信息,并揭示了不同成员在不同生长阶段和套袋处理下的基因表达。这些结果可能对未来研究 AcAPX 家族成员的结构和功能有用。

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