| Genomics and post-genomics studies have shown that the redox state of plant cells is controlled by a complex genetic network.Existing studies have shown that catalase（Cat）is one of the most important antioxidant enzymes.Cat is widely found in organisms,which main function is to remove hydrogen peroxide（H2O2）produced in the process of intracellular photorespiration,mitochondrial electron transport and fatβ-oxidation in plants,thus protecting plant cells from H2O2toxicity.In recent years,it has also been shown that by increasing the activity of antioxidant enzymes in plants and enhancing the level of antioxidant metabolism,the resistance of plants can be improved by themselves.Cat is involved in the disease resistance response of many plants,and its overexpression in the plant body can improve the resistance of plants to stress,and enhance the tolerance of plants by enhancing the activity of antioxidant enzymes and the metabolism of reactive oxygen species.The disease resulting from Areca palm necrotic ringspot virus（ANRSV）is a devastating disease which threatens the Areca catechu industry seriously in tropical and subtropical regions.The characteristics of Areca catechu Cat were preliminarily understood by cloning Cat gene of Areca catechu and its bioinformatics analysis.At the same time,the interaction between Cat and ANRSV was studied to lay a good foundation for further study of the biological function and disease resistance mechanism of Areca catechu.In order to understand the relevant characteristics of Areca Cat gene,this study used RNA extracted from Areca leaves,obtained the full length cDNA of Areca Cat gene by homologous cloning and RACE technology,and conducted bioinformatics analysis and subcellular localization.The results showed that the total length of the cloned ArCAT1（MN692600）,ArCAT2（MN692601）,ArCAT3（MN692602）gene sequences were all 1476 bp,which could encode 492 amino acids.Genetic evolution analysis showed that ArCats had a high similarity to amino acid sequences of oil palm（Elaeis guineensis）,jujube（Phoenix dactylifera）and other plants of the same palm family,among which the relationship with oil palm（Elaeis guineensis）was closest.Subcellular localization results indicated that ArCat1 proteins were localized in peroxisomes,while ArCat2 and ArCat3were localized in both peroxisomes and nuclei.To investigate the interaction between ArCats and ANRSV,ANRSV-VPg was selected as the viral target protein.VPg,a multifunctional protein encoded by ANRSV,plays an important role in viral replication,translation,and motility by interacting with multiple host factors.The interaction between ANRSV-VPg proteins and ArCat1-3 were investigated using BiFC bimolecular fluorescence complementary techniques and yeast two-hybrid methods.The results showed that ANRSV-VPg and ArCat2 interactions occurred in yeast and tobacco cells,defining the domains of ANRSV-VPg and ArCat2interactions,and that the interaction occurred in the nucleus and cytoplasm.The above ArCat characteristics and the results of the interaction between ANRSV-VPg proteins and ArCat should be beneficial to the formulation of prevention and treatment strategies for Areca yellow necrotizing ring spot disease,and also provide theoretical support for the study of the pathogenic mechanism of Areca yellow necrotizing ring spot disease. |
