Molecular Characteristics and Biochemical Functions of VpPR10s from Vitis pseudoreticulata Associated with Biotic and Abiotic Stresses

Abstract

Grapes are one of the world’s oldest and most important fruit crops. They are of high economic value in many countries, but the susceptibility of the dominant winegrape species Vitis vinifera to fungal disease is a significant problem. The Chinese wild grape species are a rich source of disease-resistance genes and these can be used to discover how disease resistance in V. vinifera grapevines might be enhanced. Pathogenesis-related (PR) 10 proteins are involved in the disease-response. Here, we use the genomic DNA of the Chinese wild species Vitis pseudoreticulata accession “Baihe-35-1” as the template to design specific primers based on VvPR10s sequences. We used overlap extension PCR to obtain the sequences: VpPR10.4, VpPR10.6, VpPR10.7 and VpPR10.9. The coding sequences of the VpPR10s were then cloned into the pGEX-4T-1 vector. The purified proteins VpPR10.4, VpPR10.6, VpPR10.7 and VpPR10.9 were used to analyse nuclease activity. Meanwhile, functional analysis of VpPR10s under different biotic and abiotic stresses was carried out to further clarify the disease-resistance mechanisms of the Chinese wild grapevine VpPR10 genes. The analysis of protein structure indicates that VpPR10.4 and VpPR10.7 had the P-loop domain and the Bet v 1 motif, which are a consistent feature of plant PR10. However, there was no P-loop domain or Bet v 1 motif in VpPR10.9 and we could not find the Bet v 1 motif in VpPR10.6. The results of the nuclease activity assay and of the functional analyses of VpPR10s under different biotic and abiotic stresses also confirm that VpPR10.4 and VpPR10.7 proteins have marked RNase, DNase, anti-fungal activities and respond to abiotic stresses. The VpPR10.6 and VpPR10.9 proteins do not have these activities and functions.

A Novel Pathogenesis-Related Class 10 Protein Gly m 4l, Increases Resistance upon Phytophthora sojae Infection in Soybean (Glycine max [L.] Merr.)

“…Some of PR10 proteins have previously been shown to possess RNase or DNase activity and performed a link to plant defense strategy repertoire, such as JIOsPR10 from Oryza sativa [ 39 ], VpPR10.2 from Vitis pseudoreticulata [ 65 ], JcPR10a from Jatropha curcas [ 78 ], VpPR10.1 from Vitis pseudoreticulata [ 16 ], VpPR10.4 and VpPR10.7 from Vitis pseudoreticulata [ 79 ]. On the basis of these findings, it supports the hypothesis that RNase activity was potentially crucial for plant defense, such as activating the death of infected cells to limit pathogen invasion, regulating and controlling mRNA transcription upon pathogen infection or other stresses, and directly degrading pathogenic RNA [ 40 , 48 ].…”

The Grape VlWRKY3 Gene Promotes Abiotic and Biotic Stress Tolerance in Transgenic Arabidopsis thaliana

“…et al, 2013 ; Yokokani et al, 2013 ), wheat ( Wang C. et al, 2013 ), tomato ( Bhattarai et al, 2010 ; Atamian et al, 2012 ; Liu et al, 2014 ), and cotton ( Yu et al, 2012 ; Shi et al, 2014 ). We have been studying WRKY genes from grape ( Vitis vinifera L.), one of the most economically important fruit crops in many countries ( Wang et al, 2014 ), and in previous study, we identified 59 WRKY genes in the grape genome have been identified ( Guo et al, 2014 ), however, only a few of these have been functionally characterized to date. VvWRKY1 was reported to be involved in defense against downy mildew ( Marchive et al, 2007 , 2013 ), while over-expression of VvWRKY2 in tobacco ( Nicotiana tabacum cv.…”

Biotechnological Approaches: Gene Overexpression, Gene Silencing, and Genome Editing to Control Fungal and Oomycete Diseases in Grapevine

“…Further studies in this field suggested that some thaumatin-like protein ( TLP ) genes, perform better against biotrophs rather than necrotrophs, like TLP29 gene of V. quinquangularis ( VqTLP29 ) expressed in A. thaliana , where an increased susceptibility to B. cinerea was detected [ 37 ]. PR10 proteins are highly expressed after pathogen invasion, and their antifungal capability seems to also be associated with their RNase/DNase activity, and to their role in the control of flavonoid biosynthesis [ 81 , 82 ]. The VpPR10.1 gene inserted through Agrobacterium -mediated transformation in Thompson Seedless cultivar led to reduced hyphal growth of Plasmopara viticola , through callose deposition around hyphae and haustoria, and hydrogen peroxide accumulation compared to non-transgenic lines [ 39 ].…”