Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi
| Autor: | Rodrigo Costa, Sofia G. Meirinho, Ivone M. Martins, Altino Choupina, Alfredo Cravador |
|---|---|
| Přispěvatelé: | Universidade do Minho |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Phytophthora npp1 Phytophthora cinnamomi Fagaceae Plant Roots Pichia pastoris Microbiology 03 medical and health sciences 0302 clinical medicine HE-TAIL PCR Gene expression Genetics Amino Acid Sequence Cloning Molecular Pyrophosphatases Molecular Biology Gene Pathogen Plant Diseases Oomycete Science & Technology biology Phosphoric Diester Hydrolases Castanea sativa RT-qPCR General Medicine Sequence Analysis DNA biology.organism_classification Engenharia e Tecnologia::Biotecnologia Ambiental In vitro 3. Good health Molecular Weight 030104 developmental biology Gene Expression Regulation 030220 oncology & carcinogenesis Biotecnologia Ambiental [Engenharia e Tecnologia] |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| Popis: | The soil-borne oomycete Phytophthora cinnamomi is a highly destructive Phytophthora species associated with the decline of forest. This pathogen secretes a novel class of necrosis-inducing proteins known as Nep1-like proteins (NLPs). In this work, we report the sequencing and molecular characterization of one of these proteins, more specifically the necrosis-inducing Phytophthora protein 1 (NPP1). The ORF of the npp1 gene (EMBL database AM403130) has 768 bp encoding a putative peptide of 256 amino acids with a molecular weight of approximately 25 kD. In order to understand its function, in vitro gene expression was studied during growth in different carbon sources (glucose, cellulose, and sawdust), and at different times of infection, in vivo by RT-qPCR. The highest expression of the npp1 gene occurred in glucose medium followed by sawdust. In vivo infection of Castanea sativa roots with P. cinnamomi revealed a decrease in npp1 expression from 12 to 24 h; at 36 h its expression increased suggesting the existence of a complex mechanism of defense/attack interaction between the pathogen and the host. Expression of recombinant npp1 gene was achieved in Pichia pastoris and assessed by SDS-PAGE analysis of the protein secreted into the culture supernatant, revealing the presence of the NPP1 protein. Project COMBATINTA/SP2.P11/02 Interreg IIIA—Cross-Border Cooperation Spain-Portugal, financed by The European Regional Development Fund and by national funding from the Portuguese Ministério da Ciência e do Ensino Superior (MCES) (PTDC/AGR-AAM/67628/2006). info:eu-repo/semantics/publishedVersion |
| Databáze: | OpenAIRE |
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