Isolation and Sequencing of Actin1, Actin2 and Tubulin1 Genes Involved in Cytoskeleton Formation in Phytophthora cinnamomi

Martins, Ivone M.; López, M Carmen; Domínguez, Ángel; Choupina, Altino

doi:10.4172/2157-7471.1000194

Cited by 2 publications

(2 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, two simple and one more complex carbon sources were selected for P. cinnamomi growing: 2% glucose, 0.2% cellulose and 0.2% sawdust respectively as described in "Materials and methods". The expression level was measured over 8 days of growth relative to the expression of the constitutive gene actin2 [26,27]. The choice of actin mRNA as a stable endogenous control to normalize the amount of sample RNA was validated by evaluation of the oomycete actin mRNA levels in in vitro and in planta conditions.…”

Section: Quantification Of Npp1 Transcriptsmentioning

confidence: 99%

Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi

et al. 2019

View full text Add to dashboard Cite

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.

show abstract

Section: Quantification Of Npp1 Transcriptsmentioning

confidence: 99%

Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Our group, using asymmetric PCR, have identified and characterized the structural proteins Actin1, Actin2 and Tubulin1 (GenBank: AM412175.1, AM412176.1, AM412177.1) (Martins et al, 2013), and some other proteins involved in mechanisms of infection by P. cinnamomi namely endo-1,3-beta-glucanase (GenBank: AM259651.1) (complete cds) (Meirinho et al, 2010); endo-glucanase (GenBank: AJ964942.1) (partial cds) (unpublished results); glucanase inhibitor protein (GIP) (GenBank: AM259384.1) (complete cds) (Martins et al, 2014a); necrosisinducing Phytophthora protein 1 (NPP1) (GenBank: AM403130.1) (complete cds) (unpublished results) (Gijzen and Nurnberger, 2006) and transglutaminase (GenBank: AM403129.1) (unpublished results) (Martins et al, 2014b;Yu et al, 2008). By standard PCR with DNA of P. cinnamomi and primers designed by homologous sequences from different Phytophthora species, our group also sequenced ribosomal RNA subunits (GenBank: AJ421265.2 and AJ457835.2) (unpublished results).…”

Section: Introductionmentioning

confidence: 99%

Genes found in partial sequencing of Phytophthora cinnamomi genome

Santos¹,

Martins²,

Maia³

et al. 2016

Rev. Ciênc. Agr.

View full text Add to dashboard Cite

A B S T R A C TMembers of the oomycete cause extensive losses in agriculture and widespread degradation in natural plant communities, being responsible for the death of thousands of trees every year. Two of the representative species are Phytophthora infestans, which causes late blight of potato, and Phytophthora cinnamomi, which causes chestnut ink disease, responsible for losses on sweet chestnut production in Europe. Genome sequencing efforts have been focused on the study of three species: P. infestans, P. sojae and P. ramorum. Phytophthora infestans has been developed as the model specie for the genus, possessing excellent genetic and genomics resources including genetic maps, BAC libraries, and EST sequences. Our research team is trying to sequence the genome of P. cinnamomi in order to gain a better understanding of this oomycete, to study changes in plant-pathogen relationships including those resulting from climate change and trying to decrease the pathogen's impact on crops and plants in natural ecosystems worldwide. We present here a preliminary report of partially sequenced genomic DNA from P. cinnamomi encoding putative protein-coding sequences and tRNAs. Database analysis reveals the presence of genes conserved in oomycetes.Keywords: DNA library, oomycetes, Phytophthora cinnamomi, tRNA. R E S U M OOs oomicetas causam enormes perdas na agricultura e degradação generalizada em áreas de crescimento natural, sendo responsáveis pela morte de milhares de árvores por ano. Duas das espécies representativas de oomicetas são Phytophthora infestans, que provoca o míldio da batateira, e Phytophthora cinnamomi, que causa a doença da tinta do castanheiro, responsável por perdas de produção de castanha na Europa. Os esforços de sequenciação de genomas de oomicetas têm estado focados no estudo de três espécies: P. infestans, P. sojae e P. ramorum. Phytophthora infestans foi considerada como espécie modelo para o género, pois possui excelentes recursos genéticos e de genómica, incluindo mapas genéticos, bibliotecas BAC, e sequências de EST. Esta equipa de investigadores procura sequenciar o genoma de P. cinnamomi, a fim de obter uma melhor compreensão deste oomiceta, para estudar mudanças nas relações plantapatogénio, incluindo as resultantes das alterações climáticas, e tentando diminuir o impacto do patogénio nas culturas e plantas em ecossistemas naturais em todo o mundo. É apresentada uma abordagem preliminar do DNA genómico parcialmente sequenciado a partir de P. cinnamomi, o qual codifica possíveis sequências de proteínas e tRNAs. As análises em bases de dados revelam a presença de genes conservados em oomicetas.Palavras-chave: Biblioteca DNA, oomicetas, Phytophthora cinnamomi, tRNA.

show abstract

Isolation and Sequencing of Actin1, Actin2 and Tubulin1 Genes Involved in Cytoskeleton Formation in Phytophthora cinnamomi

Cited by 2 publications

References 14 publications

Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi

Cloning, characterization, in vitro and in planta expression of a necrosis-inducing Phytophthora protein 1 gene npp1 from Phytophthora cinnamomi

Genes found in partial sequencing of Phytophthora cinnamomi genome

Contact Info

Product

Resources

About