RESUMO
Effector proteins in Phakopsora pachyrhizi (Pp), the causative agent of Asian Soybean rust, are involved in the infection process. A previous study identified a rust effector Egh16-like family based expression profile during the interaction with soybean. Herein, we scrutinized available the Pp genomes to validate the predicted Egh16-like family of Pp and identify new family members. We described 22 members of the Egh16-like gene family in the Pp MT2006 genome and 18 in the UFV02 and K8108 genomes, highlighting a family expansion. Family members have a small signal peptide, conserved cysteine-rich R/Y/FxC motifs in the C-terminal region, and a virulence-related Egh16-like domain and were able to suppress PTI related responses in Benthamiana. Phylogenetic analysis placed the family members into eight clusters, with members induced during the early stages of rust infection. Members of clusters VI and VII are present in different copy numbers in Pp genomes and suppressed PAMP-related responses.
RESUMO
Asian soybean rust (ASR) is one of the most destructive diseases affecting soybeans. The causative agent of ASR, the fungus Phakopsora pachyrhizi, presents characteristics that make it difficult to study in vitro, limiting our knowledge of plant-pathogen dynamics. Therefore, this work used leaf lesion laser microdissection associated with deep sequencing to determine the pathogen transcriptome during compatible and incompatible interactions with soybean. The 36,350 generated unisequences provided an overview of the main genes and biological pathways that were active in the fungus during the infection cycle. We also identified the most expressed transcripts, including sequences similar to other fungal virulence and signaling proteins. Enriched P. pachyrhizi transcripts in the resistant (PI561356) soybean genotype were related to extracellular matrix organization and metabolic signaling pathways and, among infection structures, in amino acid metabolism and intracellular transport. Unisequences were further grouped into gene families along predicted sequences from 15 other fungi and oomycetes, including rust fungi, allowing the identification of conserved multigenic families, as well as being specific to P. pachyrhizi. The results revealed important biological processes observed in P. pachyrhizi, contributing with information related to fungal biology and, consequently, a better understanding of ASR.
RESUMO
As plataformas de sequenciamento de nova geração são uma alternativa poderosa para estudos de genômica estrutural e funcional. Na genômica de plantas, os trabalhos com as novas plataformas têm sido destinados ao sequenciamento de transcritos, ressequenciamento ou sequenciamento de novo de genomas plastidiais. Neste trabalho, são detalhadas as tecnologias das plataformas mais utilizadas atualmente, bem como é revisada a aplicação dessas tecnologias na genômica estrutural e funcional de plantas.
The next-generation DNA sequencing technologies are a powerful alternative to studies in structural and functional genomics. In plant genomics studies, the work with these new platforms has been used for the sequencing of transcripts, re-sequencing, and the de novo sequencing of plastid genomes. This research details the technological principles of the next-generation DNA sequencing platforms most used and reviews its application in structural and functional plant genomics.
RESUMO
A implementação da espectrometria de massa (MS) para as análises de peptídeos (MS) e de aminoácidos (MS em tandem ou MS/MS) tornou possível a identificação de centenas de proteínas em experimentos únicos. Uma grande variedade de estratégias está disponível atualmente para o fracionamento e a purificação de amostras, a identificação de proteínas, a quantificação, a análise de modificações pós-traducionais (MPT's) e os estudos de interação. Dessa forma, a proteômica abre novas perspectivas na biologia de plantas com ênfase nos estudos de variabilidade genética, estresses fisiológicos e desenvolvimento de plantas.
The implementation of mass spectrometry (MS) for peptides (MS) and amino acids (tandem MS or MS/MS) analysis allowed the identification of hundreds of proteins in single experiments. A number of different strategies are current available for sample fractioning and purification, proteins identification, quantification, post-translational modifications (PTM) and interaction analyses. In this way, the proteomics open up new perspectives in plant biology with emphasis on studies of genetic variability, physiological stresses and plant development.