Application of cDNA array for studying the gene expression profile of mature appressoria of Magnaporthe grisea

Abstract

Appressorium is an infection structure of the phytopathogenic fungus Magnaporthe grisea. Analysis of gene expression profiles of appressorium development provides insight into the molecular basis of pathogenicity and control of this fungal plant disease. A cDNA array representing 2927 unique genes based on a large EST (expressed sequence tag) database of M. grisea strain Y34 was constructed and used to profile the gene expression patterns at mycelium and appressorium maturation stages. Compared with mycelia, 55 up-regulated and 22 down-regulated genes were identified in mature appressoria. Among 77 genes, 16 genes showed no similarity to the genome sequences of M. grisea. A novel homologue of peptidyl-prolyl cis-trans isomerase was found to be expressed at low-level in mature appressoria of M. grisea. The results indicated that the genes such as pyruvate carboxylase, phospholipid metabolism-related protein and glyceraldehyde 3-phosphate dehydrogenase involved in gluconeogenesis, lipid metabolism and glycolysis, showed differential expression in mature appressoria. Furthermore, genes such as PTH11, beta subunit of G protein and SGT1 involved in cell signalling, were expressed differentially in mature appressoria. Northern blot analysis was used to confirm the cDNA array results.

Rice blast fungus transcriptomics

“…This makes the pattern of gene expression during appressorium development important to decipher molecular mechanisms underlying appressorial morphogenesis. Transcriptomics approaches, such as suppression subtractive hybridization (SSH) (Lu et al 2005), serial analysis of gene expression (SAGE) (Irie et al 2003) and microarray (Takano et al 2003;Jin et al 2007) have been employed to identify appressorium specific genes.…”

“…Jin et al (2007) used cDNA microarray to profile the gene expression patterns of mycelium and appressorium maturation stages of M. grisea field isolate Y34. Compared with mycelia, 55 up-regulated and 22 down-regulated genes were identified in mature appressoria.…”

Genome‐wide validation of Magnaporthe grisea gene structures based on transcription evidence

“…Genome assembly version 6, which was constructed by the Broad Institute, is comprised of 30 supercontigs containing 805 contigs. In addition, expressed sequence tags (ESTs) obtained from different types of cDNA libraries were sequenced in several strains, and currently over 50 000 ESTs are available in the international nucleotide sequence databases (INSD), DDBJ/EMBL/GenBank [6–8]. However, the availability of 5′ end sequences of mRNAs from filamentous fungi is currently limited.…”

Análise da expressão gênica no dermatófito Trichophyton rubrum mimetizando a infecção in vitro: pH e diferentes fontes de carbono regulando genes

“…São necessários estímulos ambientais para ativação e consequente dissociação das subunidades (α e βγ), cada uma com função ativadora em uma ampla variedade de moléculas efetoras, como fosfolipases, canais de íons e adenilato ciclase (Kaufman et al, 2007). Na análise dos genes diferencialmente expressos no patógeno M. grisea, pesquisadores confirmaram a alta expressão da proteína G após a indução/formação do apressório maduro em comparação com o controle (micélio vegetativo) (Jin et al, 2007). ∆gna-1 exibiu vários defeitos na macroconidiação relacionados a diferenças na morfologia das hifas aéreas e baixa produção de conídios, além de conídios pouco aderentes em relação aos obtidos da cepa selvagem (Ivey et al, 1996), entre outras alterações.…”