Gene cloning, molecular modeling, and phylogenetics of serine protease P32 and serine carboxypeptidase SCP1 from nematophagous fungi Pochonia rubescens and Pochonia chlamydosporia

The fungi Pochonia chlamydosporia and Pochonia rubescens are parasites of nematode eggs and thus are biocontrol agents of nematodes. Proteolytic enzymes such as the S8 proteases VCP1 and P32, secreted during the pathogenesis of nematode eggs, are major virulence factors in these fungi. Recently, expression of these enzymes and of SCP1, a new putative S10 carboxypeptidase, was detected during endophytic colonization of barley roots by these fungi. In our study, we cloned the genomic and mRNA sequences encoding P32 from P. rubescens and SCP1 from P. chlamydosporia. P32 showed a high homology with the serine proteases Pr1A from the entomopathogenic fungus Metarhizium anisopliae and VCP1 from P. chlamydosporia (86% and 76% identity, respectively). However, the catalytic pocket of P32 showed differences in the amino acids of the substrate-recognition sites compared with the catalytic pockets of Pr1A and VCP1 proteases. Phylogenetic analysis of P32 suggests a common ancestor with protease Pr1A. SCP1 displays the characteristic features of a member of the S10 family of serine proteases. Phylogenetic comparisons show that SCP1 and other carboxypeptidases from filamentous fungi have an origin different from that of yeast vacuolar serine carboxypeptidases. Understanding protease genes from nematophagous fungi is crucial for enhancing the biocontrol potential of these organisms.

Checking the concurrence among macrobenthic organism distribution patterns at different taxonomic scales in relation to environmental factors

The interaction of both natural conditions and anthropogenic environmental impacts can lead to different soft-bottom macrobenthic distribution patterns. Soft-bottom macrobenthic community was analysed at different taxonomic scales in order to evaluate whether diverse subset of organisms respond to the variability of the environmental pressures (natural and human induced) showing or not similar distribution patterns. Therefore, this long-term survey had been focused on a heterogeneous area, where both anthropogenic and natural stress may affect the community. Three perpendicular transects to the coast were established and stations at 4, 10 and 15 m depths were sampled at each transect twice a year (summer- winter) from 2004 to 2009. Non-parametric multivariate techniques were used to analyse soft-bottom macrobenthic community distribution and its relation to the environmental factors. Similar distribution patterns between investigated taxonomic levels were detected and they were mainly related to depth.