Transcriptome analysis of Neotyphodium and Epichloë grass endophytes

Large-scale gene discovery has been performed for the grass fungal endophytes Neotyphodium coenophialum, Neotyphodium lolii, and Epichloe festucae. The resulting sequences have been annotated by comparison with public DNA and protein sequence databases and using intermediate gene ontology annotation tools. Endophyte sequences have also been analysed for the presence of simple sequence repeat and single nucleotide polymorphism molecular genetic markers. Sequences and annotation are maintained within a MySQL database that may be queried using a custom web interface. Two cDNA-based microarrays have been generated from this genome resource, They permit the interrogation of 3806 Neotyphodium genes (Nchip (TM) rnicroarray), and 4195 Neotyphodium and 920 Epichloe genes (EndoChip (TM) microarray), respectively. These microarrays provide tools for high-throughput transcriptome analysis, including genome-specific gene expression studies, profiling of novel endophyte genes, and investigation of the host grass-symbiont interaction. Comparative transcriptome analysis in Neotyphodium and Epichloe was performed. (c) 2006 Elsevier

Fungos endofíticos isolados de ápices caulinares de pupunheira cultivada in vivo e in vitro

A perda de plantas micropropagadas ocorre, principalmente, pela presença de microrganismos, responsáveis pela morte das plantas no início da cultura ou em seu estabelecimento no campo. O trabalho teve como objetivo a identificação, por taxonomia clássica, e por meio de técnicas moleculares, de fungos presentes nos ápices caulinares de pupunheiras sadias, cultivadas no campo, e a comparação com os fungos isolados, em plantas micropropagadas há dois anos. Os isolados da microbiota fúngica endofítica, das plantas cultivadas in vitro, foram: Fusarium oxysporum, Neotyphodium sp. e Epicoccum nigrum; e das plantas in vivo, foram: Fusarium sp., F. proliferatum, F. oxysporum, Colletotrichum sp., Alternaria gaisen, Neotyphodium sp. e Epicoccum nigrum. As sete espécies de fungos foram reintroduzidas in vitro na planta hospedeira, demonstrando diferentes comportamentos. Neotyphodium sp. e E. nigrum estabeleceram uma interação endofítica com a planta, e as demais comportaram-se como patógenos, diminuindo o desenvolvimento das plântulas em relação às plantas sem inoculação. As espécies endofíticas apresentam potencial para o uso no controle biológico de patógenos de pupunha.

Plant-symbiotic fungi as chemical engineers: multi-genome analysis of the Clavicipitaceae reveals dynamics of alkaloid Loci

The fungal family Clavicipitaceae includes plant symbionts and parasites that produce several psychoactive and bioprotective alkaloids. The family includes grass symbionts in the epichloae clade (Epichloë and Neotyphodium species), which are extraordinarily diverse both in their host interactions and in their alkaloid profiles. Epichloae produce alkaloids of four distinct classes, all of which deter insects, and some—including the infamous ergot alkaloids—have potent effects on mammals. The exceptional chemotypic diversity of the epichloae may relate to their broad range of host interactions, whereby some are pathogenic and contagious, others are mutualistic and vertically transmitted (seed-borne), and still others vary in pathogenic or mutualistic behavior. We profiled the alkaloids and sequenced the genomes of 10 epichloae, three ergot fungi (Claviceps species), a morning-glory symbiont (Periglandula ipomoeae), and a bamboo pathogen (Aciculosporium take), and compared the gene clusters for four classes of alkaloids. Results indicated a strong tendency for alkaloid loci to have conserved cores that specify the skeleton structures and peripheral genes that determine chemical variations that are known to affect their pharmacological specificities. Generally, gene locations in cluster peripheries positioned them near to transposon-derived, AT-rich repeat blocks, which were probably involved in gene losses, duplications, and neofunctionalizations. The alkaloid loci in the epichloae had unusual structures riddled with large, complex, and dynamic repeat blocks. This feature was not reflective of overall differences in repeat contents in the genomes, nor was it characteristic of most other specialized metabolism loci. The organization and dynamics of alkaloid loci and abundant repeat blocks in the epichloae suggested that these fungi are under selection for alkaloid diversification. We suggest that such selection is related to the variable life histories of the epichloae, their protective roles as symbionts, and their associations with the highly speciose and ecologically diverse cool-season grasses.