RESUMO
Dioscorea mosaic associated virus (DMaV) is a member of the genus Sadwavirus, family Secoviridae, that is associated with mosaic symptoms in Dioscorea rotundata in Brazil. The genome of a DMaV isolate detected in D. trifida in Guadeloupe was sequenced by high-throughput sequencing. Using an RT-PCR-based detection assay, we found that DMaV infects D. alata, D. bulbifera, D. cayenensis-rotundata, D. esculenta, and D. trifida accessions conserved in Guadeloupe and Côte d'Ivoire and displays a very high level of molecular diversity in a relatively small region of the genome targeted by the assay. We also provide evidence that DMaV is also present in D. rotundata in Benin and in D. alata in Nigeria.
Assuntos
Dioscorea , Especificidade de Hospedeiro , Secoviridae , Dioscorea/virologia , Variação Genética , Filogenia , Secoviridae/classificaçãoRESUMO
The genome of yam virus X (YVX), a new member of the genus Potexvirus from yam (Dioscorea trifida), was completely sequenced. Structural and phylogenetic analysis showed that the closest relative of YVX is nerine virus X. A prevalence study found YVX only in plants maintained in Guadeloupe and showed that it also infects members of the complex D. cayenensis rotundata. This study provides evidence for the existence of two additional potexviruses, one of which infects D. nummularia in Vanuatu and the other, D. bulbifera and D. rotundata in Haiti and D. trifida and D. rotundata in Guadeloupe. This work also shows that existing potexvirus-specific degenerate primers targeting the ORF1-encoded polymerase domain are well suited for the identification of the three potexviruses reported here.
Assuntos
Dioscorea/virologia , Variação Genética , Doenças das Plantas/virologia , Potexvirus/classificação , Potexvirus/isolamento & purificação , Análise por Conglomerados , Ordem dos Genes , Genoma Viral , Dados de Sequência Molecular , Filogenia , Potexvirus/genética , RNA Viral/genética , Análise de Sequência de DNA , Homologia de SequênciaRESUMO
Knox7, a class 2 homeobox gene has been characterized in maize. A combination of experimental (3'- and 5'-RACE) and bioinformatics approaches supported the idea that Knox7 would be transcribed into two alternative transcripts by differential initiation of transcription. Sequence differences between alternative transcripts, Knox7L the larger and Knox7S the smaller, were confined to their 5' end regions and exon 1 was only found in Knox7L transcripts. Deduced proteins shared the same homeodomain, while an Ala and Ala/Gly rich domain was found only in KNOX7L protein. We hypothesize that KNOX7L and KNOX7S might regulate (differentially) the expression of the same gene(s) by binding competitively to the same cis-acting element(s). Further expression analysis using RT-PCR to amplify cDNA portions corresponding to ORFs of both Knox7 alternative transcripts showed that seven cDNA clones were probably generated by alternative splicing of Knox7L. Alignment of these sequences showed that they are in frame suggesting the existence of the corresponding proteins. Quantitative RT-PCR experiments indicated that Knox7S and Knox7L were expressed in maize embryos during germination. In the same tissue, expression of Knox7S was stimulated by light and ABA and inhibited by GA, two hormones that control germination process.
Assuntos
Processamento Alternativo/genética , Genes Homeobox , Proteínas de Homeodomínio/genética , Sítio de Iniciação de Transcrição , Zea mays/genética , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar , Éxons , Regulação da Expressão Gênica , Proteínas de Homeodomínio/química , Íntrons , Dados de Sequência Molecular , Filogenia , Isoformas de Proteínas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Zea mays/embriologiaRESUMO
The Asp-derived amino acid pathway has been studied during the early stages of development in two maize genotypes, Io and F2, differing in germination efficiency and post-germination growth. In both genotypes expression of Ask2 (monofunctional Asp-kinase-2), Akh1 and Akh2 (bifunctional Asp-kinase-homo-Ser dehydrogenase-1 and 2), increased throughout germination and post-germination growth, suggesting a developmental regulation, whereas Ask1 (monofunctional Asp-kinase-1) was expressed constitutively. The major difference between Io and F2 concerned genes encoding bifunctional enzymes, particularly Akh2, the expression of which was dramatically low in F2. 15N-Asp labelling showed differences in in vivo Asp-kinase activities between the genotypes studied. Asp flux through the Met/Thr branches was higher in Io than in F2, while the latter exhibited a higher flux of Asp through the Lys branch. Physiological results, together with the higher Akh2 expression in Io, suggest that bifunctional enzyme activity, favourable to Met/Thr, was higher in Io than in F2 and that the monofunctional pathway was boosted in F2 because of the lower competition by the bifunctional pathway, thus allowing for higher flux of Asp through the Lys branch. In conclusion, it is suggested that F2 germination and post-germination growth might have been partially inhibited due to a limitation in Met and Thr availability. A negative physiological effect related to Lys accumulation in F2 is also discussed.
