RESUMO
A Zea mays cDNA, MZm3-4, was isolated by differential screening of a cDNA library obtained from meiotic stage anthers against a cDNA of 3-week-old seedlings. Northern blot analysis of RNA from different maize tissues and from male reproductive organs at various developmental stages demonstrated expression of a single transcript in anthers, from the pollen mother cell stage through the uninucleated microspore stage. In situ hybridization to anther sections resulted in a distinct signal only in the tapetum. The MZm3-4 cDNA is 743 nucleotides in length and has an open reading frame encoding a protein of 75 amino acids. Sequence comparisons with various databases revealed that MZm3-4 exhibits high similarities with type 2 plant metallothioneins at both the nucleotide and the amino-acid level. Primer extension analysis indicated that MZm3-4 cDNA is deleted of 13bp at the 5' end. Southern blot analysis showed that the MZm3-4 gene may be present in one or two copies in a Z. mays inbred line genome. This is the first report of the isolation of a type 2 metallothionein-like protein in maize. Moreover, the expression of this type 2 metallothionein-like gene is high in the male reproductive organs engaged in microsporogenesis.
Assuntos
Genes de Plantas/genética , Metalotioneína/genética , Proteínas de Plantas , Zea mays/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Southern Blotting , DNA Complementar/química , DNA Complementar/genética , DNA Complementar/isolamento & purificação , DNA de Plantas/genética , Dosagem de Genes , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Pólen/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Distribuição Tecidual , Transcrição Gênica , Zea mays/crescimento & desenvolvimentoRESUMO
A cDNA, MZm3-3, was isolated by differential screening of a cDNA library from Zea mays meiotic stage anthers against cDNA of 3-week-old seedlings. Characterization of this cDNA indicated that the MZm3-3 gene is expressed specifically during male gametogenesis. Its expression is highly and preferentially detected in the tapetum, from the pollen mother cell to uninucleated microspore stages. It encodes a short alkaline protein of 10.6 kDa, with a conserved pattern of eight cysteine residues. Sequence analysis showed that these features are shared with lipid transfer proteins and some male-flower-specific proteins. The presence of a putative signal peptide indicates that MZm3-3 enters into the secretory pathway to then be released into the anther loculus. Based on these features, the secretory activity of the tapetum and the temporal expression pattern of MZm3-3, a contribution to pollen coat formation is suggested. Southern blot analyses demonstrated the presence of closely related genes, indicating that MZm3-3 belongs to a multigene family.