Isolation of a type 2 metallothionein-like gene preferentially expressed in the tapetum in Zea mays

A Zea mays cDNA, MZm3-4, was isolated by differential screening of a cDNA l ibrary obtained from meiotic stage anthers against a cDNA of 3-week-old see dlings. Northern blot analysis of RNA from different maize tissues and from male reproductive organs at various developmental stages demonstrated expr ession 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 cDN A is 743 nucleotides in length and has an open reading frame encoding a pro tein of 75 amino acids. Sequence comparisons with various databases reveale d 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 13 bp at the 5' end. Southern blo t 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. (C) 2000 Elsevier Science B.V. All rig hts reserved.