Highly abundant and stage-specific mRNAs in the obligate pathogen Bremia lactucae.
Germinating spores of the obligate pathogen Bremia lactucae (lettuce downy mildew) contain several unusually abundant species of mRNA. Thirty-nine cDNA clones corresponding to prevalent transcripts were isolated from a library synthesized using poly(A)+ RNA from germinating spores; these clones represented only five distinct classes. Each corresponding mRNA accounted for from 0.4 to 9 percent by mass of poly(A)+ RNA from germinating spores and together represented greater than 20 percent of the mRNA. The expression of the corresponding genes, and a gene encoding Hsp70, was analyzed in spores during germination and during growth in planta. The Hsp70 mRNA and mRNA from one abundant cDNA clone (ham34) were expressed constitutively. Two clones (ham9 and ham12) hybridized only to mRNA from spores and germinating spores. Two clones (ham37 and ham27) showed hybridization specific to germinating spores. Quantification of the number of genes homologous to each cDNA clone indicated that four clones corresponded to one or two copies per haploid genome, and one hybridized to an approximately 11-member family of genes. A sequence of the gene corresponding to ham34 was obtained to investigate its function and to identify sequences conferring high levels of gene expression for use in constructing vectors for the transformation of B. lactucae.
Judelson
H S
HS
Department of Vegetable Crops, University of California, Davis 95616.
Michelmore
R W
RW
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal; D005800:Genes, Fungal; D015698:Genomic Library; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009868:Oomycetes / Q000235:genetics*; D010944:Plants / Q000382:microbiology; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000235:genetics*; D014170:Transformation, Genetic
D004271:DNA, Fungal; D012331:RNA, Fungal