RESUMO
Pch313 was isolated as a cDNA whose RNA accumulated during the softening period of peach (Prunus persica L. Batsch) fruit development. To better understand the role of the gene, we compared the amount of pch313-related RNA detected during fruit softening and tissue wounding between cultivars with different softening characteristics. The cultivar that softened faster, "Bailey," had a significantly higher amount of pch313-related RNA accumulate during softening than the slower-softening cultivar, "Suncrest." Pch313 was sequenced and found to be related to a tomato fruit cDNA clone, pTOM13, which has been shown to encode the ethylene-forming enzyme. The derived amino acid sequence of pch313 is 74 to 83% identical to the pTOM13-related sequences. A pch313-3' noncoding region probe was used to demonstrate that pch313 is related to both a wound-induced RNA transcript and the major fruit-softening transcript. The relationship of pch313 RNA accumulation and ethylene evolution was examined upon wounding and appeared to be both tissue and cultivar specific. When leaves were wounded, more pch313-related RNA was detected in Bailey and the rate of ethylene evolved was also higher in Bailey. When fruits were wounded, the levels of ethylene evolved were nearly identical but Suncrest accumulated more pch313-related RNA. Southern analysis of the DNA indicated a small number of related genes.
RESUMO
A cDNA library from ethephon-treated cucumber cotyledons (Cucumis sativus L. cv. Poinsett 76) was constructed. Two cDNA clones encoding putative peroxidases were isolated by means of a synthetic probe based on a partial amino acid sequence of a 33 kDa cationic peroxidase that had been previously shown to be induced by ethylene. DNA sequencing indicates that the two clones were derived from two closely related RNA species that are related to published plant peroxidase sequences. Southern analysis indicates that there are 1-5 copies in a haploid genome of a gene homologous to the cDNA clones. The deduced amino acid sequences are homologous with a tobacco (55% sequence identity), a horseradish (53%), a turnip (45%), and a potato (41%) peroxidase. The cloned sequences do not encode the 33 kDa peroxidase from which the original synthetic probe was been derived, but rather other putative peroxidases. An increase in the level of mRNA is evident by 3 hours after ethephon or ethylene treatment and plateaus by 15 hours.
Assuntos
DNA/genética , Peroxidases/genética , Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Sondas de DNA , Etilenos/farmacologia , Dados de Sequência Molecular , Compostos Organofosforados/farmacologia , Plantas/efeitos dos fármacos , Plantas/enzimologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Mapeamento por Restrição , Homologia de Sequência do Ácido NucleicoRESUMO
A tissue culture line of soybean cells (SB-1) is shown to have an unusual arrangement of mitochondrial genes. Fusion of the mitochondrial 5S gene to the distal end of an unidentified gene results in the abundant expression of an 800 nucleotide RNA with the 5S rRNA at its 5' end. Since only the fused 5S rRNA gene is found in this cell line, the functional 5S rRNA must arise by processing of the 800 nucleotide RNA or by intermittent termination of transcription. The 800 nucleotide transcript and its DNA arrangement are not detected at comparable levels in other soybean sources, including the parent plant of the SB-1 tissue culture line. The role of recombination in the origin of this gene fusion and in plant mitochondrial DNA in general is discussed.