RESUMO
A homology-based cloning strategy yielded a cDNA clone, designated Sd-cam, encoding calmodulin protein from Scoparia dulcis. The restriction digests of genomic DNA of S. dulcis showed a single hybridized signal when probed with the fragment of this gene in Southern blot analyses, suggesting that Sd-cam occurs as a sole gene encoding calmodulin in the plant. The reverse-transcription polymerase chain reaction analysis revealed that Sd-cam was appreciably expressed in leaf, root and stem tissues. It appeared that transcription of this gene increased transiently when the leaf cultures of S. dulcis were treated with methyl jasmonate and calcium ionophore A23187. These results suggest that transcriptional activation of Sd-cam is one of the early cellular events of the methyl jasmonate-induced responses of S. dulcis.
Assuntos
Calmodulina/genética , Calmodulina/metabolismo , Clonagem Molecular , Expressão Gênica , Scoparia/genética , Acetatos/farmacologia , Sequência de Aminoácidos , Southern Blotting , Calcimicina/farmacologia , Calmodulina/química , Células Cultivadas , Ciclopentanos/farmacologia , DNA Complementar , Ionóforos/farmacologia , Dados de Sequência Molecular , Oxilipinas , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/citologia , Folhas de Planta/genética , Raízes de Plantas/genética , Caules de Planta/genética , Estrutura Terciária de Proteína , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Scoparia/citologia , Transcrição GênicaRESUMO
Specific inhibitors of the MVA pathway (pravastatin) and the MEP pathway (fosmidomycin) were used to interfere with the biosynthetic flux which leads to the production of aphidicolin-like diterpene in leaf organ cultures of Scoparia dulcis. Treatment of leaf organs with fosmidomycin resulted in dose dependent inhibition of chlorophylls, carotenoids, scopadulcic acid B (SDB) and phytol production, and no effect on sterol production was observed. In response to the pravastatin treatment, a significant decrease in sterol and perturbation of SDB production was observed.
