RESUMO
Ethylene is a gaseous plant hormone involved in many physiological processes including senescence, fruit ripening, and defence. Here the effects of pollination and wound-induced ethylene signals on transcript accumulation of benzoyl CoA:benzyl alcohol/phenylethanol benzoyltransferase (PhBPBT) are shown in Petuniaxhybrida cv. Mitchell 'Diploid' (MD). In petunia, PhBPBT is responsible for the biosynthesis of both benzyl benzoate and phenylethyl benzoate from benzyl alcohol and phenylethanol, respectively. RNAi-silenced lines, with reduced PhBPBT transcript, displayed reduced benzyl benzoate emission, and increased benzyl alcohol levels. Detailed expression analysis showed that PhBPBT is regulated by both light and an endogenous circadian rhythm, while it is also differentially regulated in response to ethylene in a tissue-specific manner. Twenty-four hours following pollination of MD flowers, expression of PhBPBT decreases in the corolla, while it increases in the ovary after 48 h. This is caused by ethylene that is emitted from the flower coinciding with fertilization as this is not observed in transgenic ethylene-insensitive plants (CaMV35S::etr1-1; 44568). Ethylene is also emitted from vegetative tissue of petunia following mechanical wounding, resulting in an increase in PhBPBT expression in the leaves where expression is normally below detection levels. Indicative of this pattern of expression, we hypothesize that PhBPBT and subsequent benzyl benzoate production is involved in defence-related processes in the corolla prior to pollination, in the ovary immediately following fertilization, and in vegetative tissue in response to wounding.
Assuntos
Etilenos/metabolismo , Flores/metabolismo , Petunia/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Benzoatos/metabolismo , Ritmo Circadiano/fisiologia , Regulação da Expressão Gênica de Plantas , Luz , Petunia/genética , Proteínas de Plantas/genética , Polinização/fisiologia , Interferência de RNARESUMO
Petunia flower petals emit large amounts of isoeugenol, which has been shown to be synthesized by isoeugenol synthase (PhIGS1) from an ester of coniferyl alcohol, hypothesized to be coniferyl acetate. This paper describes the identification and characterization of a novel petunia gene encoding an enzyme belonging to the BAHD acyltransferase family whose expression correlates with isoeugenol biosynthesis. RNAi suppression of this gene results in inhibition of isoeugenol biosynthesis. Biochemical characterization of the protein encoded by this gene showed that it has acetyltransferase activity and is most efficient with coniferyl alcohol among the alcohol substrates tested. Overall, these data support the conclusion that coniferyl acetate is the substrate of isoeugenol synthase.
Assuntos
Acetiltransferases/metabolismo , Eugenol/análogos & derivados , Flores/enzimologia , Petunia/enzimologia , Acetiltransferases/genética , Sequência de Aminoácidos , DNA Complementar/química , DNA Complementar/genética , Eugenol/química , Eugenol/metabolismo , Flores/genética , Flores/metabolismo , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Estrutura Molecular , Petunia/genética , Petunia/metabolismo , Plantas Geneticamente Modificadas , Interferência de RNA , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA , Especificidade por SubstratoRESUMO
In many flowering plants, such as petunia (Petunia x hybrida), ethylene produced in floral organs after pollination elicits a series of physiological and biochemical events, ultimately leading to senescence of petals and successful fertilization. Here, we demonstrate, using transgenic ethylene insensitive (44568) and Mitchell Diploid petunias, that multiple components of emission of volatile organic compounds (VOCs) are regulated by ethylene. Expression of benzoic acid/salicylic acid carboxyl methyltransferase (PhBSMT1 and 2) mRNA is temporally and spatially down-regulated in floral organs in a manner consistent with current models for post-pollination ethylene synthesis in petunia corollas. Emission of methylbenzoate and other VOCs after pollination and exogenous ethylene treatment parallels a reduction in PhBSMT1 and 2 mRNA levels. Under cyclic light conditions (day/night), PhBSMT mRNA levels are rhythmic and precede emission of methylbenzoate by approximately 6 h. When shifted into constant dark or light conditions, PhBSMT mRNA levels and subsequent methylbenzoate emission correspondingly decrease or increase to minimum or maximum levels observed during normal conditions, thus suggesting that light may be a more critical influence on cyclic emission of methylbenzoate than a circadian clock. Transgenic PhBSMT RNAi flowers with reduced PhBSMT mRNA levels show a 75% to 99% decrease in methylbenzoate emission, with minimal changes in other petunia VOCs. These results implicate PhBSMT1 and 2 as genes responsible for synthesis of methylbenzoate in petunia.
Assuntos
Etilenos/farmacologia , Flores/fisiologia , Petunia/fisiologia , Flores/efeitos dos fármacos , Flores/genética , Cinética , Dados de Sequência Molecular , Reguladores de Crescimento de Plantas/farmacologia , Pólen/fisiologiaRESUMO
Fifteen patients having total occlusion of the proximal left anterior descending coronary artery were studied. All fifteen had normal left ventricular anterior walls. The electrocardiogram was normal in eight patients; old inferior wall infarction was evident in one; anterior ischemia in five; and left anterior hemiblock in one. Collateral circulation was found in twelve patients (80%). These findings suggest that an angiographically non-visualized left anterior descending (LAD) vessel is patent and bypassable in patients where the anterior ventricular wall is intact, thus representing an additional criterion for aortocoronary bypass surgery.
