RESUMO
Patchouli alcohol, a significant bioactive component of the herbal plant Pogostemon cablin, has considerable medicinal and commercial potential. Several genes and transcription factors involved in the biosynthesis pathway of patchouli alcohol have been identified. However, so far, regulatory factors directly interacting with patchouli synthase (PTS) have not been reported. This study was conducted to analyze the interaction between PcENO3 and PcPTS to explore the molecular regulation effect of PcENO3 on patchouli alcohol biosynthesis. PcENO3, a homologous protein of Arabidopsis ENO3 belonging to the enolase family, was identified and characterized. Subcellular localization experiments in Arabidopsis protoplast cells indicated that the PcENO3 protein was localized in both the cytoplasm and nucleus. The physical interaction between PcENO3 and PcPTS was confirmed through yeast two-hybrid (Y2H), GST pull-down, and bimolecular fluorescence complementation assays. Furthermore, the Y2H assay demonstrated that PcENO3 could also interact with JAZ proteins in the JA pathway. Enzymatic assays showed that the interaction with PcENO3 increased the catalytic activity of patchoulol synthase. Additionally, suppression of PcENO3 expression with VIGS (virus-induced gene silencing) decreased patchouli alcohol content compared to the control. These findings suggest that PcENO3 interacts with patchoulol synthase and modulates patchoulol biosynthesis by enhancing the enzymatic activity of PcPTS.
Assuntos
Arabidopsis , Pogostemon , Sesquiterpenos , Pogostemon/genética , Pogostemon/metabolismo , Arabidopsis/metabolismo , Sesquiterpenos/metabolismoRESUMO
Pogostemon cablin (Blanco) Benth., comumente conhecido como patchouli, é uma espécie aromática utilizada pela indústria de perfumes devido a fragrância amadeirada e propriedade fixadora do óleo essencial. Fatores que afetam o teor dos constituintes do óleo essencial devem ser avaliados visando obter matéria prima de melhor qualidade. O experimento foi conduzido em casa de vegetação com delineamento inteiramente casualizado em esquema fatorial 2x3 e 4 repetições, sendo dois níveis de giberelina (0 e 200 mg L-1) e três níveis de extrato de alga marinha (0, 15 e 30 mg L-1). A aplicação dos reguladores vegetais foi realizada 30 dias após o plantio das mudas, e a avaliação do experimento deu-se 45 dias após a aplicação dos tratamentos. Não houve diferença significativa para a altura das plantas, a concentração de extrato de alga marinha de 15 mg L-1 promoveu aumento no número de folhas em relação à testemunha e a concentração de 30 mg L-1 promoveu decréscimo. Em relação à área foliar e ao número de folhas houve relação inversa para os níveis de extrato de alga marinha, onde o tratamento com maior concentração do regulador resultou em aumento da área foliar e redução do número de folhas. O teor de óleo essencial foi superior após a aplicação de 15 mg L-1 de extrato alga marinha isoladamente ou combinado com GA3. A produtividade do óleo essencial também aumentou com a aplicação 15 mg L-1 de extrato de alga marinha e quando utilizado somente GA3. A utilização de extrato de alga marinha aumentou a concentração de patchoulol no óleo essencial.
Pogostemon cablin (Blanco) Benth., commonly known as patchouli, is an aromatic species used by the perfume industry due to its woody fragrance and the fixative characteristic of its essential oil. Factors affecting the content of the constituents of essential oils should be evaluated in order to obtain raw materials with better quality. The experiment was carried out in a greenhouse in a completely randomized 2x3 factorial arrangement and four replications, with two levels of gibberellin (0 and 200 mg L-1) and three levels of seaweed extract (0, 15 and 30 mg L-1). Plant regulators were applied thirty days after planting the seedlings; the experiment was evaluated forty-five days after the treatments. There was no significant difference in plant height; the 15 mg L-1 concentration of seaweed extract promoted an increase in the number of leaves in relation to the control; and the 30 mg L-1 concentration promoted a decrease. In relation to leaf area and leaf number, there was an inverse relationship for the seaweed extract levels, in which the treatment with higher a concentration of the regulator resulted in an increased leaf area, reducing the number of leaves. The essential oil content was higher after applying 15 mg L-1 of seaweed extract alone or combined with GA3. The yield of essential oil also increased by applying 15 mg L-1 of seaweed extract and when only GA3 was used. The use of seaweed extract increased the patchoulol concentration in the essential oil.