Pistil Mating Type and Morphology Are Mediated by the Brassinosteroid Inactivating Activity of the S-Locus Gene BAHD in Heterostylous Turnera Species.

Heterostyly is a breeding system that promotes outbreeding through a combination of morphological and physiological floral traits. In Turnera these traits are governed by a single, hemizygous S-locus containing just three genes. We report that the S-locus gene, BAHD, is mutated and encodes a severely truncated protein in a self-compatible long homostyle species. Further, a self-compatible long homostyle mutant possesses a T. krapovickasii BAHD allele with a point mutation in a highly conserved domain of BAHD acyl transferases. Wild type and mutant TkBAHD alleles were expressed in Arabidopsis to assay for brassinosteroid (BR) inactivating activity. The wild type but not mutant allele caused dwarfism, consistent with the wild type possessing, but the mutant allele having lost, BR inactivating activity. To investigate whether BRs act directly in self-incompatibility, BRs were added to in vitro pollen cultures of the two mating types. A small morph specific stimulatory effect on pollen tube growth was found with 5 µM brassinolide, but no genotype specific inhibition was observed. These results suggest that BAHD acts pleiotropically to mediate pistil length and physiological mating type through BR inactivation, and that in regard to self-incompatibility, BR acts by differentially regulating gene expression in pistils, rather than directly on pollen.

Evolutionary interplay between sister cytochrome P450 genes shapes plasticity in plant metabolism.

Expansion of the cytochrome P450 gene family is often proposed to have a critical role in the evolution of metabolic complexity, in particular in microorganisms, insects and plants. However, the molecular mechanisms underlying the evolution of this complexity are poorly understood. Here we describe the evolutionary history of a plant P450 retrogene, which emerged and underwent fixation in the common ancestor of Brassicales, before undergoing tandem duplication in the ancestor of Brassicaceae. Duplication leads first to gain of dual functions in one of the copies. Both sister genes are retained through subsequent speciation but eventually return to a single copy in two of three diverging lineages. In the lineage in which both copies are maintained, the ancestral functions are split between paralogs and a novel function arises in the copy under relaxed selection. Our work illustrates how retrotransposition and gene duplication can favour the emergence of novel metabolic functions.

Antioxidant responses of damiana (Turnera diffusa Willd) to exposure to artificial ultraviolet (UV) radiation in an in vitro model; part I; UV-C radiation.

INTRODUCTION: Ultraviolet type C (UV-C) radiation has higher energy than the UV-B radiation and has been less studied because it is completely absorbed by the ozone layer. However, artificial UV-C radiation can generate diverse modifications in the plants. Given that exposure to UV-C for short periods of time increases the antioxidant content, improving the appearance and shelf-life of products, its potential application in postharvest treatments to modify the antioxidant content of medicinal plants, such as damiana (Turnera diffusa), is novel and relevant. OBJECTIVE: To determine the effects of UV-C radiation on enzymatic and non-enzymatic antioxidant defenses, as well as oxidative damage levels, in damiana (Turnera diffusa) plants in vitro. RESULTS: UV-C radiation decreased superoxide dismutase (SOD, EC 1.15.1.1) and total peroxidases (POX, EC 1.11.1) activities, the concentration of chlorophylls (a and b), carotenes, vitamin C, and total antioxidant capacity. UV-C radiation increased the phenolic compound levels in damiana. Loss of antioxidant defenses was higher in damiana plants exposed to higher UV-C doses and/or for longer periods. This study suggests that UV-C radiation induces oxidative stress, evidenced as increased protein carbonyls and phenolic compound content, in damiana (T. diffusa). CONCLUSION: Low dose, short exposure to UV-C stimulates phenolic compound content in damiana. Thus, controlled UV-C treatments could be used as postharvest treatment to increase phenolic compound content in damiana plants.

Introducción: La radiación ultravioleta tipo C (UV-C) presenta mayor energía y es menos estudiada que la radiación UV-B, debido a que se considera que es totalmente absorbida por la capa de ozono. Sin embargo, la radiación UV-C artificial es capaz de generar diversas modificaciones en las plantas. Dado que la exposición a UV-C por intervalos de tiempo cortos incrementa la concentración de compuestos antioxidantes, mejorando la apariencia y vida de anaquel de los productos, su potencial aplicación en tratamientos poscosecha para modificar el contenido antioxidante de plantas medicinales, como la damiana (Turnera diffusa), es novedoso y relevante. Objetivo: Determinar el efecto de la radiación UV-C sobre las defensas antioxidantes enzimáticas y no enzimáticas, así como en los niveles de daño oxidativo de damiana (Turnera diffusa) in vitro. Resultados: La radiación UV-C disminuyó la actividad de las enzimas superóxido dismutasa (SOD, EC 1.15.1.1) y peroxidasas totales (POX, CE 1.11.1), la concentración de clorofila (a y b), carotenos, vitamina C y la capacidad antioxidante total, e incrementó el contenido de compuestos fenólicos en damiana. La disminución de las defensas antioxidantes fue mayor en plantas de damiana expuestas a dosis más altas de UV-C o por períodos más largos. Estos resultados sugieren que la radiación UV-C induce estrés oxidativo, evidenciado por el incremento del contenido de carbonilos proteicos y el contenido de compuestos fenólicos en damiana (T. diffusa). Conclusión: Dosis bajas y menor exposición a UV-C estimulan la síntesis de compuestos fenólicos en damiana. Por ello, tratamientos controlados con UV-C podrían emplearse como tratamientos poscosecha para incrementar el contenido de compuestos fenólicos en plantas de damiana.

Antioxidant responses of damiana (Turnera diffusa Willd) to exposure to artificial ultraviolet (UV) radiation in an in vitro model; part ii; UV-B radiation.

INTRODUCTION: Ultraviolet type B (UV-B) radiation effects on medicinal plants have been recently investigated in the context of climate change, but the modifications generated by UV-B radiation might be used to increase the content of antioxidants, including phenolic compounds. OBJECTIVE: To generate information on the effect of exposure to artificial UV-B radiation at different highdoses in the antioxidant content of damiana plants in an in vitro model. METHODS: Damiana plantlets (tissue cultures in Murashige- Skoog medium) were irradiated with artificial UV-B at 3 different doses (1) 0.5 ± 0.1 mW cm-2 (high) for 2 h daily, (2) 1 ± 0,1 mW cm-2 (severe) for 2 h daily, or (3) 1 ± 0.1 mW cm-2 for 4 h daily during 3 weeks. The concentration of photosynthetic pigments (chlorophylls a and b, carotenoids), vitamins (C and E) and total phenolic compounds, the enzymatic activity of superoxide dismutase (SOD, EC 1.15.1.1) and total peroxidases (POX, EC 1.11.1), as well as total antioxidant capacity and lipid peroxidation levels were quantified to assess the effect of high artificial UV-B radiation in the antioxidant content of in vitro damiana plants. RESULTS: Severe and high doses of artificial UV-B radiation modified the antioxidant content by increasing the content of vitamin C and decreased the phenolic compound content, as well as modified the oxidative damage of damiana plants in an in vitro model. CONCLUSION: UV-B radiation modified the antioxidant content in damiana plants in an in vitro model, depending on the intensity and duration of the exposure.

Introducción: Los efectos de la radiación ultravioleta tipo B (UV-B) sobre las plantas medicinales se han investigado recientemente en el contexto del cambio climático, pero las modificaciones que genera la radiación UV-B podrían emplearse para modificar el contenido de compuestos antioxidantes, incluyendo los compuestos fenólicos. Objetivo: Generar información sobre el efecto de una alta exposición a UV-B artificial en el contenido antioxidante de damiana (Turnera diffusa, Willd) en un modelo in vitro. Método: Plántulas de damiana en cultivo de tejidos (medio Murashige-Skoog) fueron irradiadas con UV-B artificial en 3 diferentes dosis: (1) 0,5 ± 0,1 mW cm-2 (alto) por 2 h diarias, (2) 1 ± 0,1 mW cm-2 (severa) por 2 h diarias, o (3) 1 ± 0,1 mW cm-2 durante 4 horas diarias por 3 semanas. Se cuantificó la concentración de pigmentos fotosintéticos (clorofilas a y b, carotenoides), vitaminas (C y E) y compuestos fenólicos totales, la actividad enzimática de la superóxido dismutasa (SOD, EC 1.15.1.1) y las peroxidasas totales (POX, EC 1.11.1), así como la capacidad antioxidante total y la peroxidación de lípidos para evaluar el efecto de la alta radiación UV-B artificial en el contenido antioxidante de damiana in vitro. Resultados: Dosis altas y severas de radiación UV-B artificial modificaron el contenido antioxidante incrementando el contenido de vitamina C y disminuyendo el contenido de compuestos fenólicos totales, además de modificar el daño oxidativo de plantas de damiana en un modelo in vitro. Conclusión: La radiación UV-B modifica el contenido antioxidante en damiana en un modelo in vitro, dependiendo de la intensidad y el tiempo de exposición.

Characterization and anti-staphylococcal activity of the essential oil from Turnera subulata Sm

The present study is pioneer in analyzing the chemical composition of the essential oil from aerial parts of Turnera subulata Sm. and evaluates their antibacterial activity against a panel of drug-resistant strains of Staphylococcus aureus. The aerial parts were kiln-dried and then powdered in mechanical mill, and was subjected to hydrodistillation in a Clevenger-type apparatus. The components were analyzed using a mass spectrometrycoupled gas chromatography (GC-MS), and the identification of substances was performed by comparison of the mass spectra obtained with the mass spectra of the database of the GC-MS and retention indices . The essential oil was evaluated using the method of broth dilution at concentrations ranging from 3.125µg/mL to 3200µg/mL. It was possible to identify 45 substances (92.1%) of the essential oils, and the major components were trans-caryophyllene (6.7%), citronellol (5.6%), sphatulenol (5.3%), α-cadinol (4.3%), n-tricosano (4.3%), geraniol (4.1%) and trans-geranilacetone (3.7%), n-pentacosano (3.5%), globulol (3.4%), caryophyllene oxide (3.2%). The essential oil of T. subulata Sm. showed effective antibacterial activity for the various strains of S. aureus tested, with the MIC values between 25 µg/mL and 1600 µg/mL. The study of the essential oil of Turnera subulata Sm. showed that it consists of a complex mixture of several classes of compounds having the sesquiterpenes as major constituents followed by monoterpenes, and showed an antibacterial activity significant front the strains tested.

O presente estudo é pioneiro em analisar a composição química dos óleos essenciais das partes aéreas de Turnera subulata Sm. e sua atividade antibacteriana frente à Staphylococcus aureus resistentes a antibióticos. As partes aéreas da planta foram secas em estufa, pulverizadas em moinho mecânico e submetidas à hidrodestilaçao em aparato tipo Clevenger. A composição dos óleos essenciais foi analisada por cromatografia gasosa acoplada a espectrometria de massas (CG-EM), e a identificação realizada por comparação dos espectros de massas com a biblioteca do CG-EM e índices de retenção. A atividade antimicrobiana do óleo essencial foi avaliada usando o método de diluição em caldo, em concentrações que variaram de 3.125µg/mL to 3200µg/mL. Foi possível identificar 45 substâncias (92,1%) do óleo essencial, sendo os componentes majoritários: trans-cariofileno (6,7%), citronelol (5,6%), espatulenol (5,3%), α-cadinol (4,3%), n-tricosano (4,3%), geraniol (4,1%) e trans-geranilacetona (3,7%), n-pentacosano (3,5%), globulol (3,4%) e óxido de cariofileno (3,2%). O óleo essencial de T. subulata Sm. mostrou atividade antibacteriana eficaz para as várias cepas de S. aureus testadas, com valores de CIM entre 25 µg/mL e 1600 µg/mL. O estudo do óleo essencial de Turnera subulata Sm. evidenciou sua complexa mistura, contendo várias classes de substâncias, tendo os sesquiterpenos como constituintes majoritários seguido dos monoterpenos, e mostrou significativa atividade antibacteriana frente as cepas testadas.

Evaluation of thin-layer chromatography methods for quality control of commercial products containing Aesculus hippocastanum, Turnera diffusa, Matricaria recutita, Passiflora incarnata, and Tilia occidentalis.

In Mexico, plant-derived products with health claims are sold as herbal dietary supplements, and there are no rules for their legal quality control. Aesculus hippocastanum, Turnera diffusa, Matricaria recutita, Passiflora incarnata, and Tilia occidentalis are some of the major commercial products obtained from plants used in this region. In this paper, we describe the effectiveness of thin-layer chromatography methods to provide for the quality control of several commercial products containing these plants. Standardized extracts were used. Of the 49 commercial products analyzed, only 32.65% matched the chromatographic characteristic of standardized extracts. A significant number of commercial products did not match their label, indicating a problem resulting from the lack of regulation for these products. The proposed methods are simple, sensitive, and specific and can be used for routine quality control of raw herbals and formulations of the tested plants. The results obtained show the need to develop simple and reliable analytical methods that can be performed in any laboratory for the purpose of quality control of dietary supplements or commercial herbal products sold in Mexico.

Turnera ulmifolia L. (Turneraceae): preliminary study of its antioxidant activity.

Turnera ulmifolia L. is used in Brazilian folk medicine as an anti-inflammatory. Since this activity may be correlated with the presence of antioxidant compounds, a leaf extract was evaluated for its radical scavenging capacity (RSC). The in vitro RSC of a 50% hydroethanolic (HE) extract was evaluated by beta-carotene/linoleic acid coupled oxidation system for the inhibition of oxidation and the lipid peroxidation inhibition in rat brain homogenates, using thiobarbituric acid reactive substances (TBARS) and chemiluminescence (CL). Results indicated, through peroxidation suppression, that this extract exhibited greater antioxidative activity (77.4% +/- 10%) than alpha-tocopherol (58.4% +/- 3.7%). TBARS and CL inhibition was concentration-dependent and Q(1/2) values were 8.2 and 6.0 microg/mL for TBARS and CL, respectively. For alpha-tocopherol these values were 7.1 microg/mL (TBARS) and 9.8 microg/mL (CL). Phenolic compounds may be responsible for this antioxidant capacity.