Assessment of chemical, biological and immunological properties of "Damiana de California" Turnera diffusa Willd extracts in Longfin yellowtail (Seriola rivoliana) leukocytes.

In Mexican herbal medicines or natural remedies, Turnera diffusa (Turneraceae) known as "Damiana de California", has ethnopharmacological relevance, including aphrodisiac, diuretic, and antimicrobial activities. To explore the immunological effect of infusion and methanolic extracts from Damiana de California, this study investigated its chemical, biological, antimicrobial and immunological properties in Longfin yellowtail Seriola rivoliana leukocytes. The analysis of chemical compounds revealed a considerable level of total phenolic and flavonoid contents in the infusion compared with methanolic extract. Furthermore, the antioxidant activity showed high hydroxyl radical scavenging activity in infusion extract compared with BHT positive control. Superoxide radical scavenging activity and ion chelation were higher in methanolic extract followed by infusion treatment. Interestingly, notable antimicrobial activity was observed in both extracts of T. diffusa against Vibrio parahaemolyticus. An in vitro study was performed using leukocytes of S. rivoliana treated with infusion or methanolic extracts at 12.5, 25 and 50 µg/mL for 24 h. Remarkably, infusion extract induced proliferation at any concentration but not the methanolic extract, which was diminished in a dose-dependent fashion. The immunostimulation study demonstrated that the phagocytosis activity increased in those leukocytes stimulated with methanolic extract but diminished the respiratory burst activity, in contrast to the activity observed in those leukocytes stimulated with infusion treatment. Finally, leukocytes incubated with the extracts and confronted with V.parahaemolyticus up-regulated the transcription of proinflammatory cytokine IL-1ß gene in a dose response relationship. These findings suggest that the infusion treatment has potential therapeutic properties, promoting the antioxidant capacity and enhancing immune parameters in Longfin yellowtail S. rivoliana.

Pollen morphology of four species of Jatropha (Euphorbiaceae), including toxic varieties, in Northwestern Mexico / Morfología del polen de cuatro especies de Jatropha (Euphorbiaceae), incluyendo variedades tóxicas, en el noroeste de México

AbstractJatropha curcas is a native Mexican plant, distributed in the forests of coastal regions; it has shown potential for the production of biofuel, and its raw protein can be used as animal feed. Nevertheless, its current varieties have low yield and production variability, as it is in domestication period. The knowledge of the pollen grains is fundamental for studies of Jatropha reproductive biology and breeding for genetic improvement to generate new hybrids and improve fruit and seed yield. This study compared pollen morphology of four Jatropha species, which include two varieties of J. curcas (one toxic from India and one non-toxic from Mexico), and two varieties of wild J. cinerea (Sinaloa and Baja California Sur); additionally, pollen was collected from wild J. platyphylla (Sinaloa) and J. vernicosa (Baja California Sur) to characterize them palynologically. We used a scanning electron microscope to describe the size and shape of pollen grains. Pollen grains were observed spheroidal. The diameter of the varieties of toxic and non-toxic J. curcas and that of J. platyphylla were 58 ± 2.3, 54 ± 1.7, and 51 ± 2.4 µm, respectively; the number of clavae was 84 ± 10, 108 ± 15, and 180 ± 15.5, respectively. Pollen grains of J.cinerea (Sinaloa) had a diameter of 50 ± 1.7 µm and 220 ± 14 clavae by area; J. cinerea (Baja California Sur) had a diameter of 45 ± 1.9 µm and 195 ± 14.7 clavae. Pollen grains of J. vernicosa had a diameter of 46 ± 2.1 µm and 231 ± 25.6 clavae. The ornamentation of the exine showed clava with capita gemma shaped for varieties of J.curcas species and clavum shaped for J. platyphylla, J. cinerea and J. vernicosa species. The studied varieties have a cross-linked endexine and ectexine intectate. Luminal bacules were observed in the reticular space of J. curcas varieties while none were present in the varieties of J. cinerea, J. platyphylla, and J. vernicosa.

ResumenJatropha curcas es nativa de México, y se distribuye en los bosques de las regiones costeras; las plantas han mostrado potencial para la producción de biodiesel, y la proteína cruda puede utilizarse para alimentación animal. Sin embargo, sus variedades actuales tienen bajo rendimiento y variabilidad en la producción, por estar en periodo de domesticación. El conocimiento de los granos de polen es fundamental para el estudio de la biología reproductiva de Jatropha y mejoramiento genético para generar nuevos híbridos y mejorar el rendimiento de frutos y semillas. Este estudio compara la morfología del polen de cuatro especies de Jatropha, que incluyen dos variedades de J. curcas, (una tóxica de India y una no tóxica de México) y dos variedades silvestres de J. cinerea (Sinaloa y Baja California Sur), por último, se obtuvo polen de J. platyphylla (Sinaloa) y J. vernicosa (Baja California Sur) silvestres para caracterizarlos palinológicamente. Para describir el tamaño y la forma de los granos de polen, se utilizó un microscopio electrónico de barrido. Los granos de polen tienen forma esferoidal. El diámetro de las variedades de J. curcas tóxica y no tóxica, y J. platyphylla fueron 58 ± 2.3, 54 ± 1.7, y 51 ± 2.4 micras, respectivamente; el número de clavas fue de 84 ± 10, 108 ± 15 y 180 ± 15.5, respectivamente. Los granos de polen de J. cinerea (Sinaloa) tienen un diámetro de 50 ± 1.7 micras y 220 ± 14 clavas por zona; J. cinerea (Baja California Sur) tiene un diámetro de 45 ± 1.9 micras y 195 ± 14.7 clavas. Los granos de polen de J. vernicosa tienen un diámetro de 46 ± 2.1 micras y 231 ± 25.6 clavas. La ornamentación de la exina mostró clavas con cápita en forma de gema y pequeños báculos en el lumen para ambas J. curcas y en forma de clavum para J. cinerea, J. platyphylla y J. vernicosa y no presentan báculos en el lumen. Las variedades estudiadas tienen una endexina reticulada y ectexina intectada.

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.

Antioxidant responses of damiana (Turnera diffusa Willd) to exposure to artificial ultraviolet (UV) radiation in an in vitro model; part I; UV-C radiation / Respuesta antioxidante en damiana (turnera diffusa willd) expuesta a radiación ultravioleta (UV) artificial en un modelo in vitro; parte I; radiación UV-C

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) plantsin 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 (AU)

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 enzimá- ticas, 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 (AU)

Antioxidant responses of damiana (Turnera diffusaWilld) to exposure to artificial ultraviolet (UV) radiation in an in vitro model; part II; UV-B radiation / Respuesta antioxidante en damiana (turnera diffusa willd) expuesta a radiación ultravioleta (UV) artificial en un modelo in vitro; parte II; radiación UV-B

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 (AU)

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 (AU)

Salinity effects on protein content, lipid peroxidation, pigments, and proline in Paulownia imperialis (Siebold & Zuccarini) and Paulownia fortunei (Seemann & Hemsley) grown in vitro

We evaluated the effects of saline stress on soluble proteins, lipid peroxidation (TBAR), chlorophyll a, chlorophyll b, beta-carotene, violaxanthin, and proline in Paulownia imperialis and Paulownia fortunei plants grown in vitro. When the propagated plants reached a determined size, they were transferred aseptically to WPM culture medium containing different sodium chloride concentrations (0, 20, 40, 60, 80, and 160 mM) and were sampled at 15 and 30 days. Proline content was determined at 30 days after transfer only. Protein concentration significantly decreased with the highest salt levels in P. imperialis compared to controls in which no sodium chloride was added. In both P. imperialis and P. fortunei, lipid peroxidation significantly increased at 15 days but decreased at 30 days. Chlorophyll a, chlorophyll b, beta-carotene, and violaxanthin significantly decreased with exposure to higher sodium chloride concentrations at 15 and 30 days in both species. Proline content in P. imperialis significantly increased in plants grown in 20 and 40 mM of sodium chloride and decreased in higher sodium chloride concentrations. In P. fortunei, this measure significantly decreased proline content at all salt concentrations in plants exposed to all levels of sodium chloride compared to controls. Our results show that P. imperialis is more tolerant to salt stress at the salinity conditions tested.

Analysis of essential oils from wild and micropropagated plants of damiana (Turnera diffusa).

Damiana is a medicinal plant with many traditional uses and a reputation as an aphrodisiac. Essential oils produced by this plant are used in traditional medicine, and for the preparation of liquors and tea. The composition of essential oils from wild damiana, plants grown with micropropagated methods involving cell suspension, and explants in solid medium, is presented. Relevant differences are observed in oils coming from wild and micropropagated plants, where micropropagated plants being more uniform with respect to quality and quantity. The most abundant constituents of the oils were caryophyllene oxide, caryophyllene, delta-cadinene, elemene and 1,8-cineol.