Effects of VALD-3,a derivative of o-vanilla Schiff base ligand,on proliferation,migration and apoptosis of colorectal cancer cells and its mechanism / 中国生物制品学杂志

@#Objective To the effects of VALD-3,a derivative of o-vanilla Schiff base ligand,on proliferation,migration and apoptosis of colorectal cancer cells and evaluate its mechanism.Methods HT-29 and HCT116 cells were cultured in vitro,and the inhibitory effects of VALD-3(5,10,20 and 40 mg/L) on proliferation of the two kinds of cells were detected by MTT assay;The effects of VALD-3(10,20 and 40 mg/L) on the morphological changes of the cells were observed by inverted microscope,while the effects on the migration ability of HT-29 cells were detected by cell scratch test,and the effects on the apoptosis of HT-29 cells were detected by flow cytometry and Hoechst 33258 fluorescence staining;The effects of VALD-3(5,10,20 and 40 mg/L) on the expression of apoptosis-related proteins in HT-29 cells were detected by Western blot.Negative control groups were set up(with no VALD-3).Results Compared with the negative control group,the survival rates of HT-29 and HCT116 cells in 10,20 and 40 mg/L VALD-3 treated groups significantly decreased(t=7.717～2 006.148,each P <0.05);the number of HT-29 and HCT116 cells in 10,20 and 40 mg/L VALD-3groups decreased significantly with the increase of VALD-3 concentration,the cells appeared irregular morphology and gradually became round and smaller,and cell fragments increased;In 10,20 and 40 mg/L VALD-3 treated groups,the migration rate of HT-29 cell scratches decreased significantly(t=7.596～73.780,each P <0.01),the apoptosis rate increased significantly(t=7.092～8.057,each P <0.01),and the number of apoptotic cells increased significantly with strong bright blue fluorescence,chromatin concentration and nuclear fragmentation;The levels of cleaved caspase-3 and Bax protein in HT-29 cells treated with 5,10,20 and 40 mg/L VALD-3 significantly increased(t=2.998～24.901,each P <0.05),the level of Bcl-2 protein in 40 mg/L VALD-3 group decreased significantly(t=10.035,P <0.05),and the levels of cleaved caspase-8 in 20 and 40 mg/L VALD-3 group significantly increased(t=12.630 and 8.064 respectively,each P <0.01).Conclusion VALD-3 inhibited the proliferation and migration of colorectal cancer cells and induced apoptosis by regulating the expression of cleaved caspase-3,cleaved caspase-8,Bax and Bcl-2 proteins.

Vanilla bahiana Hoehne (Orchidaceae): studies on fruit development and new perspectives into crop improvement for the Vanilla planifolia group / Vanilla bahiana Hoehne (Orchidaceae): estudos sobre o desenvolvimento dos frutos e novas perspectivas para o melhoramento de culturas do grupo Vanilla planifolia

Abstract: Vanilla is the most well-known and economically important genus of Orchidaceae in the world. Vanilla bahiana is restricted to Brazil and its conservation status was recently assessed as 'Endangered'. The species is phylogenetically closely related to V. planifolia, whose pods are the main natural source of vanillin. The primary desirable traits for vanilla production are high fruit set and great resistance to abiotic stress. High temperatures (> 32ºC) and heavy rain favor fungal infection, and thus are indirectly responsible for fruit drop by V. planifolia in cultivation. The aim of the present study was to follow the process of pod ripening of V. bahiana in a restinga fragment in an Environmental Protection Area of Bahia State, and to highlight implications for vanilla crop improvement. Fieldwork was carried out from April 2016 to January 2017. A total of nine manual self-pollinations and nine manual cross-pollinations were successfully carried out. Fruit length, diameter and girth were recorded every two weeks. Fruit permanence in the infrutescence was measured as consecutive days from pollination. Descriptive statistics were calculated and Pearson correlation analysis performed among all attributes per pollination method. The influence of temperature and rainfall was also analyzed. Pod measurements and fruit permanence in the infrutescence were higher for cross-pollination than self-pollination. Higher rainfall results in fruit drop. The use of V. bahiana in breeding efforts for the genus is here strongly suggested and supported by previously published studies that allowed us to highlight several desirable traits: vanillin production, its wide geographical distribution and occurrence in a variety of habitats; substantial populations; resistance to conditions of drought and high luminosity and temperature; extensive blooming period; high number of flowers per raceme; possible synchronised flowering with V. planifolia, and fruiting throughout the year. These prospects are promising and should be further evaluated, not only for V. bahiana but for other vanilla crop wild relatives. Studies of pollination ecology can be used to set guidelines for the conservation of the V. planifolia group and to encourage the development of strategies to increase the production of fruit and, consequently, vanillin for its culinary uses and medical applications.

Resumo: Vanilla é o gênero mais bem conhecido e economicamente importante de Orchidaceae no mundo. A espécie Vanilla bahiana é restrita ao Brasil e seu estado de conservação foi recentemente avaliado como "Em Perigo". A espécie é filogeneticamente próxima de V. planifolia, cujos frutos são a principal fonte natural de vanilina. As principais características desejáveis para a produção de baunilha são o elevado número de frutos e grande resistência ao estresse abiótico. Altas temperaturas (> 32 º C) e fortes chuvas favorecem a infecção fúngica, sendo, portanto, indiretamente responsáveis pela queda de frutos de V. planifolia em cultivo. O objetivo do presente estudo foi acompanhar o processo de maturação do fruto de V. bahiana em um fragmento de restinga em uma Área de Proteção Ambiental do estado da Bahia e destacar as implicações para o melhoramento da cultura da baunilha. O trabalho de campo foi realizado de abril de 2016 a janeiro de 2017. Um total de nove autopolinizações manuais e nove polinizações cruzadas manuais foram realizadas com sucesso. O comprimento, o diâmetro e a circunferência dos frutos foram registrados a cada duas semanas. A permanência do fruto na infrutescência foi medida através de dias consecutivos a partir da polinização. Medidas estatísticas descritivas dos atributos morfométricos foram calculadas e a análise de correlação de Pearson foi realizada para verificar a associação entre os pares dos atributos por método de polinização. Também foi analisada a influência da temperatura e da precipitação pluviométrica. As medidas do fruto e a permanência do fruto na infrutescência foram maiores na polinização cruzada do que na autopolinização. O aumento dos índices pluviométricos resulta em queda de frutos. O uso de V. bahiana em esforços de melhoramento para o gênero é fortemente sugerido aqui e suportado por estudos publicados anteriormente, que nos permitiram destacar várias características desejáveis: a produção de vanilina; ampla distribuição geográfica e ocorrência em diversos domínios fitogeográficos; formação de populações substanciais; resistência às condições de seca, alta luminosidade e temperatura elevada; extenso período de floração; elevado número de flores por racemo; possível florescimento sincronizado com V. planifolia; e frutificação ao longo do ano. Essas perspectivas são promissoras e devem ser avaliadas não apenas para V. bahiana, mas também para outras espécies de Vanilla restritas ao Brasil. Estudos de ecologia de polinização podem ser usados para estabelecer diretrizes para a conservação do grupo V. planifolia e encorajar o desenvolvimento de estratégias para aumentar a produção de frutos e, consequentemente, de vanilina para usos culinários e aplicações médicas.

Chemical constituents of Gelsemium elegans / 中草药

Objective: The chemical constituents were isolated and identified in order to find the bioactive natural products from Gelsemium elegans. Methods: Silica gel, sephadex LH-20 and HPLC column chromatographic techniques were used for separation and purification of the compounds and extensive spectral analysis spectrum were employed for structural elucidation. Results: Fifteen compounds were separated from G. elegans, identified by physicochemical properties, spectral analysis and other means. They are koumine (1), gelsemine (2), gelsevirine (3), gelsemicine (4), xanthotoxin (5), bergapten (6), isopimpinellin (7), imperatorin (8), osthol (9), p-hydroxybenzoic acid (10), vanilla acid (11), β-sitosterol (12), β-daucosterol (13), gelsemamide (14), and 16-epivoacarpine (15). Conclusion: Compounds 5-9 are first isolated from the plants in genus Gelsemium Juss.

Distribución potencial y características geográficas de poblaciones silvestres de Vanilla planifolia(Orchidaceae) en Oaxaca, México / Potential distribution and geographic characteristics of wild populations ofVanilla planifolia(Orchidaceae) Oaxaca, Mexico

ResumenLos especímenes silvestres de Vanilla planifolia G. Jack forman parte del acervo genético primario, los cuales solo se han reportado en Oaxaca, México. Por ello se evaluó la distribución de esta especie con el objetivo de ubicar y describir características ecológicas en zonas potenciales de distribución. La metodología empleada consistió de cuatro etapas: 1) Elaboración de una base de datos con registros de herbario;2) Construcción de la distribución potencial basado en los registros históricos de herbario para la especie, mediante el modelo de máxima entropía (Maxent), con el uso de 22 variables bioclimáticas como predictoras; 3) Realización de búsquedas sistemáticas de individuos in situ con base en los registros de herbario y las áreas de distribución potencial en 24 municipios, para conocer la situación y la distribución del hábitat actual, y 4) Descripción mediante factores ambientales de los nichos ecológicos potenciales generados por MaxEnt. La revisión de las colecciones de herbarios reportó un total de 18 registros de V. planifolia, comprendidos entre 1939 y 1998.La búsqueda sistemática de individuos en campo ubicó 28 plantas distribuidas en 12 sitios sobre 95 364 Km2. Las variables que contribuyeron con mayor valor porcentual para determinar la estimación del modelo de distribución potencial en vainilla son precipitación del periodo más lluvioso (61.9 %), régimen de humedad del suelo (23.4 %) y precipitación del cuatrimestre más lluvioso (8.1 %). El hábitat potencial de la especie se distribuyó en cuatro zonas; trópico húmedo del golfo de México, templado húmedo, trópico húmedo y templado húmedo del pacifico. La precipitación anual osciló de 2 500 a 4 000 mm, con lluvias en verano y porcentaje de precipitación invernal de 5 a 10 %. El régimen de humedad y clima predominantes fueron údico tipo I (330 a 365 días de humedad) y cálido húmedo (Am/A(C) m). Las plantas se ubicaron en altitudes de 200 a 1 190 msnm, en laderas accidentadas, que por lo general están al pie de sistemas montañosos de 1 300 a 2 500 metros de altitud. En condiciones naturales la distribución de la especie no se limita a selva alta perennifolia, dado que se ubicó en bosque mesófilo de montaña y bosque tropical perennifolio. La ubicación de nuevos especímenes de V. planifolia en condiciones silvestres reduce un 66 % del área potencial de distribución, y la fragmenta, al pasar de ser una zona continua a convertirse en tres zonas geográficamente separadas. La reducción del hábitat se debió a un aumento en el número de plantas ubicadas, lo que define las condiciones ambientales a un nivel más exacto. Por lo anterior, se pueden emprender o diseñar acciones de conservación enfocadas a áreas más específicas dentro del estado de Oaxaca, México.

AbstractWild specimens of Vanilla planifolia represent a vital part of this resource primary gene pool, and some plants have only been reported in Oaxaca, Mexico. For this reason, we studied its geographical distribution within the state, to locate and describe the ecological characteristics of the areas where they have been found, in order to identify potential areas of establishment. The method comprised four stages: 1) the creation of a database with herbarium records, 2) the construction of the potential distribution based on historical herbarium records for the species, using the model of maximum entropy (MaxEnt) and 22 bioclimatic variables as predictors; 3) an in situ systematic search of individuals, based on herbarium records and areas of potential distribution in 24 municipalities, to determine the habitat current situation and distribution; 4) the description of the environmental factors of potential ecological niches generated by MaxEnt. A review of herbarium collections revealed a total of 18 records of V. planifolia between 1939 and 1998. The systematic search located 28 plants distributed in 12 sites in 95 364 Km2. The most important variables that determined the model of vanilla potential distribution were: precipitation in the rainy season (61.9 %), soil moisture regime (23.4 %) and precipitation during the four months of highest rainfall (8.1 %). The species potential habitat was found to be distributed in four zones: wet tropics of the Gulf of Mexico, humid temperate, humid tropical, and humid temperate in the Pacific. Precipitation oscillated within the annual ranges of 2 500 to 4 000 mm, with summer rains, and winter precipitation as 5 to 10 % of the total. The moisture regime and predominating climate were udic type I (330 to 365 days of moisture) and hot humid (Am/A(C) m). The plants were located at altitudes of 200 to 1 190 masl, on rough hillsides that generally make up the foothills of mountain systems, with altitudes of 1 300 to 2 500 masl. In natural conditions, distribution of the species is not limited to high evergreen forests, since it was also found in mountain mesophyll and tropical evergreen forests. The location of new specimens of V. planifolia in its wild condition reduces the potential distribution area by 66 %. This area is fragmented into three geographically separated areas. Habitat reduction was due to the increased number of located plants that define the environmental conditions into a more accurate level. Conservation actions can thus be designed and implemented, focusing on more specific areas within the state of Oaxaca, Mexico.

Vainillas colombianas y su microbiota. II. Diversidad, cultivo y microorganismos endófitos / Colombian Vanilla and its microbiota. II. Diversity, culture and endophytic microorganisms Abstract / Baunilhas colombianas e sua microbiótica. II. Diversidade, cultivo e microrganismos endófitos

La vainilla es la única especie como producto comestible entre las orquídeas. Su cultivo en Colombia se está promoviendo como alternativa a los tradicionales; especialmente entre pequeños y medianos agricultores. El cultivo actual adolece de una angosta base genética y por lo tanto es susceptible al estrés biótico, especialmente el causado por virus y hongos. Las vainillas silvestres son posible fuente de variabilidad que podría ayudar a ampliar su acervo genético y a su vez, podrían aportar microorganismos endófitos naturalmente asociados a ellas que tuviesen propiedades como antagonistas de los patógenos de la vainilla cultivada. En este estudio, se evidenció el conocimiento del cultivo de la vainilla como la diversidad del género Vanilla en Colombia. Como resultado de esta primera fase de investigación, se encontraron: veinte especies de vainillas colombianas y más de 60 morfoespecies de microorganismos endófitos. Se discutió la relevancia de estos hallazgos y su posible impacto en la promoción del cultivo de la vainilla, así como también se planteó el uso de estas plantas como un grupo modelo en investigación básica.

Among orchids, vanilla is the only species that is an edible product. Its farming is being promoted in Colombia as an alternative to traditional crops, especially among small and medium farmers. Existing crops have a limited genetic base and are therefore susceptible to biotic stress, particularly those caused by viruses and fungi. Wild vanilla can be a source of variability that could help increase its gene pool and could also provide naturally associated endophyte microorganisms with antagonistic properties to fight pathogens of farmed vanilla. In this study we assessed the understanding of vanilla cultivation as well as the diversity of the genus Vanilla in Colombia. As a result of this first phase of research, we found twenty species of Colombian vanilla and 60 morphospecies of endophytic microorganisms. We discuss the relevance of these findings and their potential impact on promoting vanilla farming, and also discussed the use of these plants as a model group for basic research.

A baunilha é a única espécie de produto comestível entre as orquídeas. O seu cultivo em Colômbia está sendo promovido como alternativa aos tradicionais; especialmente entre pequenos e médios agricultores. O cultivo atual carece de uma forte base genética, portanto é suscetível ao estrese biótico, especialmente causados por vírus e fungos. As baunilhas silvestres são uma possível fonte de variabilidade que poderia ajudar a ampliar o acervo genético e a sua vez, poderiam contribuir com microrganismos endófitos naturalmente associados a elas que tivessem propriedades como antagonista dos patógenos da baunilha cultivada. Neste estudo, se evidenciou o conhecimento do cultivo da baunilha como a diversidade do género Baunilha em Colômbia. Como resultado desta primeira fase de investigação, se encontraram: vinte espécies de baunilhas colombianas e mais de 60 morfoespecies de microrganismos endófitos. Discutiu-se a relevância destes resultados e o seu possível impacto na promoção do cultivo da baunilha, assim como também se evidenciou o uso destas plantas como um grupo modelo em investigação básica.

Identificación oolecular de microorganismos asoiados a la rizosfera de plantass de vainilla en Colomiia / Molecular Identification of Microorganisms Associated to the Rhizosphere of Vanilla Plants in Colombia

El cultivo de vainilla es altamente promisorio en Colombia, pero se requiere mayor conocimiento de su manejo agronómico y de los microorganismos que crecen asociados a su rizosfera, de los cuales depende esta planta para su nutrición y crecimiento. En este trabajo se realizaron aislamientos de microorganismos de la rizosfera de plantas de vainilla en un cultivo piloto ubicado en el municipio de Sopetrán (Antioquia, Colombia). Los microorganismos se aislaron en medios selectivos de crecimiento para evaluar su capacidad para descomponer celulosa, proteínas, solubilizar fosfato inorgánico y orgánico (fitato) y fijar nitrógeno en forma asimbiótica. Una vez aislados y purificados, se obtuvieron un total de 109 aislamientos, de los cuales se seleccionaron 52 morfotipos para su identificación molecular por secuenciación de las regiones ITS y 16S del ADN ribosomal para hongos y bacterias, respectivamente. Se encontró una alta variedad de microorganismos en la rizosfera de plantas de vainilla, destacándose las bacterias Bacillus megaterium, Pseudomonas koreensis y Acinetobacter sp. y el hongo Plectosphaerella sp., por su potencial para ser utilizados como biofertilizantes destinados a mejorar la nutrición y el crecimiento de estas plantas.

The cultivation of vanilla (Vanilla planifolia) is highly promising in Colombia, but more research is needed on its agronomical management and beneficial microorganisms that grow associated to its rhizosphere, on which the plant depends for its nutrition and growth. This study involved the identification of microorganisms associated to the rhizosphere of vanilla plants in a crop located in Sopetrán, Colombia. The microbes were isolated in selective media for functional groups such as cellulolytic, proteolytic, inorganic and organic phosphate (phytate) solubilizers, and asymbiotic nitrogen fixing bacteria. After isolation and purification, 109 microbial isolates were obtained. DNA was extracted from 52 selected isolates for molecular identification based on ITS and 16S rDNA sequencing, for fungi and bacteria, respectively. The diversity of rhizosphere microorganisms found was significant. Bacteria such as Bacillus megaterium, Pseudomonas koreensis and Acinetobacter sp., and the fungus Plectosphaerella sp., may have a high potential to be used as biofertilizers to improve vanilla plant nutrition and growth.

Analysis on chemical constituents in ethanol extract from vanilla pod in various habitats and quantitative determination of vanillin by GC-MS / 中草药

Objective: To analyze the chemical constituents in ethanol extracts from vanilla pod in Hainan (China) and Papua-New Guinea and to assay the content of vanillin. Methods: Comparing various pretreatment methods, GC-MS analysis method was selected and the column was packed with Agilent DB-5MS capillary column (30 m × 0.32 mm, 0.25 μm); column temperature: the column initial temperature was 60 °C, kept for 1 min, rose to 260 °C at the rate of 10 °C/min, kept for 6 min, rose to 290 °C at the rate of 6 °C/min, kept for 8 min; carrier gas: helium for comparing the difference of the constituents from the above two habitats. Results: Because of the different habitats, 34 and 32 compounds were respectively identified from vanilla pod and 18 compounds were common components. Vanillin had a high content in vanilla pod and presented a good linearity with the peak area at the concentration of 0.04-1.10 mg/mL (r = 0.999 8, n = 5). The average recovery was 100.05% with the RSD of 1.88%. Conclusion: GC-MS is quick, simple, convenient, and effective to analyze the constituents in the ethanol extracts from vanilla pod, and it is also reliable and reproducible to determine the content of vanillin.

Germinación in vitro de híbridos de Vanilla planifolia y V. pompona / In vitro germination of Vanilla planifolia and V. pompona hybrids

El cultivo de la vainilla se lleva a cabo ampliamente por medio de propagación clonal, de manera asexual por cortes de tallos y la producción de frutos se realiza por autopolinización. Esta práctica inhibe la variación genética y la emergencia de nuevos individuos por recombinación sexual. Por lo anterior, se considera necesario realizar cruzas por propagación sexual entre especies para obtener nuevos individuos con características deseables para el cultivo. En el presente trabajo se obtuvieron híbridos de dos especies de vainilla como parte de un programa de mejoramiento genético. En general, las semillas híbridas obtenidas que presentaron mayor porcentaje de germinación fueron las cruzas interespecíficas interespecíficos de V. planifolia y V. pompona (85%), seguido de la cruza inversa V. planifolia y V. pompona (57,9%). Las semillas producto de V. pompona autopolinizada obtuvieron valores muy bajos de germinación (10,8%), mientras que las obtenidas de V. planifolia autopolinizada no presentaron germinación. El medio de cultivo más eficiente en todos los tratamientos fue el Murashige & Skoog (MS) adicionado con 400 mg/L-1 de glutamina y 80 mg/L-1 de sulfato de adenina.

The cultivation of vanilla is extensive clonal asexual propagation by cuttings and fruit production by artificial self-pollination. This feature tends to inhibit the genetic variation and the emergence of new individuals by sexual recombination. Therefore, sexual propagation between species it is considered necessary to obtain new individuals with desirable characteristics for cultivation. In this paper we were able to obtain hybrids of two species of vanilla. In general, the hybrids seeds obtained whit the higher germination percentage were interspecific crosses of V. planifolia and V. pompona (85%), followed by reverse cross V. planifolia and V. pompona (57.9%). Seeds obtained of V. pompona self-pollination were very low germination (10.8%) while those obtained from V. planifolia self-pollination showed no germination in any media culture. The most efficient medium for all treatments was the Murashige & Skoog (MS) supplemented whit 400 mg/L-1glutamine and 80 mg/L-1 of adenine sulfate.