Morpho-anatomy of in vitro germination and cryopreservation of the orchid Cattleya crispa (Orchidaceae Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae) / Morfoanatomía de la germinación in vitro y criopreservación de la orquídea Cattleya crispa (Orchidaceae)

Introduction: Cattleya crispa is an ornamental epiphytic orchid with geographic distribution restricted to the Brazilian Atlantic Forest. Due to predatory extractivism and human-induced habitat loss, this species appears on the Red List of Brazilian Flora. Objective: To characterize morpho-anatomical aspects regarding germination and post-seminal development from C. crispa seeds; as well as studying the effect of cryopreservation on these seeds. Methods: We used light microscopy and electron microscopy to describe the microstructure of a 100 ripe seeds. We evaluated seed viability, seed germination, survival rate and protocorm weight in cryopreserved and non-cryopreserved material, with four replicas per treatment using 20 mg of plant material. Results: The seeds are fusiform, whitish yellow with a length from 700 to 900 µm and a water content of 5 %. Germination began seven days after sowing, the formation of the globular protocorm at 30 days and the formation of the seedling occurred 150 days. The persistent seed coat can compress the protocorm and cause it to collapse. The cryopreserved seeds presented 87.15 % viability, 78.32 % germination, 8.48 % survival and protocorms with 104.27 mg five months after sowing. Data wasn't different to non-cryopreserved seeds. Conclusions: The cryocapability of the seeds shows that cryopreservation can be used for long-term conservation. The results of this work contribute to the overall biology of C. crispa and to the propagation and storage of genetic material for conservation purposes.

Introducción: Cattleya crispa es una orquídea epífita ornamental con distribución geográfica restringida a la Mata Atlántica brasileña. Debido al extractivismo depredador y a la pérdida de hábitat inducida por el hombre, esta especie aparece en la Lista Roja de la Flora Brasileña. Objetivo: Caracterizar aspectos morfoanatómicos de la germinación y desarrollo inicial de semillas de C. crispa; así como estudiar el efecto de la criopreservación de estas semillas. Métodos: Utilizamos microscopía óptica, microscopía electrónica de barrido y microscopía electrónica de transmisión para describir la microestructura en 100 semillas maduras. Evaluamos la viabilidad de la semilla, la germinación de la semilla, la tasa de supervivencia y el peso de los protocormos en el material criopreservado y no criopreservado, con cuatro réplicas por tratamiento de 20 mg de material vegetal. Resultados: Las semillas son fusiformes, amarillo blanquecinas, con una longitud de 700 a 900 µm y un contenido de agua del 5 %. La germinación comenzó siete días después de la siembra, la formación del protocormo globular a los 30 días y la formación de la plántula a los 150 días. La cubierta de semilla persistente puede comprimir el protocormo y provocar su colapso. Las semillas criopreservadas presentaron 87.15 % de viabilidad, 78.32 % de germinación, 8.48 % de supervivencia y protocormos con 104.27 mg a los cinco meses de la siembra. Los datos no fueron diferentes a las semillas no criopreservadas. Conclusiones: La capacidad criogénica de las semillas muestra que la crioconservación puede utilizarse para la conservación a largo plazo. Los resultados de este trabajo contribuyen a la biología general de C. crispa y a la propagación y almacenamiento de material genético con fines de conservación.

Orchidists' profile, management and perception of the main pests in orchid cultivation

This study investigated the profile of orchidists, their cultivation practices, and their knowledge about pests in orchid cultivation in the state of Bahia. There were applied 74 questionnaires to producers (amateurs and commercials) in 23 municipalities. The data were submitted to univariate statistics using the chi-square test, the Mann­Whitney U-test and the Kruskal­Wallis H-test. The survey shows that orchid cultivation is mostly practiced by women (67.6%) and those with higher education (48.7%). Most respondents were amateurs (87.8%). Insects (39.9%) and diseases (32.5%) were the main organisms associated with phytosanitary problems. The presence of insects was reported to occur mainly in the leaves (47%), mainly mealybugs (46.3%). Correspondence was significant between commercial and amateur groups according to the pairwise Mann­Whitney U-test(1) = 370.04 and p < 0.01. The Kruskal­Wallis H-test(27) = 924.78 and p < 0.01 discriminated categories between the groups. Despite the growing interest in orchid cultivation in the state of Bahia, most orchidists are amateurs. The study warns about the risk of pest dispersion between crops since exchanging plants and purchasing specimens in open markets are common practices.

Advances in the research of microRNA in Orchidaceae / 生物工程学报

As a class of small non-coding RNAs, microRNA (miRNA) is widely present and plays important regulatory roles in plant growth, development and stress response. Based on the mechanism of miRNAs in plants, we review the identification of miRNAs in some genera of Orchidaceae, the specific functions of several miRNAs and other relevant studies on miRNAs in the last decade, in order to provide a reference for better understanding function and regulatory network of small RNAs in orchids.

Abelhas das orquídeas em fragmentos de floresta ripária e de terra-firme em uma matriz urbana no sudoeste da Amazônia brasileira / Orchid bees in riparian and terra-firme forest fragments in an urban matrix in southwestern Brazilian Amazonia

As florestas ripárias no Brasil são importantes ecossistemas que sustentam uma enorme biodiversidade. Apesar de protegidas pela legislação brasileira, elas têm sofrido grandes impactos decorrentes da fragmentação florestal. As abelhas das orquídeas constituem um grupo chave de polinizadores na região Neotropical, porém, pouco se sabe sobre suas assembleias em florestas ripárias. Nós avaliamos o papel de fragmentos de floresta ripária e de terra-firme na conservação e manutenção da fauna de abelhas das orquídeas em uma paisagem urbana no sudoeste da Amazônia. Especificamente, avaliamos se assembleias de abelhas de fragmentos ripários e de terra-firme diferem significativamente em abundância, riqueza e composição de espécies. Também avaliamos se a abundância e a riqueza de espécies variam em função do tamanho do fragmento. Machos de abelhas foram atraídos por iscas odoríferas e coletados com redes entomológicas em 10 fragmentos florestais. Não houve diferença significativa entre fragmentos ripários e de terra-firme quanto à abundância, riqueza e composição de espécies, mas houve uma correlação positiva entre o tamanho do fragmento e a riqueza e abundância de espécies. Nossos resultados sugerem que, em uma paisagem urbana, os fragmentos de floresta ripária e de terra firme ainda podem manter 62,7% do número de espécies de abelhas das orquídeas conhecido para a região, reforçando o valor da conservação desses remanescentes florestais. Nossos dados indicam que esses fragmentos fornecem um habitat potencialmente importante para a manutenção das populações locais de abelhas na paisagem. (AU)

Progress in application of Bletilla striata polysaccharide in novel drug delivery systems and biomaterials / 中国中药杂志

As the main active ingredient of the orchidaceous herb Bletilla striata, B. striata polysaccharide(BSP) has pharmacological activities such as promoting coagulation, anti-inflammation, anti-oxidation, promoting wound healing, anti-tumor, and immunomodulation, and is biodegradable and non-toxic. Additionally, it has the material properties of suspension thickening, film-forming adhesion, coating and solubilizing, targeting and slow releasing, effect-enhancing and toxicity-reducing, etc., playing the role of unification of medicines and excipients. Therefore, BSP has a wide application prospect in the fields of drug delivery system and trauma repair. This paper reviews the research progress of BSP application in new drug delivery systems and biomaterials based on the related li-terature in recent years, with the aim of providing reference for the further research and application of BSP.

Improving the yield of Anoectochilus roxburghii by Bacillus licheniformis cultured in Agaricus bisporus industrial wastewater

BACKGROUND: There is a large amount of industrial wastewater produced by the mushroom industry during the canning processing each year, which could provide abundant carbon, nitrogen and inorganic salts for microbial growth. The aim of this study was to optimize the culture conditions for Bacillus licheniformis cultured in the Agaricus bisporus industrial wastewater to produce the agricultural microbial fertilizer. RESULTS: In this work, the maximal biomass of B. licheniformis could be obtained under the following culture conditions: 33.7°C, pH 7.0, 221 rpm shaking speed, 0.5% wastewater, 2 (v:v, %) inoculum dose, loading liquid of 60 mL/250 mL and a culture time of 24 h, and the average experimental value obtained was 1.35 ± 0.04 × 109 Obj/mL, which was within the 95% confidence interval of the predicted model (1.29­1.38 × 109 Obj/mL), and met the national microbial fertilizers' standard in China. Furthermore, the field experiment results showed that the fermentation broth of B. licheniformis could significantly improve the yield of Anoectochilus roxburghii. CONCLUSIONS: Agaricus bisporus industrial wastewater can be used to produce agricultural microbial fertilizer.

Does soaking time during disinfestation affect germination rates in Dendrobium? / O tempo de imersão durante a desinfestação influencia nas taxas de germinação de Dendrobium?

Asymbiotic germination is considered an efficient and viable technique that can increase germination rates. The effect of type and concentration of disinfestants, and the exposure time to disinfestants may differ according to the plant species. Therefore, species-specific standardization of disinfestation agent and procedure is necessary to achieve optimal germination rates. The objective of this study was to determine a disinfestation methodology to increase in vitro germination rates and the early development of seedlings of Dendrobium nobile and Dendrobium phalaenopsis, using different times for seed disinfestation and different culture media. Seeds were disinfected by soaking in a 0.8% sodium hypochlorite solution for 5 or 15 min under aseptic conditions, after which seed suspensions were either washed with water or left unwashed. Next, they were seeded in culture flasks containing four different culture media (MS, ½MS, K, and VW). The flasks were then transferred to a growth room under controlled photoperiod and temperature, where they remained under an irradiance of 20 µmol m-2 s-1. Germination rates of the species were evaluated 45 days after placement in the culture flasks. A higher germination rate was observed when the seeds were triple washed, regardless of the culture medium or soaking time. Seed soaking disinfestation for 5 min is also recommended. MS and ½MS media were the most effective culture media in promoting in vitro germination of the species under study.

A germinação assimbiótica é considerada uma técnica eficiente e viável resultando em elevados percentuais de germinação. Apesar do sucesso dessa técnica, o tipo, a concentração e o tempo de exposição do agente desinfestante diferem, necessitando padronização para cada espécie. Assim, a padronização do agente desinfestante e do procedimento são necessários para o aumento das taxas germinativas. Objetivou-se com este trabalho determinar metodologia que aumente a taxa de germinação in vitro e favoreça o desenvolvimento inicial de plântulas de Dendrobium nobile e Dendrobium phalaenopsis, utilizando diferentes métodos de desinfestação de sementes e diferentes meios de cultura. Sementes, sob condições assépticas, foram desinfestadas em solução de hipoclorito de sódio a 0,8%, por cinco ou quinze minutos. Após esses períodos, as suspensões de sementes receberam ou não a tríplice lavagem com água. Em seguida, foram semeadas em frascos de cultivo que continham quatro diferentes meios de cultura (MS, MS½, K e VW). Posteriormente, foram transferidos para sala de crescimento com fotoperíodo e temperatura controlados, onde permaneceram sob irradiância de 20 µmol m-2 s-1. Quarenta e cinco dias após a semeadura foi avaliada a porcentagem de germinação das espécies estudadas. Os resultados neste trabalho indicam que, independentemente do meio de cultura ou do tempo de desinfestação, as sementes quando submetidas à tríplice lavagem apresentaram porcentagem de germinação superior a àquelas que não receberam este procedimento. Recomenda-se a desinfestação das sementes por 5 minutos. Os meios MS e MS½ foram os mais efetivos em promover a germinação in vitro dessas espécies.

Morphological Developments of Orchis purpurea Huds. Salep Orchids

Abstract This article aims to monitor the development of Orchis purpurea Huds., salep orchids, of different sizes over a period of two years, and to investigate the relationship between the parameters studied. In the first step, the measurements taken at the time of planting and harvesting of tubers divided into eight different groups according to their size were subjected to variance and Duncan's test. In the second step, the relationship between the parameters was investigated by ignoring seedling groups. The relationship between the two variables was determined by correlation analysis. The significance of the relationships between planting and harvest data sets, and variable contributions were determined by canonical correlation analysis. Finally, leaf area prediction modeling was performed by applying multiple regression analysis. In variance analysis all parameters were significant. The canonical correlation between the first pair of canonical variables was 0.988 (p<0.01). The data obtained from the tubers made the greatest contribution to the explanatory power of the canonical variables. The leaf area model was formulized as LA (mm2) = -1237.0204 + 57.7912 × LW + 16.6211 × LL where LA is leaf area, LW is leaf width, LL is leaf length and a, b, and c are coefficients.

Development of Growth Models Based on Years for Salep Orchid Planted in Natural Environment

Abstract The orchids that produced tuber are known as salep orchids and have been collected from nature for centuries as a medicinal and aromatic plant. These orchids are endangered species because of their limited vegetative and generative production possibilities. Salep orchids, seed germination has been achieved with in-vitro studies, but no source has been found regarding the development of germinated seeds over the years. In this study, Dactylorhiza euxina (Nevski) Czerep. was used as research material. Seeds that were determined to germinate in nature were observed to develop in their own environment for five years and some morphological characteristics (plant height, number of leaves, leaf length, leaf width, total weight, tuber width, and tuber weight) were determined at the end of each vegetation year. Plants that start to develop with seed germination produce one leaf and a tuber of 0.115 g in the first year. When the tubers obtained from the first year were planted again, the plants was obtained 2-3 leaves and they were formed 0.465 grams of tubers. In the third year plants bloom for the first time. The period until flowering in the plant was called maturity period and Dactylorhiza euxina (Nevski) Czerep. was found to have a 3-year maturity period. Plants produced tuber 0.850 g in the third year, 1.585 g in the fourth year and 2.522 g in the fifth year. According to the mathematical modeling, the relationship between year (Y) and fresh tuber weight (FTW) was found Y = 1.22 + (1.61 × FTW) and there was a significant relationship among year, fresh tuber weight, and total biomass (TBM). This equation Y = 1.05 + (0.76 × FTW) + (0.14 × TBM) shows us the mathematical relations of year, fresh tuber weight, and total biomass. The mathematical relations among to fresh tuber weight, total biomass, and plant height (PH) was produced Y = 1.74 - (0.11 × FTW) + (0.57 × TBM) - (0.09 × PH). R2 values were found 0.95-0.99. All R2 values and standard errors were found to be significant at the p < 0.01 level.

In vitro conservation and acclimatization of epidendroideae (Orchidaceae) from Sergipe, Brazil / Conservação in vitro e aclimatização de Epidendroideae (Orchidaceae) de Sergipe, Brasil

Orchids are known for the beauty, exuberance, and color of their flowers and therefore represent one of the most coveted ornamental plants. Due to their high commercial value, several illegal practices of removal and commercialization have led to the verge of extinction. This study aimed to develop protocols for in vitro conservation under slow growth and acclimatization of species of the subfamily Epidendroideae (Orchidaceae), from the state of Sergipe, Brazil. Two experiments were conducted: the first one tested four concentrations of MS (MURASHIGE and SKOOG, 1962) (25%, 50%, 75%, 100%), three orchid species (Catasetum macrocarpum, Oeceoclades maculata, and Polystachya estrellensis), and two temperatures (18 and 25ºC); the second one tested three combinations of carbon sources and osmotic regulators (20 g L-1 sucrose; 10 g L-1 sucrose + 5 g L-1 de mannitol; 10 g L-1 sucrose + 5 g L-1 sorbitol) and the same three species and two temperatures. For the in vitro conservation of C. macrocarpum, O. maculata, and P. estrellensis over 450 days, the use of 25% of MS salts or 20 g.L-1 sucrose at 25ºC is recommended. C. macrocarpum seedlings were acclimatized using sand + shredded pine bark + worm castings at a ratio of 2:2:1 (w:w:w). The use of sand alone is recommended for the acclimatization of O. maculata

As orquídeas são conhecidas pela beleza, exuberância e cores de suas flores e por isso representam uma das plantas ornamentais mais cobiçadas. Devido ao alto valor comercial, muitas são as práticas ilegais de retirada e comercialização de orquídeas, o que leva as espécies à beira de extinção. O objetivo deste estudo foi desenvolver protocolos para a conservação in vitro sob crescimento lento e aclimatação de espécies da subfamília Epidendroideae (Orchidaceae) do Estado de Sergipe, Brasil. Dois experimentos foram realizados: o primeiro ensaio testou quatro concentrações dos sais MS (MURASHIGE e SKOOG, 1962) (25, 50, 75 e 100%), três espécies (Catasetum macrocarpum, Oeceocladesmaculata e Polystachya estrellensis) e duas temperaturas (18 e 25ºC); o segundo ensaio testou três combinações de fontes de carbono e reguladores osmóticos (20 g L-1 de sacarose; 10 g L-1 de sacarose + 5 g L-1 manitol; 10 g L-1 de sacarose + 5 g L-1 sorbitol), três espécies (C. macrocarpum, O. maculata e P. estrellensis) e duas temperaturas (18 e 25ºC). Para executar a aclimatação, oito substratos foram testados para as espécies C. macrocarpum e O. maculata. Para realizar a conservação in vitro de C. macrocarpum, O. maculata e P.estrellensis ao longo de 450 dias, recomenda-se a utilização 25% dos sais de MS ede 20 g L-1 de sacarose a 25°C. Mudas de C. macrocarpum foram aclimatizadas utilizando areia + casca de pinus + húmus de minhoca (2:2:1). Recomenda-se a utilização de areia para aclimatização de O. maculata.

Acetoacetate extract of pleione bulbocodioides (franch. ) rolfe induces apoptosis of human leukemia thp-1cells through a mitochondria-regulated intrinsic apoptotic pathway / Extrato de acetoacetato de Pleione bulbocodioides (Franch. ) Rolfe induz a apoptose de células THP-1 de leucemia humana através de uma via apoptótica intrínseca regulada por mitocôndrias

The tubers of three orchidaceous plants, includingPleione bulbocodioides (Franch.) Rolfe, have been used as 'Shan-Ci-Gu' in traditional Chinese medicine for the treatment of bacterial infections and cancers for thousands of years. In this study, the effects of an acetoacetate (EtOAc) extract of P. bulbocodioides on the cell viability and apoptosis of THP-1 (human acute monocytic leukemia cell line) cells and its interaction with possible apoptotic pathways were investigated. THP-1 cells were treated with the EtOAc extract of P. bulbocodioides at different concentrations. The results showed that THP-1 cell viability was significantly inhibited by the EtOAc extract ofP. bulbocodioides with an IC50 of 51.37±2.68 µ g/ mL at 24 h. The examination of cytotoxic effects on healthy cells showed that the EtOAc extract of P. bulbocodioidesdid not show any effect on healthy Vero cells. Selectivity indexes were greater than 15.57, suggesting that the EtOAc extract of P. bulbocodioides had selective toxicity against THP-1 cells. The results of annexin V-FITC/PI and DAPI staining showed that the EtOAc extract of P. bulbocodioides induced cell apoptosis in a dose-dependent manner. The apoptotic rate was increased in the treatment groups compared with that in the control group (P<0.05). The distribution of cells in the G2 phase of the cell cycle increased along with typical cell apoptosis-induced morphological changes. The levels of the pro-apoptotic proteins Bax, cleaved PARP and cleaved caspase-3 increased with increasing concentration of acetoacetate extract of P. bulbocodioides, while the anti-apoptosis protein Bcl-2 was downregulated. Cyt c and AIF, which are characteristic proteins of the mitochondria-regulated intrinsic apoptosis pathway, also increased in the cytosol with increasing concentrations of the EtOAc extract of P. bulbocodioides. These results showed that the EtOAc extract of P. bulbocodioidessignificantly inhibits cell viability and induces cell apoptosis in the human leukemia cell line THP-1 through a mitochondria-regulated intrinsic apoptotic pathway

Os tubérculos de três plantas orquidáceas, incluindo Pleione bulbocodioides (Franch.) Rolfe, têm sido usados como "Shan-Ci-Gu" na medicina tradicional chinesa para o tratamento de infecções bacterianas e cânceres por milhares de anos. Neste estudo, os efeitos de um extrato de acetoacetato (EtOAc) de P. bulbocodioides na viabilidade celular e apoptose de células THP-1 (linhagem celular de leucemia monocítica aguda humana) e sua interação com possíveis vias apoptóticas foram investigados. As células THP-1 foram tratadas com o extrato EtOAc de P. bulbocodioides em diferentes concentrações. Os resultados mostraram que a viabilidade das células THP-1 foi significativamente inibida pelo extrato EtOAc de P. bulbocodioides com IC50 de 51,37 ± 2,68 µ g/mL às 24 h. O exame dos efeitos citotóxicos em células saudáveis mostrou que oextrato de EtOAc de P. bulbocodioides não mostrou nenhum efeito sobre células Vero saudáveis. Os índices de seletividade foram maiores que 15,57, sugerindo que o extrato de EtOAc de P. bulbocodioides teve toxicidade seletiva contra as células THP-1. Os resultados da coloração da anexina V-FITC/PI e DAPI mostraram que o extrato de EtOAc de P. bulbocodioides induziu a apoptose celular de maneira dose-dependente. A taxa de apoptose foi aumentada nos grupos de tratamento em comparação com o grupo controle (P <0,05). A distribuição de células na fase G2 do ciclo celular aumentou juntamente com alterações morfológicas típicas induzidas pela apoptose celular. Os níveis das proteínas pró-apoptóticas Bax, PARP clivada e caspase-3 clivada aumentaram com o aumento da concentração do extrato acetoacetato de P. bulbocodioides, enquanto a proteína anti-apoptose Bcl-2 foi regulada negativamente. Cyt c e AIF, que são proteínas características da via de apoptose intrínseca regulada por mitocôndrias, também aumentaram no citosol com concentrações crescentes do extrato de EtOAc de P. bulbocodioides. Estes resultados mostraram que o extrato de EtOAc de P. bulbocodioides inibe significativamente a viabilidade celular e induz a apoptose na linha celular de leucemia humana THP-1 através de uma via apoptótica intrínseca regulada por mitocôndrias.

Distribución geográfica de angiospermas epífitas de la región terrestre prioritaria Cerros Negro-Yucaño, Oaxaca, México / Geographical distribution of epiphytic angiosperms of the priority terrestrial region of Cerros Negro-Yucaño, Oaxaca, Mexico

Resumen El conocimiento sobre la riqueza y la distribución de las plantas epifítas vasculares es aún incompleto en muchas áreas de México. Un ejemplo es la región terrestre prioritaria (RTP) Cerros Negro-Yucaño, que se ubica en el noroeste del estado de Oaxaca y pertenece a la región mixteca alta (Ñuu Savi Sukun, Ñuu vixi). Con base en la revisión de material de herbario en 12 colecciones institucionales mexicanas y en la recolección de especímenes en algunas localidades, principalmente cubiertas por bosque de encino, en 17 de los 18 municipios que incluye la RTP, se compilo un listado de las angiospermas epífitas. Se registró la presencia de 40 especies, distribuidas en 13 géneros y cinco familias; 28 taxa son endémicos de México y tres de ellos se conocen solamente de Oaxaca. Los géneros Tillandsia (18 spp.) y Peperomia (4) fueron los mejor representados en el área. El bosque de encino fue el tipo de vegetación en el cual se encontraron más epífitas (32 spp.) y también el intervalo altitudinal de 1 900 a 2 100 m (19 spp.). Plantas en floración de Artorima erubescens, Laelia albida, L. furfuracea y Prosthechea karwinskii, son utilizadas por pobladores de la región como ornamentales, durante las celebraciones religiosas de Día de Muertos y Semana Santa. Este impacto humano podría representar una amenaza para las poblaciones silvestres de estas especies en el futuro.(AU)

Abstract The knowledge on richness and distribution of epiphytic vascular plants is still incomplete in many areas of Mexico. An example is the terrestrial priority region (RTP) Cerros Negro-Yucaño, which is located in the Northwest portion of Oaxaca and belongs to the Mixteca Alta Region (Ñuu Savi Sukun, Ñuu Vixi). Based on herbarium material revision of 12 Mexican institutional collections and the collection of specimens in some localities, mainly covered by oak forest, in 17 of the 18 municipalities included in the RTP, we compiled a list of epiphytic angiosperms. The presence of 40 species, distributed in 13 genera and five families was recorded; 28 taxa are endemic to Mexico and three of them are only known from Oaxaca. The genera Tillandsia (18 spp.) and Peperomia (4) were the best represented genera in the area. Oak forest was the type of vegetation in which the most of the epiphytes were found (32 spp.), and also the altitudinal range between 1 900 and 2 100 m (19 spp.). Flowering plants of Artorima erubescens, Laelia albida, L. furfuracea, and Prosthechea karwinskii are used by the local people as ornamentals during the religious celebrations of Day of the Dead (Mexico) and Holly Week. This human impact might be a threat to the wild populations of these species in the future.(AU)

Comparison of contents of three active ingredients in Bletilla striata from different sources / 中国中药杂志

In order to understand the difference of contents of coelonin,batatasin Ⅲ and 3'-O-methylbatatasin Ⅲ in 60 different sources of Bletilla striata planted under the same conditions. UPLC method was used and the analysis was performed on a Waters ACQUITY UPLC BEH C18 column( 2. 1 mm×100 mm,1. 7 μm),eluted with acetonitril-0. 1% formic acid solution by gradient. The flow rate was 0. 208 m L·min-1,the detection wavelength was 270 nm,the column temperature was 35 ℃ and the injection volume was 4μL. Under the above chromatographic conditions,the three components can be separated well with good linearity in the range of 0. 156-5. 000 mg·L-1. The average contents of coelonin,batatasin Ⅲ and 3'-O-methylbatatasin Ⅲ were( 0. 116 ± 0. 071) %,( 0. 386 ±0. 185) % and( 0. 086±0. 034) %,respectively. After planting for two years under the same conditions,there was no significant difference in chemical composition among different sources and varieties,but the contents of the three components had some regional differences,which indicated that the western region was higher than the eastern region,while the contents of coelonin and batatasin Ⅲ in B.sinensis were slightly higher than those in B. striata. The chromatographic method above is simple,stable and reproducible,and can be used for quantitative analysis of three components. The content analysis of different sources of B. striata can provide reference for future B. striata breeding and quality control.

Effect of cultivation substrate on growth and active component contents of Anoectochilus roburghii from three different origins / 中国中药杂志

Three different origins of Anoectochilus roburghii were used as experimental materials to study the effect of three different substrate( peat soil-river sand-peanut shell) radio on survival rate,plant height,stem diameter,plant fresh weight,root number,the longest root length,root diameter,and the contents of polysaccharide,flavonoids,and polyphenol. The results showed that when the substrate ratio was 4 ∶2 ∶2,the survival rate of A. roburghii from different origins was the highest,and the plant height,stem diameter,plant fresh weight,the longest root length and root diameter were also the largest. The cultivation substrate had no significant effect on the polysaccharide content of A. roxburghii and A. formosanus. When the substrate ratio was 4 ∶ 2 ∶ 2,the polysaccharide content of A.chapaensis was significantly lower than that of the other two combinations. When the substrate ratio was 4 ∶2 ∶1,the flavonoid content of A. formosanus was higher than that of the other two combinations. When the substrate ratio was 4 ∶2 ∶2,A. formosanus and A. chapaensis had higher polyphenol content.

Absorptive characteristic of Bletilla striata extract in Caco-2 cell monolayer / 中国中药杂志

To investigate the active fraction from Bletilla striata in Caco-2 cell monolayer,so as to explore its absorption mechanism of oral administration preliminarily.Active fraction from B.striata in Caco-2 cell monolayer was analyzed by UPLC-Q-TOF and detected by UPLC-MS/MS,and the effects of different concentrations,pH and P-glycoprotein inhibitors on Caco-2 cells Monolayer were investigated.Six compounds were isolated from the active fraction of B.striata in Caco-2 cell monolayer by UPLC-Q-TOF,and identified as B6,B12,B14,B17,B19 and B23,with concentration dependence.Within the 0-180 min,the uptake of B12 and B14 had a time dependence,while B6,B17,B19 and B23 tended to saturate after 60 min.All of the components had a good absorption in an acidic environment.B6 had a good absorption at pH 6.0,while the other components B12,B14,B17,B19 and B23 had a good absorption at pH4.0.The absorption of the 6 main components of B.striata were not be affected by P-glycoprotein inhibitors(verapamil/cyclosporin A).Compared with the control group,there was no difference in the absorption of B6 and B12,and the absorption of B14,B17,B19 and B23 increased,but with no significant difference.The absorption characteristic of B.striata extract across the Caco-2 cell monolayer is probably passive diffusion,and the absorption process was not affected by P-glycoprotein.

Rizobacterias promotoras de crecimiento en Guarianthe skinneri (Orchidaceae) / Growth-promoting rhizobacteria from Guarianthe skinneri (Orchidaceae)

Resumen La orquídea Guarianthe skinneri está incluida en la norma NOM-059-ECOL-2010 de México como una especie amenazada. Con el fin de estudiar las BPCV (bacterias promotoras del crecimiento vegetal) en esta orquídea, se recolectaron 10 raíces de diferentes plantas para aislar bacterias asociadas a las raíces, que se analizaron mediante pruebas in vitro como: producción de AIA, fijación de nitrógeno, interacción con el hongo micorrízico Thanatephorus sp. cepa RG26 y solubilización de fosfato. De los 71 aislados bacterianos se caracterizaron 10 cepas mediante secuenciación con el marcador 16s rADN y se identificaron seis cepas: Sphingomonas sp., Sinorhizobium sp., Bacillus sp., Nocardia cerradoensis, Bacillus megaterium y Burkholderia phytofirmans. Se observó que la bacteria Sinorhizobium sp. produjo mayor cantidad de AIA (69.189 µg/ml) y Bacillus sp. presentó mayor reducción de acetileno (10.251 nmol cultivo/96 h). En las interacciones de las bacterias y el hongo RG26 se presentaron cuatro categorías (sumamente positivo, positivo, antagonismo 50-50 e inhibición). En relación a la solubilización de fosfato, la bacteria Burkholderia phytofirmans presentó mayor IS a las 48 y 96 hr con IS de 3.11 y 3.48, respectivamente. Los resultados indican que Bacillus sp. pudiera tener las mejores características para promover el desarrollo de la orquídea G. skinneri mediante la inoculación de semillas y plántulas.

Abstract The Guarianthe skinneri orchid is included in NOM-059-ECOL-2010, Mexico standard as an endangered species. In order to study PGPR (promoting growth plant rhizobacteria) from this orchid, 10 roots were collected from different plants to isolate bacteria associated with the roots, which were analyzed by in vitro tests such as: production of AIA, nitrogen fixation, interaction with the mycorrhizal fungus Thanatephorus sp. strain RG26 and phosphate solubilization. We obtain 71 bacterial isolates, 10 strains of them were characterized by sequencing with the 16d rDNA marker identifying six bacteria: Sphingomonas sp. Sinorhizobium sp. Bacillus sp. Nocardia cerradoensis, Bacillus megaterium and Burkholderia phytofirmans. We observed that the bacterium Sinorhizobium sp. produced a greater amount of AIA (69.189 μg/ml) and Bacillus sp. performed greater acetylene reduction (10.251 nmol cultivo/96h). In the interactions of the bacteria and the fungus RG26, four categories were presented (extremely positive, positive, antagonism 50-50 and inhibition). In relation to the solubilization of phosphate, Burkholderia phytofirmans presented higher IS after 48 and 96 hr with an IS of 3.11 and 3.48, respectively. The results indicate that Bacillus sp. it could have the best characteristics to promote the development of the G. skinneri orchid by inoculating seeds and seedlings. Rev. Biol. Trop. 66(3): 953-968. Epub 2018 September 01.

Agrobacterium-mediated transformation of the wild orchid Cattieya maxima Lindi / Transformación mediada por Agrobacterium de la orquídea silvestre Cattleya maxima Lindl / Transformação da orquídea silvestre Cattleya maxima Lindl mediada por Agrobacterium

Abstract Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the propagative nature of PLBs and the easiness by which they can be generated, make these structures an attractive alternative to seed-mediated production for growing large numbers of plants. To increase somatic embryogenesis and optimize the procedure, PLBs of Cattleya maxima were transformed using the Agrobacterium tumefaciens method. The T-DNA carried a Hygromycin-resistance gene, a visible marker (GFP5-GUSA) and a rice gene encoding the Somatic Embryogenesis Receptor Kinase, deemed to be important for somatic embryogenesis. Treated PLBs generated somatic embryos developing Hygromycin-resistant plantlets. The insertion of T-DNA was confirmed by PCR, and GFP expression was observed using a fluorescent stereomicroscope. Transformed Cattleya maxima PLBs were more efficient in forming somatic embryos (60 - 80%) than untransformed controls (45 - 57%), and this contrast was maximized in hormone-free, Murashige and Skoog (MS) medium (80% of the transformed plants compared to 57% of the untransformed ones). This finding supports the notion that SERK plays an important role in Orchid embryogenesis.

Resumen Los protocormos son estructuras anatómicas únicas: son similares a los rizoides y se forman por vástagos jóvenes de orquídeas bajo condiciones fisiológicas. Los tejidos explantados de orquídeas producen estructuras llamadas Cuerpos Similares a Protocormos (PLBs) cuando están expuestos a condiciones apropiadas de crecimiento in vitro. Tanto la naturaleza propagativa de los PLBs como la facilidad con que se generan, hacen de estas estructuras una alternativa atractiva, frente a la mediada por semillas, para la producción de gran número de plantas en crecimiento. Para aumentar la embriogénesis somática y optimizar el procedimiento, se transformaron PLBs de Cattleya maxima usando el método de Agrobacterium tumefaciens. El T-DNA portaba un gen de resistencia a la Higromicina, un marcador visible (GFP5-GUSA) y un gen de arroz que codificaba para el receptor tipo quinasa de embriogénesis somática (SERK), considerado importante en la embriogénesis somática. Los PLBs tratados generaron embriones somáticos y desarrollaron plántulas resistentes a la Higromicina. La inserción del T-DNA se confirmó por PCR, y la expresión de GFP se observó usando un estereomicroscopio fluorescente. Los PLBs transformados de Cattleya maxima fueron más eficientes en desarrollar embriones somáticos (60-80%) que los controles no transformados (45-57%) y este contraste se maximizó en medio Murashige y Skoog (MS) libre de hormonas (80% de las plantas transformadas en comparación con 57% de las no transformadas). Estos hallazgos apoyan la noción de que SERK juega un papel importante en la embriogénesis de orquídeas.

Resumo Os protocormos são estruturas anatômicas únicas: são similares aos rizoides e se formam por hastes jovens de orquídeas sob condições fisiológicas. Os tecidos explantados de orquídeas produzem estruturas chamadas Corpos Similares a Protocormos (PLBs) quando estão expostos a condições apropriadas de crescimento in vitro. Tanto a natureza propagativa dos PLBs como a facilidade com que se generam, fazem com que estas estruturas sejam uma alternativa atrativa, comparativamente a mediada por sementes, para a produção de grandes números de plantas em crescimento. Para aumentar a embriogênesis somática e otimizar o procedimento, se transformaram PLBs de Cattleya maxima utilizando o método de Agrobacterium tumefaciens. O T-DNA carregava um gen de resistencia a Higromicina, um marcador visível (GFP5-GUSA) e um gen de arroz que codificava para o receptor tipo quinasa de embriogênesis somática (SERK), considerado importante na embriogênesis somática. Os PLBs tratados geraram embriões somáticos e desenvolveram plântulas resistentes a Higromicina. A inserção do T-DNA se confirmou por PCR, e a expressão de GFP se observou utilizando um estereomicroscópio de fluorescência. Os PLBs transformados de Cattleya maxima foram mais eficientes em desenvolver embriões somáticos (60-80%) que os controles não transformados (45-57%) e este contraste se potencializou em meio Murashige y Skoog (MS) livre de hormônios (80% das plantas transformadas em comparação com 57% das não-transformadas). Estes resultados apoiam a noção de que SERK desempenha um papel importante na embriogênesis de orquídeas.

Determination of endogenous IAA and carbohydrates during the induction and development of protocorm-like bodies of Cattleya tigrina A. Richard / Determinação de AIA e carboidratos endógenos durante a indução e desenvolvimento deestruturas semelhantes a protocormos de Cattleya tigrina A. Richard

Tissue culture techniques have been employed for orchid mass propagation by means of themorphogenetic route of protocorm like-bodies (PLBs). This study aimed to analyze and compare Indoleaceticacid (IAA) and sugar endogenous levels in protocorm-like bodies (PLBs) induction and developmentin Cattleya tigrina, in order to better understand this process and to optimize micropropagation procedureprotocols. Leaves grown on MS (Murashige and Skoog) culture medium, suplemented with 9 μM TDZfor PLBs induction and development were collected after 0, 2, 7, 14, 30, 60 and 100 days of cultivation, forfurther analysis. Increase of IAA and reduction of sugar levels are strongly related to morphogeneticresponse, that is, PLBs formation over the preexisting ones and leaf primordia formation. Sucrose, fructoseand glucose presence in this study is related to cell signaling. Thus, hormonal signals and carbohydratesalter metabolism, triggering PLBs initiation and development in C. tigrina.

Técnicas de cultura de tecidos têm sido empregadas para a propagação em massa de orquídeaspor meio da via morfogenética de estruturas semelhantes a protocormos (ESPs). O estudo teve comoobjetivo analisar e comparar os níveis endógenos de ácido indol-3-acético (AIA) e de açúcares na indução eno desenvolvimento de estruturas semelhantes a protocormos (ESPs) em Cattleya tigrina, visando à melhorcompreensão deste processo e à otimização de protocolos de micropropagação. Folhas cultivadas em meiode cultura MS (Murashige & Skoog), suplementadas com 9 μM de TDZ para indução e desenvolvimentode ESPs foram coletadas após 0, 2, 7, 14, 30, 60 e 100 dias de cultivo, para posterior análise. O aumento dosníveis de AIA e a redução dos níveis de açúcar estão fortemente relacionados à resposta morfogenética, ouseja, à formação de ESPs sobre os pré-existentes e à formação de primórdios foliares. A presença desacarose, frutose e glicose neste estudo está relacionada à sinalização celular. Assim, sinais hormonais ecarboidratos alteram o metabolismo, desencadeando a iniciação e o desenvolvimento de ESPs em C. tigrina.

Expression profiles of five FT-like genes and functional analysis of PhFT-1 in a Phalaenopsis hybrid

Background: Phalaenopsis is an important ornamental flowering plant that belongs to the Orchidaceae family and is cultivated worldwide. Phalaenopsis has a long juvenile phase; therefore, it is important to understand the genetic elements regulating the transition from vegetative phase to reproductive phase. In this study, FLOWERING LOCUS T (FT) homologs in Phalaenopsis were cloned, and their effects on flowering were analyzed. Results: A total of five FT-like genes were identified in Phalaenopsis. Phylogenetic and expression analyses of these five FT-like genes indicated that some of these genes might participate in the regulation of flowering. A novel FT-like gene, PhFT-1, distantly related to previously reported FT genes in Arabidopsis and other dicot crops, was also found to be a positive regulator of flowering as heterologous expression of PhFT-1 in Arabidopsis causes an early flowering phenotype. Conclusions: Five FT homologous genes from Phalaenopsis orchid were identified, and PhFT-1 positively regulates flowering.

Antioxidant properties of different portions of organic Anoectochilus formosanus hayata with different drying treatments / Propriedades antioxidantes de diferentes porções de Anoectochilus formosanus hayata orgânicas com tratamentos de secagem diferentes

Anoectochilus formosanus HAYATA (Orchidaceae) is a valuable herb used as herbal medicine in Taiwan and other Asian countries. In order to enhance the economic efficiency of organic cultivation and understand how the processing affects plant properties, organic A. formosanus extracts were used as the investigative material in this study. The entire plant, the roots, stems and leaves from organic A. formosanus were freeze-dried or hot air-dried; all samples used methanol to extract. The antioxidant properties and active components were analyzed. The results indicated that antioxidant properties of both the freeze-dried and hot air-dried leaves of organic A. formosanus have better performance, including reducing power and ,-diphenyl--picrylhydrazyl (DPPH) radical scavenging activity. The contents of phenols and flavonoids showed positive relationships with reducing power, DPPH radical scavenging activity and trolox equivalent antioxidant capacity. The contents of carotenoids positively correlated with trolox equivalent antioxidant capacity and the contents of all active components were negatively correlated with the ferrous ion chelating ability. To summarize the results, freeze-dried leaves of organic A. formosanus have better antioxidant properties at a suitable concentration. This study may raise the economic values of A. formosanus by the organic cultural technique.

Anoectochilus formosanus HAYATA (Orchidaceae) é uma erva valiosa usada como medicina herbal em Taiwan e outros países asiáticos. A fim de aumentar a eficiência econômica do cultivo orgânico e entender como o processamento afeta as propriedades das plantas, os extratos orgânicos de A. formosanus foram utilizados como material investigativo neste estudo. A planta inteira, as raízes, hastes e folhas de A. formosanus orgânica foram liofilizadas ou secas por ar quente; todas as amostras usaram metanol para a extração. As propriedades antioxidantes e os componentes ativos foram analisados. Os resultados indicaram que as propriedades antioxidantes de ambas as folhas liofilizadas e secas por ar quente da A. formosanus orgânica apresentam melhor desempenho, incluindo o poder redutor e a atividade de eliminação do radical , -difenil-p-picrilidrazilo (DPPH). Os conteúdos de fenóis e flavonóides mostraram relações positivas com a potência redutora, atividade de eliminação de radicais DPPH e capacidade antioxidante equivalente de trolox. O conteúdo de carotenóides correlacionou-se positivamente com a capacidade antioxidante equivalente de trolox e os conteúdos de todos os componentes ativos foram correlacionados negativamente com a capacidade de quelação de íons ferrosos. Para resumir os resultados, as folhas liofilizadas de A. formosanus orgânica possuem melhores propriedades antioxidantes a uma concentração adequada. Este estudo pode elevar os valores econômicos de A. formosanus pela técnica cultural orgânica.