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.

Anatomy and micromorphometric analysis of leaf Catasetum x apolloi Benelli & Grade with addition of potassium silicate under different light sources / Anatomia e análise micromorfométrica foliar de Catasetum x apolloi Benelli & Grade com adição de silicato de potássio em diferentes fontes de luz

Abstract The aim of this study was to compare vitroplants Catasetum x apolloi grown under natural light and artificial light and different concentrations of potassium silicate, providing data on the anatomical differentiation that aids the acclimatization process of this species. Plants from in vitro seeding were used; 5 protocorms of approximately 0.5 cm were inoculated into vials with a capacity of 500 mL containing 100 mL of alternative culture medium plus potassium silicate (0.0, 0.5; 1.0 mL L–1), pH adjusted to 5.5 ± 0.5 and gelated with 4GL–1 agar before the autoclaving process. Cultures were maintained under natural light (TNE) and artificial light (TAE) for 90 days, and micromorphometric analysis was performed for polar and equatorial diameter, density and stomatal index, blade thickness in the central rib, and secondary veins. Applications in K2SiO4 alternative medium provided the following: elongation of the hypodermis, thicker mesophyll, and more prominent midrib; elipptical guard cells; formation of epistomatal chamber; and lower stomatal density and stomatal with lower equatorial and polar diameters. The conditions that favored the acclimatization were lower light intensities and lower potassium silicate doses.

Resumo O objetivo desse trabalho foi comparar vitroplantas de Catasetum x apolloi cultivadas sob luz natural e luz artificial e diferentes concentrações de silicato de potássio, fornecendo dados sobre diferenciação anatômica que auxiliem no processo de aclimatação dessa espécie. Utilizou-se plantas provenientes da semeadura in vitro, 5 protocormos de aproximadamente 0,5 cm foram inoculados em frascos com capacidade para 500 mL contendo 100 mL de meio de cultura alternativo, acrescido de silicato de potássio (0,0; 0,5; 1,0 mL L–1), pH ajustado para 5,5 ±0,5 e gelificado com 4gL–1 de ágar antes do processo de autoclavagem. As culturas foram mantidas sob luz natural (TAA) e luz artificial (TAN) por 90 dias, e feitas análises micromorfométricas (diâmetro polar e equatorial, densidade e índice estomático, espessura do limbo na nervura central e nervuras secundárias). As aplicações de K2SiO4 em meio alternativo, propiciaram: alongamento da hipoderme; mesofilo mais espesso e nervura central mais proeminente; células guardas elípticas; formação de câmaras supraestomáticas; menor densidade estomática e estômatos com menores diâmetros equatorial e polar. As condições que podem favorecer a aclimatação são menores intensidades de luz e menores doses de silicato de potássio.

Oncidiinae (Orchidaceae) on the great curve of the Xingu River, Pará state, Brazil / Oncidiinae (Orchidaceae) na volta grande do Rio Xingu, Pará, Brasil

Among the studies on Orchidaceae in the Amazon, none comprised the region of the Great Curve of the Xingu River, located in the lower Xingu river. The aim of this study was to inventory and taxonomically study the species of Oncidiinae (Orchidaceae) in the Great Curve of the Xingu River, Pará state. The floristic survey was performed in the area of the Belo Monte hydroelectric plant, in the Vitória do Xingu municipality, centrally inserted in the called Great Curve of the Xingu River. Botanical collections were accomplished between June 2011 and December 2013. A total of 27 species of Oncidiinae, distributed in 15 genera, was inventoried in the study area. Notylia Lindl. and Trichocentrum Poepp. & Endl. were the richest genera, with five and four species, respectively, followed by Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., and Ornithocephalus Hook., with two species each. The remaining eight genera are represented by a single species each in the study area. Morphological descriptions, a key for taxonomic identification, illustrations, and comments on distribution, ecology, phenology and morphology are provided for all inventoried species.

Entre os estudos com Orchidaceae na Amazônia, nenhum compreende a região da Volta Grande do rio Xingu, localizada no baixo Xingu. O objetivo deste estudo foi inventariar e estudar taxonomicamente as espécies de Oncidiinae (Orchidaceae) na Volta Grande do rio Xingu, estado do Pará, Brasil. O levantamento florístico foi realizado na área da Usina Hidrelétrica de Belo Monte, no município de Vitória do Xingu, inserido centralmente na chamada Volta Grande do Xingu. Foram realizadas coletas botânicas entre junho de 2011 e dezembro de 2013. Na área de estudo, foram inventariadas 27 espécies de Oncidiinae, distribuídas em 15 gêneros. Notylia Lindl. e Trichocentrum Poepp. & Endl. foram os mais ricos, com cinco e quatro espécies respectivamente, seguidos por Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., e Ornithocephalus Hook., com duas espécies cada. Os oito gêneros restantes estão representados na área de estudo por uma única espécie. São fornecidas descrições morfológicas, chave taxonômica para identificação, ilustrações e comentários sobre distribuição, ecologia, fenologia e morfologia para todas as espécies inventariadas.

Oncidiinae (Orchidaceae) on the great curve of the Xingu River, Pará state, Brazil / Oncidiinae (Orchidaceae) na volta grande do Rio Xingu, Pará, Brasil

Abstract Among the studies on Orchidaceae in the Amazon, none comprised the region of the Great Curve of the Xingu River, located in the lower Xingu river. The aim of this study was to inventory and taxonomically study the species of Oncidiinae (Orchidaceae) in the Great Curve of the Xingu River, Pará state. The floristic survey was performed in the area of the Belo Monte hydroelectric plant, in the Vitória do Xingu municipality, centrally inserted in the called Great Curve of the Xingu River. Botanical collections were accomplished between June 2011 and December 2013. A total of 27 species of Oncidiinae, distributed in 15 genera, was inventoried in the study area. Notylia Lindl. and Trichocentrum Poepp. & Endl. were the richest genera, with five and four species, respectively, followed by Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., and Ornithocephalus Hook., with two species each. The remaining eight genera are represented by a single species each in the study area. Morphological descriptions, a key for taxonomic identification, illustrations, and comments on distribution, ecology, phenology and morphology are provided for all inventoried species.

Resumo Entre os estudos com Orchidaceae na Amazônia, nenhum compreende a região da Volta Grande do rio Xingu, localizada no baixo Xingu. O objetivo deste estudo foi inventariar e estudar taxonomicamente as espécies de Oncidiinae (Orchidaceae) na Volta Grande do rio Xingu, estado do Pará, Brasil. O levantamento florístico foi realizado na área da Usina Hidrelétrica de Belo Monte, no município de Vitória do Xingu, inserido centralmente na chamada Volta Grande do Xingu. Foram realizadas coletas botânicas entre junho de 2011 e dezembro de 2013. Na área de estudo, foram inventariadas 27 espécies de Oncidiinae, distribuídas em 15 gêneros. Notylia Lindl. e Trichocentrum Poepp. & Endl. foram os mais ricos, com cinco e quatro espécies respectivamente, seguidos por Erycina Lindl., Ionopsis Kunth, Lockhartia Hook., Macradenia R.Br., e Ornithocephalus Hook., com duas espécies cada. Os oito gêneros restantes estão representados na área de estudo por uma única espécie. São fornecidas descrições morfológicas, chave taxonômica para identificação, ilustrações e comentários sobre distribuição, ecologia, fenologia e morfologia para todas as espécies inventariadas.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Mass propagation of Cymbidium giganteum Wall. ex Lindl. using in vitro seedlings.

In vitro seedlings were used as explants for protocorm like bodies (PLBs) production which in turn were used for regeneration purpose. PLBs were induced from the base of seedlings (1.0-1.5 cm in size) in MS + BAP (8.88 µM). After 90 days of inoculation, PLBs production rate started declining and most of the PLBs turned into plantlets. Preculture of seedlings in 1.0 µM thidiazuron (TDZ) for 7 days and transfer to BAP supplemented medium resulted in production of 16 PLBs per seedling within 90 days of culture. Increase of TDZ concentration to 2.5 µM and preculture time 15 days, resulted in induction of highest number of PLBs (19 PLBs per seedling) in the basal medium. The results emphasized the importance of thidiazuron (TDZ) concentration and preculture time for PLBs proliferation from the base of seedlings. The PLBs thus produced were used for regeneration studies. Irrespective of single, segmented or clumps of PLBs, the regeneration response was 100% in 2,4-D (4.52 µM) and KN (4.64 µM) but when KN was replaced by BAP (8.88 µM), response was observed only in clumps of PLBs, whereas in single and segmented ones it was 99 and 97%, respectively. Regenerants developed stout root system in half strength M medium supplemented with 2.84 µM of IAA and transferred to greenhouse with 90% survival. The present study holds tremendous potential as the mother plant is not destroyed and PLBs are produced as a continuous system.

In vitro regeneration of Coelogyne nervosa A.Rich. and Eria pseudoclavicaulis Blatt., threatened orchids of Western Ghats, India.

The seeds of C. nervosa and E. pseudoclavicaulis were germinated asymbiotically on Knudson C (KC) and Schenk and Hildebrandt basal medium (SH). Growth regulators such as 2,4-Dichlorophenoxyacetic acid (2,4-D) individually and in combinations with benzyladenine (BA) and kinetin were used for callus induction from the protocorm like bodies. Coelogyne nervosa showed maximum (90%) callus induction in Knudson C medium supplemented with 2,4-D (2.26 µM) and Eria pseudoclavicaulis showed 60% callus induction in Schenk and Hildebrandt medium supplemented with 2,4-D (2.26 µM). Calli developed a route of production of protocorm-like bodies and eventually developed into plantlets on transfer to growth regulator free half strength basal medium. The well rooted plants were hardened successfully in the potting mixture containing coconut husk, charcoal, and brick pieces in the ratio 2:1:1.

Evaluación de diferentes medios de cultivo in vitro en el desarrollo de híbridos de Phalaenopsis (Orchidaceae) / Evaluation of different in vitro culture media in the development of Phalaenopsis hybrid (Orchidaceae)

Los híbridos de Phalaenopsis tienen una gran importancia económica a nivel mundial, como flor cortada y planta ornamental, debido a sus flores vistosas y a la capacidad de adaptación a diferentes condiciones ambientales. Las técnicas de cultivo in vitro resultan indispensables para mejorar la eficacia germinativa, el crecimiento y desarrollo de orquídeas con fines comerciales e investigativos. En esta investigación se determinó el medio de cultivo más apropiado para la germinación in vitro de un híbrido de Phalaenopsis. Inicialmente se evaluó la viabilidad de las semillas utilizando la prueba de tetrazolio (TZ). Las semillas se desinfectaron y se cultivaron aplicando el método de la jeringuilla. El porcentaje de viabilidad en promedio fue de 92,2 % (P≤ 0,05: Tukey HSD), con un porcentaje de germinación entre todos los medios de 95,1 % (P≤ 0,05: Tukey HSD). El medio de cultivo más eficiente para la germinación de híbridos de Phalaenopsis a las 18 semanas de cultivo fue el Murashige & Skoog (MS) suplementado con agua de coco, y jugo de piña con diferencias estadísticamente significativas (P≤ 0,05: Tukey HSD), con respecto a los demás medios de cultivo, contribuyendo de esta manera al uso de componentes orgánicos con el fin de mejorar la germinación y desarrollo de Phalaenopsis.

The Phalaenopsis hybrids have a significant economic importance throughout the world, as ornamental flower or plant. It is because of its attractive flowers and its adaptation capacity into different environments. The different culture media in vitro are vital to improve the efficacy of germination, growing and development of the Orchids for commercial and research purposes. In this research, the most appropriated medium for in vitro propagation of Phalaenopsis hybrid was determined. At first, the seeds viability was evaluated by using tetrazolium test (TZ). The seeds were disinfected and cultivated by means of the syringe method. The viability percentage average was 92.2 % (P≤ 0.05: Tukey HSD), with a percentage of germination of 95.1 % (P≤ 0.05: Tukey HSD) in all the environments. The most efficient culture Medium for Phalaenopsis hybrid phenological development, at 16 weeks, was Murashige & Skoog (MS). Coconut water and pineapple juice were used as supplement showing statistically significant differences (P≤ 0,05: Tukey HSD), in comparison with the other culture media, contributing this way to the usage of organic components, which will be employed to improve the germination and development of the Phalaenopsis.

Structural adaptations of two sympatric epiphytic orchids (Orchidaceae) to a cloudy forest environment in rocky outcrops of Southeast Brazil / Adaptaciones estructurales de dos orquídeas epífitas simpátricas (Orchidaceae) en un bosque nuboso de un campo rupestre del sureste de Brasil

The survival of plants in epiphytic environments depends on vegetative adaptations capable to defraud different stresses. Based on the structural diversity of the Orchidaceae, the current study has the objective of relating the anatomical structure of Dichaea cogniauxiana and Epidendrum secundum with the distinct environments where they live. It was expected that, despite structural similarities as strategies for resource acquisition, some peculiar variations related to the distinct light microenvironments (inside or in the edge of the nebular forest, near to “campo rupestre” area) might be found. Leaves and roots of both species were collected in a nebular forest located at a “campo rupestre” area at Serra da Piedade, Brazil), in January and February 2005. D. cogniauxiana is adhered to trunks, in sites with high atmospheric humidity and shaded, while E. secundum is located at the edge of the nebular forest, in more luminous sites. The leaves of E. secundum had thicker cuticle and higher number of stomata per area than those of D. cogniauxiana, characteristics coherent with their distinct pattern of exhibition to sun light. The suprastomatic chambers formed by the thicker cuticle may function as a barrier of resistance to water evaporation. The succulence of the leaves of E. secundum propitiates organic acids storage at night, and the storage of starch may be involved in PEP-carboxylase metabolism, both propitiating CAM mechanism. Roots with larger number of cell layers of the velamen, and specialized thick walled cortical cells (both in E. secundum) help water absorption and indicate better adaptation to an environment with intense solar radiation and a probable higher water deficit. The remarkable cell wall thickening of E. secundum exodermis can confer more efficient protection against the excess of transpiration at the border of the nebular forest. On the other hand, besides D. cogniauxiana be epiphyte, it is in a low position - in a shaded environment and with high relative humidity. Its thin thickened velamen permits the entrance of the low available light, and photosynthesis, producing oxygen and helping to avoid hypoxia condition. As features registered for D. cogniauxiana and E. secundum roots, we can depict the velamen, distinct exodermis and endodermis, and specialized thick walled cortical cells as characteristic of epiphytic plants.

La sobrevivencia de las plantas epífitas a su ambiente depende de las adaptaciones vegetativas que les permiten afrontar diferentes tipos de estrés. Basado en la diversidad estructural de las Orchidaceae, el presente estudio tiene por objetivo relatar la estructura anatómica de Dichaea cogniauxiana y Epidendrum secundum con relación a los distintos ambientes donde viven. Hojas y raíces fueron recolectados en un bosque nuboso de un campo rupestre en la Serra da Piedade, Brasil. Dichaea cogniauxiana está adherida a los troncos, en sitios con alta humedad atmosférica y sombra, mientras que E. secundum está localizada en el margen del bosque nebular, en sitios más iluminados. Las hojas de E. secundum tienen una cutícula más gruesa y un mayor número de estomas por área que las de D. cogniauxiana, características coherentes con los diferentes patrones de exhibición a la luz del sol. Las cámaras supraestomáticas formadas por la cutícula pueden funcionar como una barrera de resistencia a la evaporación del agua. La suculencia de las hojas de E. secundum propicia el almacenamiento de ácidos orgánicos por la noche, y el almacenamiento de almidón puede estar implícito en el metabolismo de la PEP-carboxilasa. Las raíces con mayor velamen y especializadas paredes gruesas de las células corticales (ambas en E. secundum) ayudan a la absorción e indican una mejor adaptación al ambiente con intensa radiación solar y probable alto déficit de agua. El engrosamiento de la pared celular en la exodermis de E. secundum puede conferir una protección más eficaz contra el exceso de transpiración en el margen del bosque nebular. Por otra parte, D. cogniauxiana además de ser epífita, está en una posición baja, en un ambiente sombreado y con una alta humedad relativa. Su velamen delgado permite la entrada de la baja luz disponible y la fotosíntesis, produciendo oxígeno que ayuda a evitar la condición hipóxica.

Acclimatization of phalaenopsis and cattleya obtained by micropropagation / . Aclimatización de phalaenopsis y cattleya obtenidas por micropropagación

The quality of micropropagated plants relies on the acclimatization stage. This research intends to develop an efficient protocol to obtain the acclimatization of Phalaenopsis and Cattleya. Plants of Phalaenopsis obtained from protocorms were selected. They came from flowering stalks grown at modified Murashige and Skoog (MS) (1962) medium and classified by growth ranks and put into moss, mesquite wood shaving and perlite (1:1:1), into a humidity chamber. The protocorms were multiplied at MS from Cattleya sown in Knudson C (1946) medium; regenerated plants of 1-2 cm were selected, and implanted in humidity chamber on: moss, coal and perlite (1:1:1) MCP; mesquite wood shavings, coal and perlite (1:1:1) ACP; moss and perlite (1:1) MP; mesquite wood shaving and perlite (1:1) AP. The following results were obtained: Phalanopsis: a) Survival: 44% in R0 and 100% in RI and RII of the. b) Number of leaves: RI gave on average 1 more leaf than the range 0; c) Roots number and length: RI and RII gave on average 2 more roots than R0, and there were no significant differences in length. d) Height: RII presented greater growth than RI and Ro. Cattleya: a) The survival in MCP was 0%, MP 16 %, ACP 32% and AP 80%. b) The height in MP was significantly superior to the ones in ACP and AP. Plants from both genera need to achieve a 2 to 4 cm growth rank in vitro to endure the greenhouse conditions. MAP was the best substrate in Phalaenopsis and moss-perlite in Cattleya.

La calidad final de las plantas producidas por micropropagación depende de la etapa de aclimatización. Se intenta desarrollar un protocolo eficiente para la aclimatización de Phalaenopsis y Cattleya. Se seleccionaron plantas de Phalaenopsis, obtenidas de protocormos provenientes de estacas florales cultivadas en Murashige y Skoog modificado (MS) (1962), por rangos de crecimiento e implantadas en musgo, viruta de algarrobo y perlita (1:1:1), en cámara húmeda. De siembras de Cattleya en medio de Knudson C (1951) se multiplicaron protocormos en MS; se seleccionaron plantas regeneradas de 1-2 cm, e implantadas en cámara húmeda en los sustratos: musgo, carbón y perlita (1:1:1) MCP; viruta de algarrobo, carbón y perlita (1:1:1) ACP; musgo y perlita (1:1) MP; viruta de algarrobo y perlita (1:1) AP. Se obtuvieron los siguientes resultados: en Phalaenopsis: a) Supervivencia: para R0 de 44% y RI y RII del 100%; b) número de hojas: RI generó en promedio 1 hoja más que el rango 0; c) número y longitud de raíces: RI y RII generaron en promedio dos raíces más que R0, no detectándose diferencias significativas en longitud; d) altura: RII presentó mayor crecimiento que RI y R0 En Cattleya: a) La supervivencia en MCP fue 0%, MP 16%, ACP 32% y AP 80%; b) La altura en MP resultó significativamente superior que en ACP y AP. Ambos géneros necesitan alcanzar un crecimiento de 2 a 4 cm in vitro para tolerar las condiciones de invernáculo. El mejor sustrato fue MAP en Phalaenopsis, y la mezcla musgo-perlita en Cattleya.

Rapid in vitro propagation of the threatened endemic orchid, Ipsea malabarica (Reichb.f.) J D Hook through protocorm-like bodies.

Rapid propagation of I. malabarica (Reichb. f.) J D Hook, an endemic and endangered orchid of the Western Ghats of Kerala, India through conversion of axillary buds to protocorm-like bodies (PLBs), and subsequent plant regeneration was achieved. Growth regulators and sugar displayed significant influence in the induction of PLBs. In vitro derived shoots from field grown rhizomes of Ipsea cultured on Murashige and Skoog (MS) medium with 13.3 microM N6-benzyladenine (BA) containing 2% commercial grade sugar turned the axillary buds to PLBs within 25 days, and developed a mean of 33.1 PLBs within 50 days. Kinetin (KIN) did not induce PLBs, but facilitated axillary bud proliferation. Transfer of PLBs on medium having same concentration of BA and sugar facilitated rapid multiplication, and developed a mean of 47.5 PLBs. No decline of PLB proliferation was observed up to 10th subculture. Half strength MS medium with 6.97 microM KIN facilitated conversion of 98% PLBs to plantlets. On this media, a mean of 5.8 roots were also developed per shoot. Shoots developed bulbs during culture were grown to rhizomes. Increase of sugar to 6 or 8% hastened the development of bulbs/rhizomes. Reintroduction of PLB-derived plantlets in the natural habitat i.e. at Vellarimala (at 1300 m height) of the Western Ghats of Kerala was attempted as a means to assist in situ conservation. This is the first report of conversion of axillary buds to PLBs. The protocol enables to surmount the threat of extinction of this endemic and endangered orchid.

Las orquídeas de los cafetales en México: una opción para el uso sostenible de ecosistemas tropicales / Orchids from coffee-plantations in Mexico: an alternative for the sustainable use of tropical ecosystems

Life form, endemism, conservation status, and horticultural interest are detailed for orchid species associated to shade coffee-plantations in Mexico. About 11% of the orchid taxa (214 species) found in these agroecosystems are in the Mexican list of species requiring some form of protection. Almost 40% of the species are of horticultural interest. The importance of promoting shaded coffee plantations as an alternative to the conservation of primary plant communities in Mexican and other tropical regions is clear. Long term management plans are recommended

Se presenta el listado de las especies de orquídeas asociadas a cultivos de café de sombra en México, incluyendo datos sobre forma de vida, endemismo, status de conservación e interés hortícola. Se discute brevemente la importancia de promover el cultivo de café de sombra como una alternativa para la conservación de comunidades vegetales primarias en áreas tropicales y se presentan algunos puntos a considerar para impulsar una estrategia clara y bien definida de protección y conservación de la diversidad orquideológica en los cafetales mexicanos