Mexican Orchid Research Articles

Medicinal Orchids of Mexico: A Review.

Some species of the Orchidaceae family are used in Mexican traditional medicine. However, there are no current and critical compilations of the medicinal uses and pharmacological effects of the members of the Orchidaceae family. This review provides a current, critical, and comprehensive analysis of the traditional medicinal uses, pharmacological reports, and active compounds isolated from Mexican orchids. A total of 62 Mexican orchids with medicinal potential have been recorded, of which 14 have scientific evidence. The remaining 48 plant species have ethnomedicinal information but have not been validated with scientific studies. These orchids are distributed in 14 states of the Mexican Republic, mainly in the southern region of Mexico. The most common pharmacological activities reported are anti-inflammatory, vasorelaxant, antinociceptive, antioxidant, spasmolytic, antihypertensive, and hallucinogenic activities. It is necessary to increase the number of pharmacological, phytochemical, and toxicological studies with medicinal orchids from Mexico because there are scientific studies on only 22.5% of these species. In further studies, it will be possible to evaluate the pharmacological effects of Mexican orchids in clinical trials. In addition, the mechanisms of action by which plant extracts and their active compounds exert medicinal effects remain to be studied. Plant extracts from orchids and their active compounds show promising antinociceptive and spasmolytic effects, respectively.

Assessment of tepojal as a support medium in in vitro germination of Barkeria whartoniana (Orchidaceae) and subcultures of B. uniflora and B. scandens

Objective: We evaluated the use of tepojal as a substitute for bacteriological agar for in vitro culture of three species of Mexican orchids of the genus Barkeria. We had three objectives: (1) evaluate the use of tepojal for the in vitro germination of B. whartoniana seeds; (2) compare the growth and survival of B. uniflora and B. scandens plants in in vitro culture on bacteriological agar and tepojal, and (3) compare the early ex situ survival of the B. uniflora and B. scandens previously cultivated in vitro on bacteriological agar and tepojal. Design/Methodology/Approach: For the growth experiment, 1,050 seedlings of Barkeria scandens and B. uniflora were used. Two types of culture medium were prepared: (1) Liquid with tepojal and 40 % MS medium with dextrose, yeast, coconut water, activated carbon and refined sugar, and (2) solid at 40 %, with the same elements as the liquid one, but with 6 g of bacteriological agar. For the germination experiment of Barkeria whartoniana in tepojal we use seeds from a mature (open) capsule. After disinfection, seeds were sown in tepojal with liquid medium and in a solid (agar) culture using a syringe technique. All the seeds were sown in 100 % p668 MS medium with 100 mL/L of coconut water and 15 g/L of refined sugar and using in both the MS medium. Results: Barkeria uniflora seedlings had a greater growth than in B. scandens regardless of the type of treatment. When comparing within each species, we found that the two treatments (tepojal vs agar) did not produce differences in the growth of the shoot of both species. Roots growth was influenced by both the effect of the species and the treatment, as the longest roots were found in tepojal medium. Seed germination observed in the liquid medium with tepojal was not apparently different from that in agar. In both cases the entire surface of the jar was covered with green protocorms. Findings/Conclusions: Tepojal did function well as a substrate to germinate Barkeria whartoniana, but tests must continue to be carried out to evaluate its effectiveness. As no differences were found for aerial growth between agar and tepojal in Barkeria scandens and Barkeria uniflora, we consider this substrate as a good substitute for agar because it can produce more biomass per gram of culture medium used; therefore, if we consider the costs the use of tepojal is much cheaper and much easier to get.

Symbiotic bacteria associated to Prosthechea citrina, a Mexican endemic orchid

En el proceso de cultivo <em>in vitro</em> de tejidos de plantas, se presentan microorganismos potencialmente fitopatógenos y/o aparentemente contaminantes, que eventualmente ocasionan perdidas de material vegetal. La presente investigación tuvo como objetivo identificar a bacterias endófitas asociadas a <em>Prosthechea citrina</em>, una orquídea endémica de México, mediante el aislamiento, cultivo<em> in vitro</em> y secuenciación del gen 16S del ARN ribosomal de las cepas bacterianas. Se identificaron a <em>Aeromonas hydrophila</em> y <em>Enterobacter</em> sp. como simbiontes, no patogénicos, del bulbo y de base de hoja y a <em>A. hydrophila</em> en la parte media y el ápice foliar. Se evaluaron cuatro antibióticos para su control<em> in vitro</em> mediante el método de disco-placa, cuantificando el diámetro de crecimiento de la colonia. El mayor halo de inhibición, con significancia estadística, se obtuvo con oxitetraciclina para <em>A. hydrophila</em> y con ampicilina para <em>Enterobacter</em> sp. Se demuestra la presencia de bacterias endófitas, con especificidad de ubicación en el tejido, así como el antibiótico correspondiente que las inhibe.

Prosthechea karwinskii, an orchid used as traditional medicine, exerts anti-inflammatory activity and inhibits ROS

Ethnopharmacological relevanceProsthechea karwinskii (Mart.) J.M.H. Shaw is a Mexican orchid used in traditional medicine by some indigenous communities to treat issues related to inflammation (cough, wounds, burns, and diabetes). Pharmacological research of this orchid could validate its therapeutic uses and demonstrate its potential for treating other health conditions of high prevalence in Mexico, including those associated with oxidative stress such as diabetes, cancer, atherosclerosis, and hypertension as well as inflammation. Aim of the studyThe leaf extract from P. karwinskii was examined to identify its compounds and elucidate its inhibitory effect on reactive oxygen species as well as its anti-inflammatory activity and gastroprotective effects in an animal model. Materials and methodsCompounds were identified via ultra-high-performance liquid chromatography coupled with electrospray ionization with quadrupole time of flight-mass spectrometry. Inhibition of reactive oxygen species was determined ex vivo in peripheral blood mononuclear cells with 2′,7′-dichlorodihydrofluorescein diacetate. The anti-inflammatory activity was assessed using a carrageenan-induced paw edema model in Wistar rats; nitric oxide and tumor necrosis factor alpha levels were quantified. The gastroprotective effect was evaluated in Wistar rats with indomethacin-induced gastric injury. ResultsNine compounds were identified in the P. karwinskii leaf extract. Most compounds, such as quinic acid, malic acid, neochlorogenic acid, chlorogenic acid, rutin, embelin, pinellic acid, and azelaic acid, were reported to exhibit antioxidant and/or anti-inflammatory activity. The extract was also found to inhibit reactive oxygen species in the ex vivo model. Unlike other anti-inflammatory drugs, the extract exerted a dual effect: anti-inflammatory activity and protection of the gastric mucosa. The results showed that the extract could significantly inhibit the release of nitric oxide without a dose-response relationship. ConclusionP. karwinskii leaf extract inhibited reactive oxygen species and exerted an anti-inflammatory effect. Moreover, this extract did not induce gastric damage in the animals. The bioactivity of the species was found to support its use in traditional medicine. This orchid could be used to treat inflammatory diseases without causing the side effects associated with nonsteroidal anti-inflammatory drugs. It can also be employed to treat other pathological conditions associated with oxidative stress. The findings herein form the basis for the future discovery of natural products that may serve as safe alternative therapies for inflammatory disorders.

Synthesis of wild orchid trade and demography provides new insight on conservation strategies

AbstractIllegal wildlife trade represents a global conservation priority, but the booming illegal trade in wild plants remains understudied. We use the Mexican orchid trade to illustrate an interdisciplinary approach to provide novel insight on conservation strategies and policies. We synthesize studies of orchid markets, national orchid confiscation records, CITES registers, and global population dynamics studies to document trade patterns and potential ecological impacts. We found 333 wild‐harvested orchid taxa illegally traded in domestic markets. Clear patterns emerged: 90% were epiphytic and <4% traded in high volumes, all of which had pseudobulbs and bloomed during cultural festivals. Most sales were pseudobulbs, not whole plants. Review of demographic studies indicates whole‐plant harvest is unviable but simulations show potential for sustainable harvest of pseudobulbs. The combination of social and ecological findings suggests a novel multipronged approach to improve conservation, including selective monitoring, enforcement focused on whole‐plant harvest, and community‐based wild harvest of pseudobulbs.

Species Richness May Determine the Income from Illicit Wild Orchid Trading in Traditional Markets in Mexico

The orchid trade is an informal activity in Mexico traditionally performed by indigenous people; however, this activity, based on the extraction of wild specimens without a management plan, is sanctioned by environmental laws. Few studies have documented the extraction volumes for illicitly traded orchids, the status of the populations affected by this activity, the socioeconomic profiles of the vendors, or the factors determining income, which are essential for designing strategies to conserve this resource. We documented species richness, abundance, and prices for orchids traded during December in traditional markets in Chiapas, Mexico. The relationships among price, species showiness, and some variables determining income were evaluated. We recorded 28 traded species, all of which were extracted from neighboring, protected, and high-biodiversity forests. The most income was generated from three species, which were also the most abundant in the markets. This trade affects the conservation of two threatened Mexican orchids, but at a state level, it also impacts populations of other species that are not considered at risk. A significant relationship was found between the price of an orchid and its flower size and between the vendor’s income and the quantity of orchids traded by him. The income obtained by a vendor was significantly explained by orchid inflorescence abundance, vendor’s age, and a biodiversity index estimated here.

ESTRATEGIAS PARA EL APROVECHAMIENTO SUSTENTABLE DE LAS ORQUÍDEAS DE MÉXICO


 
 
 Las orquídeas mexicanas son uno de los grupos de plantas de mayor interés hortícola en los ámbitos nacional e internacional. Dado que no hay suficientes viveros con planes de manejo y programas de reproducción para satisfacer la demanda que existe de estos organismos, numerosos ejemplares han sido extraídos de su hábitat durante más de 200 años. Se han registrado más de 1,200 especies en México y de ellas, alrededor de 200 están catalogadas con alguna categoría de riesgo. Una de las estrategias de conservación aquí propuestas es promover su aprovechamiento sustentable en las comunidades rurales, mediante el registro de viveros como Unidades de Manejo Ambiental (UMA), lo que permitirá disminuir la presión que ejerce la recolección sobre sus poblaciones. Debido a la dificultad para que sus semillas germinen y el tiempo que se invierte en obtener plantas por división vegetativa, los viveros rurales tienen que estar en coordinación con centros de reproducción masiva, donde se pueden obtener plantas en cantidades suficientes para su comercialización mediante técnicas de micropropagación. Además, es factible propagar materiales selectos con características ornamentales atractivos, los cuales les aportan valor agregado para su venta. En el presente documento se exponen algunas experiencias y se proponen estrategias para el aprovechamiento sustentable de las orquídeas de México.
 
 

The endemic orchids of Mexico: a review

Orchidaceae is the most extensive family of flowering plants. There are approximately 30,000 - 35,000 species, belonging to 850 genera, dispersed in almost all the ecosystems on earth, except in polar zones and extreme deserts. In Mexico, there are more than 1,200 species distribuited in approximately 170 genera, of which 40% are endemic to the country. In this review, aspects related to geographical distribution, ecology, conservation, biotechnological applications and uses of orchids growing in Mexico are discussed. The geographical zones where their ecological niches are located are described and the zones that belong to priority conservation areas in Mexico are highlighted. In addition, all works on Mexican orchids related to micropropagation, medicinal uses, beneficial applications for humans, and microorganism-host interaction are discussed. In summary, this study provides perspectives for new studies and research focused on the use, propagation and conservation of Mexican endemic orchids.

Effect of Prosthechea karwinskii (Orchidaceae) on obesity and dyslipidemia in Wistar rats

BackgroundProsthechea karwinskii is an endemic Mexican orchid, it’s currently used as decorative element and in the traditional medicine to treat diabetes and some problems related to inflammatory processes. AimTo determine antioxidant activity index (AAI) and to validate by the first time and through an rat model the hydroalcoholic extract obtained from Prosthechea karwinskii, a plant used in traditional medicine for treat conditions relate to the metabolic syndrome. MethodsFor in vivo assays 25 weaned male Wistar rats were divided into a control group (CG; n=5) and a Metabolic Syndrome group (MS; n=20). The rats of the latter were induced to MS with 40% sucrose in the drink water during 13weeks. After MS induction this group was subdivided into 4 groups: MS group (n=5) received sucrose, and three groups receiving 200mg/kg of body weight of each extract pseudobulb (P, n=5), leaf (L, n=5), and flower (F, n=5). All treatments were followed for 13days. Blood was collected at the end of the study to measure glucose, cholesterol and triglycerides. AAI were measured in the extracts by the method of DPPH. The results were analyzed using MINITAB 16.1.0, and the statistical significance was determined by ANOVA and a Tukey’s test (P<0.05). ResultsLeaves (L) extract had highest values in AAI, followed by flowers (F) and pseudobulb (P) extracts. Leaves extract had highest reducing effect on glucose level, while flower extract had highest reducing effect on the cholesterol and triglycerides levels. ConclusionsThe P. karwinskii extracts evaluated here reduces the glycemic and lipidemic parameters in Wistar rats with MS induced. These effects may be attributed to the high antioxidant capacity of the extracts.

Risk of extinction and patterns of diversity loss in Mexican orchids

La norma oficial mexicana (NOM-059-ECOL-2001) lista cerca de 200 especies de orquídeas en alguna categoría de riesgo (Extintas, En Peligro de Extinción, Amenazadas y Sujetas a Protección Especial). Construimos una base de datos que incluye la información más relevante para planear las estrategias de con- servación de estos taxa (nomenclatura, descripciones e ilustraciones para identificación, datos geográficos de todas las poblaciones conocidas, clima, hábitat, refugios, historia natural y ecología, características poblacio- nales, factores de riesgo, etc.). La información estará disponible través de la Comisión Nacional para el Conocimiento y Uso de la Biodiversidad, México (CONABIO). Con esta base de datos y usando sistemas de información geográfica se detectaron las áreas de concentración de orquídeas en riesgo y como éstas se rela- cionan con las Áreas Naturales Protegidas y las Áreas Terrestres Prioritarias para la conservación. Se discuten distintos patrones de riesgo, de amenazas y de pérdida de diversidad. Es evidente que los efectos del cambio climático, combinados con el mal manejo de los sistemas en hábitats únicos, constituyen las mayores amena- zas. La erradicación de poblaciones de orquídeas en amplias zonas densamente pobladas y afectadas, espe- cialmente en Veracruz y Puebla, pueden representar una pérdida importante de la diversidad genética total de las especies. Se hizo un esfuerzo especial por determinar las tasas de extinción de orquídeas en México. Finalmente, y de manera conjunta con otros biólogos de la conservación se desarrolló un método para evaluar el riesgo de extinción en plantas que está siendo adoptado por las autoridades del país y su uso pretende ser obligatorio en el futuro para la inclusión en las distintas categorías de riesgo.

The important role that the Botanical Garden of the National Autonomous University of Mexico plays in the conservation of mexican orchids

la Ciudad de México, México; asentado en terrenos de origen volcánico en un matorral xerófito; tiene el propósito de mantener colecciones de plantas vivas representativas de la diversidad vegetal de México, que sirven de apoyo a la investigación, conservación, educación y Divulgación de la diversidad botánica. El Jardín posee una colección Viva de Orquídeas, tanto epífitas como terrestres, en la que se han llevado a cabo diferentes actividades. Con este trabajo se enfatiza el papel que juega el Jardín Botánico de la UNAM, en la conservación de la familia Orchidaceae de México.

Two additions to the Mexican orchid flora

Two species of Orchidaceae are recorded here for the first time in Mexico, namely Cryptarrhena guatemalensis and Domingoa gemma. Cryptarrhena guatemalensis is widespread in the Neotropics and can be distinguished from C. lunata, the only other species of the genus, by possessing pseudobulbs and narrowly triangular, attenuate apical labellum lobules. Domingoa gemma, previously known only from Guatemala and El Salvador, differs from the similar D. purpurea in the smooth, linear-lanceolate leaves and labellum lacking a prominent sac at the base. Both species are known in Mexico from single localities in the state of Chiapas.

First Report of Cymbidium mosaic virus and Odontoglossum ringspot virus in Orchids in Mexico

In 2010, a survey for viral diseases in commercial, orchid-producing greenhouses was carried out in Morelos, Mexico. Many symptomatic plants were observed. The most common leaf symptoms were yellow mottle, yellow streaks, and chlorotic and necrotic ringspots. Leaf samples were collected from eight symptomatic plants from the following genera: Encyclia, Oncidium, Shomburghia, Brassia, Guarianthe, Cattleya, Epidendrum, Vanilla, Xilobium, Laelia, and Brassocattleya. Samples were tested using double-antibody sandwich (DAS)-ELISA (Agdia, Elkhart, IN) with antiserum for Cymbidium mosaic virus (CymMV), Odontoglossum ringspot virus (ORSV), Cymbidium ringspot mosaic virus, and Tobacco mosaic virus (TMV) and a general antiserum for potyviruses. At least one plant from each genus was positive to CymMV and ORSV as individual or mixed infections. Encyclia and Laelia plants were the most frequently found with mixed infections by both viruses. All genera were negative for TMV and potyviruses. Total RNA extracts were obtained from all ELISA-positive samples by a modified silica capture protocol (2). Reverse transcription (RT)-PCR was carried out with general polymerase (RdRp) gene primers corresponding to the Potexvirus group (3) and specific primers for the coat protein gene (CP) of CymMV and ORSV (1). The PCR amplification from a positive sample of each genus was resolved in agarose gels. Amplification products of the expected size were obtained for CymMV and ORSV. Five CymMV RdRp gene clones from five different plants of Laelia (GenBank Accession Nos. HQ393958, HQ393959, HQ393960, HQ393961, and HQ393962), two CP gene clones of CP gene of CymMV from two different plants of Oncidium (GenBank Accession Nos. HQ393956 and HQ393957), and three CP clones of CP of ORSV from three different plants of Encyclia (GenBank Accession Nos. HQ393953, HQ393954, and HQ393955) were sequenced. The nucleotide sequences of the Mexican orchid CymMV isolates were 96 to 97% identical to CymMV sequences in the GenBank, while those of ORSV were 99 to 100% identical to deposited ORSV sequences. To our knowledge, this is the first report of CymMV and ORSV in orchids in Mexico, which are two of the most important quarantine virus in orchids in Mexico.

Acclimatization of the endangered Mexican epiphytic orchid, Laelia speciosa (H.B.K.) Schltr

In vitro propagation could be an alternative for the conservation of the endemic and endangered Mexican epiphytic orchid Laelia speciosa (H.B.K.) Schltr. The goal of this study was to develop a protocol that would enhance acclimatization of in vitro - derived L. speciosa plantlets - a critical stage in propagation and subsequent conservation. Observations of stomata opening during ex vitro acclimatization, and the time of in vitro culture (0, 5, 10, 15, 20, 25, 30 days) in greenhouse conditions (pre-acclimatization), on the survival and development of seedlings during the ex vitro acclimatization were carried out. In addition, the effect of different levels of nutrients (100%, 75%, 50%, 25% and 0% -strength salts) and sucrose (0, 10, 20, 30, 40 g l -1) in the Murashige and Skoog medium (MS) on the same parameters were measured. Plantlets incubated 20 days in greenhouse conditions before ex vitro acclimatization also displayed the best growth with a survival rate of 97.5%, related with high stomata opening. Plantlets on MS containing 100% -strength salts (with 20 days of pre-acclimatization, 40 g l -1 sucrose) had the highest rate (97.5%-100%) of survival and vigor when acclimatized. By improving micropropagation through acclimatization, the sustainable management of L. speciosa now more likely, benefitting the conservation of this endangered species.

Dos nuevos nuevos registros y un redescubrimiento de Orchidaceae de México

Se reporta para la flora mexicana la presencia de dos especies de orquídeas, Oncidium poikilostalix y Telipogon helleri, ambas previamente registradas de Centroamérica. La primera se conocía ampliamente con el nombre de Sigmatostalix poikilostalix, mientras que la segunda con el de Stellilabium helleri. También se comunica el redescubrimiento en México de otra orquídea, Plectrophora alata, la cual había sido encontrada en el país hace 75 años y no había vuelto a ser colectada desde entonces, por lo que se suponía extinta. Estas tres plantas fueron recientemente halladas al explorar los bosques húmedos de elevaciones intermedias en la región del volcán Tacaná, en Chiapas. Cada especie se describe e ilustra a partir de especímenes mexicanos y se discuten sus similitudes con taxa relacionados.

Phragmipedium warscewiczii, a new record for the Mexican orchid flora

Phragmipedium warscewiczii is reported as new record for the Mexican orchid flora from material recently collected in Chiapas. The species is described from Mexican specimens and it is compared with P. exstaminodium, its closest ally. The Mexican locality is the northernmost and most distant population for P. warscewiczii. Information on nomenclature, distribution, habitat, and factors of risk for this orchid is provided.

Oncidium cavendishianum (Bateman), NUEVO REPORTE PARA LA ESTACIÓN CIENTÍFICA LAS JOYAS. GUADALAJARA, JALISCO, MÉXICO.

A new report is presented herein of the Mexican orchid Oncidium cavendishianum (Bateman) in cloud forest areas at the Las Joyas Research Station, the Sierra de Manantlán Biosphere Reserve, Guadalajara, Jalisco, Mexico. The species has also been reported for the municipalities of Temascaltepec, Cieneguillas de González, San Lucas del Pulque, Telpintla, Carniceria and Real de Arriba. Species identification was carried out by the botany department faculty of the Centro Universitario de la Costa Sur, Autlan de Navarro. The collection was processed and preserved in the herbarium of the above-mentioned entity. This work includes a photographic image of the plant, along.

Morphological variation in Laelia anceps subsp. dawsonii f. chilapensis Soto-Arenas Orchidaceae in traditional home gardens of Chilapa, Guerrero, Mexico

Calaverita an endangered Mexican orchid is one of the most elegant and showy orchids of the genus Laelia and essential symbol within the local traditional celebrations of the Day of the Dead, in the region of Chilapa-Guerrero. We investigated morphological variation in flowers of la calaverita (Laelia anceps Lind. subsp. dawsonii (J. Anderson) Rolfe f. chilapensis Soto-Arenas, Orchidaceae) using the labellum of the specimens cultivated in traditional home gardens in the region of Chilapa, Guerrero, Mexico. The use of morphometric traits of the labellum is useful in appraising infra-subspecific morphological variation within the chilapensis form. Floral morphological variation exists among the specimens examined. These specimens, found exclusively in traditional home gardens, were grouped into four morphotypes: NAT, Topiltepec, Xulchuchio, and Terrero. The study of variation of la calaverita revealed the existence of four local morphotypes that originated from a long and complex process of selection of traits which responds to traditional preferences and knowledge about specific biotic and agroecological conditions. The cultural and aesthetic influence of traditional homegarden owners in the Chilapa region acts directly on selection, use and conservation of the morphological traits of la calaverita.

Propagation and Establishment of Three Endangered Mexican Orchids from Protocorms

Protocols for in vitro propagation from protocorms of Mormodes tuxtlensis Salazar, Cuitlauzina pendula La Llave &amp; Lex., and Lycaste skinneri (Batem. Ex. Lind.) Lind., three endangered species distributed in Mexico and highly appreciated as ornamentals, were developed. The effect of two different culture media, Murashige and Skoog (MS) and modified Knudson (KCm), combined with varying concentrations of N6-benzyladenine (0, 2.2, 4.4, 8.9, and 13.3 μM) and α-naphthaleneacetic acid (0, 0.5 and 2.7 μM), were investigated. Shoot formation and development of protocorm-like bodies were observed. For all three species, cultures in MS produced more shoots per explant than those in KCm, and those shoots were longer and more robust in appearance. Maximum number of shoots for M. tuxtlensis (1.5) and C. pendula (24.3) were obtained in media supplemented with 13.3 μM and 2.2 μM N6-benzyladenine, respectively. Conversely, for L. skinneri the greatest shoot production (16.4) was achieved in medium supplemented with 2.7 μM α-naphthaleneacetic acid. Subculturing explants in MS basal medium allowed further development and rooting of the shoots as well as growth of protocorm-like bodies. The effect of different potting mixes on ex vitro survival plantlets was also investigated; pine bark:oak charcoal:pumice (3:1:1) allowed the highest survival rates in all three species.

Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid

Two procedures for the in vitro propagation of Encyclia mariae, a threatened Mexican orchid, were developed. In the first procedure, leaves from in vitro germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with the range of 2.21–4.4 μM 6-benzylaminopurine (BA) in combination with 2.69–10.74 μM naphthalene acetic (NAA), 2.07–8.29 μM indole-3-butyric (IBA), or 2.85–11.42 μM indole-3-acetic acid (IAA) to determine the best medium for the induction of shooting. Maximum direct shoot formation from leaves was observed on MS containing 22.21 μM BA and 10.74 μM NAA (25 shoots/explant). The second procedure began with the culture of protocorms on media containing NAA, IBA, or IAA, which induced callus formation with high regenerative potential in the form of protocorm-like-bodies (PLBs) that eventually differentiated into shoots. The optimal response was attained when these structures were cultured on medium with 4.14 μM IBA (30 shoots/PLB). To promote the elongation of shoots derived from PLBs, the material was subcultured onto MS medium containing 22.21 μM BA and 5.37 μM NAA. Through the exploration of the effects of auxins and matrix on the rooting of shoots, it was determined that the optimal rooting occurred on media supplemented either with 5.71 μM IAA or 4.14 μM IBA either on agar-gelled medium or in liquid media with coir as the matrix. Rooting was found to be 20% higher in liquid media than in agar-gelled medium.