Karely, new variety of cut flower heliconia, suitable for the humid Mexican tropics

Objective: To describe Heliconia uxpanapensis x Heliconia latispatha Var. Karely, interspecific variety generated for cut flower based on commercial standards. Design/methodology/approach: Interspecific hand pollination was carried out between H. uxpanapensis Gutiérrez Báez x H. latispatha Benth. The female parent is endemic to Mexico, it presents compact growth due to its pachymorphic rhizome, red inflorescence, erect and bracts in distal position. H. latispatha has an expansive growth due to its leptomorphic rhizome, it has yellow with red inflorescence, erect and helical. The F1 hybrid was self-fertilized and produced viable seed. The mature zygotic embryos were germinated in vitro, the seedlings were acclimatized at six months and at ten months they were established in the field; Heliconia Var Karely was generated from this population. Results: Karely variety is F2 product of the interspecific cross between H. uxpanapensis x H. latispatha, it presents a compact growth habit, erect inflorescence, bracts in distal position of orange color with red border. The inflorescence is clean, it does not present watermarks that accumulate its bracts. Post-harvest life is greater than 15 days. Limitations on study/implications: Karely be cultivated under the agroforestry system in conditions of 20 to 30% shade for the adequate pigmentation of the bracts. In addition, it requires sandy-loam soil for proper development and growth of its inflorescence. Findings/conclusions: Karely has ideal characteristics for cut flowers and meets the characteristics established by the national and international market.Objective: To describe Heliconia uxpanapensis x Heliconia latispatha var. Karely, interspecific variety generated for cut flower based on commercial standards. Design/Methodology/Approach: Interspecific hand pollination was carried out between H. uxpanapensis Gutiérrez Báez x H. latispatha Benth. The female parent is endemic to Mexico; it presents compact growth due to its pachymorphic rhizome, red and erect inflorescence, and bracts in distal position. H. latispatha grows expansively due to its leptomorphic rhizome, its inflorescence, composed of orange bracts, is erect and helical. The F1 hybrid was self-fertilized and produced viable seed. The mature zygotic embryos were germinated in vitro, the seedlings were acclimatized at six months and at ten months they were established in the field; Heliconia var. Karely was generated from this population. Results: Karely variety is F2 product of the interspecific cross between H. uxpanapensis x H. latispatha, it presents a compact growth habit, erect inflorescence, bracts in distal position of orange color with red border. The inflorescence is clean and healthy. The postharvest half-life is 15 days Study Limitations/Implications: Karely must be cultivated under an agroforestry system in conditions of 20 to 30 % shade for the adequate pigmentation of the bracts. In addition, it requires sandy-loam soil for proper development and growth of its inflorescence. Findings/Conclusions: Karely has ideal characteristics for cut flower and meets the characteristics established by the market

Heliconia SCARLET: a mexican variety for cut flower and gardening

Objective: To carry out the morphological characterization of Heliconia uxpanapensis x Heliconia latispatha var. Scarlet for varietal registration purposes. Design, Methodology and Approximation: By means of rhizomes division of 71-1 plant from a plot of H. uxpanapensis x H. latispatha segregant plants (F2), twelve plants (tillers) were generated, planted, and cultivated in open field under 30% of natural shade. Morphological characterization was methodic conducted using the Technical Guide descriptors for Heliconias designed by the National Seed Inspection and Certification Service (SNICS, 2023). Results: Morphological characters of the clones from the 71-1 segregant plant, tested with the SNICS Technical Guide descriptors, were constant so that a differentiation could be make between the Scarlet and Karely, a reference variety found in the Guide. Scarlet variety’s primary distinctive characteristics are the red color, high brilliance, and the revolute-involute margins of their bracts. Their inflorescence morphological characteristic suggest they can be cultivated for cut flower and gardening.  Study limitations and implications: In order for Scarlet variety express, their characteristics of intense color and bracts brightness, they have to be cultivated under 30% of shade. Findings and conclusions: Based on the Technical Guide descriptors for heliconias varietal description (SNICS, 2023), the Scarlet variety differentiates from the Karely (reference variety) as it presents a stability-distinction-homogeneity. For this reason, the varietal registration seems appropriate.Objective: To carry out the morphological characterization of Heliconia uxpanapensis X Heliconia latispatha var. Scarlet for varietal registration purposes.Design, Methodology and Approximation: By means of rhizomes division of 71-1 plant from a plot of H. uxpanapensis X H. latispatha segregant plants (F2), twelve plants (tillers) were generated, planted, and cultivated in open field under 30% of natural shade. Morphological characterization was methodic conducted using the Technical Guide descriptors for Heliconias designed by the National Seed Inspection and Certification Service (SNICS, 2023).Results: Morphological characters of the clones from the 71-1 segregant plant, tested with the SNICS Technical Guide descriptors, were constant so that a differentiation could be make between the Scarlet and Karely, a reference variety found in the Guide. Scarlet variety’s primary distinctive characteristics are the red color, high brilliance, and the revolute-involute margins of their bracts. Their inflorescence morphological characteristic suggest they can be cultivated for cut flower and gardening.Study limitations and implications: In order for Scarlet variety express, their characteristics of intense color and bracts brightness, they have to be cultivated under 30% of shade.Findings and conclusions: Based on the Technical Guide descriptors for heliconias varietal description (SNICS, 2023), the Scarlet variety differentiates from the Karely (reference variety) as it presents a stabilitydistinction-homogeneity. For this reason, the varietal registration seems appropriate

In vitro callogenesis induction of Guarianthe skinneri (Bateman) Dressler and W.E. Higgins (Orchidaceae)

Guarianthe skinneri (Bateman) Dressler and W.E. Higgins, is a native orchid from Mexico, considered as threatened species in the NOM-ECOL-059-SEMARNAT-2010, mainly due to the disappearance of its natural habitat and illegal collection during the flowering season. The aim of this research was to induce in vitro callogenesis using different types of explants and phytoregulators, in order to establish a mass production technique as a contribution to the conservation of this species. We evaluated leaf and pseudobulb pith explants growing in semi-solid medium MS, adding BAP, 2, 4-D, Kin, the combination of BAP/2 and 4-D/Kin/Sad and a control without any type of plant growth regulators. Statistical analysis showed that pseudobulb pith explants are more suitable for in vitro callus induction in comparison to leaf explants, because of a lower percentage of contamination (18.8% in pith and 73.2% in leaves). Likewise, the pseudobulb pith explants showed increased callus formation (10.8%) in comparison to leaf explants (7.6%). Regarding the phytoregulators employed, BAP promoted increased callus formation (17%) compared to other phytoregulators (7-10%). This is the first report of the use of pseudobulb pith as an explant for callus induction in G. skinneri.Guarianthe skinneri (Bateman) Dressler and W.E. Higgins., is a native orchid from Mexico, considered as threatened species by NOM-ECOL-059-SEMARNAT-2010, mainly due to the disappearance of its natural habitat and the illegal collection during its flowering season. The aim of this research was to induce in vitro callogenesis from different type of explants, using phytoregulators, in order to look for a massive production technique to contribute to its conservation. We evaluated the leaf and pseudobulb marrow explants growing in semi-solid medium MS adding BAP, 2, 4-D, Kin, the interaction of BAP/2, 4-D/Kin/Sad and a control without any type of plant growth regulators. Statistical analysis showed the pseudobulb marrow explants are more suitable for in vitro introduction in comparison to leaf explants, since they perform a lower percentage of contamination (18.8% in marrow and 73.2% in leaves). Likewise, the pseudobulb marrow explants increased callus formation (10.8%) in comparison to leaf explants (7.6%). Regarding the phytoregulators employed, BAP have allowed to increased callus formation (17%) compared to other phytoregulators (7-10%). This is the first report, which proposes the use of pseudobulb marrow as explant for callus induction in G. skinneri

Caracterización de genotipos élite de Coffea canephora por su tolerancia a sequía.

El objetivo del presente trabajo fue identificar el grado de tolerancia a sequía en diferentes genotipos élite de Coffea canephora P. var. Robusta. El estudio se realizó del 2013 al 2015 en Tuxtla Chico, Chiapas, México. De cada genotipo (INIFAP P9, INIFAP P4, INIFAP P5, INIFAP P7, INIFAP P8) se utilizaron veinte plantas, excepto para el INIFAP P9 con doce; un grupo con la mitad de plantas de cada genotipo se mantuvo en condiciones de capacidad de campo (CC) y la otra mitad en punto de marchitez permanente (PMP). Al nalizar la etapa de estrés y después del riego de recuperación, se evaluaron variables morfológicas y siológicas, tales como parámetros de crecimiento, contenido relativo de agua, acumulación de prolina, índice y densidad estomática e índice de tolerancia a sequía. Los genotipos de C. canephora presentaron entre ellos diferente grado de tolerancia al déficit hídrico, pero compartieron entre sí la detención de sus funciones y activación de diferentes rutas para tolerar la sequía, como disminución en el contenido relativo de agua y aumento de prolina durante el periodo de estrés. Al determinar el índice de tolerancia a sequía, el genotipo más tolerante fue el INIFAP P5 de acuerdo con las variables número de hojas retenidas y totales, tasa de crecimiento relativo, número de pares de ramas plagiotrópicas y nudos totales; mientras que el INIFAP P4 se identificó como uno de los más susceptibles de acuerdo con la tasa de crecimiento relativo y el número de nudos totales, en las cuales presentó los valores más bajos.

Medio de cultivo e inhibidores de etileno en la embriogénesis somática de café

The combination of salts, hormones and the gaseous environment in vitro could affect the expression of somatic embryogenesis. The effect of the combination of two culture media and inhibitors of ethylene induction of embryogenic callus of Coffea canephora P., and their ability to proliferate and form embryos was studied. five leaf explants of C. canephora P. genotypes were established in two different media, 23A8 [Yasuda salts with BAP (1.12 mg L-1)]; and T1B/T2B sequence consisting of a first crop T1B [50% of MS salts with 2iP (2 mg L-1), 2, 4-D (0.5 mg L-1) and AIB (1 mg L-1) ]; followed by T2B [50% of MS salts with BAP (4 mg L-1) and 2, 4-D (1 mg L-1) ]. These media were supplemented with AgNO3 (6.8 mg L-1) and Na2S2O3 (6.3 mg L-1). Using T1B/T2B led greater callus proliferation (up to 248 mg), but low percentages of embryogenic callus (less than 29%) and less differentiated than those obtained in the middle 23A8 (160 mg). In the multiplication phase were obtained calluses from 1 to 2.5 mg and multiplication rate of 10 to 24 times the initial biomass they. In differentiation, embryogenic potential of 25 000 to 137 000 embryos per gram of inoculated callus was observed. At each stage the effect of ethylene inhibitors genotype dependent on interaction with the culture medium. Keywords: Coffea canephora P.; AgNO3; embryogenic potential; multiplication rate; Na2S2O3La combinación de sales, hormonas y el ambiente gaseoso in vitro podrían afectar la expresión de la embriogénesis somática. Se estudió el efecto de la combinación de dos medios de cultivo e inhibidores de etileno sobre la inducción de callo embriogénico de Coffea canephora P., y su capacidad para proliferar y formar embriones. Se establecieron explantes foliares de cinco genotipos de C. canephora P., en dos medios diferentes, el 23A8 [sales de Yasuda con BAP (1.12 mg L-1)]; y la secuencia T1B/T2B consistente en un primer cultivo en T1B [50% de sales de MS con 2iP (2 mg L-1), 2, 4-D (0.5 mg L-1) y AIB (1 mg L-1) ]; seguido del T2B [50 % de sales de MS con BAP (4 mg L-1) y 2, 4-D (1 mg L-1) ]. Estos medios se suplementaron conAgNO3 (6.8 mg L-1) y Na2S2O3 (6.3 mg L-1). El uso de T1B/T2B propició mayor proliferación de callos (hasta248 mg), pero bajos porcentajes de callo embriogénico (menores a 29 %) y menor grado de diferenciación que los obtenidos en el medio 23A8 (con 160 mg). En la fase de multiplicación se obtuvieron callos de 1 a 2.5 mg y una tasa de multiplicación de 10 a 24 veces la biomasa inicial. En la de diferenciación, se observó un potencial embriogénico de 25 mil a 137 mil embriones por gramo de callo inoculado. En cada etapa el efecto de los inhibidores de etileno dependió del genotipo en interacción con el medio de cultivo. Palabras clave: Coffea canephora P.; AgNO3; Na2S2O3; potencial embriogénico; tasa de multiplicació

In vitro callogenesis induction of Guarianthe skinneri (Bateman) Dressler & W.E. Higgins (Orchidaceae)

Guarianthe skinneri (Bateman) Dressler & W.E. Higgins, is a native orchid from Mexico, considered as threatened species in the NOM-ECOL-059-SEMARNAT-2010, mainly due to the disappearance of its natural habitat and illegal collection during the flowering season. The aim of this research was to induce in vitro callogenesis using different types of explants and phytoregulators, in order to establish a mass production technique as a contribution to the conservation of this species. We evaluated leaf and pseudobulb pith explants growing in semi-solid medium MS, adding BAP, 2, 4-D, Kin, the combination of BAP/2 and 4-D/Kin/Sad and a control without any type of plant growth regulators. Statistical analysis showed that pseudobulb pith explants are more suitable for in vitro callus induction in comparison to leaf explants, because of a lower percentage of contamination (18.8% in pith and 73.2% in leaves). Likewise, the pseudobulb pith explants showed increased callus formation (10.8%) in comparison to leaf explants (7.6%). Regarding the phytoregulators employed, BAP promoted increased callus formation (17%) compared to other phytoregulators (7-10%). This is the first report of the use of pseudobulb pith as an explant for callus induction in G. skinneri

Respuesta rizogénica en Pinus maximartinezii Rzedowski

Pinus maximartinezii Rzedowski is an endemic specie in danger of extinction due to population alteration in the wild and has survived extreme genetic restriction, it is also considered the rarest of pines. In this study rhizogenic response of Pinus maximartinezii sprouts, obtained from zygotic embryos regenerated via organogenesis subjected to in vitro rooting treatments was evaluated. The highest rooting response was obtained with a pulse of 24 h in the presence of 2.0 mg L-1 of IBA in DCR, GD and MS culture media at 100 and 50% of its original concentration plus 0.5% activated carbon per L-1. Number of shoots with roots was evaluated after eight weeks, the X² test showed that rooting is independent of culture media, concentration and dependent to the pulse times. Root formation in shoots, allow us to propose this species to be propagated in vitro.El piñón azul o maxi piñón Pinus maximartinezii Rzedowski es una especie endémica en peligro de extinción debido a la alteración de sus poblaciones en su hábitat natural y que ha sobrevivido a una restricción genética extrema, además es considerado el más raro de los pinos piñoneros. En este trabajo se evaluó la respuesta rizogénica de brotes de Pinus maximartinezii, obtenidos a partir de embriones cigóticos regenerado vía organogénesis, los cuales se sometieron a tratamientos de enraizamiento in vitro. La mayor respuesta de enraizamiento se presentó con pulso de 24 h en presencia de 2.0 mg L-1 de IBA en los medios de cultivo DCR, GD y MS, al 100 y 50% de su concentración original más 0.5% de carbón activado por L-1. Después de ocho semanas se evaluó la variable conforme al número de brotes con raíces, la prueba de X² mostró que el enraizamiento es independiente de los medios de cultivo así como la concentración de los mismos y dependiente al tiempos de pulso. La formación de raíces obtenidas en los brotes permite proponer esta especie con capacidad para ser propagada in vitro

El uso ornamental de Guarianthe skinneri (Orchidaceae), en Chiapas y Guatemala, determina parcialmente su diversidad y estructura genética

Background and Aims: Guarianthe skinneri is a native orchid from Chiapas, Mexico, with a Central American distribution, which is threatened by extraction and illegal trade. For this reason, it is classified as threatened in the Mexican rule NOM-059-SEMARNAT-2010. In the region of Soconusco (Chiapas, Mexico) and Guatemala, it is commonly called “Candelaria”, because its flowering time coincides with the dates of the virgin of the same name. In Tapachula city (Chiapas, Mexico), it is common to find plants of this species in the courtyards of pri-vate gardens. The objective of this work was to estimate the diversity and genetic structure of G. skinneri from samples obtained from an urban population of Tapachula and from its comparison with five wild populations of Guatemala, in order to determine the potential of the urban population as a source of propagules and understand if genetic distance between populations could be related with the geographic distance that separates them. Methods: Nucleotidic variation in sequences of two regions of ribosomal nuclear DNA (ITS 1-2 and ITS 3-4) was used as genetic marker. The sequences were concatenated obtaining the conventional parameters of diversity and genetic structure. Key results: We observed populations with low and high genetic diversity. A total of 38 haplotypes were observed, whose number per population was between 1 and 15. Haplotidic diversity (Hd) was between 0 and 1, the average number of polymorphic sites was between 0 and 165. The nucleotidic diversity (p) and the average number of nucleotidic differences shows that Tapachula had the greatest diversity, followed by populations of Guatemala. The genetic structure was moderate (Fst=0.083) and the genetic distance was not associated with the geographic distance of the populations. Conclusions: The diversity patterns and genetic structure between populations were attributed to anthro-pogenic factors derived from its ornamental use, especially in the city of Tapachula.Antecedentes y Objetivos: Guarianthe skinneri es una orquídea nativa de Chiapas, México, con distribución centroamericana, objeto de extracción y comercio ilícito; por esta razón, está catalogada como amenazada en la Norma Mexicana NOM-059-SEMARNAT-2010. En la región del Soconusco (Chiapas, México) y Guatemala, se le denomina “Candelaria”, debido a que su época de floración coincide con las fechas de la homónima virgen. En la ciudad de Tapachula (Chiapas, México), es común encontrar individuos de esta especie en los patios de domicilios particulares. El objetivo de este trabajo fue estimar la diversidad y estructura genética de G. skinneri a partir de muestras obtenidas de una población urbana de Tapachula y de su comparación con cinco poblacio-nes silvestres de Guatemala, para determinar el potencial de la población urbana como fuente de propágulos y averiguar si la distancia genética entre poblaciones se relaciona con la distancia geográfica que las separa. Métodos: Como marcador genético se utilizó la variación nucleotídica en secuencias de dos regiones de ADN nuclear ribosomal (ITS 1-2 y ITS 3-4). Se concatenaron las secuencias y se obtuvieron los paráme-tros convencionales de diversidad y estructura genética. Resultados clave: Se observaron poblaciones desde bajos hasta altos niveles de diversidad genética. Se registró un total de 38 haplotipos cuyo número entre poblaciones osciló entre 1 y 15. La diversidad ha-plotípica (Hd) estuvo entre 0 y 1, el número promedio de sitios polimórficos entre 0 y 165. La diversidad nucleotídica (p) y el número promedio de diferencias nucleotídicas muestran que Tapachula tuvo la mayor diversidad, seguida por poblaciones de Guatemala. La estructura genética fue moderada (Fst=0.083) y la distancia genética no se asoció con la distancia geográfica de las poblaciones. Conclusiones: Los patrones de diversidad y estructura genética entre poblaciones se atribuyen a factores antropogénicos derivados de su uso ornamental, especialmente en la ciudad de Tapachula

Remoción nutrimental del fruto de cacao (Theobroma cacao L.) en México: Remoción nutrimental en cacao

Objective: To determine the nutritional concentration of the cocoa fruit in its components: shell, mucilage, testa and seed of different genetic groups. Design / methodology / approach: The determination of the nutrient removal was from three cocoa clones: Criollo (Carmelo), Forastero (PMCT 58) and Trinitario (RIM 88). Ten fruits from each clone were collected. Each fruit was divided into its components: peel, mucilage, testa and seed. Each of the fractions were dried in the drying oven for 72 h at 65 ° C; each fraction was then ground, packaged, labeled and sent to the laboratory, where the macro and micro-nutrients of each of these fractions were determined. Results. The nutritional removal of the macro and micro nutrients (N, PK, Ca, Mg and S), of the different structures (shell, mucilage, testa and seed) and genetic group (outsider, Trinidadian and Creole), ranged in a range of acceptable values ??(0.01 <s <0.199). Study limitations / implications: Nutrient removal will be mainly influenced by the genetic group of cocoa and the conditions where it is grown. Findings / Conclusions: The nutritional removal of cocoa varies according to the different structures (seed, testa, mucilage and shell) and the concentration depends largely on the variety of cocoa (Forastero, Trinitario or Criollo). Potassium is the element most absorbed by the peel, followed by calcium and nitrogen and phosphorus in the seed. In general, the Trinitario cocoa (RIM) presented higher nutrient removal in K, N, Mg and P, followed by the Forastero (PMCT 58) and with lesser quantity the Criollo (Carmelo) and in the microelements the most absorbed were Zn, Fe and B in Trinitario, Criollo and Forastero, respectively.Objetivo: Determinar la concentración nutrimental del fruto de cacao en sus componentes: cáscara, mucílago, testa y semilla de diferentes grupos genéticos. Diseño/metodología/aproximación: La determinación de la remoción nutrimental fue de tres clones de cacao: Criollo (Carmelo), Forastero (PMCT 58) y Trinitario (RIM 88). Se colectaron diez frutos de cada clon. Cada fruto se fraccionó en sus componentes: cáscara, mucílago, testa y semilla. Cada una de las fracciones fueron secadas en el horno de secado por 72 h a 65°C; después cada fracción fue molida, envasada, etiquetada y enviadas al laboratorio, donde se determinaron los macro y micro-nutrimentos de cada una de estas fracciones. Resultados. La remoción nutrimental de los macro y micro nutrimentos (N, P, K, Ca, Mg y S),de las diferentes estructuras (cáscara, mucílago, testa y semilla) y grupo genético (forastero, trinitario y criollo), osciló en un intervalo de valores aceptables (0.01 < s < 0.199).   Limitaciones del estudio/implicaciones: La remoción nutrimental va estar influenciado principalmente por el grupo genético de cacao y las condiciones donde se cultive. Hallazgos/conclusiones: La remoción nutrimental del cacao varía de acuerdo a las diferentes estructuras (semilla, testa, mucilago y cáscara) y la concentración depende en gran medida de la variedad de cacao (forastero, trinitario o criollo). El Potasio es el elemento más absorbido por la cáscara, seguido del calcio y en la semilla fue el nitrógeno y el fósforo. En general el cacao trinitario (RIM) presentó mayor remoción nutrimental en K, N, Mg y P, seguido del forastero (PMCT 58) y con menor cantidad el criollo (Carmelo) y en los microelementos los más absorbidos fueron Zn, Fe y B en trinitario, criollo y forastero, respectivamente

Variabilidad genética de cultivo "in vitro" de aguacate raza mexicana

Explantes foliares de Persea americana Mill. var. drymifolia, se establecieron en medio DCR (Gupta y Durzan, 1985) suplementado con vitaminas y reguladores del crecimiento para producir callos, dos sub cultivos se realizaron en intervalos de 30 a 45 días entre ellos. Con el fin de determinar la variabilidad genética se generaron 341 marcadores AFLP (amplified fragment lenght polymorphism) polimórficos con 10 combinaciones de iniciadores, se analizaron 94 muestras: la planta madre, 31 callos, 31 callos del primer sub cultivo y 31 callos del segundo sub cultivo. Se determinó el contenido de información polimórfica (PIC) por combinación de iniciadores y se calculó la distancia genética entre muestras para hacer el análisis de agrupamiento. Se obtuvo la diversidad genética entre la planta madre, callos y callos de los dos sub cultivos y se realizó ei análisis de varianza molecular. Los valores PIC indicaron que las combinaciones de iniciadores fueron adecuadas para estimar la variabilidad genética de los callos. En el análisis de agrupamiento, la planta madre mostró distancias de 0.63 a 0.7 con respecto a sus callos, distancias de 0.78-0.99 se obtuvieron entre los callos y los callos de los dos sub cultivos. La diversidad genética presentó diferencias significativas, el análisis de varianza molecular demostró variabilidad genética entre la planta madre, callos y callos de los dos sub cultivos, así como dentro de los callos, callos del primer sub cultivo y callos del segundo sub cultivo. Los resultados indicaron que la variabilidad genética puede ocurrir en cualquier sub cultivo. Debido a la variabilidad genética presentada los callos no se consideraron clones.Leaf explants of Persea americana Mill. var. drymifoliawere established on DCR medium (Gupta and Durzan, 1985) supplemented with vitamins and growth regulators to produce callus, two sub cultures were made in intervals of 30 to 45 days between them. In order to determine the genetic variability 341 AFLP (amplified fragment lenght polymorphism) polymorphism with 10 primer combinations were generated, 94 samples were analyzed: the mother plant, 31 calluses, 31 calluses from the first sub culture and 31 calluses from the second sub culture. Polymorphic information content (PIC) was determined by primer combination and the genetic distance between samples was calculated for the cluster analysis. Genetic diversity between the mother plant, calluses and calluses of the two sub cultures were obtained and the analysis of molecular variance was performed. The PIC values indicated that the primer combinations were adequate to estimate the genetic variability of calluses. In the cluster analysis, the mother plant showed distances of 0.63 to 0.7 in regard to their calluses, distances of 0.78 - 0.99 were obtained between calluses and calluses of the two sub cultures. Genetic diversity showed significant differences, analysis of molecular variance demonstrated genetic variability between the mother plant, calluses and calluses of the two subcultures as well as within the calluses, calluses of the first sub culture and calluses of the second sub culture. The results indicated that genetic variability can occur in any sub culture. Due to genetic variability presented in callus clones were not considered