The effect of subculture cycles on somaclonal variation of V. planifolia using intersimple sequence repeat (ISSR) markers was analyzed. Nodal segments of 2 cm in length were established in vitro and multiplied by 10 subculture cycles in Murashige and Skoog (MS) medium supplemented with 8.86 lM BAP (benzylaminopurine). After 45 days in each culture, the length and number of shoots per explant were evaluated. For ISSR markers, ten shoots per each subculture and the mother plant were used. Ten ISSR primers were used and a total of 118 bands were obtained. The polymorphism (%) was calculated and a dendrogram based on Jaccard's genetic distance between the subcultures and the donor plant was obtained. These results show that the multiplication rate tends to increase until subculture five, whereas shoot length decreases as the number of subcultures increases. The ISSR markers revealed an increase in the polymorphism percentage after the fifth culture cycle. The dendrogram showed the formation of two groups. The first group, with less genetic variability, is the donor plant and subcultures 1-5; the second group has greater genetic distance and is formed by subcultures 6-10. The results revealed that the number of subcultures with 8.86 lM BAP is a factor that affects the somaclonal variation during in vitro regeneration of V. planifolia. In conclusion, the subculture number affects somaclonal variation and in vitro development of V. planifolia.
Sugarcane (Saccharum spp. hybrids) is moderately sensitive to salinity and the effects on plant performance vary according to stress level and genotype. This study aimed to evaluate the effect of salt stress induced by application of different NaCl levels in the irrigation solution on plant height, indirect index of chlorophylls (SPAD), and macronutrients concentration (N, P, K, Ca, Mg and S) and Na in leaves of two sugarcane varieties: CP 72-2086 and Mex 69-290. The experiment was set in a completely randomized design with a 2×5 factorial arrangement. The study factors were sugarcane variety (CP 72-2086 and Mex 69-290) and NaCl concentration (0.0, 71.8, 143.6, 215.4 and 282.7 mM NaCl). Salinity as a single factor negatively affected plant height, SPAD units and N and P concentration in leaves; Ca concentrations increased, while K, Mg and S remained unaffected by the tested NaCl levels. Mex 69-290 grew higher and concentrated greater levels of N and K. Interactions of factors showed that salinity reduced growth in both varieties, but this reduction was more pronounced in CP 72-2086. SPAD units were also significantly reduced by salinity in both varieties. Concentrations of N and P in leaves decreased in both varieties in response to NaCl, while those of K and Ca increased in Mex 69-290. Concentration of Na was higher in Mex 69-290 which exhibited better performance than CP 72-2086. Sodium concentrations in leaves increased in direct relation to the tested NaCl concentrations. Mex 69-290 reached higher concentrations of Na in leave tissues but displayed better health than CP 72-2086. Thus, the variety Mex 69-290 showed more efficient Na-tolerance mechanisms related to Ca and K concentrations, and an indirect chlorophyll index better than CP 72-2086.
Objective: To analyze the genetic uniformity of MSXJ hybrid papaya in vitro plants, obtained by direct organogenesis.Design/Methodology/Approach: The MSXJ papaya hybrid demonstrates quality characteristics for the national and exports market. In vitro culture of plant tissues represents a useful tool for their multiplication and conservation, but somaclonal variation can diminish their genetic and agronomic uniformity. In order to analyze the genetic uniformity of in vitro plants of this hybrid, ten ISSR primers were used for in vitro plants micropropagated during nine subcultures. DNA was extracted using the CTAB method. Data were analyzed using the program PopGene v 1.3.1.Results: Eighty-five loci of 200 to up to 2000 pb were generated, with 37 polymorphic loci. In the cluster analysis, three groups were observed which separate subculture one, subcultures two to eight, and subculture nine; the Gst value of 0.87 indicated genetic uniformity as far as subculture eight.Study Limitations/Implications: Papaya is one of the most important tropical fruits worldwide; however, these plants need to be healthy and genetically uniform to guarantee commercial success. In vitro propagation allows obtaining healthy and uniform plants, but it is necessary to study genetic uniformity during their micropropagation.Findings/Conclusions: The in vitro multiplication of the MSXJ papaya hybrid permitted the regeneration of vigorous plants in 30 d. Molecular profiles indicate that as far as subculture eight, there is genetic uniformity. As such, no more thaneight subcultures are recommended during micropropagation.
La producción agrícola es afectada negativamente por estrés biótico y abiótico, siendo responsables de grandes pérdidas económicas en el mundo. La caña de azúcar es la materia prima para obtener jugo de caña que se transforma en sacarosa y en la producción de etanol de segunda generación. En el presente estudio se evaluó el número de brotes y hojas, la concentración de prolina, clorofila a, b y total y la concentración de azúcares como respuesta al estrés hídrico y salino en dos variedades de caña de azúcar in vitro. Brotes individualizados de 5 cm de longitud de las variedades MotzMex 91-207 y SP 71-6180 de caña de azúcar se cultivaron in vitro bajo estrés hídrico (PEG 6000) y salino (NaCl). Las dos variedades de caña de azúcar analizadas in vitro presentaron respuestas diferentes al estrés osmótico. La variedad MotzMex 91-207 fue mejor que la variedad SP 71-6180 para responder al estrés hídrico con una mayor acumulación de prolina (82.34 mg g-1 PS), no presentó disminución en el contenido de clorofilas a, b y total y generó 20.8 brotes por explante aun en condiciones de estrés. Por otro lado, la respuesta al estrés salino con 50 mM NaCl in vitro la presentó la variedad SP 71-6180 al generar una mayor acumulación de carbohidratos como galactosa, glucosa y manosa.
Objective: To identify the sex of in vitro plants of papaya (Carica papaya L.) MSXJhybrid obtained via somatic organogenesis, through SCAR type molecular markers. Design/Methodology/Approach: Eight-month old MSXJ papaya hybrid plants in thefructification stage were collected in Cotaxtla, Veracruz, Mexico. They weresuperficially disinfected with abundant running water, detergent during 30 min, andthen alcohol at 70% was added for one minute, commercial chlorine at 30% for 30min, and they were rinsed with sterile distilled water; then the meristems werecultivated in MS medium and after 30 d a subculture was made. The DNA extractionwas made with the CTAB method, and the DNA PCR was done with the Deputy et al.(2002) method, and the primers T1, T12 and W11 were used.Results: The T1 primer was the positive control and the T12 and W11 primersallowed the amplification of fragments that identify hermaphrodite, feminine and maleplants, while the T12 and W11 primers were specific for hermaphrodite plants.Study Limitations/Implications: It is required to standardize the method for it to beinexpensive.Findings/Conclusions: The sexuality of papaya plants can be differentiated until thestage of flowering, which is why the implementation of molecular markers wouldfacilitate plant selection if it is implemented at a large scale. Costs, maintenance timeand elimination of plants of unwanted sex are reduced this way.
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