Organogenesis is influenced by factors like genotype, type of explant, culture medium components, and incubation conditions. The influence of ethylene, which can be produced in the culture process, can also be a liming factor in recalcitrant species like pepper. In this work, bud induction was achieved from cotyledons and hypocotyls -from eight pepper cultivars -on Murashige and Skoog (MS) medium supplemented with 22.2 µM 6-benzyladenine (6BA) and 5.7 µM indole-3-acetic acid (IAA), in media with or without silver nitrate (SN) (58.9 µM), a suppressor of ethylene action. In the SN-supplemented medium, the frequencies of explants with buds and with callus formation were lower in both kinds of explant, but higher numbers of developed shoots were isolated from explants cultured on SN. Bud elongation was better in medium with gibberellic acid (GA3) (2.9 µM) than in medium free of growth regulators or supplemented with 1aminocyclopropane-1-carboxylic acid (ACC) at 34.5 µM. However, isolation of shoots was difficult and few plants were recovered. The effect of adding SN following bud induction (at 7 d) and that of dark incubation (the first 7 d of culture) was also assessed in order to improve the previous results. When SN was added after bud induction, similar percentages of bud induction were found for cotyledons (average frequency 89.4% without SN and 94.4% with SN) whereas they doubled in hypocotyls (50 % without SN and 87.7% with SN). In addition, in both kinds of explant, the number of developed plants able to be transferred to soil (developed and rooted) was greatly increased by SN. Dark incubation does not seem to improve organogenesis in pepper, and hypocotyl explants clearly represent a better explant choice -with respect to cotyledonary explants -for the pepper cultivars assayed. REGENERATION IN PEPPER3
BackgroundSomatic embryogenesis is the preferred method for cell to plant regeneration in Vitis vinifera L. However, low frequencies of plant embryo conversion are commonly found. In a previous work we obtained from cut-seeds of a grapevine infected with the Grapevine leafroll associated viruses 1 and 3 (GLRaV-1 and GLRaV-3), high rates of direct regeneration, embryo plant conversion and sanitation. The aim of this study is to evaluate the usefulness of this procedure for regeneration of other grapevine varieties which include some infected with one to three common grapevine viruses (GLRaV-3, Grapevine fanleaf virus (GFLV) and Grapevine fleck virus (GFkV)). As grapevine is highly heterozygous, it was necessary to select from among the virus-free plants those that regenerated from mother tissues around the embryo, (true-to-type).ResultsSomatic embryogenesis and plant regeneration were achieved in a first experiment, using cut-seeds from the 14 grapevine varieties Airén, Cabernet Franc, Cabernet Sauvignon, Mencía, Merlot, Monastrell, Petit Verdot, Pinot Blanc (infected by GFLV and GFkV), Pinot Gris, Pinot Meunier, Pinot Noir, Syrah, Tempranillo (infected by GFLV), and Verdil. All regenerated plants were confirmed to be free of GFkV whereas at least 68% sanitation was obtained for GFLV. The SSR profiles of the virus-free plants showed, in both varieties, around 10% regeneration from mother tissue (the same genetic make-up as the mother plant). In a second experiment, this procedure was used to sanitize the varieties Cabernet Franc, Godello, Merlot and Valencí Blanc infected by GLRaV-3, GFkV and/or GFLV.ConclusionsCut-seeds can be used as explants for embryogenesis induction and plant conversion in a broad range of grapevine varieties. The high regeneration rates obtained with this procedure facilitate the posterior selection of true-to-type virus-free plants. A sanitation rate of 100% was obtained for GFkV as this virus is not seed-transmitted. However, the presence of GLRaV-3 and GFLV in some of the regenerated plants showed that both viruses are seed-transmitted. The regeneration of true-to-type virus-free plants from all infected varieties indicates that this methodology may represent an alternative procedure for virus cleaning in grapevine.Electronic supplementary materialThe online version of this article (10.1186/s12870-017-1159-3) contains supplementary material, which is available to authorized users.
-Introduction. Interest in grafting onto rootstocks resistant to soil-borne pathogens has risen since the phasing-out of methyl bromide, an efficient soil disinfectant. In this study, we have evaluated the putative effect of rootstock-scion combinations on pepper fruit nutritional quality in relation to grafting, an effect that is sometimes overlooked. Materials and methods. As the scions, we used two representative sweet pepper cultivars, 'Almuden' and 'Coyote', and as rootstocks 'Foc' and 'Charlot', both resistant to Phytophthora capsici and Meloidogyne incognita. Dry matter, soluble solids, proteins, phenolics, and vitamin C content, as well as eight minerals (P, K, Ca, Mg, Na, Fe, Cu, and Zn) were measured in fruits from non-grafted, self-grafted, and grafted plants. Results and discussion. Differences in the fruit composition were found, depending on the genotype, rootstock, or rootstock-scion combination. The main effects were as follows. Higher values of• Brix, phenolics, P, Mg, and Na were found in the fruit of 'Coyote' whereas the vitamin C and Fe levels were higher in 'Almuden' fruit. The dry matter, protein, and K concentrations were higher in the fruit from grafted plants with respect to the values from non-grafted plants, especially in the fruit from plants grafted onto 'Charlot' and 'Foc' rootstocks (increases of 11.8%, 17.6%, and 9.6%, respectively; when average values of the four combinations of these rootstocks). Fruit from 'Almuden' and 'Coyote' plants grafted onto the rootstock 'Foc' had lower• Brix values (about one degree less than the control) and lower values of phenolics were also obtained when used this rootstock (14.4% less than for the control). On the other hand, the P concentration was higher in the fruit of 'Coyote' grafted onto 'Charlot' (17.2% higher than for the control). This combination also exhibited the highest fruit K concentration (226.70 mg 100 g −1 fresh weight). Conclusion. From a nutritional point of view it is important to evaluate and select the best rootstock-scion combinations: in our assay, this was 'Charlot'-'Coyote' the fruit of which showed the highest• Brix and K, P, and protein concentrations.Keywords: pepper / Capsicum annuum / fruit quality / mineral composition / rootstockscion combination Résumé -Effets des combinaisons de greffage sur la composition nutritionnelle des fruits du poivron. Introduction. L'intérêt du greffage sur des porte-greffes résistants aux pathogènes du sol a augmenté depuis l'interdiction du bromure de méthyle, un efficace désinfectant des sols. Dans cette étude, nous avons évalué l'effet putatif de combinaisons porte-greffe-greffon sur la qualité nutritionnelle des fruits de piment qui peut être touchée par greffage et qui est parfois négligée. Les principaux effets que nous avons observés sont les suivants : de plus hauts• Brix et de plus fortes teneurs en composés phénoliques, P, Mg et Na ont été trouvés dans les fruits 'Coyote', tandis que, les teneurs en vitamine C et en Fe étaient plus élevées dans les fruits d'Almuden'....
Background Contamination−free culture is a prerequisite for the success of in vitro − based plant biotechnology. Aseptic initiation is an extremely strenuous stride, particularly in woody species. Meanwhile, over−sterilization is potentially detrimental to plant tissue. The recent rise of machine learning algorithms in plant tissue culture proposes an advanced interpretive tool for the combinational effect of influential factors for such in vitro − based steps. Results A multilayer perceptron (MLP) model of artificial neural network (ANN) was implemented with four inputs, three sterilizing chemicals at various concentrations and the immersion time, and two outputs, disinfection efficiency (DE) and negative disinfection effect (NDE), intending to assess twenty−seven disinfection procedures of Pistacia vera L. seeds. Mercury chloride (HgCl2; 0.05–0.2%; 5–15 min) appears the most effective with 100% DE, then hydrogen peroxide (H2O2; 5.25–12.25%; 10–30 min) with 66–100% DE, followed by 27–77% DE for sodium hypochlorite (NaOCl; 0.54–1.26% w/v; 10–30 min). Concurrently, NDE was detected, including chlorosis, hard embryo germination, embryo deformation, and browning tissue, namely, a low repercussion with NaOCl (0–14%), a moderate impact with H2O2 (6–46%), and pronounced damage with HgCl2 (22–100%). Developed ANN showed R values of 0.9658, 0.9653, 0.8937, and 0.9454 for training, validation, testing, and all sets, respectively, which revealed the uprightness of the model. Subsequently, the model was linked to multi−objective genetic algorithm (MOGA) which proposed an optimized combination of 0.56% NaOCl, 12.23% H2O2, and 0.068% HgCl2 for 5.022 min. The validation assay reflects the high utility and accuracy of the model with maximum DE (100%) and lower phytotoxicity (7.1%). Conclusion In one more case, machine learning algorithms emphasized their ability to resolve commonly encountered problems. The current successful implementation of MLP–MOGA inspires its application for more complicated plant tissue culture processes.
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