Increasing gland number and red pigments in St. John’s wort in vitro

In order to develop a protocol for increasing the gland number and red pigments of Hypericum perforatum, this study was carried out to evaluate the effect of hydrolyzed casein (0.0 and 500 mg l-1), mannitol (0.0, 5 and10 g l-1) and sucrose (20 and 30 g l-1) on the synthesis of these pigments and glands on the produced leaves. Leaf discs of in vitro plantlets, were prepared and cultured on MS medium with 0.5 mg l-1 BAP to induce the shoot. All the cultures were incubated in the dark at 25 ± 2°C for 1 month. In all of the treatments, callus and shoot induction were observed. Percentage of calli and leaves containing red pigments, number of glands and percentage of leaves containing gland were noted as indicating the presence of hypericin and pseudohypericin pigments. Percentage of calli and leaves containing red pigments were significantly influenced by different concentrations of the hydrolyzed casein, mannitol and sucrose. The highest percentage of calli containing red pigments was observed in the culture medium which had 500 or 0.0 mg l-1 hydrolyzed casein and 20 g l-1 sucrose, without mannitol. Glands were observed on all the produced leaves. Number of glands and percentage of leaves containing gland were significantly influenced by the different concentrations of mannitol and sucrose and their interaction. The highest number of gland and percentage of leaves containing gland was achieved when explants were cultured in medium that included 30 g l-1 sucrose with 5 or 10 g l-1 mannitol and in medium containing 20 g l-1 sucrose, with 5 g l-1 mannitol. Morphological changes induced by carbon source and hydrolyzed casein were observed and described in detail. The obtained results will be applied in experimental botany and in the technology of H. perforatum cultivation for pharmaceutical applications.Key words: Hydrolyzed casein, hypericin, Hypericum perforatum, mannitol, pseudohypericin, sucrose

Effect of Glycine Betaine Nanocomposite Coated with Chitosan and Moderate Salinity Stress on In vitro Microtuberization of Potato (Solanum tuberosum L.) cv. Agria

Introduction Potato (Solanum tuberosum L.) is an important food and cash crop having the first rank in the world from non grain crops to ensure food security. The tubers produced through the conventional propagation are characterized by low multiplication rate and susceptibility to pathogens. Microtubers are an ideal propagating material for producing high quality seed potatoes. Nowadays, the production, application, and biological risk assessment of nano-scaled products have attracted global concerns in various fields such as agricultural, biotechnological, medicinal, and plant sciences. Chitosan, a biocompatible polymer, has been widely utilized to improve the production of nano-chemicals, thereby improving crop growth, productivity, and immunity. Nanotechnology plays an important role in modern agriculture to address global challenges such as climate change, severity of plant diseases and the limited availability of important plant nutrients. Polymer-based nano-formulations have recently received the greatest attention with the key objectives of developing less harmful, plant growth promoting and protective agents of biodegradable and natural origin. Use of chitosan-based nanoparticles in agriculture field is still in a budding phase. Significant outcomes have been reported in in vitro and a few in vivo studies in plant growth and protection by chitosan-based nanomaterials.   Materials and Methods MS medium containing 80 g of sucrose, containing glycine betaine at concentrations of 20 and 40 mg/l, chitosan at a concentration of 240 mg/l, glycine betaine 20 and chitosan 240 mg/l, glycine betaine 40 and chitosan 240 mg/l, 120 mg/l, glycine betaine nanocomposite coated with chitosan 120 and 240 mg/l and control treatment, as well as nonsalinity treatments with the same compounds and concentrations mentioned for salinity were cultured. The study was performed in randomized complete block design with three replications and sixteen treatment. Glycine betaine nanocomposite coated with chitosan were prepared at Maragheh University as follows. Chitosan, glycine betaine, and triphosphate are major consumables. First, chitosan was dissolved well with acetic acid under the influence of temperature. Then, a certain amount of glycine betaine was dissolved in distilled water and added to the chitosan. Tri-polyphosphate (TPP) was dissolved in distilled water at a certain volume ratio and added dropwise to the mixture of chitosan and glycine betaine. The precipitate obtained under the freeze-drying process lead to the preparation of its powder.     Results and Discussion According to the comparison of the means, treatment of glycine betaine coated with chitosan nanocomposite at a concentration of 120 mg/l produced the highest number of microtuber during the experiment from the first month to the final month. The nanocomposite was more capable of improving growth and biomass than the bare ZnONPs in pepper. The application of the nanocomposite increased the concentration of chlorophylls (51%), carotenoids (70%), proline (2-fold), and proteins (about 2- fold). The supplementation of culture medium with the nanomaterials upregulated enzymatic antioxidant biomarkers (catalase and peroxidase) (Asgari-targhi et al., 2021). The highest mean microtuber weight was related to the nanocomposite treatment of 120 mg/l with a weight of 29 mg. In the microtuber diameter, this treatment had the highest value. The results of analysis of variance in Table 1 indicate that the effect of moderate salinity and the interaction of moderate salinity and nanocomposite treatments are not significant and the effect of experimental treatments is significant at the level of 5% probability. Due to the fact that the most important trait in the potato microtuberization is microtuber yield, so in the experiment, the highest microtuber yield was earned nanocomposite treatment with 131 mg and the lowest in control treatment with 87 mg. It seems that by using stress-reducing compounds such as glycine betaine and chitosan and nanocomposites, these compounds increase the amount of genes responsible for the formation of microtuber, and as a result, increasing the proteins involved in stresses induce more microtuberization. Also, nanocomposite materials, more assimilated materials may be transferred from the roots to microtuberization processes.   Conclusion  The microtuber produced in the glycine betaine coated with chitosan nanocomposite treatment produced the highest number of microtubers in the first, second, third, and final months. In treatments with moderate salinity in the first, second, third and final months, the number of microtuber and eyes and sprouted microtubers had the highest amount compared to the treatment without moderate salinity. Also, plants treated with glycine betaine voated with chitosan nanocomposite in the microtuber trate showed a greater effect than chitosan and glycine betaine with chitosan. According to the findings of this study, it seems that the use of nanocomposite materials in increasing the microtuber and reducing the vegetative growth of potato shoots has been made in Agria cultivar

Dehydrin profiles of some Iranian melon varieties (Cucumis melo L. Merr) under drought stress conditions

Dehydrins are proteins that play a role in the mechanism of drought tolerance. This study aimed at establishing dehydrin profile and accumulation in four local melon varieties of Iran: Mino, Dargazi, Saveii, and Semsori, as well as in a commercial variety Honeydew. Plants were treated with drought stress by adjusting the soil water content to 75, 50, 40, 30 and 20% of field capacity (FC) by withholding water. Water status of plants was monitored based on the seedling fresh weight (FW) and relative water content of leaves (RWC). Total protein content was extracted, then heat-stable protein (HSP) fraction was isolated for each variety and water stress treatment. After SDS-PAGE of HSP, Western blotting analysis was carried out with Anti-dehydrin rabbit (primary) and Goat anti rabbit (secondary) antibodies. ANOVA results showed that with decreasing FC below 75%, FW and RWC decreased, but these changes significantly varied among genotypes. On the basis of FW and RWC data under different drought stress treatments, the following drought-tolerant ranking was established: Mino > Dargazi > Saveii and Honeydew > Semsori, from tolerant to sensitive order. Results of Western blot analysis showed that expression of some proteins with molecular weights of 19– 52 kDa was induced in the studied varieties under drought stress (% FC). Expression level of the dehydrin proteins in different varieties was variable and also depending on the drought stress level applied. However, dehydrin proteins (45 and 50 kDa) showed strong expression levels in all varieties under severe drought stress (20% FC). The abundance of dehydrin proteins was higher in tolerant varieties (Mino and Dargazi) than in moderate and drought sensitive genotypes. Consequently, dehydrins represent a potential marker for selection of genotypes with enhanced drought tolerance

Effect of Addition of Nutritional Supplements to Substrate on Yield and Protein Content of Oyster Mushroom (Pleurotu florida)

In order to investigate the effect of addition of nutritional supplements to substrate on yield of oyster mushroom (Pleurotus florida(, an experiment was carried out as completely randomized design in three replicates in 2010 . In this study, three types of nutritional supplements including alfalfa meal, soybean meal and vermicompost, each at three levels (2.5, 5 and 7.5% of wet substrate), were added to wheat straw substrate, and were compared to control. Traits such as preliminary and total yield, biological efficiency, protein content and mineral phosphorus and potassium concentrations were determined. Results indicated that all traits were significantly affected by nutritional supplements (P<0.01). Preliminary and total yields, biological efficiency and protein content were highest in the substrate of nutritional supplements containing 7.5% vermicompost (831.19 g/kg wet substrate), 2.5% soybean meal (1231.4 g/kg wet substrate), 2.5% soybean meal (171.1%) and 7.5% soybean meal (23.14%), respectively. While, the highest concentrations of phosphorus and potassium were obtained in the substrates containing 5% vermicompost (5.86 mg/g dry matter) and 7.5% vermicompost (24.05 mg/g dry matter), respectively

Nerinine and homolycorine, amaryllidaceae alkaloids from the bulbs of Galanthus transcaucasicus Fomin

Background and objectives: Many members of the Amaryllidaceae are regarded as toxic. The toxic constituents that occur in the whole family are referred to as the Amaryllidaceae alkaloids. The main aim of this study was the identification of alkaloid compounds from Galanthus transcaucasicus Fomin, a medicinal plant from Amaryllidaceae. Methods: Planar and column chromatography techniques were used for isolation of alkaloid components. GC/MS analysis was carried out for the identification of alkaloid compounds. Results: Silica gel column chromatography of the alkaloidal extract of G. transcaucasicus bulbs afforded seven fractions. Preparative thin layer chromatography of these fractions led to the isolation of compounds 1 (nerinine)and 2 (homolycorine). Galantamine was not detected in any of these fractions. Conclusion: Our findings showed that G. transcaucasicus could be a new source of bioactive alkaloids for possible applications in pharmaceutical industries

Ability of Callus Induction and Plant Regeneration in White Kashan and Red Rey Onions Cultivars (Allium cepa L.) Using Root - tip Culture under Invitro Conditions

This present study has been carried out in order to evaluate the effect of culture medium and genotype on callus induction and regeneration of two onion varieties through root tip culture. For callus induction, micro samples from 1-3 mm root tip of 2 day old in vitro germinated plants laied on four culture media (1mg/l 2, 4-D, 0/5mg/l 2, 4-D, 0/5mg/l 2, 4-D with 0/5mg/l kinetin, 1mg/l 2,4-D with 1mg/l kinetin) in darkness conditions. Subculture of samples was done after four weeks. The  produced calluses were laied in embryogenic culture medium for 4 weeks and after formation of embryoes, they were put in plant regeneration culture medium for eight weeks. Result showed that callus induction percentage is affected significantly by cultivar and culture medium, but reciprocal effect of both factors was not meaningful on callus induction percentage. Plant regeneration percentage and number of embryo per callus is affected meaningfully by cultivar, but type of culture medium and reciprocal effect of both factors was not meaningful on plant regeneration percentage and number of embryoes per callus. Embryogenic callus percentage is affected meaningfully by cultivar, but type of culture medium had not meaningful effect on embryogenic callus percentage. Callus induction percentage, plant regeneration percentage, number of embryo per callus and embryogenic callus percentage, are higher in white Kashan cultivar in relation to red Rey one and in culture medium involving hormonal combination of 2,4-D and kinetin, application of 0/5mg/l 2, 4-D can be usful. Considering the callus induction percentage, embryogenic and plant regeneration percentage, white Kashan cultivar and  culture medium containing 0/5mg/l 2, 4-D is appropriate as compared with other cultures