bap

Microbial contamination is one of the main challenges and influential factors in the response of explants under in vitro cultures, especially in woody plants where the explants are prepared from natural habitats. In the present study, to investigate the effects of benomyl on microbial contamination and growth of Vaccinium arctostaphylos L. node explants, MS containing different concentrations of benomyl (0-2000 mg.L-1) was used. Furthermore, to investigate the interaction of benomyl, basal medium, and hormonal composition on the growth response of explants, different basal media (MS, AN, and WPM) containing IBA (0.1 mg.L-1), cytokinin (Zeatin, BAP, and TDZ, each at concentrations of 0, 0.5, 1, and 2 mg.L-1), and benomyl (0 and 150 mg.L-1) were studied. The results showed that benomyl in MS medium had no significant effect on the explants contamination, but use of lower benomyl concentrations (400 mg.L-1 or less) could provide better growth conditions. Interaction of treatments significantly affected the percentage of survival and explants foliation. MS contained benomyl showed higher percentage of survival and foliation than MS without benomyl, but in AN and WPM media, the percentage of survival and foliation were significantly decreased in the presence of benomyl. Furthermore, in AN and WPM media contained cytokinin, the presence of benomyl significantly reduced the foliation percentage, while in MS medium, the effect of benomyl was different depending on the concentration and cytokinin type.

Purslane (P. oleracea) is considered as valuable plant due to its high antioxidant compounds and important fatty acids such as omega-3 and 6. Phenolic and flavonoid compounds are one of the most important constituents in the purslane. Phenolics are a large group of natural plant compounds with antioxidant and Anti-inflammatory properties. Flavonoids, as a subset of phenolic compounds, have a wide range of effects on plants, including antioxidant activity and improve resistance to environmental stresses. Callus culture is one of the important strategies for the production secondary metabolites, which are difficult to produce chemically. Plant growth regulators including auxins and cytokinins play a crucial role in the stages of plant growth. Various combinations of these two hormones are used to make the desired changes in the cultures. Studies suggest that the accumulation of secondary metabolites can be increased by the application of different elicitors in medium. Researchers reported an increase in the content of secondary metabolites such as phenol and flavonoid compounds in calli treated with elicitors such as yeast extract. The purpose of this study was to determine the best explant, medium and hormonal treatment for calli induction of purslane. The effect of different levels of yeast extract on total phenol and flavonoid content and antioxidant capacity of purslane calluses was also investigated.

Seeds of purslane plant were cultivated in a solid 1/2MS medium for the preparation of sterile seedlings. The explants from sterile seedlings including to leaves, 1 cm stem specimens and terminal buds, were placed on MS and 1/2MS medium containing 0, 0.1, 0.3, and 0.5 mg L-1 BAP and NAA. After five months, calluses were evaluated for callogenesis and some morphological traits such as color, texture, and size, fresh and dry weight. This experiment was conducted based on completely randomized design with three replications.  In the second experiment, the calluses obtained from the previous stage were transferred to MS medium with selected hormone treatment of the first experiment (0.5 mg L-1 NAA and BAP) and different levels of yeast extract (0, 125, 250, and 500 mg L-1). Total phenol and flavonoid contents of the calluses were determined by Folin-Ciocalteau and aluminum chloride methods, respectively. Furthermore, Ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays were used to determine the antioxidant activities.

The results showed that 1/2MS medium was suitable for sterile seedling production from purslane seeds. Based on the present study, only stem explants in a medium containing BAP and NAA, produced durable calluses. The color of the resulting calluses were green and had a constant and firm texture. The highest callus percentage (90.46%), the size (21.6 mm), and fresh (1826.5 mg) and dry weight (75.33 mg) of calluses belong to MS medium containing 0.5 mg L-1 BAP and NAA. Results of the second experiment showed positive and significant effects of yeast extract on the total phenol, flavonoid contents and antioxidant activities. The highest content of total phenol (664.12 mg GAE 100g-1 DW), flavonoid (42.25 mg QE 100g-1 DW) and FRAP data (787 µmol Fe g-1 DW) were obtained from the calli treated with 500 mg l-1 yeast extract. The maximum DPPH IC50 (2.45 mg ml-1) was also observed in control. The formation of callus associated with plant species, hormonal composition, the stage of development, and the type of explants. Auxin and cytokinin as plant growth regulators are key factors for controlling cell division in tissue culture. In most studies, callus formation in purslane plant were induced in medium containing auxin and cytokinin. The presence of green calluses derived from purslane explants can be due to the formation of chloroplastids in the cells of the callus tissue that rapidly produce chloroplasts under light conditions. In the second experiment, increased phenolic and flavonoid compounds with yeast extract treatment probably resulted in increased antioxidant activity.

In the present study, 1/2MS medium is suitable for the production of sterile seedlings from purslane seeds. MS medium containing 0.5 mg l-1 BAP and NAA is the best treatment for calli induction from stem specimens. The concentration of 500 mg L-1 of yeast extract is introduced as the most effective concentration for increasing the phenolic and flavonoid content and antioxidant activity in the purslane calluses.

In order to achieve maximum proliferation in culture media, different amounts of salts and growth regulators could be used regarding to plant species. Sometimes, interaction effects of these materials could results in different results on explant’s proliferation in vitro condition. In the present study the effect of two basic culture media including MS and QL on Micro propagation of Pyrus betulifolia (pear vigorous rootstock) were studied. The highest amount of proliferation was achieved in QL culture medium and highest amount leaf expansion was in MS culture medium. In order to produce more leaf expansion in QL culture medium and as for existence of difference in ions pure amount including NO3-, NH4+, Cl-and Ca2+ in two culture media, NH4NO3 in three concentrations including 6.25, 12.5 and 18.75 mM and CaCl2 in two concentrations including 0.9 and 1.8 mM were added to QL culture medium. As for the result, increasing NH4NO3 and CaCl2 amounts had no effect on proliferation of explants but by increasing NH4NO3 and CaCl2 leaf expansion increased in QL culture medium. In the other experiment for increasing of proliferation rate in MS culture medium, by increasing BAP amount, from 0.5 to 2 mg/l in MS culture medium, proliferation in this medium was increased. Apparently in MS culture medium with more concentration of macro elements compared to QL culture medium, it needs to use more growth regulators for achieving suitable proliferation. Totally, Results of this study showed that to succeed micro propagation of Pyrus betulifolia in both two basic culture media requires suitable concentrations of BAP, NH4NO3 and CaCl2.

In order to investigate effects of plant regulators on callus and embryo induction in cucumber anther culture, 25 media for callus induction and 4 methods for embryo induction were tested. The experimental layout was conducted in factorial arrangement in randomized completely design with four and three replications for callus and embryo induction, respectively. Factors for callus induction were two cucumber genotypes (Esfahani and Beta Alfa) and different concentrations of BAP (0, 0.225, 0.45, 0.68 and 0.91 mg/l) and 2, 4-D (0, 0.25, 0.5, 0.75 and 1 mg/l). Also factors for embryo induction were different concentration of BAP (2, 3 and 4 mg/l) and NAA (0.05 and 0.1 mg/l) and different solid and liquid media. Results indicated that there are statistical significant effect at 1% among PGR concentrations and 5% among interaction between genotype and PGR concentrations. The anthers, which were cultured on basal MS medium and media supplemented with individual concentration of plant regulators did not show any response and they became necrotic after 4 weeks of culture, but combination of plant regulators had different effect on callus induction. According to results of means comparison M24, M22 and M15 media with average of 98, 97.75 and 96.75 percent and M10 medium with average of 77.5 percent had highest and lowest callus induction for Esfahani genotype, respectively. The highest and lowest callus induction for Beta Alfa cultivar obtained in M13 (98.45%) and M1 (59.7%) media, respectively. There was no response for embryo induction in Solid-Solid, Solid-Liquid and Liquid-Liquid methods, but high rate of embryo induction obtained in Liquid-Solid method. Results indicated that there was statistical significant effect at 5% among media. Means comparison for tested media showed that M6 (60.58%) and M4 (20%) medium had highest and lowest embryo induction, respectively.

Iran is the third producer of Melon (Cucumismelo L.) after China and Turkey in the World.Melon is one of the main crops in Khorasan province in the term of cultivation area. The cultivar of Khatooni, among many varieties of melon, can be considered as a breeding cultivar because of its great qualitative traits including sweetness and flavor. This cultivar is also economically important and has many attractions for export. However, this cultivar is susceptible to some of plant diseases caused by different bacterial, viral and fungal pathogens which among them vascular wilt caused by Fusariumoxysporum f. sp. melonis (Leach &Currenu) Snyder & Hansen (FOM) is one of the most destructive soil-borne diseases in melon in Iran particularly in temperate and cold regions. The control of melon diseases has depended increasingly on the extensive use of toxic chemicals (pesticides). Many of these chemicals have been shown to be toxic to non-target microorganisms and animals and may be toxic to humans. Another problem with using chemicals to control plant pathogens is that a pathogen may become resistant to the chemicals.
The most promising control approaches are included conventional breeding and genetic engineering of disease-resistant plants. However, the conventional breeding method of melon is very complicated therefore; genetic engineering could be an effective and quick tool for producing new cultivars. In vitro direct regeneration is one of the most crucial step in gene transferring programs (20). In the present study, the effects of explant type and medium were considered in order to optimize the regeneration condition for melon (cv. Khatooni).
Material and The Khatoonicv.was provided by the Seed Bank of Plant Sciences Research Center from Ferdowsi University of Mashhad. The seeds were disinfected by NaClO in 1 % concentration, washed three times by distilled water and cultivated on MS medium (Murashigo and Skoog) for germination. The explant types were; lateral buds of cotyledon and the true leaves and hypocotyls. The selected media were: M1(0.1 mg.l-1 BAP (6-Benzyl amino purine)), M2 (0.25 mg.l-1 BAP), M3 (0.5 mg.l-1 BAP), M4 (0.75 mg.l-1 BAP),M5 (1 mg.l-1 BAP)and M6(including 0.5 mg.l-1 BAP and 0.01 mg.l-1 NAA (1-Naphthalene acetic acid))The experiment was repeated three times. The experimental –was conducted factorial based on completely randomized design and statistical analysis was performed using the SAS and JMP software. The corresponded graphs were drawn by Microsoft excel 2007.
The greatest amount of shoot regeneration in cotyledons was observed in M5 with 1 mg.l-1 BAP while minimum shoot regeneration was observed in M1with 0.1 mg.l-1 BAP. Cotyledon leaves showed the best regeneration efficiency among the other evaluated explants as it has also been reported by many other researchers (12, 8, 20, 21, and 2). The observations showed that in all BAP concentration, hypocotyls only formed callus tissue and did not produce any stem. In all three explants (true leaves, cotyledon and hypocotyls) rooting was observed in a treatment combination including 0.5 mg.l-1BAP and 0.01 mg.l-1 NAA 20 days after cultivation while no stem was formed at the same time. In three weeks after exposure , a weak root system was formed on the rooting mediumin without hormonetreatment . In MS medium enriched with 0.01 and 0.05 mg.l-1 NAA an appropriate root system was formed after 8 to 10 days. The medium containing 0.01mg.l-1 NAA was more appropriate for root regeneration (rooting).
Many studies have been shown that, the manipulation of different phytohormones ratio such as auxin and cytokinin in cultural medium is a suitable strategy to manage differentiation and organogenesis programs in plant (9). In the present study, the effects of explant type and medium were investigated in order to optimize the regeneration condition for melon (cv. Khatooni). As a result, in all BAP concentration, hypocotyls only formed callus tissue and did not produce any stem. However, by increasing of BAP hormone concentration in the cultivated media, shoot regeneration will also increase. The result of our study showed that, cotyledon leaves were more effective on shoot regeneration compared to the true leaves and hypocotyls as it has also been reported by other researchers.

Stevia (Stevia rebaudiana Bertoni) is a cross-pollinated plant whose seed vigor is poor. Therefore, the vegetative propagation approaches like in vitro culture techniques could be the best choice for mass and true-to-type production of stevia plantlets. This study was aimed to evaluate the effects of basal media (MS, B5 and LS) and physical parameters such as light (55 and 110 μmol m-2 s-1) and temperature (22, 25 and 27ºC) on in vitro culture of stevia. The nodal segments were cultured on basal media containing various combination of BAP and IAA or IBA. The ANOVA showed significant differences (P≤0.05) among treatments on micropropagation of stevia. In another experiment, the effect of mT (0.5, 1, 1.5 and 2 mg/l) was also compared with the best treatment from the first experiment. It was found that mT at 1 mg/l was the best concentration. The genetic stability of micropropagated plantlets relative to the mother plant was achieved by AFLP marker using 12 selected primers. In this study, 231 reproducible bands around 50-500bp were scored, indicating similarity of in vitro plantlets with the donor.

Echinacea is one of the most important medicinal plants and an important source of cichoric acid. In this research, callus induction, indirect shoot regeneration, and shoot rooting of Echinacea angustifolia DC. were investigated. The effects of explant (leaf, petiole and root) and NAA concentrations (0.5, 1 and 1.5 mg l-1) on callus induction as well as the effects of explant (leaf, petiole and root) and BAP concentrations (3, 4 and 5 mg l-1) on the indirect shoot regeneration of callus were evaluated in two separated experiments. The study was conducted as factorial in a completely randomized design (CRD). The effects of IBA concentrations (0.5, 1, 1.5 and 2 mg l-1) on shoot rooting were also investigated in a completely randomized design. The effects of the factors studied on the callogenesis were not significant while these factors had significant effects on the shoot regeneration and rooting at 1% probability level. In general, the results indicated that the highest indirect shoot regeneration (72%) was obtained from the leaf explant and by the use of 4 mgl-1 BAP. In addition, the highest shoot rooting (38% and 40%) was obtained using 1 and 1.5 mgl-1 IBA. The good potential of this species for indirect shoot regeneration was shown in this research. These results can be used in micropropagation programs, gene transfer, and transgenic plants production.

Rosa damascena is one of the most important Aromatic plants in the world. The extracted oil from the petals of the flowers is used in the pharmaceutical, cosmetic and food industries. Traditional methods to propagate this plant have problems such as transfer of pollution agents and limited native plants. Due to more advantages of the in vitro proliferation in compared to traditional methods, this study designed to evaluate impression different medium cultures with diverse hormone levels to improve the proliferation and control of shoots chlorosis. In this research we used medium cultures of MS and mMS (mMS0, mMS1, mMS2, mMS3, mMS4 and mMS5). In order to establishment in MS medium, nodal segments containing 1 or 2 lateral buds were cultured. For regenerating the shoots, mediums with different concentrations of 1 and 2 mg/l BAP and 0/2 mg/l IBA were supplied, then plantlets were transferred to the rooting medium (1/2 MS and LS) with different levels of IBA (1 and 2 mg/l). The best treatment to increase the number of rose shoots was mMS medium containing 1.5 mg/l of BAP and 0.2 mg/l of IBA. The highest chlorophyll content of leaves was observed in mMS medium. Gallica plantlets produced the highest percentage of rooting on 1/2 MS with 2 mg/l of IBA.

Satureja bachtiarica L., belonging to the Lamiaceae family, is rich in essential oils, used for various purposes including pharmaceutical, food, and health applications. In vitro culture of Satureja, in order to callus induction and optimization of cell suspension, provides rapid production and easy extraction of secondary metabolites. In the present study, callus induction was conducted by using different explants and concentrations of growth regulators. The most suitable calli were selected to induce suspension cultures and were transferred to liquid media supplemented with different combination of BAP and NAA. The experiment was performed in a factorial completely randomized design by using three replications. The cell biomass (cell number) was examined on different days. The mean comparison results for the explant origin showed that the internode explant produced the highest cell number. In addition, evaluation of interaction of explants with media showed that the maximum cell biomass was obtained by internode explant and the medium containing 2mg.L-1 NAA plus 0.5mg.L-1 BAP in the fifth day. On the other hand, study of growth process and cell proliferation during the days after the establishment of suspension cultures, also revealed that this hormonal composition had the highest utility to generate the maximum number of cells in all explants tested.

In order to investigate the in vitro micropropagation Hibiscus Rosa-sinensis three tests in a completely randomized design with 10 replications were performed in tissue culture laboratory Shahid Chamran University in 2014.The first experiment showed the perfect media.Second tests best consentration BAP for multiplication and third tests showed the best type and concentration rooting hormone. Compare average of first experiments showedthe highest average shoot length, number of leaves, fresh weight and dry weight was observed in VS than other media. Compare average of second experiments showedthe concentration of 5.0 mg per liter hormone BAP highest average shoot length, number of leaves, fresh weight and dry weight than other concentrations was observed.Compare third experiment showed that the average concentration of 2.0 mg per liter of IBA hormone highest percentage of rooting, number of roots and root length was compared to other concentrations.

Micropropagation is a proper approach to rapid and large-scale propagation of rootstocks and rose cultivars for huge demand of flower market. Proliferation rate of shoot is decreased drastically following several subcultures. Growth regulators have remarkable effects on the key phase of proliferation in micropropagation of this popular crop. In this research the effects of BAP and antiauxin of TIBA on quality and quantity of developed shoots in Rosa hybrida cv. Full House were studied. BAP and TIBA were applied at three concentrations of 0، 2. 2 and 8. 8 µmol in proliferation phase of micropropagation. The experiment was conducted based on factorial and completely randomized design with four replications. After two months، the percentage of proliferated explants، survived main and lateral shoot number، length of the main and lateral shoots، number of green leaves on the shoots، the average number of shoots with chlorotic and necrotic leaves، the average axillary shoot base diameter، fresh weight of shoots and number of shoots with necrotic tip were recorded. Analysis of variance indicated that BAP was ineffective on the number of the main shoot green leaves and decreasing number of shoots with necrotic tip، but enhanced other traits. The concentration of 8. 8 µmol of BAP had greater effect than 2. 2 µmol of this growth regulator on mentioned traits. The higher concentration of TIBA resulted to more shoot with necrotic tip. This antiauxin had anegative impact on shoot fresh weight، but the other parameters were not significantly affected.

Lemon balm (Melissa officinalis L.) is an herbaceous perennial plant of the Lamiaceae family that has a wide edible and medicinal uses. In this research the potential of five different explants (nodal segments، shoot tip، cotyledon، hypocotyl and leaf segments) on MS mediums supplemented with different concentrations of 6-Benzylaminopurine (BAP) (2. 2، 4. 4 and 8. 8 µm) alone or in combination with 1µm Indoleacetic acid (IAA) for direct regeneration of Lemon balm was investigated. The experiment was conducted using a factorial based on completely randomized design with three replications. The maximum mean number of shoots (23. 59 and 16. 90 shoots in shoot tip and nodal segment explants، respectively) was observed on the MS medium with 8. 8 µm BAP. The highest regeneration rate (99%) in shoot tip and nodal segment explants was achieved on the MS medium supplemented with 8. 8 and 2. 2 µm BAP، respectively. No shoot regeneration occurred in other explants. The rooting of regenerated shoots was assessed on MS، ½MS and MS medium supplemented with 1، 2. 5، 4. 92 and 9. 84 µm Indole butyric acid (IBA). The highest mean number of roots (9. 06 roots per explants) was observed on the MS medium supplemented with 4. 92 µm IBA.

Melon (cv. ‘Semsuri’) is widely cultivated in Iran and therefore increasing the fruit yield and quality is necessary. Some growth regulators including 6-Benzylaminopurin (BAP) may affect plant growth and development. In two field trials, impact of BAP was studied on growth and fruit yield of melon. In first experiment, growth parameters of plants were recorded after 10 days of foliar treatment (6-7 leaf stage) with 0, 10, 20 and 40 mg l-1 BAP. In second experiment, plants were foliar sprayed with BAP at two growth stages (6-7 leaf and after fruit set stages). In both experiments, 40 mg l-1 BAP was most effective when compared to control, one spray increased relative water content, chlorophyll index, plant fresh weight, rind firmness and fruit yield by 16, 24, 26, 5 and 22%, respectively. High correlation coefficients were observed between fruit yield and other growth parameters recorded, so it may be concluded that fruit yield can be improved by promotion of growth parameters.

In this research callus induction under in vitro condition in medicinal plant, lemon balm (Melissa officinalis L.) has been considered as a preliminary method for modification of this plant. For this purpose factorial expriment based on completely randomize design has been used with factors, ecotype (Isfahan and Karaj), origin of explant (leaf, petiol, internode), Lighting conditions (light and dark), 2, 4-D (0, 0.5, 1, 2 mg/l) and BAP (0, 1, 2 mg/l) with three replications. Results showed that the best hormonal treatment for callus induction in lemon balm was 1mg/l 2, 4-D + 1mg/l BAP for internodes and petiole in light condition. After that, use of leaf explants with 0.5mg/l 2, 4-D + 1mg/l BAP in dark and then light condition was the best.