Callus Dry Weight Research Articles

Effect of plant growth regulators in inducing Vitisvinifera Callus and increased production of flavonoids in vitro

This study was aimed to use of plant tissue culture technology to induce callus from grapevine Vitisvinifera and stimulate it to increase the production of flavonoids. The study was carried out in two stages after the sterilization process was carried out: The first included establishing callus farms by cultivating cuttings containing a single node after completing their sterilization and cultivating them on MS medium containing 2,4-D and BA at different concentrations (0, 1, 2, 3,4) mg L -1and BA in concentrations (0, 0.1, 0.2, 0.4) in independent experiments. The second phase included studying the effect of growth regulators on the concentrations of carbohydrates and flavonoids in the callus induced from the first experiment. The results of the study showed the superiority of auxien 2,4-D at a concentration of 2mg L -1as it achieved the highest average fresh and dry weight of callus, reaching (3.32 and 1.03) mg respectively, while BA at a concentration of 0.2 mg L -1 achieved the highest average fresh and dry weight of callus was (1.91 and 0.51) mg respectively. The results also showed that the MS medium prepared with a concentration of 4 mg L -1of 2,4-D was significantly superior in the average concentration of the compounds Hesperdine, Nargnine, Proanthocyanin, and Rutin, reaching (64.94, 62.56, 55.63, and 71.22)% respectively.While the medium prepared with a concentration of 3 mg L -1outperformed 2,4-D achieved the highest concentration of Quercetin compounds reaching 75.36% compared to the neutral treatment, which achieved the lowest concentrations.

Induksi Kalus dari Hipokotil Belimbing Merah (Baccaurea angulata) dengan Penambahan 2,4-D (Dichlorophenoxy Acetic Acid) dan BAP (6-Benzyl Amino Purin)

Red starfruit is one of the endemic plants of Kalimantan which produces secondary metabolites which has been empirically used as antioxidant, antibacterial, and anticancer. This plant has a long reproductive period and seedless fruit is often found during the fruiting season, which is a problem for the sustainability of the red starfruit in nature. Conservation of red star fruit for propagation and production of secondary metabolites can be done in vitro through callus culture. This research aims to determine the effect of adding the growth regulators dichlorophenoxy acetic acid (2,4-D) and 6-benzyl amino purine (BAP) on callus induction time, percentage of callus forming explants, callus color and texture, and callus growth. This research used a Completely Randomized Design (CRD) with a factorial pattern consisting of two treatment factors, namely 2,4-D (0; 0.25; 0.5; 0.75; 1 ppm) and BAP (0; 0.25; 0.5; 0.75; 1 ppm). The percentage of explants forming callus (%), the time the callus appeared (days after planting), the color and texture of the callus, the wet weight and dry weight of the callus (g) were research observation parameters.The results showed that the single factor 2,4-D, single factor BAP, and the interaction of 2,4-D and BAP had a significant effect to callus appearance time. Single administration of 2,4-D and BAP had a significant effect on callus dry weight. The color variations of the callus produced are white, green and brown with a compact texture. The fastest callus emergence time was at 11.00 days after planting (DAP), namely in the combination treatment of 1 ppm 2,4-D and 0.75 ppm BAP, single treatment 0.75 ppm and 1 ppm 2,4- D. The highest average dry weight of 0.024 g was obtained in the single treatment of 0.75 ppm BAP.

<em>In Vitro</em> Effect of PEG and Proline on Callus Growth and Minerals Values in Basmati Rice (<em>Oryza sativa</em>)

Purpose: Drought is a universal issue that disturbs photosynthetic system, adversely affecting quality and crop yield. The study was conducted to determine response of polyethylene glycol (PEG) as stressing agent alone or combination with proline on four fine rice genotypes for callus components as well as biochemical parameters like Chromium (Cr), Magnesium (Mg), Manganese (Mn), Iron (Fe), Phosphorus (P).Research Method: Callus was induced on media having 4.43g/L MS + 4mg/L 2, 4-D (2, 4-dichlorophenoxy acetic acid) + 30g/L sucrose + 1.76g/L phytagel with different levels of PEG “1.25g/L and 2.5g/L” with and without 1ml/L proline.Findings: Callus was used for different analyses after 30 days and results showed that PEG treatments have a negative effect on callus fresh weight, callus dry weight, days to callus induction, callus percentage, and mineral nutrients like K, Cr, Mg, Mn, Fe, P. PEG 2.5g/L showed a greater decrease as compared to PEG 1.25g/L. Proline significantly raises callus growth alone or in combination with PEG but less than control whereas callus dry weight, K, and Ca contents increased more than control when proline applied alone or with PEG combination.Originality/value: Exogenous proline application improves callus growth rate as well as macro- micro elements by mitigating the effect of PEG.

Secondary metabolite profiles, antimicrobial and antioxidant activities of callus, and leaves extract of Piper sarmentosum Roxb.

This research was aimed to investigate the effect of different combination concentrations of 2,4-diclorophenoxyacetic acid (2,4-D) and benzylaminopurine (BAP) on callus induction, profiles of secondary metabolites, antimicrobial, and antioxidant activities from Piper sarmentosum leaves. These leaves explants were cultured on Murashige and Skoog (MS) medium supplemented with different combination concentrations of 2,4-D and BAP (0.5; 1.0; 1.5; 2.0; 2.5 mg/L) during 6-week culture callus period. Result showed that the morphology of callus grown during this study was compact in various colors appearance, such as light brown, brownish, and dark brown. The fastest mean of callus induction time was 0.5 mg/L 2,4-D and 1.5 mg/L BAP (10 days). The highest callus fresh weight was 1.5 mg/L 2,4-D and 2.0 mg/L BAP (0.350 g). The combination concentration of 1.5 mg/L 2,4-D and 1.5 mg/L BAP was the highest callus dry weight (0.078 g), this treatment was contained 11 different compounds, which were dominated by cyanoacetamide (25.08%) and diisooctyl-phthalate (21.81%). Myristicin and propanamide compounds were also identified in the methanol extract of P. sarmentosum callus. Methanolic extract of callus P. sarmentosum from the optimum treatment had the highest antioxidant activity (26.709 μL/mL) and antimicrobial activity against three test microbes.

Induction of Callus of Two Cultivars from Rice Seed Embryos by the Effect of Abiotic Factors for Drought Tolerance ex Vivo

The experiment was conducted at the Plant Tissue Culture Laboratory, College of Agricultural Engineering Sciences, University of Baghdad, during the period 2021-2023, with the aim of inducing Callus from embryos of two rice cultivars: black rice and Amber 33, to enhance drought tolerance in vitro. The Completely Randomized Design (CRD) was applied for experimental arrangements. Different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 1.5, 3, 4.5 mg L-1) were used to stimulate Callus, along with UV radiation of type UV-C with a wavelength (<280 nanometers), It does not affect the environment because it is absorbed by the atmosphere’s oxygen and ozone, for various durations of 15, 30, 45, and 60 minutes. A control treatment was also used for comparison. The experimental stress conditions were induced using different concentrations (0, 1.5, 3, 4.5, and 6%) of polyethylene glycol (PEG). The fresh and dry weights of Callus were measured for all treatments, in addition to assessing the drought tolerance index. The results showed significant differences for most of the studied traits. The highest fresh and dry weight of Callus was achieved at a concentration of 3 mg L-1 of 2,4-D, at 258.9 mg and 26.06 mg, respectively. The control treatment with UV-C radiation outperformed with the highest fresh and dry weight of Callus, reaching 356.8 mg and 37.81 mg, respectively, compared to other irradiation doses. The control treatment with other PEG concentrations yielded the highest fresh and dry weight of Callus for the cultivars treated with UV-C radiation, reaching 356.4 mg and 38.81 mg, respectively. The drought tolerance index in terms of dry weight, the treatment with 1.5% PEG yielded the highest rate at 0.73 mg, while the treatment with 6% PEG resulted in the lowest rate at 0.21 mg. additionally; the black rice cultivar treated with UV-C radiation outperformed the Amber 33 rice cultivar in terms of both fresh and dry weights of Callus.

Effect of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin on callus induction and growth of Physalis angulata L. leaf explants

Physalis angulata L. is an annual herbaceous plant used as traditional medicine. The roots, stems and leaves are used to treat diabetes, hypertension, fever and headaches. The fruit contains lots of provitamin A and has antitumor, antihyperglycemic, anti-inflammatory, antidiabetic and immunomodulatory activities. P. angulata leaves also contain secondary metabolites of alkaloids, flavonoids, saponins, steroids, tannins, physalin A, physalin B, glycosides and stearic acid. The use of plants as a source of medicine is not only very dangerous for the survival of plants, but also not standardized. Secondary metabolites can be produced in callus by in vitro culture induced with appropriate growth regulators. Through callus culture, secondary metabolites or chemical substances from plants can be produced. This study aims to determine the effect of growth regulators 2,4-D and kinetin on callus induction time, the percentage of explants forming callus, fresh weight, dry weight, and callus morphology. This study was a laboratory experimental with a completely randomized design and 5 treatments (0.0 mg/L 2,4-D + 2.0 mg/L kinetin; 0.5 mg/L 2,4-D + 1.5 mg/L kinetin; 1.0 mg/L 2,4-D + 1.0 mg/L kinetin; 1.5 mg/L 2,4-D + 0.5 mg/L kinetin; 2.0 mg/L 2,4-D + 0.0 mg/L kinetin), each treatment consisted of five replications. The data obtained has been analyzed qualitatively and quantitatively. Qualitative data were obtained from callus morphological (color and texture) descriptions. Quantitative data obtained from callus induction time, the percentage of explants forming callus, fresh weight and dry weight of callus were analyzed using SPSS 24 with a significance value of 0,05. Data analysis on the effect of 2,4-D and kinetin concentrations used the ANOVA test. Difference between treatments were tested using the Duncan´s multiple range test. The results showed that the suitable growth regulator for inducing callus growth in leaf explants of P. angulata L was 2.0 mg/L 2.4D + 0.0 mg/L kinetin. Growth regulator 2.0 mg/L 2,4-D + 0.0 mg/L kinetin induced callus in a short time i. e. 10.60 days after culture, resulting in the highest fresh and dry weight of callus i.e., 0.35 g and 0.03 g, as well as the percentage of explants forming callus 100%. Callus has a compact texture with a variety of colours.

Effect of copper, nickel and lead on callus growth dynamics of Solanum lycopersicum

Solanum lycopersicum cv. (Punjab Kesar Cherry) (Tomato) belonging to family Solanaceae has been priced for the presence of lycopene, low sugar content and curative properties. However, the productivity of tomatoes has been observed to be low due to both biotic as well as abiotic stress. Considering the toxicity of copper (Cu), lead (Pb) and nickel (Ni) as environmental contamination and the economic importance of S. lycopersicum, the present study was undertaken to investigate the effects of these heavy metals on various growth parameters of callus cultures and plantlet regeneration. Callus induced through nodal segments was inoculated on Murashige and Skoog medium containing different concentrations (0 µM, 50 µM, 100 µM, 200 µM, 300 µM, 400 µM and 500 µM) of copper, lead and nickel. Decrease in different growth parameters was observed as 91.66-16.66 %, 76.38-11.11 %, 81.94-13.88 % for percent callus survival, 90.90-25 %, 72.72-37.50 %, 79.66-20 % for percent callus multiplication, 79.54-0 %, 82.75-33 %, 63.8-50 % for percent root regeneration and 73.33-33 %, 72.50-33 %, 79.6- 20 % for percent shoot generation with increasing concentrations of Cu, Pb and Ni, respectively. Decrease in the average fresh and dry weight of callus was observed for all the metals. The order of toxicity was observed as Pb &gt; Ni &gt; Cu at the highest concentrations used for treating S. lycopersicum. The present study revealed that all three metals induced stress in the studied plant and need attention to developing methods to mitigate the consequences of metal toxicity in crop plants.

PERTUMBUHAN KULTUR KALUS YANG DIINDUKSI DARI EKSPLAN HIPOKOTIL LAKUM (Causonis trifolia (L.) Mabb. &amp; J.Wen) DENGAN PENAMBAHAN NAA (Naphthalene Acetic Acid) DAN BAP (6-Benzyl Amino Purin)

Lakum plant (Causonis trifolia (L.) Mabb. &amp; J.Wen) contains secondary metabolites with potential medicinal ingredients. It can treat various diseases, such as antidiabetic, antibacterial, antiprotozoal, antitumor, and anti-cancer. Propagation and production of secondary metabolites in plants can be carried out in vitro through callus culture and are influenced by the concentration of growth regulators. This study aimed to determine the effect of NAA (Naphthalene Acetic Acid) and BAP (6-Benzyl Amino Purin) on callus growth of hypocotyl lakum (C. trifolia) explants and to determine the concentration of addition of NAA and BAP that could produce the best callus growth. Completely randomized design (CRD) factorial with two factors adding NAA (0, 0.45, 0.9, and 1.4 µg/l) and BAP (0, 0.23, and 0.56 µg/l). The results showed that adding a single NAA and a combination of NAA and BAP significantly affected callus emergence. The fastest callus emergence was found in a combination of 0.45 µg/l NAA + 0.23 µg/l BAP, 13 days after planting. Both NAA and BAP alone significantly affect the weight of dry and wet callus. The administration of a single NAA and a single BAP had a significant effect on the wet weight and dry weight of callus, with the highest average wet callus weight at a concentration of BAP 0.56 µg/l, which was 4.431 g, and the highest dry weight callus concentration of BAP 0.56 µg/l was 0.192 g.

Phytochemicals and Antioxidant Activities of Conventionally Propagated Nodal Segment and In Vitro-Induced Callus of Bougainvillea glabra Choisy Using Different Solvents

Bougainvillea, popularly known as ‘Bunga kertas’ in Malaysia, is thoroughly explored for nutritional and medicinal purposes. Bougainvillea has been shown to possess alkaloids and flavonoids which are widely used in folk medicine to treat different illnesses such as inflammatory, diarrheal, ulcer, and diabetic. Despite its major conventional therapeutic importance, only limited attempts have been made to investigate this species’ chemical and pharmacological properties in relation to its medicinal uses. Therefore, this study was conducted to determine the effect of in vitro-induced callus under different light conditions and plant growth regulators on phytochemical and antioxidant activities using different extraction solvents. Based on the results, the maximum days (17.67) to callus initiation were recorded when nodal was cultured on woody plant medium (WPM) supplemented with 7.5 µM 2,4-D + 0.5 µM BAP under light condition. On the contrary, the minimum days (7) to callus initiation were obtained when nodal was treated with 2.5 and 5 µM 2,4-D + 1 and 1.5 µM BAP under dark conditions. However, higher fresh and dry weight of callus was obtained when nodal was cultured on woody plant medium fortified with 7.5 µM 2,4-D + 1.5 µM BAP under dark and light conditions. In the analysis of the phenolics content and antioxidant activities, aqueous extract of conventionally propagated nodal part exhibited the highest phenolic content and antioxidant activities. However, the highest iron (II) chelating activity was produced from the aqueous extract of the calli induced under a dark condition. Hence, it can be concluded that the callus culture of Bougainvillea produced plant secondary metabolites and antioxidant activities comparable to the mother plants.

Impact on Growth and Secondary Metabolites in White Poplar (Populus alba L.) Callus using SeNPs@Moringa

Background: Green synthesis of nanoparticles has provided a cost-effective, environmentally friendly method, as well as raised safe strategies for the synthesis of nanomaterials. Methods: In this study, we investigated the synthesis of Selenium nanoparticles from the leaf aqueous extract of Moringa oleifera (MO-SeNPs) and studied the effect of MO-SeNPs and Moringa extract with various concentrations on the growth, active constituents, and antioxidant activity (scavenging DPPH) of Populus alba callus. MS medium supplemented with (2, 4-D at 2 mg/l + 0.5 mg/l BA) took the maximum callus induction percentage (100%), increasing the callus fresh weight, dry weight, and degree of callus formation compared with (2.0 mg/l 2,4-D + 0.5 mg/l Kin). Results: The highest values of callus fresh and dry weight were recorded on MS containing 20% MO-SeNPs. Regarding the active constituents, the GC-MS chromatogram revealed the presence of 23 major compounds identified in the chloroform extract. The phytoconstituents with maximum peak area in callus extract are Tetradecane, Hexadecen-1-ol, Trans-9, 5-Octadecene, (Benzene, 1- pentyloctyl), (Benzene, 1-butylnonyl), Cycloeicosane, and 10-Heneicosene in all treatments. Moringa extract and MO-SeNPs stimulate the increase of compounds in callus compared to control. In this regard, MO-SeNPs 5% led to a higher increase in the area percent of active substances compared to the treatment of Moringa extract and control. Concerning the antioxidant activity, the results indicated that Moringa extract and MO-SeNPs had a positive effect on scavenging DPPH compared with the control. The highest percentages of scavenging DPPH were recorded when Populus alba L. callus treated with MO-SeNPs 20%, 10%, and Moringa extract 20%, percentages reached (84.65, 75.35, and 71.94%), respectively. Conclusion : Green synthesis of MO-SeNPs improved secondary metabolites and antioxidant activity in Populus alba L. callus.

In Vitro Culture of Eremurus spectabilis (Liliaceae), a Rare Ornamental and Medicinal Plant, through Root Explants

Eremurus spectabilis M. Bieb, a perennial herbaceous wild species, is commonly used in the horticultural, ornamental, and pharmaceutical markets. Studies on the tissue culture systems for this species would be beneficial for mass multiplication as well as for future breeding programs. An in vitro propagation technique was established here using tuberous root explants as unique and responsive starting materials for culture initiation. The proliferated calli were sub-cultured on shoot proliferation media and regenerated microshoots were assessed. The shoot proliferation rate, leaf number, leaf length, and chlorophyll and carotenoid contents were recorded. The highest callus induction per explant (76.67%), callus dry weight (10.25 mg), callus firmness ratio (3.97), and callus color intensity ratio (2.83) were observed in explants inoculated on Murashige and Skoog (MS) medium supplemented with 10.0 mgL−1 6-benzylaminopurine (BAP). The highest shoot proliferation rates were obtained when calli were sub-cultured on MS or Schenk and Hildebrandt (SH) basal media supplemented with 2.0 mgL−1 BAP. The half-strength MS medium fortified with 4.0% sucrose + 2.0 mgL−1 indole butyric acid (IBA) + 200 mgL−1 activated charcoal was a superior combination for root emergence and rooting parameters. Regenerated plantlets were then successfully adapted to ex vitro conditions. The reported protocol can be exploited at a commercial scale following minor modification, or could be beneficial in the production of secondary metabolites in bioreactors where callus is required as fresh plant material.

In vitro salinity stress mediates grass pea genotypes' (Lathyrus sativus L.) responses

This study was carried out to determine the tolerance of grass pea genotypes to salinity stress at callus and seedling stages under in vitro conditions. The calli and seedlings of six selected tolerant genotypes based on the primary screening in the field were separately exposed to salinity treatments (0, 125 mM) in vitro. Salinity was imposed with NaCl during in vitro culture, and it significantly affected all seedling traits. Genotype of Iran had the lowest seedling dry weight and therefore was more sensitive to salinity stress. According to salinity tolerance indices for seedlings, genotype Greece-III was characterized as high-yield and relatively high?salt-tolerant genotype. Salinity significantly affected callus size, callus RWC, callus RGR, and callogenesis index. Calli fresh and dry weights were not affected by the treatments. For callus dry weight, genotype Greece-III had the highest mean; and the lowest mean belonged to Greece-I. The stress tolerance indices showed that the highest values belonged to genotype Greece-III, which showed high yield and yield stability and so reasonable salinity tolerance. Cluster analysis divided the genotypes into two separate groups. The first cluster consisted of Iran, Greece-II, and Greece-III genotypes, and the second cluster consisted of Bangladesh, Canada, and Greece-I genotypes. Cluster analysis potentially separated the tolerant and sensitive genotypes to salinity in terms of callus dry weight. Grass pea callus and seedlings were able to survive at 125 mM salinity. Salinity did not affect callus dry and fresh weights, but its effect was remarkable on seedling dry and fresh weights (55% less than control). Therefore, calli were reasonably salinity tolerant. The present study suggests that grass pea was reasonably tolerant to salinity and can survive under salinity conditions during the seedling and callus stages.

Preparation of Readily-to-Use Stilbenoids Extract from Morus alba Callus Using a Natural Deep Eutectic Solvent

The consumer and cosmetic industries have recently placed a greater emphasis on ecofriendly solvents for botanical extraction, including natural deep eutectic solvents (NADES). In this study, NADES were prepared for Morus alba callus extraction. The efficiency of extraction from the NADES and methanol was investigated by comparison of the stilbenoids yield and anti-melanogenesis activity. Prior to testing the irritability of a suitable NADES on the reconstructed human epidermis (RhE), the effect of the selected NADES on stilbenoids stability was determined. The results showed that the highest yields of stilbenoids were obtained from choline chloride-glycerol mixtures (Ch1G2) and methanol extracts, with no significant difference in yields (5.06 ± 0.05 and 6.32 ± 0.40 mg/g callus dry weight, respectively). The NADES extracts of M. alba callus showed comparable anti-melanogenesis activity compared to methanol. In term of stability, stilbenoids in Ch1G2 remained stable after six months of storage at 4 °C except resveratrol. Furthermore, Ch1G2 had no irritation effect on RhE. Thus, based on the findings of this study, Ch1G2 is an intriguing green solvent alternative for the extraction of M. alba callus and may be advantageous for the preparation of skin-lightening cosmetics.

Nigella sativa callus treated with sodium azide exhibit augmented antioxidant activity and DNA damage inhibition

Nigella sativa L. (NS) is an herbaceous plant, possessing phytochemicals of therapeutic importance. Thymoquinone is one of the active phytochemicals of NS that confers noteworthy antioxidant properties. Sodium azide, an agent of abiotic stress, can modulates antioxidant system in plants. In the present investigation, sodium azide (0, 5 µM, 10 µM, 20 µM, 50 µM, 100 µM and 200 µM) doses administered to the in vitro NS callus cultures for production/modification of secondary metabolites with augmented activity. 200 µM sodium azide treated NS callus exhibited maximum peroxidase activity (1.286 ± 0.101 nanokatal mg−1 protein) and polyphenol oxidase activity (1.590 ± 0.110 nanokatal mg−1 protein), while 100 µM sodium azide treated NS callus for optimum catalase activity (1.250 ± 0.105 nanokatal mg−1 protein). Further, 200 µM sodium azide treated NS callus obtained significantly the highest phenolics (3.666 ± 0.475 mg g−1 callus fresh weight), 20 µM sodium azide treated NS callus, the highest flavonoids (1.308 ± 0.082 mg g−1 callus fresh weight) and 100 µM sodium azide treated NS callus, the highest carotenes (1.273 ± 0.066 mg g−1 callus fresh weight). However, NS callus exhibited a decrease in thymoquinone yield/content vis-à-vis possible emergence of its analog with 5.3 min retention time and an increase in antioxidant property. Treatment with 200 µM sodium azide registered significantly the lowest percent yield of callus extract (4.6 ± 0.36 mg g−1 callus fresh weight) and thymoquinone yield (16.65 ± 2.52 µg g−1 callus fresh weight) and content (0.36 ± 0.07 mg g−1 callus dry weight) and the highest antioxidant activity (3.873 ± 0.402%), signifying a negative correlation of the former with the latter. DNA damage inhibition (24.3 ± 1.7%) was recorded significantly maximum at 200 µM sodium azide treatment. Sodium azide treated callus also recorded emergence of a new peak at 5.3 min retention time (possibly an analog of thymoquinone with augmented antioxidant activity) whose area exhibits significantly negative correlation with callus extract yield and thymoquinone yield/content and positive correlation with antioxidant activity and in vitro DNA damage inhibition. Thus, sodium azide treatment to NS callus confers possible production of secondary metabolites or thymoquinone analog (s) responsible for elevated antioxidant property and inhibition to DNA damage. The formation of potent antioxidants through sodium azide treatment to NS could be worthy for nutraceutical and pharmaceutical industries.

Effect of 2, 4-D and BAP on Callus Induction of Piper retrofractum Vahl.

Piper retrofractum Vahl. in Indonesia has potential to be one of the medicinal plants to treat infertility problems. Several secondary metabolite compounds contained in Piper retrofractum Vahl. are essential oils, alkaloids, piperins, saponins, flavonoids, triterpenoids, steroids and glycosides. Plant tissue culture method to induce callus media requires proper formulation to get the optimum result. The purpose of this research was to know the effect of growth regulators combination of 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and 6-Benzylaminopurine (BAP) on time of induction, percentage, fresh weight, dry weight, and morphology on callus culture of Piper retrofractum Vahl.'s leaf explant. Leaf explant of Piper retrofractum Vahl. cultured on Murashige and Skoog (MS) medium with combination of 2, 4-D and BAP with concentrations of 0.0mg/L, 0.5mg/L, 1mg/L, 1.5mg/L, 2mg/L. This research was a laboratory experimental research with the complete randomized design method. The data obtained were analyzed quantitatively and qualitatively. Quantitative data consisted of time of induction, percentage, fresh weight, dry weight of callus, while qualitative data was obtained from the morphology description of Piper retrofractum Vahl. The results showed that the combination of growth regulators 2, 4-D and BAP added in culture medium influenced the growth of Piper retrofractum Vahl. leaf explant. Both at the time of induction, percentage, fresh weight and dry weight of callus. The best combination of growth regulators 2, 4-D 1.0mg/L and BAP 1.0mg/L showed the average time in inducing callus 15 days after planting. The combination of growth regulators 2, 4-D 1.0mg/L and BAP 2.0mg/L was the best combination resulted in an average of fresh weight and dry weight of callus 31.20mg and 5.92mg. Morphology of callus was produced by explant mostly textured friable greenish-white which cultured in medium with combination of growth regulators 2, 4-D 1.0mg/L and BAP 2.0mg/L in all parts of edge and surface explant.

FTIR, GC-MS and HPLC analysis of Baliospermum montanum (Willd.) Muell. Arg.

The present work is aimed to determine the chemical constituents in Baliospermum montanum methanolic extracts. An in vitro regenerated procedure was developed for the induction of callus from stem explant cultured on Murashige and Skoog (MS) medium fortified with various concentration and permutations of 2, 4-dichloro phenoxy acetic acid, 1-naphthalene acetic acid, 6-benzyl amino purine and gibberellic acid. FTIR &amp;amp; GC-MS analysis was done according to standard procedure. The quantitative estimation of β-sitosterol was done by HPLC method. Maximum fresh and dry weight of callus was estimated in the combination of GA3 (0.5 mg/L) + NAA (2 mg/L) compared to other concentration. The FTIR analysis showed various functional compounds with different characteristic peak values in the extracts. Major bioactive constituents were recognized in the GC-MS analysis. Root extract revealed the existence of 1-hexadecanol, pentanoic acid, 2-(aminooxy)- and 1-hexacosanol. Leaf extract showed the presence of propanoic acid, 2-oxo-, trimethylsilyl ester, 9,12-octadecadienoic acid (z,z)-, trimethylsilyl ester, docosane, 1,22-dibromo- and pentatriacontane. Stem and stem derived callus exhibit the presence of 1,6,3,4-dihydro-2-deoxy-beta-d-lyxo-hexopyranose, n-hexadecanoic acid and pentanoic acid, 2-(aminooxy). The methanolic extract of leaf exhibited 0.2149 % of β-sitosterol content. There were no peaks observed in the root, stem and stem derived callus. Further studies are necessary for the isolation and characterization of bioactive compounds from B. montanum.

Production of ascorbic acid, total protein, callus and root in vitro of non-heading Chinese cabbage by tissue culture

The objective of the present work was the selection of cultivar, suitable medium and explant type for callus, root production, ascorbic acid, total ascorbic acid, dehydroascorbic and total protein of non-heading Chinese cabbage in two cultivars 'Caixin' and 'Suzhouqing'. We compared 10 types of MS media supplemented with 0.0, 1.0, 2.0 and 3.0mg/l TDZ; 0.0, 0.25, 0.50 and 1.0mg/l NAA and 0.0, 5.0, 7.5 and 9.0mg/l AgNO3 and 5 kinds of explants as embryo, leaf, root, cotyledon and hypocotyl. Maximum frequency of callus fresh weight was recorded with hypocotyl explant, which were cultured on MS + 2.0mg/l TDZ + 1.0mg/l NAA + 9.0mg/l AgNO3 in 'Suzhouqing', optimum callus dry weight was obtained on the same media. The highest result for root fresh and dry weight recorded with 'Caixin' with MS + 3.0mg/l TDZ + 1.0mg/l NAA + 9.0mg/l AgNO3 when we used embryo as explant. The highest ascorbic acid content was found with callus cultured on MS + 1.0mg/l TDZ + 0.25mg/l NAA + 5.0mg/l AgNO3, when used leaf explant in 'Caixin' or root in 'Suzhouqing', andthere were no significant difference between them. While the highest value of total AsA content was registered with callus cultured on MS + 2.0mg/l TDZ + 0.25mg/l NAA + 5.0mg/l AgNO3 extracted from cotyledon in 'Caixin'. The highest content of DHA was registered with MS + 2.0mg/l TDZ + 0.25mg/l NAA + 5.0mg/l AgNO3 with cotyledon in 'Caixin'. Also, in 'Caixin' MS + 3.0mg/l TDZ + 0.25mg/l NAA + 5.0mg/l AgNO3 recorded the highest value of total protein content with embryo explant.

Pigment elicitation and sun protection factor of callus induced from Cassia tora seedling explants

Natural pigments are derived from sources such as plant, animal, insects, and fungi. The anthraquinones are natural pigments that have pharmacological activities. They have applications in several domains such as the pharmaceutical, cosmetic and textile industries. Anthraquinones occur naturally in many plants including Cassia tora of the Fabaceae family. The present study focused on the production of pigments (anthraquinones) from callus of Cassia tora cultured on medium supplemented with 6-benzylaminopurine and thidiazuron. The elicitor, salicylic acid, increased anthraquinone production in callus cultures whereas ethrel had minimum effect. Anthraquinone production increased twofold in callus inoculated in medium containing 10 mM salicylate. The yield obtained was 3709 ± 269.65 mg/kg dry weight of callus in Fraction B. The sun protection factor of the callus extracts was also assessed. The maximum sun protection factor observed was 38. Growth analysis of Cassia tora callus, modulating callus growth and pigment (anthraquinones) production with the elicitors Ethrel and salicylic acid. Potential application of callus extract as Sun protectant.

In Vitro Assessment of Kurdish Rice Genotypes in Response to PEG-Induced Drought Stress

Rice (Oryza sativa L.) is productively affected by different environmental factors, including biotic and abiotic stress. The objectives of this research were to evaluate the genetic distinction among Kurdish rice genotypes using the simple sequence repeats (SSRs) molecular markers and to perform in vitro tests to characterize the drought tolerance of six local rice genotypes. The polymorphic information content (PIC) varied from 0.38 to 0.84 with an average of 0.56. The genetic distance ranged from 0.33 to 0.88. Drought stress had a significant impact (p ≤ 0.05) on callus growth parameters. Enzymatic antioxidant systems were predicted and exhibited a significant variation. The findings revealed that proline levels increase in proportion to polyethylene glycol (PEG) concentrations. Kalar and Gwll Swr genotypes showed the worst performances in phenotypic and biochemical traits, while Choman and Shawre exhibited the best phenotypic and biochemical performances. A positive and substantial relationship between callus fresh weight (CFW) and callus dry weight (CDW) was found under stressful and optimized conditions. Callus induction (CI) was positively and significantly associated with the catalase activity (CAT) in all stressed treatments. Based on the results for callus growth and the biochemical parameters under stress conditions, a remarkable genotype distinction, based on the tolerance reaction, was noted as follows: PEG resistant &gt; susceptible, Choman &gt; Shawre &gt; White Bazyan &gt; Red Bazyan &gt; Gwll Swr &gt; Kalar. The CI and CAT characteristics were considered as reliable predictors of drought tolerance in rice genotypes.

Callus growth kinetics and accumulation of secondary metabolites of Bletilla striataRchb.f. using a callus suspension culture.

Bletilla striata is an endangered traditional Chinese medicinal plant with multiple uses and a slow regeneration rate of its germplasm resources. To evaluate the callus growth kinetics and accumulation of secondary metabolites (SMs), a callus suspension culture was proven to be a valuable approach for acquiring high yields of medicinal compounds. An effective callus suspension culture for obtaining B. striata callus growth and its SMs was achieved with the in vitro induction of calluses from B. striata seeds. The callus growth kinetics and accumulation of SMs were analyzed using a mathematical model. The resulting callus growth kinetic model revealed that the growth curves of B. striata suspension-cultured calluses were sigmoidal, indicating changes in the growth of the suspension-cultured calluses. Improved Murashige and Skoog callus growth medium was the most favorable medium for B. striata callus formation, with the highest callus growth occurring during the stationary phase of the cultivation period. Callus growth acceleration started after 7 days and thereafter gradually decreased until day 24 of the cultivation period and reached its highest at day 36 period in both the dry weight and fresh weight analyses. The coelonin concentration peaked during the exponential growth stage and decreased afterward during the stationary stage of the callus suspension culture. The maximum content of coelonin (approximately 0.3323 mg/g callus dry weight) was observed on the 18th day of the cultivation cycle, while dactylorhin A and militarine reached the highest concentrations at day 24, and p-hydroxybenzyl alcohol at day 39. This investigation also laid a foundation for a multimathematical model to better describe the accumulation variation of SMs. The production of SMs showed great specificity during callus growth and development. This research provided a well-organized way to increase the accumulation and production of SMs during the scaled-up biosynthesis of calluses in B. striata callus suspension cultures.