Highest Callus Fresh Weight Research Articles

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.

Callogenesis of Dayak Onion (Eleutherine palmifolia) Bulb in response of Picloram, 2,4-D, and Kinetin

Dayak onion contains bioactive compounds that are traditionally used for medicine. The production of bioactive compounds can be increased through callus culture. This study aims to analyze the effect of the combination of plant growth regulator on the growth of Dayak bulb callus. The design of this study used a completely randomized factorial design with 2 factors. The first factor is the concentration of auxin. While the second factor is the concentration of kinetin. The parameters observed include the percentage of callus, the time of callus formation, fresh weight, dry weight, and callus morphology. In this study, a callus growth curve was also made to determine the best harvest time. The results showed that highest callus fresh weight was obtained in the P3K1 treatment. The percentage of callus formation in the picloram + kinetin treatment was higher than in the 2,4-D + kinetin treatment. The time of callus formation occurred in the combination of picloram + kinetin faster than the 2.4-D + kinetin treatment. The callus color is diverse, while the texture of the entire callus is compact. The best callus induction medium for Dayak onion bulbs is to use picloram 2-4 ppm + kinetin0.025-0.5 ppm. The results of this study provide optimal growth regulatory information for the induction of dayak onion callus that has never been reported before. This information can be the basis for the development of methods of production of bioactive compounds from dayak onions through callus culture.

Improving the Production of Secondary Metabolites via the Application of Biogenic Zinc Oxide Nanoparticles in the Calli of Delonix elata: A Potential Medicinal Plant.

The implementation of nanotechnology in the field of plant tissue culture has demonstrated an interesting impact on in vitro plant growth and development. Furthermore, the plant tissue culture accompanying nanoparticles has been showed to be a reliable alternative for the biosynthesis of secondary metabolites. Herein, the effectiveness of zinc oxide nanoparticles (ZnONPs) on the growth of Delonix elata calli, as well as their phytochemical profiles, were investigated. Delonix elata seeds were collected and germinated, and then the plant species was determined based on the PCR product sequence of ITS1 and ITS4 primers. Afterward, the calli derived from Delonix elata seedlings were subjected to 0, 10, 20, 30, 40, and 50 mg/L of ZnONPs. The ZnONPs were biologically synthesized using the Ricinus communis aqueous leaf extract, which acts as a capping and reducing agent, and zinc nitrate solution. The nanostructures of the biogenic ZnONPs were confirmed using different techniques like UV-visible spectroscopy (UV), zeta potential measurement, Fourier transform infrared spectra (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). Adding 30 mg/L of ZnONPs to the MS media (containing 2.5 µM 2,4-D and 1 µM BAP) resulted in the highest callus fresh weight (5.65 g) compared to the control and other ZnONP treatments. Similarly, more phenolic accumulation (358.85 µg/g DW) and flavonoid (112.88 µg/g DW) contents were achieved at 30 mg/L. Furthermore, the high-performance liquid chromatography (HPLC) analysis showed significant increments in gallic acid, quercetin, hesperidin, and rutin in all treated ZnONP calli compared to the control. On the other hand, the gas chromatography and mass spectroscopy (GC-MS) analysis of the calli extracts revealed that nine phytochemical compounds were common among all extracts. Moreover, the most predominant compound found in calli treated with 20, 30, 40, and 50 mg/L of ZnONPs was bis(2-ethylhexyl) phthalate, with percentage areas of 27.33, 38.68, 22.66, and 17.98%, respectively. The predominant compounds in the control and in calli treated with 10 mg/L of ZnONPs were octadecanoic acid, 2-propenyl ester and heptanoic acid. In conclusion, in this study, green ZnONPs exerted beneficial effects on Delonix elata calli and improved their production of bioactive compounds, especially at a dose of 30 mg/L.

In Vitro Effects of Different Combinations of Phytohormones on Callus Induction from Different Explants of Cabbage Brassica Oleracea Var. Capitata L. Seedlings

The leading purpose of this work is the development of efficient culture conditions to induce calli from cabbage (Brassica oleracea var. capitata L.) under in vitro conditions. The mature seeds were surface sterilized with combinations of different concentrations of ethanol and NaOCl in different time durations and were germinated on MS basal medium. The results revealed that the best sterilization method of cabbage seeds was by using 70% ethanol for one minute, followed by 15 min in 2% (NaOCl). Seedlings were used as donor sources for hypocotyls, cotyledon leaves, true leaves, and shoot tip explants. These explants were cultured on different combinations of cytokinins (TDZ, BAP, Ad) and auxins (IAA, NAA, 2, 4-D) then implanted in Murashige and Skoog (MS) media. 4 weeks after culturing, a significant difference was found among the explants in response to plant hormones. The maximum percentage of callus induction (100%) was using the combinations of 1 BAP + 1 2, 4-D, 1 BAP + 1 NAA, and 1 BAP + 2 2,4-D mg. l-1. In addition, explants responses varied and the hypocotyls showed a superior result (85.71 %) as compared to other explants. For callus fresh weight, the combination of 0.22 TDZ + 79.9 Ad mg. l-1 had a significant effect, causing the highest fresh weight (0.2745g), while control treatment gave the lowest mean of 0.0066 g. Data showed that cotyledon explants were significantly superior in giving highest callus fresh weight with the mean of 0.1723 g. On the other hand, hypocotyl explants gave the lowest mean, reaching 0.1542 g.

Spectral Lights Based Treatment Enhanced Biomass Accumulation and Secondary Metabolites Production in Callus Culture of Citrus reticulata

Citrus reticulata is a common fruit, belongs to the family Rutaceae. It contains terpenes, phenolics, flavonoids, and vitamin C in conjugation with sugars, amino acids, minerals, organic acids and other biologically important compounds. Application of light elicitation strategies in in vitro plant cultures is acquiring more interest for their potential in feasible biomass accumulation and therapeutically important secondary metabolites production. The present study was designed to investigate the effects of different monochromatic lights (white, red, blue, green, and yellow) on the proliferation and secondary metabolites accumulation in callus cultures of Citrus reticulata raised in response to treatment with AgNPs and TDZ. Silver NPs were synthesized by using green method, followed by characterization of these NPs. Nucellus tissues obtained from seeds of C. reticulata and inoculated on MS medium supplemented with TDZ (1 mg/L) + AgNPs (20 ppm) for callus development. To examine the effects of different light spectrum, the developing calli was sub-cultured on similar media and incubated in growth chamber under five different light treatments (white, red, blue, green, and yellow). Total protein content, chlorophyll content, phenolic and flavonoid content, along with activity of antioxidant enzymes on all samples was also analyzed. Highest callus fresh weight (730.21 mg) was recorded in callus cultures proliferated under white light condition. Phytochemical analysis of callus cultures showed significantly higher values for TPC, TFC and antioxidant activity under red light. However, maximum production of anti-oxidative enzymes (SOD and POD) was recorded under blue light. White light is considered more favorable for accumulation of total protein content. To our knowledge, this is the first study on production of biomass and phytochemicals in response to different monochromatic lights in callus cultures of Citrus reticulata. The present research outcomes clearly exhibited the interest to apply monochromatic light as elicitor of in vitro callus cultures to promote the production of biomass and important bioactive metabolites.

Plant tissue culture of Nicotiana tabacum cv. TAPM 26 and its minimum inhibition against herbicide-Dalapon

Current study is to establish a basic plant tissue culture of Nicotiana tabacum TAPM 26 and test the plant tissue on resistancies against 2,2 DCP an active ingredient in herbicide-Dalapon. During micropropagation, the surface sterilization method was ascertained on seeds of tobacco. HgCl2 was used to disinfect tobacco seeds at different concentrations (0.05 gL-1, 0.2 gL-1, 0.5 gL-1 and 1.0 gL-1) within three minutes. About 70% seeds were survived when exposed to 0.05 gL-1 of HgCl2, whereas, no seeds were germinated when sterilized at concentrations above 0.05 gL-1 of HgCl2. To optimize an efficient protocol of shoots and callus formation during in vitro regeneration, explant types and plant growth were studied. Growth regulators NAA (0.1 mgL-1, 0.2 mgL-1, 0.5 mgL-1, 1.0 mgL-1 and 2.0 mgL-1) and BAP (1.0 mgL-1, 2.0 mgL-1, 3.0 mgL-1 and 4.0 mgL-1) were used. The explants types were one month old leaves and two weeks old cotyledons. The maximum numbers of shoots per explants were obtained from cotyledon with combination 0.1 mgL-1 NAA and 1.0 mgL-1 BAP. The highest callus fresh weight was achieved when NAA 0.5 mgL-1 with BAP 1.0 mgL-1 after four weeks. Thus, the highest number of shoots produced per explants from leaves culture on the MS media containing 0.2 mgL-1 NAA and 4.0 mgL-1 BAP. The best callus fresh weight was obtained with combination of 1.0 mgL-1 NAA and 1.0 mgL-1 BAP by using leaves explant. Finally, Dalapon (5 gL-1, 10 gL-1, 15 gL-1 and 20 gL-1) were applied onto leaves and cotyledon cultures of N. tabacum to check on the minimum concentration of inhibition. The minimum concentration of inhibition of leaves and cotyledon cultures of N. tabacum was at 5 gL-1 of 2,2DCP but not at 10 gL-1, 15 gL-1 and 20 gL-1. This investigation will shed alight for future studies on transgenic tobacco resistant against Dalapon

Establishment of a Protocol for in vitro Seed Germination and Callus Formation of Ficus religiosa L., an Important Medicinal Plant

Background: Ficus religiosa is known as a pure source in traditional medicine for the treatment of diabetes, asthma, diarrhea, gastric problems, epilepsy, sexual, infectious, and inflammatory disorders. Despite the fact that many studies have authorized its traditional medicinal uses, yet these utilized raw extracts have not been yet characterized. Therefore, there is a necessity for standardizing its phytochemical features and recognizing bioactivity, guided by bioactive metabolites. Objectives: In this study, the effect of light and dark condition, different strengths of Murashige and Skoog (MS) medium, and their interactions on seed germination and the effect of plant growth regulators on callogenesis of F. religiosa via leaf, petiole, root, and internode explants were investigated. Results: The results demonstrated the highest seed germination percentage was achieved at one-tenth strength of MS medium under the light condition. Also, the highest callus fresh weight was obtained from media supplemented with 0.5 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.05 mg/L 6- benzyl amino purine (BAP) in all explants. The MS medium containing indole-3-butyric acid (IBA) produced greenish and compact calli while yellowish and compact calli was achieved in α-naphthalene acetic acid (NAA) and finally 2,4-D produced yellow-brownish calli. Conclusions: To the best of the author’s knowledge, this study was the first report of seed germination and callus induction through immature explants of F. religiosa. The seed germination and callogenesis system established in this study could be applied in the future for the enrichment of certain secondary metabolites as well as the production of new secondary metabolites, with the purpose of understanding and use of medicinal properties for this valuable germplasm.

In Vitro Adventitious Shoot Regeneration through Direct and Indirect Organogenesis from Seedling-derived Hypocotyl Segments of Ficus religiosa L.: An Important Medicinal Plant

Ficus religiosa is an important industrial, medicinal, and ornamental plant, so in vitro regeneration is of high paramount in this valuable germplasm. Two efficient protocols were developed for indirect and direct shoot organogenesis through hypocotyl explants. In the first experiment, different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and indole butyric acid (IBA) (0.5, 1.0, and 1.5 mg·L−1) in combination with 6-benzyl amino purine (BAP) (ratio 10:1, respectively) were used for callus formation. Two types of callus were obtained from different concentrations of plant growth regulators (PGRs). Also, 2,4-D produced yellow-brownish and friable callus (Type I), whereas the green and compact callus (Type II) was achieved in IBA. The highest callus fresh weight (2.43 g) was observed in Murashige and Skoog (MS) medium containing 0.5 mg·L−1 2,4-D plus 0.05 mg·L−1 BAP. In the later experiments, various concentrations of thidiazuron (TDZ), 6-furfuryl amino purine (KN), and BAP (0.25, 0.5, 1.0, and 1.5 mg·L−1) in combination with IBA (ratio 10:1, respectively) were applied for shoot regeneration (direct and indirect organogenesis). In shoot regeneration from callus, the highest regeneration frequency (86.66%) and shoot number per callus (4.13) were achieved in MS medium supplemented with 1.5 mg·L−1 BAP plus 0.15 mg·L−1 IBA from type I calli. However, no regeneration was observed in type II calli. In direct shoot regeneration, the highest regeneration frequency (96.66%) and shoot number (6.26) were obtained in the medium mentioned previously. In root induction experiment, different concentrations of naphthalene acetic acid (NAA) and IBA alone or in combination were applied, and MS medium containing 2.0 mg·L−1 IBA along with 0.1 mg·L−1 NAA was the best hormonal balance for root induction. The rooted plantlets’ survival rate was more than 95% in the acclimatization stage. These results demonstrated that the direct regeneration method provides more shoot regeneration frequency and take a less time for shoot organogenesis than the indirect regeneration method. Based on our knowledge, this study is the first report of direct and indirect shoot organogenesis of F. religiosa via hypocotyl from in vitro–grown seedling.

THE USE OF EMBRYO CULTURES FOR STUDYING SALT STRESS TOLERANCE OF FOUR DIFFERENT BREAD WHEAT VARIETIES AND THEIR PROMISING MUTANTS

The present study was carried out to determine the genetic response of four different bread wheat varieties and their mutants for callus induction using mature embryo culture and salt stress tolerance and to assess the genetic diversity between them using ISSR markers. The results of the ability of callus induction revealed that differences among genotypes, callus induction media (CI) and their interactions were highly significant for callus induction frequencies (CIF). Also in relation to the callus fresh weight (CFW), the differences between genotypes and their interactions with media were highly significant, whereas insignificant differences were found between media after 14 at 28 days of culture. The best medium for CIF was CIM2 containing 2 mg/l of 2,4-D for all genotypes except Gemiza 9-1 and Sakha 93-3 had the best CIF of CIM3 containing 3 mg/l of 2,4-D. Seds 12-7 had the highest CIF, whereas Sakha 93-3 had the lowest CIF of CIM2. Gemiza 9-1 had the highest CFW, whereas Sakha 93-3 had the lowest CFW after 28 days of culture. The fresh callus produced after month was transferred to salt stress media at 10000ppm NaCl level. Giza 168, Gemiza 9 and Sakha 93 showed increase in CFW, whereas other genotypes strongly decreased. Sakha 93 and Gemiza 9 gave the lowest value of callus necrosis (CN) and the highest value of callus in vitro tolerance (CINTOL) at the 10000ppm NaCl level after 28 days on salt stress media. The salt stressed callus was tested for shoot induction ability; callus of Gemiza 9 and Sakha 93 showed the best shoot induction frequencies at 10000 ppm level. Days to shoot induction were decreased with the increase of salt levels compared to normal conditions. Transfer of shooty callus on plant regeneration media to be grown and development failed. The callus turned brown and some of the resulting growths were weak and white. The use of ISSR technique illustrated the genetic similarities between the wheat genotypes, which ranged from 0.638 to 0.979 with an average of 0.809, suggesting a high genetic diversity between the wheat genotypes. Cluster analysis performed to generate dendrogram showed that wheat genotypes had been clustered into two main clusters. From the previous results, we recommended that Gemiza9 and Sakha93 are good materials for salt stress tolerance of bread wheat breeding programs.

In vitro propagation, genetic stability, and secondary metabolite analysis of wild lavender (Lavandula coronopifolia Poir.)

Lavenders (Lavandula species) are important aromatic ornamental medicinal plants with wide ranging applications in perfume and pharmaceutical industries. We developed an in vitro propagation protocol for Lavandula coronopifolia. Murashige and Skoog (MS) medium supplemented with 0.5 mg·L-1 N 6 -benzyladenine was the best medium for the proliferation of microshoots, while the highest rooting frequency was obtained using MS medium supplemented with 1.0 mg·L-1 indole-3-butyric acid. Inter-simple sequence repeat analysis revealed that the in vitro-propagated microshoots were highly genetically stable, even after subculture. The highest callus fresh weight (667.9 mg) was obtained by propagating on medium supplemented with a combination of 1.0 mg·L-1 naphthaleneacetic acid and 0.5 mg·L-1 butyric acid. Using the Folin-Ciocalteu method, methanolic extracts of wild L. coronopifolia revealed total phenolic content of 4.9 mg expressed in gallic acid equivalents (GAE) (mg GAE·g-1 dry matter). Radical scavenging activity was estimated at 85% using the free radical 2,2-diphyenyl-picrylhydrazyl assay. Using the brine shrimp assay for cytotoxicity, the methanolic extract was found to be nontoxic. Finally, liquid chromatography tandem mass spectrometry with standard reference compounds was used to quantify the key phenolic compounds in both in vitro and in vivo-grown L. coronopifolia. Six major phenolic compounds were identified: caffeic acid, rosmarinic acid, rutin, quercetin, luteolin, and hesperidin. Levels of these phenolic compounds were highest in wild plant extracts.

ROLE OF PLANT GROWTH REGULATORS IN CALLUS INITIATION AND PLANTLETS REPRODUCING OF TWO ALFALFA CULTIVARS

An experiment was conducted at the central lab of Graduate studies - College of Agriculture, University of Baghdad during 2015-2016. The objective was to study the effect of growth regulator on inducing callus from embryos of two alfalfa cultivars (PAC-78001 and local) variety by tissue culture technique using factorial experiments according to CRD. Seeds sterilized by NaOCl at 4.5% for 15 minutes, then, cultured on MS media free of growth regulators to embryo induced. The embryos were cultured on MS media at different concentrations of 2,4-D (0, 1, 2, 3 and 4) mg Li-1. Induced callus from embryos was cultured on MS media supplementing with Kin. at 0, 0.25, 0.50, 0.75 and 1 mg Li-1 to determine the best concentration. The results showed that best concentration of 2,4-D and Kin were 2 and 0.5 mg L-1 respectively which gave highest callus fresh weight and callus dry weight (204.65 and 21.50) mg Li-1, respectively. BA at 2 mg Li-1 gave the highest plantlets reproducing from callus amounted 5.5 vegetative growth tube-1.

CALLUS INDUCTION AND PLANT REGENERATION IN ECHINOCACTUS PLATYACANTHUS

The aim of this research was to establish the optimal conditions for micropropagation of E. platyacanthus, a cactus that is used as a source of water and fodder for livestock during the dry season. To this end, two experiments were carried out. In the first, seed germination on Murashige and Skoog (MS) medium with different concentrations of mineral salts (50, 75 and 100%) and sucrose (10, 20, 30 and 40 g L-1) was evaluated. From the first experiment, we selected three-month-old seedlings, approximately 1.0 cm in length, and from each one of them three explants were obtained; the explants, in turn, were placed on MS medium with different concentrations of benzyladenine (BA) (0.0, 0.5, 1.0 and 2.0 mg L-1) and naphthaleneacetic acid (NAA) (0.0, 0.2, 0.5 and 1.0 mg L-1). In both experiments the factors and levels were combined in a factorial arrangement and established in a completely randomized design. In all cases, the culture medium was supplemented with myo-inositol (100 mg L-1), thiamine (0.4 mg L-1) and agar (6 g L-1), and its pH was adjusted to 5.7. In the first experiment, we began recording the number of germinated seeds 15 days after sowing (DAS); in the second, 60 DAS the variables callus fresh weight, callus diameter, shoot number and length and root number and length were recorded. The data were subjected to analysis of variance and mean separation tests (Tukey, p≤0.05) using the SAS Version 9 statistical package (SAS, 2003). Seed germination began at 16 DAS and ended 37 days later (53 DAS). Salt proportions under or equal to 75% were associated with germination rates of 70%, statistically higher than those obtained with MS medium with 100% mineral salts. The highest callus fresh weight was obtained in BA concentrations at or above 1.0 mg L-1. NAA concentrations at or above 0.5 generated callus diameters two to three times greater than without the hormone (0.59 cm). The NAAA—BA interaction was significant in shoot height, with the highest value (0.68 mm) being obtained with 0.2 mg L-1 NAA and 0.5 mg L-1 BA. Root number and length were negatively affected by the BA concentration.

English

The present study investigates in vitro regeneration of three garlic cultivars; that is, Balady, Sids 40 and VFG 180 (3 - 1) as well as a garlic wild type through adventitious shoot organogenesis. Shoot and root apices were subjected to eight callus induction treatments. A combination of 1 mg L -1 2,4-D + 5 mg L -1 BA + 5 mg L -1 NAA produced a 100% of callus induction from root apices for all garlic cultivars tested. Shoot apices showed higher frequency of callus induction than root apices. Balady cultivar showed the highest frequency of callus induction while Sids 40 showed the lowest values. Shoot apices had higher callus fresh weight than root apices in Balady and Sids 40 cultivars as well as wild type. The cultivar Balady had the highest callus fresh weight whereas the wild type had the lowest values. There were large variations in the regeneration efficiency among the eight callus clones as well as different garlic cultivars. The wild type failed to regenerate shoots. The highest shoot number per root induced callus (48.8) was obtained from C7 callus line cultured into B5 medium supplemented with 10 mg L -1 Kin and 2 mg L -1 IAA. The cultivar Sids 40 showed the lowest regeneration efficiency among other cultivars with 1.5 shoots per explant. For callus induced from shoot apices, the eight callus lines for both Sids 40 cultivar and the wild type failed to regenerate shoots and the cultivar VFG 180 (3 -1) showed very low regeneration efficiency. The Balady cultivar showed the highest regeneration efficiency with 39 shoots per explant. The regenerated garlic shoots were in vitro rooted and acclimatized in greenhouse prior to their cultivation in open field. Garlic plantlets derived through tissue culture required three vegetative generations to produce bulbs of commercial size. Keywords: Acclimatization, garlic, organogenesis, tissue culture African Journal of Biotechnology , Vol 13(38) 3892-3900

Establishment of an efficient callus induction and plant regeneration system in some wheat (Triticum aestivum L.) cultivars grown in Sudan

A procedure for callus induction and plant regeneration was developed for wheat ( Triticum aestivum ) cultivars grown in Sudan. Mature embryos of cultivar Khaleefa removed from surface sterilized seeds were used as explants. For callus induction, explants were cultured on Murashige and Skoog medium (MS) supplemented with different levels (1.5 to 4.0 mg/l) of 2,4-dichlorophenoxy acetic acid (2,4-D), indole-3- butyric acid (IBA) and indole-3- acetic acid (IAA). After 4 weeks of culture, the highest callus fresh weight (0.05 ± 0.01 g) was obtained on MS medium supplemented with 2.0 mg/L of 2,4-D. For shoot regeneration, the developed calli were transferred to MS medium without growth regulators or with different levels (0.1 to 3.0 mg/l) of 5-phenylcarbamoylamino-1,2,3-thiadiazole (thidiazuron) (TDZ), benzyladenine (BA) and kinetin (KN) alone or in combination with 2.0 mg/L of 2,4-D. After 6 weeks of culture, the best result for regeneration percentage (40%) was obtained on MS medium without growth regulators. Callus derived shoots were rooted most effectively in half-strength MS medium without or with different levels of IBA. To determine the genotypic effect, the developed procedure was used to evaluate the regeneration ability of other 5 Sudanese wheat cultivars. Significant differences were observed between genotypes for plant regeneration ability, indicating that these criteria are genotype dependent. The acclimatized regenerated plants were morphologically uniform with normal leaf shape and growth pattern under greenhouse conditions. Key words : Triticum aestivum , callus induction, regeneration, 2,4-dichlorophenoxyacetic acid (2,4-D), genotypes.

English

Callus induction from cotyledon explants was studied in indigenous chick pea ( Cicer arietinum L.) cultivars KK-1 and Hassan-2K on MS and B 5 media containing different combinations and concentrations of growth regulators. Different MS and B 5 callusing media containing varying level of 2, 4-D (2 and 4 mg/l), NAA (0.50 and 1 mg/l), BAP (5 and 10 μM) and their combinations were tested for callus induction response. Percent callus and callus fresh weight (g) were recorded after two and four weeks of culture for both genotypes. For KK-1 cultivar, the maximum callus frequency (71 and 97%) followed by (65 and 96%) were observed on 4 mg/l 2,4-D+5 μM BAP in MS and 4 mg/l 2,4-D in B 5 media, respectively after two and four weeks of culture. Similarly, the highest callus fresh weight (0.411 and 0.787 g) were also recorded for MS+4 mg/l 2,4-D+5 μM BAP in contrast to B 5 where the highest callus weight (0.401 and 0.693 g) was achieved on 4 mg/l 2,4-D only. In Hassan -2K, the highest callus % (68 and 96) and fresh weight (0.572 and 0.821 g) were recorded on MS+4 mg/l 2,4-D+0.50 mg/l NAA after two and four weeks of culture, respectively. In B 5 medium, 2,4-D+BAP combination produced average callus induction response for both cultivars. Key words : Callus induction, chick pea cotyledons, growth regulators.