Percentage Of Callus Induction Research Articles

Genotypic effects on accelerated propagation of oil palm breeding materials selected (Elaeis guineensis jacq.) using somatic embryogenesis

Vegetable oil production from oil palm (Elaeis guineensis Jacq.) is an important industry due to the rising demand every year. The somatic embryogenesis culture can propagate oil palm duplicate as parent plant, which can be selected as breeding material to produce new planting germplasm with high production or disease resistance. This study aims to evaluate the genotypic effect of somatic embryogenesis, while immature leaflets were employed as explants. The culture used embryo induction medium based on Murashige and Skoog (MS) modifications that contained 5 ​mg/L Naphthalene Acetic acid (NAA) and 0.5 ​mg/L Benzyl Amino Purine (BAP). The genotypic effect was statistically significant in the percentage of callus induction, producing somatic embryos, and germination embryos. In this study, we successfully cloned thirteen oil palm genotypes (GE-02, GE-03, GE-06, GE-07, GE-09, GE-23, GE-24, GE-27, GE-28, GE-32, GE-33, GE-34, and GE-35), with the highest number of somatic embryos formed on GE-27 with a percentage of 70.1%. The cloning was successful in accelerating the propagation of oil palm for materials breeding programs to create new varieties with high production and disease resistance. It is necessary to observation the performance of these clones in the field in terms of mantle flower appearance.

Studies on In vitro Surface Sterilisation and Antioxidants on Pomegranate (Punica granatum L.) cv. Bhagwa

Pomegranate, regarded as the “Fruit of Paradise”, is one of the important fruit crops of tropical and subtropical regions. Pomegranate fruits are delicious and possess significant nutritional and medicinal benefits. Due to the increase in demand, the proliferation of pomegranate through tissue culture is essential to get high-quality planting material. However, microbial contamination in plant tissue culture is one of the bottlenecks for establishing aseptic cultures. Although multiple explant sterilisation methods have been developed for pomegranate, explant sterilisation methods can indeed vary from region to region due to the local environment and the mother plant. In the present study, 1-2 drops of tween 20 (20 min) + 500 mg/L carbendazim (30 min) + 500 mg/L streptomycin sulphate (10 min) + 100 mg/L citric acid (30 min) + 100 mg/L tebuconazole (folicur) (30 min) + 0.1% HgCl2 (1 min) + dipping explant after cutting the edges in sterile water (30 min) + dipping in 500 mg/L PVP (5 min) was found to be optimum in the prevention of microbial contamination with minimal cell death. Nodal segments-initiated callus at 54 days after culture, showing a callus induction percentage of 55.55% and a calli weight of 80 mg/explant.

High Efficiency In vitro Whole Plant Regeneration via Desiccated Callus in Oryza sativa cv. MTU1010

For most people across the planet, rice is also considered as most beneficial crops in terms of fulfilling every day's energy and nutritional needs. Worldwide food safety is seriously threatened since severe limitations on rice output caused by both abiotic and biotic variables. Even though rice features much more developed regeneration mechanism than other agricultural crops, majority of desi cultivars continue to stay as impervious to genetic alterations and regeneration. Therefore, among preliminary steps in the transgenic plant generation necessitates to improve tissue culture procedures to produce viable plants via embryogenic calli mediated regeneration. In this study, we reporting that enhanced rate of plant regeneration was observed from mature seed-derived embryogenic calli of the indica rice cultivar MTU1010 following partially desiccation procedure. We studied the effects of several plant growth regulators on the efficiency of inducing embryogenic callus and regeneration of whole plantlets. The MS-CIM medium with 2.0 mg-1 2,4, D showed the highest callus induction percentage. Maximum rate of shoot regeneration (92%) was observed from 48hr partially dehydrated calli, when enriched with high cytokinin (2 mg/l kinetin) and lower auxin (0.5 mg/l NAA) concentrations. Calli that had been moderately dehydrated showed significantly greater regeneration rates than undehydrated calli. Upon being transferred onto a rooting medium, the well-rooted plantlets were shifted to shaded glass house for hardening. Hence, this approach is rather simple to apply and can be employed in genetic modification studies alongside other biotechnological endeavours.

Effect of Media and Photoperiod on Embryogenic and Organogenic Callus Induction from Mature Seed in Oryza sativa L.

Rice is the staple food for billions of people and a source of income for hundreds of millions. This food security crop production needs to be improved to face the projected demands of the exponentially augmenting world’s population and overcome biotic and abiotic stress in the fields. Optimized in vitro culture protocols are a prerequisite to genetic engineering which appears to be the best potential way to improve the rice plant. In this study, ‘Nerica 3’ and ‘Nerica L36’ seeds were in vitro culture tested into 4 media (m1-4), in dark and photoperiod environments. The percentage of callus induction was calculated and callus weight was recorded. Results show that callus induction was influenced by variety × environment and medium × environment interactions, with a strong influence of the environment used. ‘Nerica 3’ showed the highest mean (88.25%) callus induction after six weeks of incubation on different media. m1 and m2 media showed greater mean callus induction (more than 86%). Higher callus induction came from m1 (82% and 91%) and m2 (81% and 93%) media, while lower rates came from m3 (73%) and m4 (69%) media for embryogenic and organogenic calli respectively. Culturing the explant in dark environment to produce embryogenic callus, resulted in greater callus weight for ‘Nerica 3’ (1.03 g) and ‘Nerica L36’ (0.79 g). This study is a contribution to the rice plant genetic improvement by proposing protocols for somatic embryogenesis of two Nerica rice varieties.

Seed germination and vegetative and in vitro propagation of Hieracium lucidum subsp. lucidum (Asteraceae), a critically endangered endemic taxon of the Sicilian flora.

Hieracium lucidum subsp. lucidum is a critically endangered endemic taxa of the Sicilian flora. It is a relict of the Tertiary period surviving on the cliffs of Monte Gallo (NW-Sicily). This research focused on finding the best protocols for seed germination and vegetative and in vitro propagation to contribute to ex situ conservation. Seed germination tests were carried out using constant temperatures of 15 °C, 20 °C and 25 °C in continuous darkness and an alternating temperature of 30/15 °C (16 h/8 h, light/dark). The seeds had no dormancy, and a high germination capacity (70-95%) was obtained at all tested thermoperiods. The possibility of vegetative propagation of the taxon was evaluated through the rooting capacity of stem cuttings treated or not treated with indole-3-butyric acid (IBA). All cuttings were treated with IBA rooted within 2 months, while only 50% of the untreated cuttings were rooted within a longer time. An efficient protocol for rapid in vitro propagation from leaf portions was developed. The response of explants was tested on hormone-free Murashige and Skoog (MS) basal medium and MS enriched with different types of cytokinins: 6-Benzylaminopurine (BAP) and meta-Topolin (mT) in combination with naphthaleneacetic acid (NAA) and 2,4-Dichlorophenoxyacetic acid (2,4-D) at the same concentration. The combination of mT (2 mg L-1) and 2,4-D (1 mg L-1) in the medium was the most effective and showed the highest percentage of callus induction and the mean number of regenerated shoots. The maximum rate of root regeneration and the maximum number and length of roots were obtained on hormone-free MS and MS enriched with IBA at concentrations of 1 mg L-1. From the results obtained, it can be concluded that H. lucidum subsp. lucidum can be successfully propagated using one of the tested techniques, subject to the availability of the material for reproduction.

EFFECT OF CHEMICAL MEDIA COMPONENTS AND SILVER NANOPARTICLES ON THE INDUCTION OF CALLUS CULTURE AND ACCUMULATION OF SECONDARY COMPOUNDS IN A Trigonella foenum-graecum L.

This study was investigate the effect of components media and silver nanoparticles (Ag-NPs) synthesized in the laboratory on the induction of biomass and secondary metabolites of calli cultures induced from the cotyledons of Trigonella foenum-graecum L. The culture media LS and B5 were tested with the growth regulators 2,4-Dichlorophenoxyacetic acid (2,4-D) at a concentration of 1.0, 2.0 and 3.0 mg l-1 and 6-benzyladenine (BA) at a concentration of 0.0, 0.5, 1.0, 1.5 and 2.0 mg l-1, with the aim of studying its effect on callus induction. In comparison, Ag-NPs were used at a concentration of 0.0, 0.5, 1.0, 1.5 and 2.0 mg l-1 to study its effect on the production of some secondary compounds. The results showed that the mixture consisting of the culture media LS with 3.0 mg l-1 of 2,4-D and 1.0 mg L-1 of BA the highest percentage of callus induction and the minimum days to complete callus formation. While, the same media with 2.0 mg l-1 2,4-D and 1.0 mg l-1 of BA recorded the highest dry weight (DW) and relative water content (RWC). The combination B5×2.0 mg l-1 2,4-D×1.5 or 2.0 mg l-1 showed the highest fresh weight (FW) and the lowest time required to start callus induction, respectively. As for the production of SC, the B5 media with 2.0 mg l-1 of Ag-NPs achieved the highest concentration of alkaloid compounds represented by trigonelline and diosgenin, and flavonoids expressed by qurcetine, vitxein, rutin, apigenin, isovitexin, and kaemferol.

Impact of Radiation on Rice Calli and Subsequent In vitro Regeneration

Aromatic rice has high commercial importance with greater economic return. Among the aromatic rice varieties, Kataribhog is a famous local variety that is mostly cultivated in the Dinajpur district northern part of Bangladesh. The yield performance of Kataribhog is lower (<2.50 t/ha) and has shown lodging tendency. Under these circumstances, a research program was taken to improve the existing problem through in vitro culture including nuclear irradiation techniques. Gamma irradiation (Co60) is a powerful tool in crop improvement, and its effects on callus development from kataribhog rice were explored in this research. Three types of media, PT-100G, PT-011G, and PT-100, were utilized to induce callus, which were then subjected to three different doses of radiation (10Gy, 12Gy, and 15Gy). Among the tested media, PT-100G was the most effective, boasting an impressive average of Callus initiation of 79.27% and embryogenic callus induction percentage of 32.93%. In contrast, PT-011G and PT-100 lagged with 52.46% and 44.44% callus initiation; 27.87% and 11.11% embryogenic callus induction, respectively. The radiation dose of 10 Gy was optimal for calli survival, showing an average percentage of 80.00%. 10 Gy is the most effective dose for promoting plant regeneration.

Efficient callus induction from various explants of Jatropha curcas

Jatropha curcas L. is a tropical plant of the Euphorbiaceae family, characterized by its high oil content and its ability to grow in a variety of conditions, which made it the most promising non-food biofuel crop in the world. The current study was directed to develop effective methods of tissue culture using this plant. The study was able to induce callus from cotyledons of sterile seedlings, as well as from true leaves taken from the field and from the embryo, using MS medium supplemented with interfering concentrations of Naphthalene acetic acid (NAA) with Benzyl adenine (BA), as the results indicated that The medium containing 0.5 mgL-1 for each of them gave the highest percentage of callus induction for the cotyledonous leaves, true leaves and embryonic peduncle, which reached 100%, 80% and 100%, respectively.

Effect of 2,4-D and NAA in Callus Induction and Differentiation from Different Explants of Moringa Oleifera Lam

This study was carry out in the Laboratory of Plant Cells and Tissue Culture / Department of Forestry Sciences in the College of Agriculture and Forestry / University of Mosul during the year 2021-2022 to study the effect of adding different concentrations of 2,4-Dichloro-Phenoxyacetic acid (2,4-D) or Naphthalene Acetic Acid (NAA) to the MS medium in the induction of callus from different parts of the Moringa oleifera Lam. and then study the effect of different concentrations of 6-Benzyladenine (BA) and Kinetin (Kin) on the differentiation of callus into buds and embryogenesis. Concerning callus induction, the results showed that 2,4-D or NAA in the nutrient medium was necessary after three weeks of cultivation 2,4-D was superior to NAA in the percentage of callus induction, as callus induction occurred by 100% of the explants cultured in the media supplemented with 0.1 mg.l−1 2,4-D. On the other hand, the parts (apical buds, single node, parts of the leaf blade, epicotyl, cotyledon, hypocotyl) differed significantly among themselves in the percentage of their ability on callus induction, as the cotyledons gave the highest rate of induction, amounting to 80%. The shoots resulting from the callus rooted after four weeks of cultivation in media supplemented with different concentrations of IBA at a rate of 100%, regardless of the concentration of IBA. The study proved the existence of a genetic match between the mother plants and the plants produced from the callus induction from them.

Role of phytohormone on in vitro regeneration of tea [Camellia sinensis (L.) O. Kuntze

Phytohormones are small molecules present in very low concentrations in plants and provide a promising strategy to improve mass culture biotechnology due to their intrinsic role in growth, development, and survival. This study is aimed at evaluating the effect of different plant growth regulators on in vitro callus induction and shoot regeneration in tea. The experiment was conducted at Complete Randomize Design (CRD) with three replications. Callus induction and shoot regeneration was done by using Murashige & Skoog (MS) media supplemented with various concentrations and combinations of Naphthalene Acetic Acid (NAA), Benzyl Adenine (BA), and 2,4-Dichlorophenoxyacetic Acid (2,4-D). Shoot tips and nodal segments were used as explant. The highest percentage (86.51%) of callus induction was observed in the combined treatment of 2.00 milligram per litre (mg/L) BA + 1.50 mg/L 2, 4-D in 2.12 weeks. The same treatment produced the highest weight of calli, which was 0.18, 0.56, and 1.11 grams (g) for 6, 10, and 14 weeks after inoculation (WAI), respectively. The highest percentage (84.39%) of shoot induction was recorded with 2.00 mg/L BA + 2.00 mg/L NAA in 32.43 days. The maximum number (1.42, 2.46, and 4.49) and the highest length of the shoot (1.41, 3.28, and 4.61 cm) at 60, 90, and 120 DAI (days after inoculation) were also recorded from the same treatment. No callus or shoot induction was recorded in the controlled treatment. The protocol developed from the present study might be useful for the large-scale production of healthy, disease-free planting material for tea.

Molecular profiling of some blast resistance genes and analysis of physiological variability in rice plants produced by anther culture and hybridization

For genetic improvement of rice plants against blast fungus disease caused by Magnaporthe oryzae, eight rice genotypes including four Egyptian and four foreign genotypes which represented a broad range of variation for blast resistance and other several traits were used in this study through line X tester mating design to produce F1 hybrids used as source of anther culture. Results of analysis of variance showed highly significant differences for all studied traits indicating overall genetic variations among parents and hybrids suggested that specific combining ability referred heavily to the expression of studied traits. Data of anther culture experiment for parents (8 parents) and hybrids (15 crosses) showed highly significant differences for all studied traits which illustrate genetic variations through the used materials. The highest callus induction percentage was found for CR7 (18.3%) and P6 (14.0%) while, the lowest callus induction percentage was found for CR5 (2.0%) and CR1 (4.0%). On the other hand, nine out of 15 hybrids recorded regeneration percentage more than the highest parental genotypes. Anther culture-derived plants and their parental genotypes were evaluated for blast disease resistance according to artificial infection with 10 blast races. The results showed that resistance percentage varied between 10% and 100%. While all anther culture-derived plants showed high level of resistance (100%) against all tested blast races. For enzyme activities for catalase, peroxidase and polyphenol oxidase in anther culture-drived plants and their parent, results showed that line TC9 derived from CR9 gave the highest value for catalase activity (301). Also, CR11 and P4 recorded the high level of peroxidase and polyphenol oxidase activities, respectively. Moreover, genetic diversity among anther culture-derived plants and their parental genotypes using five SSR primers related to Pi5(t), Pikh, Pi-b, Pi-1 and Pi-ta blast resistance genes showed heterozygous bands in all tested genotypes. The highest PIC value recorded for RM 247 and RM 206 markers which had PIC values of 0.778 and 0.761, respectively. Out of the used five SSR markers, RM 247 was the most informative marker with amplicon size ranged from 134 to 242 bp. These markers are therefore useful markers for studying polymorphism concerning blast resistance in rice and hence can be used in the marker-assisted selection for blast resistance. While development of resistant genotypes is the most effective way to control rice blast disease, anther culture technique used in this study helpful in production of resistant double haploid plants used in rice preeding programe in Egypt.

Factors Affecting Callus Induction from Anther and Ovary of Okra (Abelmoschus esculentus L.)

Background: Okra [Abelmoschus esculentus (L.) Moench.] is a nutrient-rich vegetable crop widely grown in the tropics and sub-tropics mainly for its edible pods. The haploid technique has been used in plant breeding for the improvements of plants and to develop new varieties in relatively a short time. Hence, we have optimized several factors such as plant growth regulators (PGR), sucrose concentration, cold treatment, type of media and culture conditions for callus induction from the anther and ovary of okra (557 F1 hybrid). Methods: The flower buds of different sizes were collected to determine various stages of development and then subjected to cold pre-treatments. The explants were then cultured on various combinations of PGRs i.e., naphthyloxy acetic (NOA), Indole acetic acid (IAA), 2, 4-Dichlorophenoxy acetic acid (2, 4-D), Benzyl amino purine (BAP), isopentenyl adenine 2iP, Kinetin (KIN) and Thidiazuron (TDZ). Result: The optimum developmental stage of microspore for callus initiation was achieved from flower buds of okra and its size was about 11 mm long. Flower buds that emerged one week after the flowering showed significantly higher percentage of callus induction. The optimum stage for ovary and ovule culture were one or two days prior to anthesis and the flower buds stage was 35±1mm. In conclusion, our study investigated the effect of several factors that affects callus induction in okra and optimized cultured conditions for future haploid development for okra.

Indirect organogenesis of Aceh patchouli leaf explants (Pogostemon cablin Benth) by in vitro

The Aceh patchouli plant (Pogostemon cablin. Benth) is a plantation plant that produces essential oils with great potential, known as patchouli oil. The use of in vitro culture techniques on patchouli plants needs to be developed to meet the needs of patchouli seeds. This study aimed to analyze the effect of plant growth regulators (PGRs) cytokinins 6-Benzyl Amino Purine (BAP) and Thidiazuron (TDZ) and auxin 1-Naphthaleneacetic Acid (NAA) on the organogenesis ability of Aceh patchouli leaf explants var. Lhokseumawe. The treatments analyzed were P1 = MS; P2 = BAP 0.75 mg.l−1, P3 = BAP 1.0 mg.l−1, P4 = TDZ 0.75 mg.l−1, P5 = TDZ 1.0 mg.l−1, P6 = NAA 0.5 mg.l−1, P7 = NAA 1.0 mg.l−1, P8 = BAP 0.75 mg.l−1 + NAA 0.5 mg.l−1, P9 = BAP 0.75 mg.l−1 + NAA 1.0 mg.l−1, P10 = BAP 1.0 mg.l−1 + NAA 0.5 mg.l−1, P11 = BAP 1.0 mg.l−1 + NAA 1.0 mg.l−1, P12 = TDZ 0.75 mg.l−1 + NAA 0.5 mg.l−1, P13 = TDZ 0.75 mg.l−1 + NAA 1.0 mg. l−1, P14 = TDZ 1.0 mg.l−1 + NAA 0.5 mg.l−1, P15 = TDZ 1.0 mg.l−1 + NAA 1.0 mg.l-1. The percentage of callus induction observed, percentage of shoot induction, percentage of browning, percentage of contamination and percentage of live explants at 28 DAI. The results showed that the percentage of explants forming callus was 56.4%, and the percentage of explants forming organ was 54.6%. In these two parameters the treatment of P14 and P15 gave the best results of 6.6% respectively. The percentage of explants experiencing browning was 26.1%, treatment P1 gave the highest response for this parameter, namely 6.6%. The parameter percentage of live explants was 71.3%. The best response was found in treatment P2, P3, P12 and P15, which were 6.6% each. The fastest bud formation occurred at 14 Days After Initiation (DAI) in P4 treatment. The resulting shoots look quite vigor and have formed leaves. In the combination treatment P14, showed the best results, the addition of auxin was able to increase the initiation of embryogenic callus.

Micropropagation of Endemic Endangered Taxa of the Italian Flora: Adenostyles alpina subsp. macrocephala (Asteraceae), as a Case Study.

The conservation of endangered, rare, and endemic plant species is based on in situ and ex situ conservation strategies. When in situ conservation alone is not sufficient to guarantee the survival of the species, ex situ techniques are adopted in support. This study aimed to develop an efficient micropropagation protocol for Adenostyles by evaluating the effect of different plant growth regulators on leaf explants. Adenostyles alpina subsp. macrocephala (Asterace) is a perennial herbaceous plant endemic to Calabria (Southern Italy). The genus Adenostyles includes three species confined to the mountains of the Mediterranean and southern Europe. For callus induction, media supplemented with different concentrations of Benzylaminopurine (BAP) (0.5, 1, 2, and 3 mg L-1), Naphthaleneacetic Acid (NAA) (1 mg L-1), and 2,4-Dichlorophenoxyacetic Acid (2,4-D) (1 mg L-1) were tested. Shoot regeneration and proliferation were obtained in media supplemented with BAP (1, 2, and 3 mg L-1) and NAA (1 mg L-1). Root induction was obtained in media supplemented with IBA (0.25, 0.50, and 1 mg L-1) and NAA (0.25, 0.50, and 1 mg L-1). Statistically significant differences in callus induction and shoot regeneration were observed between the various media tested. The medium containing Murashige and Skoog (MS) supplemented with 3 mg L-1 of BAP and 1 mg L-1 of NAA showed the highest percentage of callus induction and increased shoot regeneration. The regenerated shoots showed more effective root induction in the hormone-free MS medium and in the presence of Indole-3-Butyric Acid (IBA) at concentrations of 0.25, 0.50, and 1 mg L-1. These results can be used as a basis for the preparation of a micropropagation protocol for different taxa of Adenostyles, as well as other species of Asteraceae specialized to the Mediterranean mountain habitat.

In vitro propagation of Geranium wallichianum: An indigenously threatened medicinal plant in Pakistan

Geranium wallichianum is a well-known medicinal plant and has numerous ethnobotanical usages but its population in Pakistan is severely reducing due to overexploitation and over-harvesting. Moreover, the conventional propagation systems are not enough to sustain the population and meet the ever-increasing demand for this plant. Therefore, an efficient propagation system is necessary to conserve and increase the population of this plant. To control browning, citric acid, ascorbic acid and activated charcoal were added to Murashige and Skoog (MS) medium. The 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthaleneacetic acid (NAA) were used in different concentrations to induce the callus while shoot proliferation and root induction were achieved with media supplemented with 6-benzyl aminopurine (BAP), and indole-3-butyric acid (IBA), respectively. Callus was successfully induced in 5.1 days in a medium containing 15 ​μM NAA while the maximum percentage of callus induction (77.78%) was obtained from petiole segments in a medium containing 10 ​μM NAA. Benzyl aminopurine at a concentration of 15 ​μM produced 1.33 shoots per nodal segment compared with 1.0 in the control. Indole butyric acid at a concentration of 10 ​μM increased root length (2.67 ​cm). The results showed a decrease in the browning intensity and an increase in the percentage of survivals in both leaf and petiole cultures (38.88%) and (61.11%) at 0.3 ​g/L activated charcoal compared to control (0% and 5.5%), respectively. Activated charcoal also reduced the total phenolic content of the leaf segment (9.51 ​mg/g) and petiole segment (2.06 ​mg/g). The in vitro produced rooted plantlets were successfully acclimatized in small pots containing peatmoss.

Effects of 2,4-Dichlorophenoxyacetic Acid on The Induction of Callus from Cotyledon and Hypocotyl Explants of Butterfly Pea (Clitoria ternatea)

Clitoria ternatea (Butterfly pea) is a tropical medicinal and fodder legume from the Fabaceae family possessing various beneficial phytochemical compounds linked to the mammalian neuroprotective mechanism. Callus and cell suspension cultures are excellent alternatives for harnessing secondary metabolites from medicinal plants. The current study aims to induce callus from cotyledon and hypocotyl explants of C. ternatea for the establishment of cell suspension cultures. Cotyledon and hypocotyl explants from two-weeks-old seedlings were subjected to half-strength MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at different concentrations (0.5 mg/L to 2.5 mg/L) and callus scoring and morphology were assessed at week 8 of culture. Results revealed that the treatment of 0.5 mg/L 2,4-D resulted in the highest percentage of callus induction (100%) and the highest callus scoring for both cotyledon and hypocotyl explants with friable callus morphology. Cotyledon explants exhibited a higher callus scoring with a relative value of 3.03 ± 0.20 compared to hypocotyl explants at 1.80 ± 0.12. This study thereby provides a basis for future studies on callus induction studies and the establishment of cell suspension cultures of C. ternatea for the production of valuable secondary metabolites linked to the memory enhancing properties of the plant.

The effect of plant growth regulators on the antioxidant enzyme activity and secondary metabolite production in the cell suspension cultures of Melia azedarach L.

ABSTRACT In the present study, we investigated the effect of plant growth regulators and explant type on callus induction, cell suspension culture growth, antioxidant enzyme activity, and accumulation of rutin, quercetin, and kaempferol in M. azedarach L. cell cultures. Our studies showed that the inflorescence and petiole explants had a high percentage of callus induction compared to the leaf explants, whereas the highest callus growth was obtained from the leaf explants cultured on the MS medium containing 5 mg l−1 NAA and 5 mg l−1 BAP and 5 mg l−1 NAA and 3 mg l−1 kinetin Kin. Furthermore, the highest cell growth was obtained from the leaf and inflorescence calli transferred to liquid MS medium supplemented with 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D)+1 mg l−1 Kin/BAP. The highest activity of antioxidant enzymes and the lowest protein content was related to the leaf cell suspension cultures in liquid MS medium containing 3 mg l−1 NAA and 3 mg l−1 BAP. The inflorescence cell suspension cultures in the MS medium containing 3 mg l−1 NAA+1 mg l−1 BAP exhibited the highest rutin accumulation (47.536 mg g−1 FW). However, the highest amount of quercetin (8.570 mg g−1 FW) and kaempferol (5.420 mg g−1 FW) accumulation were obtained from the petiole cell suspension cultures in the MS medium with 1 mg l−1 2,4-D +1 mg l−1 Kin.

Stem cell induction and plant regeneration are affected by medium components in maca (Lepidium meyenii Walp)

In medicinal plants, selection, reproduction and preservation of important genotypes are very necessary. Nowadays, using tissue culture and regeneration techniques of medicinal plants under in vitro conditions has been able to proliferate medicinal plants widely, which is much higher than traditional methods of vegetative propagation. Maca (Lepidium meyenii), is an industrial plant whose root is the usable part. Maca has valuable medicinal effects such as sexual enhancement and reproductive power, infertility treatment, improved sperm count and quality, anti-stress, osteoporosis prevention and more. This study was conducted to induce callus and regeneration of Maca. First, MS medium supplemented with different concentrations of Kinetin, Naphthaleneacetic acid and 2,4-Dichlorophenoxyacetic acid [0.5, 1 and 2 µM respectively] and control were compared for callus induction from root and leaves. After 38 days of incubation, the first callus appeared, after 50 days of callus induction and after 79 days regeneration occurred. The callus induction experiment was performed for the study of the effect of three explants (leaf, stem and root) and seven hormone levels. The regeneration experiment was carried out by studying the effect of three explants (leaf, stem and root) on eight levels of the hormone. The results of data analysis on callus induction showed that the effects of explants, hormones and their interactions on callus induction percentage were highly significant but not significant on callus growth rate. The results of regression analysis showed that explants, hormones and their interactions had no significant effect on regeneration percentage. Based on our results, the best medium for inducing callus was Hormone 2,4-D [2 µM] and Kinetin [0.5 µM], in which the highest percentage of callus induction was in leaf explants (62%). And the lowest were in stem (30%) and root (27%) explants. According to the comparison of the mean, the best environment for regeneration of the environment was 4 µM 6-Benzylaminopurine 2.5 + Thidiazuron, in which the highest percentage of regeneration was in leaf explant (87%) and stem (69%) and the lowest in root explant (12). %).

In Vitro Propagation of Three Date Palm (Phoenix dactylifera L.) Varieties Using Immature Female Inflorescences.

Immature female inflorescences are promising materials for use as explants for the tissue culture of date palm. Four types of MS media were used in this study during the four micropropagation stages-starting media (SM), maturation media (MM), multiplication media (PM) and rooting media (RM)-to micropropagate three elite date palm varieties, Amri, Magdoul and Barhy using the immature female inflorescences as explant. The highest percentage of callus induction in all the varieties studied was obtained on the SM1 (9 µM 2,4-D + 5.7 µM IAA + 10 µM NAA). Culturing on the MM1 (4.5 µM 2,4-D + 9.8 µM 2-iP + 1.5 AC) allowed us to obtain the best value in terms of callus weight. After culturing on the PM1 (4.4 µM BA + 9.8 µM 2-iP) produced the highest numbers of somatic embryos and shoots. The explants on RM2 (0.5 µM NAA + 1.25 µM IBA + 3 g AC) showed the highest root numbers and root lengths, while the highest shoot length was achieved on RM3 (0.5 µM NAA + 0.5 µM IBA + 3 g AC). The Amri variety presented the best response among the three varieties in all parameters, followed by the Magdoul and Barhy varieties. In all the stages of micropropagation, the analysis of variance revealed highly significant variations among varieties and culture media, and a significant difference in the number of roots during the rooting stage. The results also showed non-significant differences in the interaction between varieties and culture media, except for shoot length in the rooting stage. The results also reveal the broad sense heritability ranging from low to high for the measured parameters. It can be concluded that the immature female inflorescences can be used as a productive explant source for successful date palm micropropagation using the SM1, MM1, PM1 and RM2 culture media. It can also be concluded that the success of date palm micropropagation not only depends on the concentrations of growth regulators, but also on their types.

Effect of different concentrations and combinations of some plant growth regulators on Punica granatum anther culture

Plant growth regulators (PGR), also known as plant hormones, are a variety of chemical compounds that have a significant impact on the development and differentiation of plant cells. This work aimed to study the effect of various combinations of Plant Growth Regulators (PGRs) for example [6-Benzyladenine (BA), 2,4-dichlorophenoxyaceticg acid (2,4-D), Naphthalene acetic acid (NAA) and indol-3-ylbutyric acid (IBA)] on callus induction and shoot regeneration from anther culture of S1 progenies resulted from Manfalouty cultivar open pollinated. The results showed that highest value of percentage of callus induction was 10.6% on MS medium supplemented with 1 mg / l of NAA and 2 mg / l of BA also highest value of percentage of shoot formation 60 % on MS medium supplemented with 3 mg/ l of NAA and 2 mg /l of BA.