Highest Percentage Of Callus Induction Research Articles

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.

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.

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.

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.

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.

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.

EVALUATION OF SOMACLONES OF POTATO (SOLANUM TUBEROSUM L.)

Evaluation of somaclones in the field of three potato varieties (Diamant, Multa and Cardinal) were examined through callus culture using node, internode and leaf segments. Multiple shoot regeneration from callus at various frequencies was observed using different concentrations and combinations of growth regulators. The highest percentage of callus induction from internode was observed in MS medium containing 3.0 mg l-1 of 2,4-D + 150 mg l-1 of L-asparagine. In this combination, the percentage of callusing was 90% for Diamant, 80% for Multa and 70% for Cardinal. The best response for multiple shoot formation was observed in MS medium supplemented with 4.0 mg l-1 of KIN + 0.5 mg l-1 of NAA. Diamant responded better than Multa and Cardinal for shoot regeneration from callus. Best rooting was observed from node or internode derived plantlets of primary calliclone in MS medium supplemented with IBA. Rooted calliclones were transplanted in the field and evaluated somaclones variation using parameters viz. plant height, number of leaves/plant, number of tuber/plant and tuber weight/plant. The result of statistical analysis on individual character supports existence of significant variation among the different somaclones.

In vitro Regeneration of Strawberry Plant from Leaf Explants via Callus Induction

The present investigation was carried out to develop an efficient in vitro regeneration protocol for strawberry plant using leaf explants. The highest percentage of callus induction was obtained on MS medium supplemented with 2.0 mg/l BAP and 0.5mg/l IBA. The highest numbers of shoots were regenerated from the callus on MS medium supplemented with 2.0 mg/l BAP and 1.5mg/l IBA. In case of root induction, MS medium supplemented with 1 mg/l IBA was found to be most effective. Following the development of roots, the in vitro regenerated plantlets were successfully transplanted into soil. Plant Tissue Cult. & Biotech. 32(1): 67-75, 2022 (June)

An efficient protocol optimization for callus induction, regeneration and acclimatization in sugarcane cv. CO 94012

Sugarcane's vast and complex genome, photosensitivity, low seed setting, prolonged crop duration, and complex environmental effect make it difficult to create diverse genetic variability by conventional breeding. However, the in vitro culture system can play a vital role in boosting yield, quality, and making the crop climate smart through in vitro approaches. In the present study in vitro culture using immature leaf sheath of Sugarcane (Saccharum officinarum L. cv- CO 94012) as an ex plant was used to standardize the protocol for callus induction and whole plant regeneration. MS medium supplemented with 2.0 mg/l, 2-4 D, recorded the highest percentage of callus induction. The highest number of multiple shoots was produced on MS medium fortified with KIN 1.0 mg/l. Rooting was more profuse when in vitro regenerated shoots were inoculated on MS basal medium supplemented with 3.0 mg/l NAA. Rooted shoots were hardened in the greenhouse before being transplanted to the field, where 90% survival rate.

Contenido total de fenoles en frutos y cultivos in vitro de Bromelia karatas L.

Bromelia karatas L. is a species native from America, used in traditional medicine as an alternative in the treatment of diabetes mellitus. Overexploitation and change in land use have led to a drastic decline in species. In the present work, in vitro cultures were established from B. karatas seeds and their total phenolic content was determined. To promote the germination of the seeds several scarification treatments were tested. Furthermore, the seeds were cultured on Murashige & Skoog (MS) media containing different concentration of N-6 benzyladenine (BA) with 2,4-dichlorophenoxyacetic acid (2,4-D). Obtaining a higher percentage of germination (100%) when exposing the seeds to H2SO4 (98%, 5 min) was obtaining. Additionally, in in vitro cultures, the germination time was considerably reduced (36 times), compared to the data reported in the field. Highest percentage of callus induction of leaf sections (1.5 cm2) of adult specimens and plants germinated in vitro in MS media containing BA (1.0 mg • L-1) + 2,4-D (0.0, 0.5, 1.0, and 2.0 mg • L-1) was established. Mature fruits of B. karatas resulted in the greatest phenolic content (1,110 mg GAE • 100 g-1, biomass, dry weight (DW)) measured by the Folin–Ciocalteu reagent method, while the smallest content was recorded for callus and the leaves of in vitro seedling germinated recorded 205 and 741 mg GAE • 100 g-1 DW, respectively. These results provide novel data, which can be part of programs for sustainable use and conservation of this species of ethnobotanical importance.

Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

Melia azedarach L. is a valuable source of antioxidants and secondary metabolites. This study is a first extensive report about the effect of different serialization protocols and plant growth regulators (PGRs) on explant disinfection efficiency, callus induction and secondary metabolites production and accumulation in callus cultures of M. azedarach L. In this regard, the effect of plant growth regulators on callus induction and secondary metabolites production were examined. In addition, different sterilization agents were evaluated for disinfection of chinaberry leaf explants. The results showed that the lowest percentage of explant contamination and browning with the highest percentage of callus induction and callus growth obtained with explants pretreated with benomyl (2 g/L) for 2 h and sterilized with 7% H2O2 for 10 min and NaOCl 2% (without pH adjustment) for 12 min. Although adjusting the pH of NaOCl to pH = 7 and 10 significantly reduced the microbial contamination and increased the percentage of contamination-free cultures of M. azedarach L., adversely influenced the explant viability and callus induction and growth. The highest percentage of callus induction obtained on the MS medium containing 3 mg/L NAA/2,4-D and 1 or 3 mg/L Kin/BAP, and the highest callus yield (1804.833 mg/explant) belonged to the MS medium supplemented with 5 mg/L 2,4-D and 5 mg/L Kin. The callus cultures grown on the MS medium supplemented with 3 mg/L NAA and 1 mg/L Kin produced the highest amount of Quercetin (2.06 mg/g fresh weight), Rutin (5.56 mg/g fresh weight) and Kaempferol (1.84 mg/g fresh weight).

The Role of Growth Regulators in The Multiplication of Stevia Rebaudiana Bertoni Shoot and Callus Induction in Vitro

The research was conducted to investigate the effect of plant growth regulators on the vegetative multiplication of the Stevia plant shoot and the induction of callus from it. The results indicated that the interaction between 2.0 mg. L-1 BA with 0.5 mg. L -1 Kin gave the highest rate number of shoot with 6.32 branches and the highest average of leaves number Were 9.60 leaves compared to the lowest average for the number of shoot and leaves Were 1.40 shoot and 3.80 leaves respectively. As for the length of the shoot, the interaction between (1.0 mg. L-1 BA and 0.5 mg. L-1 Kin) gave the highest average shoot length 4.31 cm compared to the control which gave 2.20 cm. in connection with the callus induction, the concentration 3 mg. L-1 of NAA gave the highest percentage of callus induction from leaves 100%, and the lowest mean of days that for callus initiation was 9 days compared to the control which reached to 20 days. As for the wet and dry weights of callus tissue, the interaction between 2.0 mg. L-1 NAA and 0.5 mg. L -1BA gave the highest wet weight rate 3.68 g and the average dry weight was 0.31 g compared to the control which gave the lowest rate 0.95 g, 0.08 g respectively.

Effect of 2,4-Dichlorophenoxy Acetic Acid and Activated Charcoal on Callus Induction of Cocos Nucifera L. Hybrid MATAG Inflorescence

Cocos Nucifera Linn. Var. MATAG is a Dwarf coconut variety that had high demand in Malaysia but low supply. Vegetative propagation of high-yielding MATAG coconut by using in vitro cloning must be considered in contributing to increase coconut productivity. Thus, attempts were made to develop a protocol that would enhance callogenesis as a first preliminary step towards a protocol for mass propagation of C. nucifera L. var. MATAG. The anther isolated from immature inflorescence was used as explants and cultured on modified Eeuwens Y3 media in different concentrations of 2,4-dicholorophenoxy acetic acid (2,4-D) and activated charcoal. The highest callus induction percentage (31.25 ± 12.18) was observed in 20 mg/L 2,4-D. However, 2,4-D at any level tested were not statistically significant. Callus induction media supplemented with 0.5 mg/L activated charcoal gave the highest callusing percentage (25.89 ± 13.59 %) indicating a positive effect of activated charcoal on callusing even though the result obtained not significant compared to control (15.95 ± 6.76 %). But, activated charcoal supplemented in media produced a significant effect compared to control in reducing the percentage of browning. In conclusion, media supplemented with activated charcoal produced a higher rate on callus induction and preventing tissue browning in explant. Besides that, the anther and ovule explant may serve as an efficient explant to study the callus induction of C. nucifera L. var. MATAG and as a basis to screen the potential useful plant growth regulators for somatic embryogenesis.

In vitro Callus Induction of Tomato and Evaluation of Antioxidant Activity of Aqueous Extracts and Enzymatic Activities in Callus Cultures

Background: Tomato (Solanum lycopersicum L.) is an important plant rich in many vitamins and antioxidant enzymes. Methods: Tomato leaves of two cultivars (‘Peto 86’ and ‘Strain B’) were used as explant sources for callus induction. The antioxidant activity of the calli ethanol (ET) and methanol (ME)extracts were determined. The enzymatic activities were evaluated in callus cultures. Callus induced from leaf explants of tomato cultivars on the Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of cytokinins and auxins such as BAP (6-benzylaminopurine), NAA (1-naphthalene acetic acid), 2,4-D (2,4-dichlorophenoxyacetic acid), IAA (indole-3-acetic acid) and Kin (Kinetin) for rapid induction of callus and biomass growth. Results: The medium (M2) containing 3 mg L-1 BAP with 1 mg L-1 IAA produced the highest percentage of callus induction (PCI) (100%) in two cultivars. The relative fresh weight growth (RFWG) was reported by the fresh callus weighed after four weeks of culture and again weighed after one month of sub-culture. In both cultivars cultured on M2 medium the RFWG was (1.60) in ‘Peto 86’ and (1.47) in ‘Strain B’. The results showed that PCI and RFWG differed with the cultivars tested. The scavenging DPPH free radical activity in callus (ET) extracts exhibited a significant increase in (P < 0.05) than the activity in callus (ME) extract. The peroxidase and polyphenol oxidase activities were found in calli of both tomato cultivars. The enzymatic activities were higher in callus of ‘Peto 86’ cultivar than in callus of ‘Strain B’ cultivar. Conclusions: Calli had antioxidant and enzyme activities that can be beneficial for extracting important components or for plant regeneration.

A Comparative Study between Plant and Callus Extracts of Abutilon indicum (L.) Sweet: Antioxidant, Antibacterial, Antidiabetic and Anti-Proliferative Activity

Abutilon indicum is consider to be used in the traditional system of medicine. It is found in tropical and subtropical regions of the world. It is used to treat various diseases. This plant does not cause any side effects to humans. As the plant has wide variety of medicinal properties, the present study aimed to comparative between plant and callus extract of Abutilon indicum (L.) sweet for antioxidant, antibacterial, antidiabetic and anti- proliferative activity. The highest percentage of callus induction (89.50%) and callus weight (1.26 g) was observed in T5 (MS + 2, 4-D (2.5 mg/l) + BAP (2 mg/l) and T8 [IBA (4 mg/l)] respectively. Phytochemical analysis of aqueous and ethyl acetate extracts of A. indicum in vivo plant and in vitro grown callus showed the presence of alkaloids, flavonoids, phenols, carbohydrates, glycosides, protein, terpenoids, saponins, tannins and coumarin. The total phenolic content was high in aqueous extract of callus (30.68 mg TAE/g). Maximum DPPH radical scavenging activity was found in aqueous extract of callus (86%) with IC50 value of 68.49 µg/ml. FT-IR analysis of aqueous extract of A. indicum plant and callus showed the presence of characteristic stretching at 2930.28 and 2927.75 indicating the presence of C-H stretching respectively. GC-MS analysis revealed the presence of 17 compounds in ethyl acetate plant extract, whereas 7 compounds in ethyl acetate callus extract such as tetradecane, 1-chloro, Sulfurous acid 2-prophytridecyl ester and 1- ethyl-3-[2-(octadecylthio) ethyl] thiourea. The ethyl acetate extracts of callus and plant and was found to be effective against Bacillus subtilis (3.1 mm) and Staphylococcus aureus (2.9 mm). Maximum α-amylase inhibitory activity was observed in aqueous callus extract (32.65%) with IC50 value of 833.61 µg/ml. HeLa cell viability was found to be 26.8% and 21.8% in plant and callus extract respectively.

Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo (SE) yield, unsynchronized SE development, and a high percentage of deformed SEs. We aimed to improve F. mandshurica SE production by synchronizing SE development, improving SE quality, and inducing root formation to obtain complete regenerated plants. Cotyledons of immature zygotic embryos of F. mandshurica were induced to form callus and then SEs. The SE induction percentage from explants differed among 32 mother trees, and the one with the highest SE induction percentage (29.8%) was used for further experiments. The highest callus induction percentage was 94.2% on ½-strength Murashige and Skoog medium (MS½) supplemented with 0.15 mg·L−1 naphthalene acetic acid. The highest callus proliferation coefficient (240.5) was obtained on McCown’s Woody Plant Medium containing 0.1 mg·L−1 6-benzyl adenine and 0.15 mg·L−1 2, 4-dichlorophenoxyacetic acid. The highest number of SEs (1020.5 g−1 fresh weight) was obtained on MS½ medium supplemented with 1 mg·L−1 6-benzyladenine. The highest number of cotyledon embryos (397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L−1 abscisic acid and then applying a drying treatment. The cotyledon embryos were milky white, uniformly sized (average length 4.7 mm), and 80% of them were normal. The SE rooting percentage on ½MS medium containing 0.01 mg·L−1 NAA was 37.5%. Overall, the germination percentage of SEs was 26.4%, and complete regenerated plants were obtained after transplanting and acclimation. These results provide more possibilities for the preservation and breeding of F. mandshurica.

In vitro regeneration and molecular characterization of Jatropha curcas plant

Background and objectiveA simple, rapid, efficient, and reproducible protocol for callus induction and regeneration of plantlets from callus of Jatropha curcas plant was established.Materials and methodsRandomly amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses were used to determine the genetic variation between the regenerated, micropropagated, and mother plants.ResultsThe highest callus induction percentage from leaf explant was recorded with MS medium containing 2.5 mg/l BA (6-benzylaminopurine) + 1.0 mg/l NAA (1-naphthaleneacetic acid). Leaf-derived callus was grown on medium containing 2.0 mg/l BA + 0.2 mg/l IBA (indole-3-butyric acid) for adventitious shoot regeneration. In addition, using five random RAPD primers with the tested samples produced 117 amplified products out of which 25 were polymorphic bands resulting in 21.37% polymorphism, whereas the five ISSR primers used yielded 116 scorable bands out of which 22 were polymorphic bands producing a polymorphism pecentage of 18.96.ConclusionAn optimized protocol for large-scale production of J. curcas plants using plant biotechnology tools was achieved. RAPD and ISSR techniques would introduce an alternative system for large-scale production and establishment of genetically stable plants.

Identification of DNA variation in callus derived from Zingiber officinale and anticoagulation activities of ginger rhizome and callus

BackgroundThe medicinal plants have been used as alternative treatments for many diseases in many countries. Thus, the possibility of the alteration of some naturally relatively cheap sources into highly valuable products for pharmaceutical and biological importance via tissue culture is investigated in this study. Special attention is needed to estimate the molecular genetic variation between the studied plant and the callus.ResultsGinger (Zingiber officinale Roscoe) rhizome was affected by various concentrations of cytokinin and auxin for the induction of callus. The highest percentage of callus induction and maximal callus fresh weight was achieved when Murashige and Skoog (MS) medium was supplemented with 2 mg/l 2, 4-D + 1mg/l BA. The genetic variations accompanied with in vitro conditions of callus induction was evaluated by four primers of inter-simple sequence repeat (ISSR) that amplified 36 bands. The highest readings for clotting times were found by using the sulfated neutral extract of ginger rhizome at a concentration of 400 μg/ml, and the extract of sulfated alkaline plant callus had an anticoagulation activity at (200 μg/ml) comparable to that of a standard preparation of heparin sodium.ConclusionThese results showed that ginger (Zingiber officinale Roscoe) rhizome was affected by various concentrations of cytokinin and auxin for induction of callus especially when MS was supplemented with 2 mg/l 2, 4-D + 1 mg/l BA. Also, the results of ISSR markers confirmed the occurrence of genetic variations during callus induction process. The results indicated that the sulfated alkaline of ginger rhizome and sulfated aqueous extracts of ginger rhizome and callus exhibited anticoagulant activity. So, it was clear that the addition of sulfate group into the investigated extracts enhances the anticoagulation activities.