Callus Growth Research Articles

Unraveling the interaction of copper, cadmium, calcium, and nitrate on phenolics, flavonoids, and shikonin contents of Onosma dichroantha calli by statistical modeling.

Increased anthropogenic activities have led to the accumulation of certain minerals to ecotoxic levels in the environment, which could influence the secondary metabolism of plants. Shikonin, an exudate from the roots of Onosma dichroantha, is a secondary metabolite involved in plant defense and invasion success; however, the interactive effects of copper (Cu), cadmium (Cd), calcium (Ca), and nitrate (NO3) in shikonin biosynthesis and accumulation are not known. Here, the individual, curvilinear, and pairwise effects of these elements on shikonin biosynthesis in callus culture of O. dichroantha have been investigated by means of a statistical modeling approach and multivariate regression analyses. Although the main effects of the examined minerals seemed to be suppressive, their combined interactions could enhance callus growth and secondary metabolism of O. dichroantha. Accordingly, maximum values were recorded for the callus growth index (6.85 at 23.25μM Cu, 70mM NO3, 1mM Ca, 27.50μM Cd), total phenolics (24.83mg gallic acid equivalent at 9.75μM Cu, 70mM NO3, 1mM Ca, 62.50μM Cd), total flavonoids (6.12mg quercetin equivalent at 30μM Cu, 80mM NO3, 1.5mM Ca, 45μM Cd), and shikonin (24.33μgg-1 FW at 9.75μM Cu, 70mM NO3, 2mM Ca, 27.5μM Cd). Overall, these data show that increasing concentrations of the examined minerals in culture medium can markedly influence the secondary metabolism of O. dichroantha cells and suggest that a comparable phenomenon may exist in a wider range of medicinal plants, grown on polluted environments, which may affect their invasive capabilities.

IBA Delivery Technique and Media Salts Affected In Vitro Rooting and Acclimatization of Eight Prunus Genotypes

Difficult-to-root plants often perform poorly during acclimatization and in vitro rooting can increase the survival and quality of plants. The influence of auxin application and mineral nutrition on in vitro rooting and subsequent effects on plant quality in eight Prunus genotypes were investigated. Microshoots were rooted in vitro on Murashige and Skoog (MS), ½ MS, Driver and Kuniyuki (DKW), or New Prunus Medium (NPM) media formulations in combination with 15 µM indole-3-butyric acid (IBA), 4-day 15 µM IBA pulse, 1 mM 30 s quick-dip, or IBA-free treatments. Shoots were observed pre- and post-acclimatization to determine rooting methods to maximize quality and minimize labor. A genotype-specific response to auxin application was observed with seven of eight genotypes achieving 100% survival when paired with the recommended IBA treatment. Peaches performed best when treated with 4-day IBA pulse or 30 s quick-dip. Rooting of P. cerasifera, it's hybrid to P. persica, and P. munsoniana all benefitted from IBA application. Shoots rooted with 15 µM IBA were smaller and lower quality in most genotypes. DKW maximized size and quality in six genotypes. Better shoots and larger root systems during in vitro rooting produced better plants in the greenhouse with no detrimental effect of callus growth. Rooting techniques to maximize plant quality while reducing labor are specified.

THE EFFECT OF SEVERAL CONCENTRATIONS OF GLYCINE AMINO ACID ADDITIONS IN MS MEDIUM TOWARS CALLUS GROWTH OF KONJAC (Amorphophallus muelleri) BY in vitro CULTURE

Konjac (Amorphophallus muelleri) is a plant that can be a source of carbohydrates and has high economic value. To accelerate the fulfillment of porang seeds in order to support maximum economic potential, it is necessary to develop alternative propagation techniques,which is in vitro culture. This study aimed to determine the effect of adding the glycine amino acid to MS medium on the growth of konjac callus and to determine the best concentration for callus growth in konjac plant tissue culture. This research was using frog bulb (bulbil) for the explant. In this study used a completely randomized design (CRD) with glycine amino acid concentrations from 0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L, 4 mg/L, dan 5 mg/L with 3 replications. Data were analyzed using SPSS software with one way ANAVA and further test using DMRT 5%.The results showed that the addition of the glycine amino acid to MS medium affected the callus growth of konjac plant tissue culture based on the analysis of variance (ANAVA) test. The best concentration used based on the Duncan Multiple Range Test (DMRT) 5%, was 4 mg/L with the average callus appearance time of 3.06 wac and the average bending time of 2.1 wac.

Propagation of Gembili (Dioscorea esculenta. L) Accession from West Papua by In-Vitro Callus Induction

Diversification of tuber foods is expected to help food security and improve public health. Gembili tuber glucomannan contains high fiber. Gembili seeds are becoming scarce, causing gembili plants to be very rarely found in the community. In the West Papua region, local people still rely on conventional supplies which are feared to threaten the gembili plant population. Tissue Culture Technology is very necessary in multiplying gembili seeds quickly and without knowing the season, by growing callus, plant propagation does not require a lot of explants to produce a large number of seeds. The research was conducted at the Politeknik Negeri Lampung Tissue Culture Laboratory, from May to July 2022. The research used a randomized block design, with each treatment repeated three times. The results showed that the composition of the media with the addition of 2.4 D 2 mL.L-1 produces an average of 8.00 days of callus growth, however, for callus growth the culture media with the addition of 2.4 D mL.L-1 produces a value best. 1.73 cm for callus diameter and a value of 0.22 g for callus weight.

Effects of Early Weight-Bearing Treadmill Training Combined with Pre-Emptive Analgesia on Femoral Fracture Recovery.

The effect of pre-emptive analgesia plus early weight-bearing treadmill training (EWBTT) on healing and motor function recovery of femoral shaft fracture is not clear. A total of 60 SD male rats were randomly allocated into 4 groups: group A (pre-emptive analgesia with EWBTT), group B (pre-emptive analgesia with delayed weight-bearing treadmill training, DWBTT), group C (pre-emptive analgesia with no weight-bearing), and group D (EWBTT with no pre-emptive analgesia). All rats were molded by internal fixation with Kirschner wire after right femoral shaft fracture. In groups A, B, and C, tramadol was intramuscularly injected 15 minutes before surgery. EWBTT was performed at day 1 postoperatively in groups A and D, and DWBTT was performed at day 14 postoperatively in group B. Oblique plate test was accomplished to assess hindlimb motor function recovery of rats in each group. Status of fracture healing was assessed through digital radiography (DR). Hematoxylin-eosin (HE) staining and immunohistochemistry of bone morphogenetic protein-2 (MBP-2) and vascular endothelial growth factor (VEGF) in callus were performed to explore fracture healing. The expression of BMP-2 and VEGF protein in quadriceps femoris muscle was detected by Western blot technique and mRNA expression of BMP-2 and VEGF in callus ascertained via reverse transcription-polymerase chain reaction (RT-PCR) technique. For oblique plate test, rats in group A outperformed those in groups B and C at all time points after operation. DR image revealed that large numbers of callus growth, blurred fracture line, and obvious continuous callus passing through the fracture line can be found in group A at day 28 postoperatively, which is the best healing status among all groups. HE staining of callus confirmed the optimal effect of healing for rats in group A. VEGF and BMP-2 expression by immunohistochemistry showed a significantly higher positive score for callus in group A while those in group C being the lowest at all time points postoperatively. Significantly higher expression level of VEGF and BMP-2 protein was detected in quadriceps femoris muscle from group A, which exceeded those in all other groups at all time points. RT-PCR testing proved the highest expression of BMP-2 and VEGF mRNA in callus of rats from group A, significantly higher than those of other groups. Both pre-emptive analgesia and EWBTT can effectively invoke the expression of VEGF and BMP-2 and promote recovery of hindlimb locomotor function in rats with femoral fracture, and the combination of them leads to more superior results.

In vitro Induction and Proliferation of Callus in Piper longum L. through Leaf Culture

Piper longum L. (Family Piperaceae) is a well- known health promoter used to treat cough, chronic bronchitis, asthma, and diabetes mellitus. The study is aimed to develop a protocol for callus induction and proliferation in P. longum. The leaf explants from mature plants were cultured on an MS basal medium supplemented with various concentrations of plant growth hormones, viz. 2,4-Dichlorophenoxyacetic acid (2, 4-D), Kinetin (KN), 6-Benzylaminopurine (BAP), and α-Naphthalene acetic acid (NAA), as well as 10% coconut water. In primary culture, the callus was compact and light white. The best callus induction and growth were observed in the MS basal medium containing1.0 mg/L 2,4-D+ 2.0 mg/L KN at 12 weeks of primary culture. At eight weeks of secondary culture, the MS medium containing 2.0 mg/L BAP alone and 0.5 mg/L NAA + 2.0 mg/L BAP and 10% coconut water had the best callus proliferation. Compared to 2,4-D and KN alone, BAP alone supported rapid callus growth in the MS medium. In P. longum, large-scale callus formation from leaf explants could be exploited to produce, isolate, and increase bioactive secondary metabolites for therapeutic purposes.

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

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

THE GROWTH OF PIMPINELLA ALPINA HOST CALLUS AT VARIOUS TREATMENTS OF PLANT GROWTH REGULATOR CONCENTRATIONS OF NAA. 2,4 D AND ITS COMBINATION WITH BAP

Purwaceng (Pimpinella alpina) is an herbal plant that has aphrodisiac, diuretic, and tonic properties. The imbalance between the growth and the exploitation of this plant makes it included in Appendix I (endangered), so its conservation will be crucial. One method of the conservation of this plant is tissue culture. This method is considered one way to induct the callus. The research objective is to find the effect of various treatments of plant growth regulator concentrations of NAA. 2,4 D and its combination with BAP on the growth of purwoceng callus. Concentration of plant growth regulator NAA (0.5 mg L-1, 1 mg L-1, 2 mg L-1), 2,4-D (0.4 mg L-1, 0.8 mg L-1, 1.2 mg L-1 ), a combination of NAA with BAP (0.5 mg L-1 NAA: 1 mg L-1 BAP, 1 mg L-1 NAA: 2 mg L-1 BAP, mg L-1 NAA: 3 mg L-1 BAP) and a combination of 2, 4- D with BAP (1 mg LL 2,4-D : 0.5 mg L BAP, 1 mg L-1 2,4-D : 1 mg L-1 BAP, 2 mg L-1 2,4-D : 1 mg L-1 BAP) into Murashige Skoog medium. Then purwaceng leaf explants were planted on the medium until the growth was seen. This research shows that the plant growth regulator concentration has a different influence on the growth of callus that is recognized by the day of callus induction, colour and callus texture. MS media with growth regulator 1.2 mg L-1 2,4-D can grow callus faster (17 days) with friable texture.

Uptake of Copper Oxide Nanoparticles by Hypocotyl Callus Tissues of Helianthus annuus L. Seedlings

This study gives a special indication about the ability of callus tissues of sunflower (Helianthus annuus L.) seedling stems in uptake of copper oxide Cu2O nanoparticles with concentration (30,60,90,150) µg ml-1 which reflect their important role in phytoremediation, and in the same time the nanoparticles have role in improve callus growth and fresh weight. The positive role of growth regulators NAA and BA it become clear in induction the callus of sunflower (Helianthus annuus L.) plant. The result showed superiority of 150 µg ml-1 in encourage the increase of the fresh weight of the callus which developing on MS mediums that supported with growth regulator 0.5 mg ml-1 NAA and 1.0 mg ml-1 BA. Where the average fresh weight of the callus was 7.1 gm, 12.9 gm after 20 and 40 days of the growth respectively. Compared with the fresh weight of callus developing on same medium without nanoparticles where the average of the fresh weight was 4.1 gm, 6.2 gm after 20 and 40 day of the growth respectively, similarly the same concentration of nanoparticles above encourage the highest fresh weight when added alone in nutritional medium where it was 2.2 gm, 3.6 gm after 20 and 40 day of the growth respectively in compared with the other treatment. The Scanning Electron microscopy (SEM) pictures explained the ability of callus tissues in uptake of nanoparticles on the surface of their cells in variant quantity according to the different concentration that used.

Morfogenesis Kalus Sorgum Pada Berbagai Media Secara In Vitro

Sorghum has great potential to be developed as a food source in Indonesia because it has many benefits. Sorghum has various potentials and uses, such as as a source of functional food, industrial raw materials and also animal feed. All parts of the sorghum plant can be utilized. Genetic transformation will be successful and beneficial if a plant regeneration system has been obtained in vitro culture. In its implementation, the selection of explants is an important first step to support the success of in vitro culture. Generally, the explants used for tissue culture are shoot tips, lateral shoots, and epicotyls. This study aims todetermine the morphogenesis of sorghum in vitro, from tissue culture. The analysis method in this study used 3 treatments, namely Murashige & Skoog (MS), Vacint & Went (VW) and Nagata & Takebe (NT) media with repeated 5 times. The method of analysis with data on the percentage of living callus and observing the morphogenesis of growing shoots and calluses. The result of this study is that the treatment of Murashige & Skoog (MS) media affects the morphogenesis of buds in the form of the number of perexplant shoots, the number of leaves, and the length of the shoots with the best treatment. The number of shoots per growing explant by 85%. The treatment of Vacint & Went (VW) and Nagata & Takebe (NT) media did not have enough effect on callus induction in the form of growth and development of callus planted in-vitro. So that the morphagenics of sorghum plants are good in the growth of shoots on Murashige & Skoog (MS) media.

Thiol, volatile and semi-volatile compounds alleviate the stress of zinc oxide nanoparticles of the pomegranate callus

Pomegranate trees are tropical and subtropical shrubs with nutritional benefits and pharmaceutical and therapeutic uses. Antioxidative systems protect the structure and function of cellular membranes. This study demonstrated the connection between oxidative stress generated by excess nanoparticles ZnO (ZnO-NPs) accumulation in pomegranate calli and the involvement of thiol groups and volatile and semi-volatile compounds in alleviating this stress. The effect of the non-enzymatic antioxidant system was studied using callus treated with three levels of ZnO-NPs or bulk particles (ZnO-BPs). With rising ZnO levels in the media, callus growth was gradually decreased by ZnO in both forms (NPs and BPs). Malondialdehyde (MDA) measurements revealed that different concentrations of both forms promoted lipid peroxidation. The supply of both forms had a considerable stimulatory influence on the cysteine (Cys) content in calli. Raised ZnO-NP concentrations increased glutathione (GSH) and non-protein thiols (NPTs) content in calli, but higher ZnO-BP concentrations lowered their content. Conversely, ZnO-NP levels reduced the protein thiols (PTs) content in calli, but ZnO-BP concentrations increased their content. GC-MS analysis was employed to investigate the volatile and semi-volatile chemical profiles within calli following exposure to 0 and 150 μg mL−1 of ZnO in both forms. GC-MS analysis detected 77, 67, and 83 compounds in ZnO-treated calli, of which 14, 16, and 20 with a similarity value greater than 70%, based on a NIST library, were recognized as metabolites for ZnO untreated and NPs- and BPs-treated calli, respectively. Six substances, including five alkanes and one morphinan, showed similarities in metabolite composition between control and NPs- or BPs-treated calli. ZnO-NPs-treated calli contained two alkane compounds only similar to the control, but ZnO-BPs-treated calli had six metabolites, including four alkanes, one carboxylic acid, and one ester. However, eight alkanes were similar within the callus treated with NPs and BPs.

Comparative Analysis on the Role of 2,4-Dichlorophenoxyacetic Acid in the Expression of Bioactive Compounds in Callus of Capsicum frutescens

Plant cell culture technology serves as an effective alternative system for in vitro production of bioactive molecules, as it allows for the exploration of valuable compounds under a controlled environment. The present study was conducted to evaluate the effect of plant growth regulator (PGR); 2,4-dichlorophenoxyacetic acid (2,4-D) on the expression of compounds in coloured callus of Capsicum frutescens, a vital spice in various cuisines worldwide. The differential accumulation of compounds in the callus was analysed using liquid chromatography-mass spectrometry (LCMS) and the PGR concentration that resulted in the highest accumulation of the valuable compounds was identified. In this study, calli of various colours (cream, yellow and green) were successfully produced from C. frutescens through plant tissue culture. The increase in 2,4-D concentrations was found to increase callus growth index (GI) and specific growth rate (Sg), where the highest GI (0.5690) and Sg (0.6348 mg/week) were observed in callus produced in media supplemented with 0.5 mg/L 2,4-D. LCMS data analyses showed that 19 compounds were detected in the callus, with 8 compounds (fatty acids and phenolics) were successfully identified, while the remaining 11 compounds were reported as unknowns. Yellow-coloured callus was observed to contain the highest number of compounds (18 compounds), while green callus contained the least (14 compounds). This analysis provides valuable information on the application of biotechnological tools such as plant tissue culture as an alternative for sustainable production of compounds with high bioactivity in Capsicum frutescens.

Radioelectronic wireless systems for non-invasive diagnostics of tubular bone fractures during body osteosynthesis

During the rehabilitation of the patient, accidental and shock loads are the main cause of aseptic loosening and damage to the elements of the fixing structure (FC). The lack of control over the formation of cartilaginous callus in the fracture zone causes an increase in the period of incapacity for work and an increase in material costs associated with the patient's stay in the hospital.The partial load on the bone is simulated, which is 25% of the calculated value and is equal to the average body weight of the patient. On the basis of the obtained data, a new method and device were developed for non-invasive detection of the initial moment of weakening or fracture of the fixing structure, detection of improper fracture fusion, diagnosis of the formation and control of cartilage callus growth, detection of the appearance of the initial moment of the inflammatory process in the patient's body and diagnosis of the causes of this process. The requirements for a possible method of non-invasive diagnosis of the condition of a tubular bone fracture are formulated. Criterion dependencies of the controlled parameters were established. The effect of random loads, which are the cause of failures of the fixing structure, has been proven. Wireless transmission of electricity and information made it possible to develop a method of non-invasive control and diagnosis of fracture of tubular bones and to establish criterion dependencies between controlled parameters. For a comprehensive diagnosis of the patient's recovery, it is sufficient to monitor the pressure at three points, the bending moment at the site of the bone fracture, the temperature of the periosteum and the temperature of any other point of the body.

Influence of Ag nanoparticles on physiological and biochemical aspects of callus of Thymus species and Zataria multiflora Boiss.

Thymus species have found remarkable importance in food and medicine industries. The present study investigates the potential effect of Ag nanoparticle elicitors on proliferation of callus, and production of carvacrol and thymol in Zataria multiflora and three Thymus species. Firstly, callus was induced on Murashige and Skoog (MS) medium containing 2 mg l−1 of 2, 4-dichlorophenoxy acetic acid (2,4-D) and 1 mg l−1 of kinetin (Kin)). Secondly, the effects of two different concentrations of Ag nanoparticles (4 and 8 mg l-1) were studied on callus growth and its secondary metabolites production. Results elucidated that after elicitation by 8 mg l-1 ofAg NPs, significantly the highest callus growth rate (CGR) (0.02 mm day-1), callus fresh mass (CFM) (0.99 g), and carvacrol (0.68 mg l-1) and thymol (11.09 mg l-1) content was achieved. Comparing different Thymus species, notably the greatest carvacrol and thymol amount was obtained in .kotschyanus Boiss. & Hohen. and T. Daenesis Čelak. at 8 mg l-1 concentration ofAg NPs. Hence, it is evident that the stimulation by NPs is dose-dependent. This study has potential to be commercially applied for the enhancement of pharmaceutical compounds in different species of Thymus.

PpyABF3 recruits the COMPASS-like complex to regulate bud dormancy maintenance via integrating ABA signaling and GA catabolism.

Bud dormancy is essential for perennial trees that survive the cold winters and to flower on time in the following spring. Histone modifications have been reported to be involved in the control of the dormancy cycle and DAM/SVPs are considered targets. However, how the histone modification marks are added to the specific gene loci during bud dormancy cycle is still unknown. Using yeast-two hybrid library screening and co-immunoprecipitation assays, we found that PpyABF3, a key protein regulating bud dormancy, recruits Complex of Proteins Associated with Set1-like complex via interacting with PpyWDR5a, which increases the H3K4me3 deposition at DAM4 locus. Chromatin immunoprecipitation-quantitative polymerase chain reaction showed that PpyGA2OX1 was downstream gene of PpyABF3 and it was also activated by H3K4me3 deposition. Silencing of GA2OX1 in pear calli and pear buds resulted in a similar phenotype with silencing of ABF3. Furthermore, overexpression of PpyWDR5a increased H3K4me3 levels at DAM4 and GA2OX1 loci and inhibited the growth of pear calli, whereas silencing of PpyWDR5a in pear buds resulted in a higher bud-break percentage. Our findings provide new insights into how H3K4me3 marks are added to dormancy-related genes in perennial woody plants and reveal a novel mechanism by which ABF3 integrates abscisic acid signaling and gibberellic acid catabolism during bud dormancy maintenance.

Molecular characteristics of humic substances from different origins and their effects on growth and metabolism of Pinus laricio callus

Humic susbtances (HS) are increasingly applied as biostimulants in agriculture, though their mechanism of action is not yet completely understood due to their complex and heterogeneous composition. Here, we isolated thirteen different humic and fulvic acids from different sources, such as soils, composts, peat, leonardite and lignite, and characterized the distribution of their carbon components by 13C-CPMAS–NMR spectroscopy. Callus of Pinus Laricio, as a model plant species, was treated with different humic extracts and its growth and content of carbohydrates, phenols, and enzyme related to the nitrogen metabolism (invertase, glutamine synthetase, glutamate synthase, phosphoenoly pyruvate carboxylase, malate dehydrogenase) and stress resistance (catalase) were monitored. While a multivariate statistical analysis of NMR results well-separated the HS characteristics based on their origin, humic materials generally increased callus growth, as expected, with largest effects being exerted by the mostly polar humic acids from composts. However, the rest of measured parameters were not linearly related to hydrophobicity and aromaticity of humic isolates as well as their origin, but their conformational dynamics had to be advocated to explain their effects on callus cellular components. This work confirms that HS of various origin can act as sustainable biostimulants of plant growth, though the comprehension of their effects on plants biochemical activities requires further research and additional understanding of their molecular composition and conformational behaviour.Graphical

Light Quality Modulates Growth, Triggers Differential Accumulation of Phenolic Compounds, and Changes the Total Antioxidant Capacity in the Red Callus of Vitis davidii.

Light quality is one of the key elicitors that directly affect plant cell growth and biosynthesis of secondary metabolites. In this study, the red callus of spine grape was cultured under nine light qualities (namely, dark, white, red, yellow, blue, green, purple, warm-yellow, and warm-white light). The effects of different light qualities were studied on callus growth, accumulation of phenolic compounds, and total antioxidant capacity of the red callus of spine grape. The results showed that blue and purple light induced increased red coloration in the callus, whereas yellow light induced the greatest callus proliferation. Among all of the light quality treatments, darkness treatment downregulated the contents of phenolic compounds, whereas blue light was the treatment most conducive to the accumulation of total phenolics. White, blue, and purple light induced increased anthocyanin accumulation. Mixed-wavelength light was beneficial to the accumulation of flavonoids. Blue and purple light were conducive to the accumulation of proanthocyanidins. A further study showed that cyanidin 3-glucoside (Cy3G) and peonidin 3-glucoside (P3G) were the main anthocyanin components in the callus, and blue, purple, and white light treatments promoted their accumulation, whereas flavan-3-ols and flavonols were the main components of non-anthocyanin phenolics, and their accumulation changed in response to not only light quality but also culture duration. The total antioxidant capacity of the callus cultures changed significantly in response to different light qualities. These results will provide evidence for an abiotic elicitor strategy to stimulate callus growth and enhance the accumulation of the main phenolic compounds in the red callus of spine grape.

Electrical Induction as Stress Factor for Callus Growth Enhancement in Plumular Explant of Coconut (Cocos nucifera L.)

The study was carried out at the Biotechnology Laboratory, Division of Crop Improvement, ICAR- Central Plantation Crops Research Institute, Kasaragod during 2019−2021 to investigate the effect of electric current induction in coconut plumular callus obtained from WCT variety of coconut. Callus induction was observed when the plumules extracted from the embryo were inoculated into Eeuwens Y3 media supplemented with 16.5 mg l-1 2,4-D and 1 g l-1 activated charcoal. An electrical induction set up was developed to induce minute quantity of electric current to the calli. Electric current of 1µA and 2µA was applied to the system continuously as well as for an interval of 1 h day-1 for a month and differences in callus mass before and after treatment were observed. Callus induced with electric current of 2 µA continuously for a month had maximum weight gain followed by callus induced with 1µA current continuously for a month. Here, anode and cathode were inserted into media and callus respectively. In other two treatments where, 1 µA and 2 µA current was inducted into callus for an interval of 1 hday-1 for a month as well as in control where no current was induced significant weight gain was not observed. The study implicates the effect of electric current in cell division of plants and the results suggests that continuous application of weak electric current in range of 1 and 2 µA enhances callus growth in plumular explants of coconut and could be used as a novel strategy for callus multiplication of coconut, a monospecific crop recalcitrant to in-vitro culture.

Influence of Plant Growth Regulators and Artificial Light on the Growth and Accumulation of Inulin of Dedifferentiated Chicory (Cichorium intybus L.) Callus Cells.

Chicory (Chicorium intybus L.) is a perennial herb of the family Asteraceae, widely distributed in Asia and Europe, commonly used industrially as a raw material for extracting inulin because of a high content of inulin and biologically active compounds. Light conditions and plant growth regulators (PGRs) are two of many factors that affect the growth and inulin content of chicory callus. The aim of this work is to study the effect of PGRs and light conditions on proliferation and accumulation of inulin of chicory callus in vitro. In this study, we used semi-solid MS medium supplemented with different auxins (including Indole-3-acetic acid (IAA), naphthylacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D)) at a concentration of 5.5–9.5 mg/L in combination with 2.0 mg/L 6 benzylaminopurine (BA) to determine induction and proliferation of callus. The increasing value of callus fresh weight was used to assess the growth of the callus in treatments. The results showed that a steady increase in callus fresh weight and inulin content in callus cells was obtained when they were cultured on MS medium supplemented with a combination of 2.0 mg/L BA with 7.5 mg/L IAA in lighting conditions with radiation equalized by the flux density of photosynthetic photons and ratios of radiation levels in the region of FR—far red > R—red. Increasing demand for organic inulin sources in production practice can be met by our finding.

Evaluation of the Anti-Cancer Potential of Rosa damascena Mill. Callus Extracts against the Human Colorectal Adenocarcinoma Cell Line.

Chemotherapy is an aggressive form of chemical drug therapy aiming to destroy cancer cells. Adjuvant therapy may reduce hazards of chemotherapy and help in destroying these cells when obtained from natural products, such as medical plants. In this study, the potential therapeutic effect of Rosa damascena callus crude extract produced in vitamin-enhanced media is investigated on colorectal cancer cell line Caco-2. Two elicitors, i.e., L-ascorbic acid and citric acid at a concentration of 0.5 g/L were added to the callus induction medium. Callus extraction and the GC–MS analysis of methanolic crude extracts were also determined. Cytotoxicity, clonogenicity, proliferation and migration of Caco-2 colorectal cancer cells were investigated using MTT cytotoxicity, colony-forming, Ki-67 flow cytometry proliferation and Migration Scratch assays, respectively. Our results indicated that L-ascorbic acid treatment enhanced callus growth parameters and improved secondary metabolite contents. It showed the least IC50 value of 137 ug/mL compared to 237 ug/mL and 180 ug/mL in the citric acid-treated and control group. We can conclude that R. damascena callus elicited by L-ascorbic acid improved growth and secondary metabolite contents as well as having an efficient antiproliferative, anti-clonogenic and anti-migratory effect on Caco-2 cancer cells, thus, can be used as an adjuvant anti-cancer therapy.