6-benzylaminopurine Research Articles

ОПТИМИЗАЦИЯ СОСТАВА ПИТАТЕЛЬНОЙ СРЕДЫ И УСЛОВИЙ КУЛЬТИВИРОВАНИЯ ДЛЯ МИКРОРАЗМНОЖЕНИЯ IN VITRO THYMUS SERPYLLUM L. И THYMUS CAUCASICUS WILLD.

The study was conducted to optimize the cultivation conditions for the second stage of clonal micropropagation of Thymus caucasicus Willd. and Thymus serpullum L. Stem segments with one node (8…10 mm), obtained by microcutting of shoots, were cultured on 10 different Murashige and Skoog (MS) nutrient media containing 2% sucrose and 0.8% agar-agar, with the addition of kinetin, thidiazuron, benzylaminopurine (BAP), indoleacetic and gibberellic acids. Various culture vessels (jars, flasks, test tubes) were used for micropropagation. The duration of the cultivation cycle varied from 40 to 70 days. The highest reproduction coefficient of T. serpyllum was noted on the MS medium containing 1.0 mg/l BAP and was 6.7, while that of T. caucasicus was on the MS medium containing 1.0 mg/l kinetin (16.1). The highest efficiency of culturing both thyme species was achieved in jars, with the use of which the reproduction coefficient was 1.4…2.1 times higher than when grown in test tubes or flasks. It is advisable to cultivate the studied thyme species with a standard cycle of 40 days. The best combinations of various thyme reproduction factors contributed to the maximum manifestation of the morphogenetic potential in in vitro experiments. The greatest influence on the multiplication coefficient was exerted by the type of culture vessel, the interaction of the composition of the nutrient medium and the genotype, as well as the composition of the nutrient medium (the shares of the influence of factors ranged from 20.0% to 25.3%). The results of the studies served as the basis for the development of a protocol that can be used for accelerated micropropagation of T. caucasicus and T. serpullum.

Induction and Characteristics of Callus Cultures of the Medicinal Plant Tussilago farfara L.

Tussilago farfara L. is a traditional medicinal plant valued for its potentially health-promoting metabolites. Its herbal raw material has been recognized and used since ancient times and continues to be widely used in traditional medicine. Introducing this plant species to in vitro cultivation is a challenging task, but once the protocol is developed, such cultures can provide an abundant and inexhaustible source of plant material. In this study, we report the successful induction and growth of vigorous T. farfara callus in vitro. Callus induction was achieved on MS solid media with the combination of indole-3-acetic acid (3 mg/L) and benzyl aminopurine (2 mg/L) in darkness, whereas it appeared inefficient under light conditions and in suspension culture. We present a detailed description of callus growth kinetics, morphological analysis, photosynthetic activity, and biochemical parameters (including protein content and photosynthetic pigments) supported by histological studies. Furthermore, we observed the potential for organogenesis and somatic embryogenesis. This method for the in vitro propagation of T. farfara, along with callus culture maintenance, offers a wide range of applications in pharmacy for the production of valuable metabolites. Moreover, it could benefit the environment by reducing the depletion of natural populations of this species and may serve as an alternative strategy for species conservation in light of global warming.

Effect of Different Nitrogen Levels and Plant Growth Regulators on Stomatal Resistance, Leaf Temperature and Transpiration Rate and Yield of Banana cv Ney Poovan

The present Study was carried out to understand the effects of certain plant growth regulators under different nitrogen levels on different gas exchanging parameters viz., stomatal resistance, leaf temperature, transpiration rate and yield of banana cv. neypoovan. Different levels of nitrogen viz.,150 g Nplant-1 (45g + 75g +30g N plant-1 at 3,5 and 7 MAP, respectively), M1 + Urea 2% foliar spray, 200 g N plant-1 (60g + 100g + 40g N plant-1 at 3,5 and 7 MAP respectively), M3 + Urea 2% foliar spray and foliar spray of salicylic acid 100 ppm, mepiquat chloride 500 ppm, chlormequat chloride 1000 ppm, nitrobenzene 50 ppm, benzyl adenine 20 ppm and 25 ppm of 2, 4-D at 3rd , 5th and 7th month after planting were given and compared with untreated control.Salicylic acid treated leaves showed high stomatal resistance with low transpiration. The yield components were favourably influenced by higher level of N application. Soil application of 200 g N plant-1 and giving two per cent urea spray on 3rd, 5th,and 7th month after planting distinctly enhanced all the yield components. Salicylic acid spray at 100 ppm improved the number of hands, fingers and finger size. Benzyl adenine at 20 ppm spray also was found beneficial in increasing yield components. More number of fingers per bunch were observed in benzyl adenine treatment. With maximum fruit weight, salicylic acid spray surpassed benzyl adenine by recording maximum bunch weight of 12.58 kg (13.9 % increase over control), while benzyl adenine recorded 12.52 kg (13.4 % increase).

In vitro clonal micropropagation of selected Malus niedzwetzkyana Dieck forms demonstrating high resilience when introduced into urban environments

Background. Currently, the use of an interdisciplinary approach based on a combination of traditional introduction methods and clonal micropropagation techniques makes it possible to solve one of the key problems of introduction – the establishment of bioresource collections consisting of selected plant accessions with valuable agronomic traits and resistance to unfavorable urban environments. Materials and methods. A plant introduction study resulted in identifying seven specimens of Malus niedzwetzkyana Dieck with resilience to urban environments, high rate of crown development, and longevity. They served as source material for the development of a clonal propagation protocol and in vitro preservation of selected genotypes of this species. Results. It was shown for M. niedzwetzkyana that the most favorable time for taking its plant material for introduction into in vitro culture is the beginning of the active growth of vegetative shoots after flowering. The most optimal sterilization technique for such plant material was a stepwise regime using alcohol, sodium hypochlorite, and silver nitrate: it provided from 50 to 70% of sterile explants and the maximum percentage of meristem proliferation. Combining 0.8 mg/L of benzylaminopurine (BAP) with 0.14 mg/L of indole-3-butyric acid (IBA) at the stage of microclonal propagation ensured a significant increase of the reproduction coefficient (on average up to 5.3 ± 0.7) and an improvement in morphometric parameters of microshoots; the maximum frequency of shoot proliferation was 100%. The yield of shoots adapted to ex vitro conditions was 90%. Conclusion. The developed clonal micropropagation protocol made it possible to introduce selected M. niedzwetzkyana forms into in vitro culture and reproduce them in order to set up a resource base for further fundamental and applied research into the system of the genus Malus Mill.

Effect of Benzylaminopurine (BAP) and Coconut Water on the Growth of Bucephalandra sp. in Vitro

Tissue culture is useful as a way to obtain seeds in large quantities of seedlings in a short time as an effort to preserve Bucephalandra sp. in nature. The use of PGR has problems such as expensive prices and difficult to find, therefore its use can be replaced by natural PGR that are easy to obtain and have relatively cheap prices such as coconut water. The aim of this research was to determine the effect of Benzylaminopurine (BAP) and coconut water on the growth of Bucephalandra sp. in vitro. This research is a 2-factor factorial experiment using a completely randomized design. The factors are concentration of BAP (0; 1; 3; 5 mg/l) and concentration of coconut water (0; 50; 100 ml/l). The results showed Murashige and Skoog (MS) media with the addition of BAP 1 mg/l + coconut water 50 ml/l (4.11 shoots), BAP 5 mg/l + coconut water 50 ml/l (4.33 shoots), and BAP 3 mg/l (6.22 shoots) were able to provide the number of shoots with high results. The addition of coconut water did not affect the growth of Bucephalandra sp. MS media can grow shoots with the best results in measurements of shoot emergence time, number of roots, shoot and root length. Keywords: Aquatic plant, Endemic plant, Plant growth regulators, Tissue culture.

The effect of UV-C radiation and cytokinin on pea plants

Aim. The effect of ultraviolet C (UV-C) radiation and cytokinin benzylaminopurine (BAP) on the growth and content of photosynthetic pigments in leaves of pea plants (Pisum sativum L.) was studied. Methods. Pea plants cultivar Aronis were irradiated by UV-C at a dose of 15 kJ/m2 with a power of 7 W/m2. Part of the non-irradiated plants was treated with BAP, part of the plants was treated with BAP one day before irradiation and part of plants were treated with BAP in one day after UV-C irradiation. Length and mass of plant shoots and roots were measured during the experiment. Content of photosynthetic pigments in leaves were determined during all time the experiment. Results. It was shown that pea plants growth delayed content of carotenoids in leaves reduced after the UV-C radiation of pea plants. Treatment of plants with BAP after the end of the UV-C radiation accelerated the restoration of photosynthetic pigments content. Conclusions. It was shown that UV-C radiation of pea plants by dose of 15 kJ/m2 caused inhibition of growth, decreasing photosynthetic pigments content in leaves. The BAP treatment after radiation stimulated the restoration of photosynthetic pigments content in the leaves.

IN VITRO PROPAGATION COLLA AROMATIC ROXB

This experiment was tested the propagation of Colla aromatica Roxb with tissue culture, the study objective to study of propagation possibility of Colla aromatica Roxb (Homalomena aromatica Schott. Araceae), this study found the possibility of disinfecting Colla aromatica Roxb using sodium hypochlorite solution in contains of 15 and 20 for 5 and 10 minutes, bacteria contamination was 0%, fungal contamination was 14.29%, the survival rate was 85.71% At the same time without Chemicals stimulate growth of plant, can't cause callus form callus in the case . After rearing young shoots of for 30 days, feeding Benzyl Adenine Pure (BAP) at a contains of 3 mg/l combined with Naphthalene Acetic Acid (NAA) at a contain of 1 mg/l, there was a high number of shoots 1.88 shoots, but at the same time, giving NAA only was found to be statistically different. after culture the shoots in Agar media for 60 days, it was found that the result of giving NAA combined with BAP for the elongation of the shoot is not statistically different, from the observation that giving NAA combined with BAP with a high contains, will have the characteristics of the shoot elongation that is better than giving NAA combined with low contains

The Elimination of Viroids through In Vitro Thermotherapy and a Meristem Tip Culture from a New Limonime Hybrid (Citrus x limon var. limon (L.) Burm. f. x Citrus latifolia var. latifolia)

Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) from a new limonime hybrid (Citrus x limon var. limon x Citrus latifolia var. latifolia). The elimination success was confirmed by RT-PCR assays. The in vitro elimination rate for CEVd during the shoot proliferation stage (43%) was higher than for HSVd (21%). Accordingly, in the subsequent rooting stage, the in vitro elimination rate for CEVd (50%) was higher than for HSVd (33%). Successful CEVd and HSVd eradication at a 100% rate was confirmed in the ex vitro acclimatized plants in the greenhouse. The study also established an efficient micropropagation protocol. The optimal treatment for in vitro shoot induction was 0.5–2 mg L−1 benzyladenine (BA) + 0.5 mg L−1 gibberellic acid (GA3) + 0.25 mg L−1 naphthalene acetic acid (NAA), while for shoot elongation, it was 0.5 mg L−1 BA + 0.5 mg L−1 kinetin (KIN) + 0.5 mg L−1 GA3 + 0.25 mg L−1 NAA. Rooting was best promoted by 1 mg L−1 NAA. This study provides valuable insights for the mass production of viroid-free propagation material in this new lemon x lime hybrid, contributing to the conservation of genetic resources in citrus breeding programs through the combined application of in vitro thermotherapy and an in vitro meristem tip culture, a novel and highlighted achievement reported for the first time in this study.

Effects of plant hormones and genotypes on anther culture response of safflower (Carthamus tinctorius L.)

Safflower (Carthamus tinctorius L.) belongs to the composite family with various uses, from the stem to the seeds. In vitro flowering and embryo rescue are common techniques for improving safflower hybrid fertility by overcoming breeding constraints. However, anther culture expedites the production of superior types. The aim of the study was to evaluate the effect of plant hormones and genotypes on anther culture responses and to develop a suitable protocol. In both safflower genotypes, callus, shoot, and root formation were investigated using ten factorial hormone treatments. To promote callus induction and shoot regeneration, variable doses of Thidiazuron (TDZ) at 1 mg/l indole butyric acid (IBA) and variable benzylaminopurine (BAP) at 0.5 mg/l NAA were added to the MS medium, respectively. ½ MS media with varying IBA concentrations at a fixed 0.5 mg/l NAA were used for root regeneration. Results showed that Turkan was superior in all aspects of callus induction, and the highest degree of callus formation (46 %) was observed at 0.5 mg/l TDZ. On the other hand, the local safflower performed better for shoot and root regeneration. The highest shoot regeneration capacity (20 %) was shown at 2.0 mg/l BAP, while shoot rooting was reached (21 %) at 1.0 mg/l IBA of culture media. Despite successful in vitro regeneration, acclimated plantlets did not survive in the glasshouse. In this study, we demonstrated the higher capability for both shoot and root regeneration of calli produced from local safflower anthers. More effort is required to improve shoot and root development for both genotypes.

Effect of Foliar-applied Plant Growth Regulators on Coriander (Coriandrum sativum L.) to Regulate Growth and Yield Attributes

This investigation was conducted to obtain the effect of plant growth regulator on growth, yield and quality. The experiment utilized a randomized block design with three replications and nine treatments, including a control. The four different plant growth regulators (PGRs) used were Benzyl adenine (BA, 10 and 20 ppm), Brassinosteroid (BR, 0.5 and 1.0 ppm), Jasmonic acid (JA, 50 and 100 ppm) and Salicylic acid (SA, 50 and 100 ppm), applied as foliar sprays at 30 and 60 days after sowing (DAS). The experiment was conducted during the Rabi season of 2023-24 at the Vegetable Research Centre, Maharajpur, Department of Horticulture, College of Agriculture, Jawaharlal Nehru Krishi Vishwa Vidyalaya, Jabalpur, Madhya Pradesh. The investigation revealed that BR at 0.5 ppm resulted in the maximum plant height (103.67 cm). The highest number of primary (15.53) and secondary (26.50) branches at harvest was achieved with JA at 50 ppm. Phenological observations indicated that JA at 50 ppm led to the earliest 50% flowering at 36.67 days. For yield attributes, the highest number of umbels per plant (42.13) and umbellets per umbel (6.31) were found with JA at 50 ppm. BA at 20 ppm showed the maximum number of seeds per umbel (36.40), test weight (10.84 g), harvest index (9.92), and seed yield (13.65 q ha-1). Economically, BA at 20 ppm provided the highest net return (Rs. 170735.81), gross return (Rs. 232117.39), and benefit-cost ratio (2.78). Therefore, it can be concluded that foliar sprays of BR, JA, and BA are effective on improving phenological traits, yield attributes, and economic returns in coriander cultivation.

Stimulating action of sodium nitroprusside and vinasse on salicin and direct regeneration in Salix Safsaf Forssk.

The present study aimed to enhance salicin and direct regeneration inwillow (Salix safsaf Forssk) using the sodium nitroprusside (SNP) regulation of nitric oxide (NO) and vinasse for its nutrition effect in culture medium. Internodes of Salix safsaf were cultured on Murashige and Skoog (MS) medium supplemented with benzyl adenine (BA) (0.25 mg L-1) and different concentrations of SNP (0, 5, 10, 15, and 20 mg L-1) or vinasse (0, 5, 10, and 20%) to examine shoot regeneration, antioxidant defense enzyme activity, total phenolic compounds, flavonoids, and salicine contents. The reported data revealed that application of SNP at15 mg L-1 and vinasse at10% induced asignificant effectin vitro Salix safsaf shoot regeneration. To confirm that, nitric oxide is required for auxin-mediated activation of cell division in a dose-dependent manner. A concentration of 15 mg L-1 SNP promotes regeneration and salicin accumulation (3162.16mg/100g) during signaling action. On the other hand, the cross talk effect of nitric oxide and vinasse combination inSalix safsaf significantly induced a synergistic effect on direct propagation more than vinasse alone. SNP significantly stimulates salicylate accumulation in a dose-dependent manner, but the data on the association of vinasse and SNP on salicylate up-regulation showed a significant reduction in salicin accumulation when SNP was combined with 10% vinasse, which directly affected the signaling action of SNP as secondary product stimulators. Vinasse's phenolic compounds affect directly on thereduction activity ofSNP to suppress its signaling action, or indirectly by inhibiting the sequence cascade of the SNP signaling transduction process to decrease the accumulation of salicin contents. Data confirmed that vinasse andSNP stimulatedtheantioxidant enzymes activity throw quenching thestimulated reactive oxygen species that produced via SNP. Results show that modified media with SNP administration at 15 mg L-1 and the combination of vinasse at 10% and SNP at 15 mg L-1 are recommended for modifying tissue culture media for induced direct regeneration and salicin accumulation in tissue culture applications, which will be very useful for commercial salicin overproduction as a biological active ingredient in willows.

Plant biotechnology for the conservation of Argania spinosa (Skeels L): optimising propagation and sustainable valorisation

ABSTRACT Argania spinosa, an endangered species from southwestern Morocco, is studied for preservation through plant biotechnology. The research aims to optimise argan tree propagation to boost production, preserve heritage, and use residual species. In vitro seedlings are transferred to a modified Murashige and Skoog medium with various phytohormones. Six combinations of auxins and cytokinins are tested for germination. Results show that a liquid medium with activated charcoal from argan shells promotes stem elongation. Meta-topoline at 0.5 mg L−1, combined with naphthalene acetic acid (NAA) at 0.2 mg L−1, enhances shoot growth, leaf formation, and callogenesis. Adding benzylaminopurine (BAP) at 0.1 mg L−1 and indole-3-butyric acid (IBA) at 0.02 mg L−1 to the medium elongates stems by 4.3 cm after 21 days. A 2000 ppm IBA concentration is key for a 90% rooting rate. These findings underscore plant biotechnology’s role in preserving and propagating argan trees, improving resource management, and supporting sustainability.

Cytotoxic activity of callus extract from Vachellia farnesiana (L) Wight & Arn.

The in vitro cultures of Vachellia farnesiana (L) Wight & Arn. have demonstrated cytotoxic activity through callus extract on the HeLa cell line. Explants excised from in vitro-grown seedlings from seeds of two different locations were inoculated on Murashige and Skoog (MS) culture media containing various concentrations of N-6 benzyladenine (BA) or kinetin with 2,4-dichlorophenoxyacetic acid (2,4-D). Optimal efficiency in friable callus induction (100%) was achieved in leaf explants cultured on MS media containing 2.32µM BA + 13.57µM 2,4-D. Plant tissues (callus and leaf) were extracted and subjected to quantitative phytochemical analysis, revealing the highest total alkaloid and phenolic content in leaf extracts from Queretaro adult specimens (339.5 ± 20.9mg atropine equivalents (AE) per g dry extract (DE) and 158.4 ± 12.5mg gallic acid equivalents (GAE) per g DE, respectively). In contrast, callus cultures exhibited significantly higher total triterpene content (356-381mg ursolic acid equivalents (UAE) per g DE) compared to leaf extracts (208-243mg UAE/g DE). Both leaf and callus extracts displayed cytotoxic activity against the HeLa cell line, with a significantly lower half-maximal inhibitory concentration (IC50) for leaf extracts (28-32µg/mL) compared to callus cultures (43-66µg/mL), suggesting that alkaloids were primarily responsible for the cytotoxic activity. Furthermore, this study provides valuable insights into the controlled production of bioactive compounds with cytotoxic activity, with callus serving as a rich source.

An Overview of Pharmaceutical Applications and In vitro Micropropagation Techniques for Rare and Endangered Plant Species

Many rare and endangered plant species possess valuable secondary metabolites with pharmacological applications. These bioactive compounds are often integral to traditional medicine systems, highlighting the cultural significance of these plants. The health benefits of many medicinal species are not fully validated by contemporary scientific research, and some may be facing extinction due to habitat loss, overharvesting or climate change. This situation highlights the urgent need for effective conservation strategies of the species and sustainable cultivation methods. Micropropagation is a valuable technique for producing large numbers of plants from a single explant, significantly aiding in the conservation and commercial cultivation of rare species. Among the various types of explants, shoot tips and nodal segments have been identified as the most effective explants for micropropagation. These explants can be induced to generate multiple shoots in Murashige and Skoog (MS) medium containing Benzyl aminopurine (BAP). Thidiazuron (TDZ), Kinetin (KIN), or 2,4-Dichlorophenoxyacetic acid (2,4-D) are commonly used in MS medium to promote shoot and root development in both direct and indirect organogenesis processes. Rooting of the plantlets was typically achieved using MS medium either supplemented with Indole-3-butyric acid (IBA) or devoid of auxins, depending on the species and the specific requirements for rooting.

Optimization of embryo rescue technique for development of hybrid plants in stenospermic grapes

The development of seedless cultivars is a primary goal in grapevine breeding. Since grapes are stenospermic and tend to abort their embryos before development, traditional breeding methods often yield seedless cultivars at a low frequency. Therefore, embryo rescue has emerged as a promising approach for creating seedless grape cultivars. This study aimed to optimize the ideal sampling time and protocol for efficient embryo rescue in grapes. Ovules from immature berries collected at various days after pollination (20DAP, 30DAP, 40DAP, and 50DAP) were cultured on Nitsch and Nitsch (NN) medium with different concentrations of Benzyl aminopurine (BAP) (0.1, 0.2, and 0.5 mg/L) and Activated Charcoal (AC) (1.5, 2, and 2.5 g/L). Several parameters were assessed, including maximum ovule growth, the percentage of enlarged ovules, percentage of collapsed ovules, callus formation percentage, and embryo germination percentage. The study’s results indicated that berries collected at 40 DAP yielded the best outcomes across all parameters. Regarding treatments, the most favorable results were achieved when ovules were cultured on NN medium supplemented with 0.5 mg/L BAP and 2 g/L AC. In conclusion, the study underscores the significance of choosing the right sampling time and treatments to ensure efficient embryo rescue in grapes. The protocol standardized from this research is recommended for effectively rescuing embryos and developing seedless hybrids.

An effective in vitro propagation of Coelogyne mossiae Rolfe: An endangered endemic orchid of Western Ghats, India

Coelogyne mossiae Rolfe, an unexplored taxon given its red-listed status by IUCN was selected for in vitro propagation studies due to its conservational significance. The mature pods harboring viable seeds were subjected to inoculation on eight different nutrient media, revealing full-strength Murashige and Skoog (MS) media as highly efficient for seed germination (96.65 %) and subsequent stages of seedling development. Protocorm explants are used for micropropagation on MS media supplemented with various plant growth regulators (PGRs). α- Naphthaleneacetic acid (NAA) in the concentration of 0.5 mg/L facilitated optimal shoot induction resulting in an average of 2.39 multiple buds and 2.46 cm shoot length while 1.0 mg/L 6-Benzyl amino purine (BAP) augmented cultures provided 2.32 multiple shoot buds with an average 1.92 cm height. Half macro-MS augmented with 0.5 mg/L BAP promoted pseudobulb elongation (0.73 cm) while 0.5 mg/L NAA supported root formation (8.33 roots with an average length of 1.12 cm). The resultant plants were acclimatized in pots containing coco peat and brick pieces, exhibiting a survival rate of 63.23 %. The anatomical characteristics of the hardened plants provided insight into factors conducive to their adaptability. Also, the 3-month-old hardened plants demonstrated significantly greater total photosynthetic pigment (0.4915±0.00 mg g-1), total carbohydrate content (72.33±1.69 mg SSE/g), and total protein content (40.26±0.53 mg BSAE/g) when compared to in vitro plants. These findings underscore the potential of in vitro propagation techniques in the conservation and sustainable utilization of Coelogyne mossiae.

Micropropagation of three ginger cultivars and the subsequent growth and rhizome yield assessment and phytochemical composition analysis

Farmers have turned to biotechnology techniques to produce ginger, due to disease issues. Micropropagation is a proven and effective means for mass production of disease-free plants. Our research focused on ginger cultivar specific reactions (from culture initiation to rhizome harvest) to plant growth regulators (PGRs) to determine if plant response to PGRs is cultivar specific. Ginger (Zingiber officinale Rosc.) cultivars studied were: Chinese White (CW), Hawaii Yellow (HY), and Khing Yai (KY) under benzylaminopurine (BA) versus kinetin (KT) treatments using MS basal medium to micropropagate ginger seedlings and subsequent growth and yield performance and phytochemical composition analysis. In vitro culture data were collected every month for number of buds and shoots (and shoot length) produced per initiating bud and shoot growth. Mature seedlings were subsequently transplanted in a greenhouse with a randomized complete block design (RCBD) with 4 replications, 18 plants per replication (Main Plot = Cultivar, Sub Plot = PGR) for a total of 72 pots/plants. Growth data included number of stems per seedling, stem length, and stem diameter. Rhizome yield data included number of pieces of rhizome per seedling, and rhizome weights (biological, edible, and total rhizome weights respectively). Phytochemical composition analysis of ginger rhizome/PGR treatment included average amount of 6-gingerol, 8-gingerol, 10-gingerol, 6-shogoal, 8-shogoal, and 10-shogoal. Data was analyzed using SAS OnDemand for Academics with PROC GLM at α=0.05 level of significance. As ginger tissue culture seedlings progressed in culture, KT was more suitable for multiplication of buds, BA more suitable for proliferation of shoots, and KT matured ginger tissue culture seedlings at a higher rate than BA. CW had the highest rhizome yield and least amount of biological roots in comparison to other cultivars. CW produced the least amount of 8-gingerol, 10-gingerol, 8-shogoal and 10-shogoal of all three cultivars and with no significant change between two PGR treatments. In comparison, HY produced the highest amount of 6-gingerol, 8-gingerol, 10-gingerol, 6-shogoal, and 8-shogoal, followed by KY and CW, when micropropagated with BA. KY produced the highest amount of 10-shogoal when micropropagated with BA and the highest amount of 6-gingerol, 6-shogoal, 8-shogoal, and 10-shogoal when micropropagated with KT.

Effect of BA on Callus Induction, Shoot and Root Development from Callus of Nepenthes ampullaria through In Vitro Culture

Nepenthes ampullaria Jack. or pitcher plant is a carnivorous plant in the Nepenthaceae family. They are in danger of extinction because of the less forest area, volatile weather, and pollution. This study aimed to investigate the influence of BA (6-benzyl adenine) on callus induction of Nepenthes sp. through In vitro culture. The experiment was divided into three parts; (1) Callus induction; shoot of a pitcher plant were cultured on solid MS medium (Murashige & Skoog, 1962) supplemented with BA (6-benzyladenine) at 0, 1, 2, and 4 mg/l for 70 days. The experiment design was completely Randomized Design (CRD) (4 treatments X 10 replicates). It was found that MS medium supplemented with 4 mg/l BA proved to be a suitable medium for callus induction (70%). (2) Shoot induction from callus; callus were cultured on MS medium supplemented with 0, 2, and 4 mg/l BA (CRD, 3 treatments X 10 replicates) for 45 days. The highest average number of regenerated shoots (19.7±0.3 shoots) was observed on MS medium without BA. (3) Rooting induction; the young shoots, 1-2 cm in length, were cultured on MS medium containing 0, 1, 3, and 5 mg/l NAA combination with 0 and 3 mg/l IBA for 40 days (CRD, 8 treatments X 10 replicates). The results showed that the MS medium supplemented with 1 mg/l NAA and 3 mg/l IBA was suitable for root induction with the average of 7.4±1.6 roots.

FORMULATION OF NANOSTRUCTURED LIPID CARRIER GEL FROM CALLUS EXTRACT OF MULBERRY LEAF (MORUS ALBA L.) WITH 2, 4-DICHLORO PHENOXY ACETIC ACID AND BENZYL AMINO PURINE AS CALLUS GROWTH FACTOR

Objective: This research aimed to formulate the callus extract of mulberry leaf in the form of a Nanostructured lipid Carrier (NLC) gel. Methods: Dichlorophenoxyacetic acid (2,4–D) and Benzyl Amino Purine (BAP) was used as a callus growth factor. Callus leaf extracted with ethanol 96% by maceration-sonication method. An amount of 0.5% callus leaf extract was formulated into NLC. The NLC is then evaluated for its particle size and polydispersity index. The NLC gel is evaluated for its organoleptic, homogeneity, viscosity, flow ability, and pH. The callus extract and the NLC gel were also evaluated for their tyrosinase inhibitor activity. Results: The best formulation of NLC showed a particle size of 189.8 nm and a polydispersity index of 0.578. The NLC is a semi-solid, yellowish, odorless, homogeneous gel, with viscosity of 26,666.67 cPs, plastic-thixotropic type, pH of 5.26. The evaluation of tyrosinase inhibitory activity of the callus extract and the NLC gel showed IC50 value of 217.64 and 248.12 ug/ml. Conclusion: It can be concluded that leaf callus extract of mulberry can be formulated into an NLC gel that is physically and chemically stable and has good skin-lightening activity.

Effects of different growth hormones on Solanum tuberosum L. callus induction and regeneration

This study was carried out to develop a protocol for callus induction, regeneration, shoot, and root induction in two potato cultivars (Riviera and Almonda) that can be further used to develop genetically modified plants. Callus was induced from leaf and internodal explants on Murashige and Skoog's medium (MS) supplemented with different combinations of hormones. The highest percentage of callus induction was obtained from the internodal explants of Riviera cultivar was 100% in Callus Induction Media I (CIM I) supplemented with 2 mg/L Benzyl Adenine (BA), 0.2 mg/L ?-Naphthalene Acetic Acid (NAA), 2 mg/L 2,4-Dichlorophenoxy acetic acid (2,4-D), 0.2 mg/L Kinetin (Kin) and 0.2 mg/L gibberellic acid (GA3). After one-month calli were shifted to Shoot Induction Media I and II (SIM I and SIM II), the highest percentage for shoot induction was 33.3% with the leaf explants of Riviera cultivar on SIM II supplemented with 2 mg/L BA, 0.2 mg/L NAA, 0.5 mg/L Kin and 0.2 mg/L GA3. The root induction percentage was 100% for both cultivars. Regenerated potato plants had normal leaf shapes with uniform morphology. These findings provide valuable insights for developing tissue culture protocols for potato regeneration, which are essential for genetic engineering and potato breeding programs.