Concentrations Of Growth Regulators Research Articles

In Vitro Propagation of Ruta Chalepensis Through and Callus Culture

Plant tissue culture is a field that that enables culturing of various plants and parts of plants usually treated under a nutrient medium and in highly sterile conditions. Out of them callus culture is one of the very interesting arenas of plant biotechnology that encompasses many pivotal benefits. The study focuses on such callus enrichment using different hormones that there by enhance its biological activities. The plant namely Ruta chalepensiswas chosen upon wherein the callus growth was noticed. Ruta chalepensis has multiple medicinal activities like anti-cancer, anti-ulcer, anti-diabetic and many more pharmacological properties that yields in treating and curing of illness. Ruta chalepensis the leaf and internode were taken for the study and to analyse which of those at what concentration of plant growth regulators showed a better callus induction. The MS medium as well as Various hormone concentrations was used for the study like auxin and cytokinin from 0.5mg to 2 mg (2,4 – D, NAA, IAA, IBA). Increased concentration of 2,4-D (1.0 mg/L) alone in the MS medium showed profused callus growth. Both the explants used such as leaf and Internode were also tested in the MS medium which was devoid of hormones/plant growth regulators which was treated as control for comparison. From the data obtained, MS medium supplemented with hormones showed better growth rate and callus induction when compared with that of MS medium without hormones/plant growth regulators. Among the plant growth regulators 2,4-D (1.0 mg/L) showed maximum callus initiation from both leaf and internode explants. Further work was carried out in single and combination of the plant growth regulators for callus proliferation and accumulation. Further analysis is being done to study the growth pattern on combination of hormones and fix the hormone concentrations for the mass propagation of callus from the explants.

Cell culture of Euphorbia fischeriana and enhancement of terpenoid accumulation through MeJA elicitation

Euphorbia fischeriana Steud is a perennial medicinal plant belonging to the Euphorbiaceae family, but its clinical application has been limited due to the scarcity of natural raw materials. Therefore, this study attempted to provide a new method for producing plant materials of E. fischeriana by cell culture. During cell culture, the effects of concentrations of plant growth regulators and sucrose were investigated to improve the culture efficiency. The results showed that the combination of 1.5 mg/L 6-benzylaminopurine and 0.2 mg/L naphthaleneacetic acid, together with a sucrose concentration of 20 g/L in the medium was appropriate for cell growth and terpenoid accumulation. Furthermore, this study applied an elicitation strategy with methyl jasmonate (MeJA) to enhance terpenoid production and investigated the involvement of signaling molecules, such as nitric oxide (NO) and hydrogen peroxide (H2O2). The results indicated that terpenoid accumulation depended on the concentration and treatment duration of MeJA. The maximum total terpenoid content was obtained when the cells were treated with 100 μM MeJA for 4 days. The levels of NO and H2O2 in the cells increased after MeJA treatment and peaked at 8 h and 18 h, respectively, suggesting the involvement of NO and H2O2 in MeJA-induced terpenoid synthesis. The findings of this study demonstrated the potential of cell culture as a method for obtaining plant materials of E. fischeriana.

Pengaruh Ekstrak Bawang Merah (Allium cepa L.) sebagai Zat Pengatur Tumbuh pada Benih Kakao (Theobroma cacao L.)

Cocoa (Theobroma cacao L.) is a beverage crop with high economic value. Provision of natural PGR, namely red unions, which will be extracted. This study aimed to determine the effect of red onionxtract (Allium cepa L.) as a growth regulator on cocoa seeds. The research was conducted at a screen house at the Polytechnic of Agricultural Development Manokwari (Politeknik Pembangunan Pertanian Manokwari). The study method used was a completely randomized design (CRD), using five treatments and 20 replications, so the number of samples was 100 seeds. The treatments tested on cocoa plant seeds P0 (control), P1 (25% concentration of red onion extract), P2 (50% concentration of red onion extract), P3 (75% concentration of red onion extract), and P4 (100% concentration of red onion extract). The measurement variables included germination percentage, plant height, stem diameter, number of leaves, and seedling growth percentage. Differences in the concentration of growth regulators of red onion extract had a significantly different effect on seedling percentage and seed growth percentage and had a significant effect on plant height, stem diameter, and number of leaves of cocoa seedlings. Soaked cocoa seeds in red union extract with a concentration of 25% give optimum results.

Callus Induction of Dendrobium discolor Through The Thin Cell Layer (TCL) Technique Added with 2,4-Dichlorophenoxyaceticacid

Thin Cell Layer (TCL) is a plant propagation technique using thin explants (1-2 mm) through tissue culture. Callus induction is a crucial step in establishing plant regeneration. The type and concentration of growth regulators influence callus formation. 2.4 D is a growth regulator that is commonly used for callus induction in various plants. The research aims to determine the effectiveness of thin explants and 2,4-D in inducing Dendrobium discolor callus. The method used a Completely Randomized Design (CRD) with four treatments (1, 2, 3, and 4 mg/L). Adding 1, 2, and 3 mg/L 2,4-D can induce 100% calluses through the TCL explant. The callus has a friable to compact texture. The result showed that 2 mg/L 2,4-D was the best concentration for callus induction of D. discolor through TCL technique, indicating the greenish color, friable texture, and meristematic tissue.

Plant regeneration from embryogenic callus-derived from immature leaves of Momordica charantia L

Bitter melon (Momordica charantia L.), a widely cultivated food and medicinal plant native to the world's subtropics and tropics, is a Cucurbitaceae rich in carotenoids. However, the low seed germination frequency and progeny variability associated with the production of this plant have a substantial impact on its growth and yield. These constraints affect the availability and exploitation of this crop, especially the fruits, which are rich in secondary metabolites such as β-carotene and α-carotene. In vitro regeneration would help overcome the obstacle linked to the germination of this plant and increase its yield and utilization. A reproducible in vitro organogenesis protocol was established using bitter melon embryogenic callus derived from immature leaf explants of in vivo grown seedlings and in vitro plantlets. Regeneration via callus was conducted on MSB5 media augmented with different plant growth regulator concentrations. The maximum frequency of callus formation (95.09 %) was produced in MSB5 media incorporated with 1.2 mg L−1 NAA augmented with 0.5 mg L−1 TDZ. MSB5 medium with no growth regulators was observed to be the most suitable for the shoot and root formation from the callus, producing a significantly high shoot percentage of 90.91 % and 21.53 shoots per explants, and the highest rooting frequency and root number of 88.92 % and 6.23 roots per explant, respectively, from leaf-derived callus of in vitro plantlets. The elongated plantlets had grown to a significantly higher average height of 12.20 cm on media added with 0.75 mg L−1 GA3. This reproducible method for regenerating bitter melon plantlets could facilitate mass multiplication, conservation, and commercial field production.

The Effect of Shallot Extract as Natural Plant Growth Regulator and Cuttings Materials on The Growth of Water Apple (Syzygium aqueum l.) Stem Cuttings

The propagation of water apple by cuttings can meet the needs of the seeds in a short time and in large quantities. This study aimed to examine the effect of shallot extract as a natural plant growth regulator (PGR) and cutting material on the growth of water apple stem cuttings. The research was carried out from July to September 2022 at Greenhouse and the Physiology and Plant Breeding Laboratory, Faculty of Animal Husbandry and Agriculture, Diponegoro University. This study used a factorial Completely Randomized Design (CRD) with two factors. The first factor was the PGR concentration of shallots at four levels : K0 = 0%, K1 = 30%, K2 = 60%, and K3 = 90%. The second factor is the material of the water apple cuttings, with three levels: A1 = tip, A2 = middle, and A3 = base. The results of this study showed that the PGR concentration of shallots had significant effect on the age of shoots. The material of cuttings had a significant effect on age of shoots and the number of shoots. The natural growth regulator concentration of shallots 30% (v/v) in the shoots gave the best results for the growth of water apple cuttings

In vitro propagation of Valuable succulents: A review.

Succulents, popular group of terrestrial plants known for their unique appearance and ability to thrive in tropical arid environments. They can be vegetatively propagated through cuttings and seeds, also through in vitro propagation mass plantlets can be produced. Tissue culture raised plantlets have a number of advantages over traditional propagation methods, such as ability to produce large numbers of plants from a small amount of starting material, plants with free of pests, diseases and the ability to produce plants that are resistant to adverse environmental conditions. In this review, various results and discussion regarding different concentrations of plant growth regulators for multiplication, rooting of succulents with effective growth condition are described.

Effect of sodium hydrosulfide on physicochemical characteristics and melatonin mediated stress mitigation in bitter gourd (Momordica charantia L.) grown under lead stress

Lead (Pb) is a toxic heavy metal that contaminates soil and water resources after its discharge from different industries. This research study was held to check the consequence of Pb stress and NaHS growth regulator on physicochemical features of Mamodica charantia. A pot experiment was conducted to determine the effect of NaHS (1 Mm and 1.5 Mm) for the screening of M. charantia exposed to severe Pb (500 ppm,1000 ppm and 2000 ppm) stress. Our results depicted that maximum growth of plants was recorded at 1.5 mM concentration, while inhibition was observed in the plants of M. charantia at highest concentration of lead chloride (2000 ppm). Significant positive change in agronomical structures of bitter melon like leaf, root, shoot, branches, root length and shoot length recorded at high concentration of NaHS growth regulator (1.5 mM). In case of lead stress study, plants show reduction in growth and development due to production of ROS species. ROS species reduced the concentration of divalent cations such Zn2+, Mn2+, Mg2+, Ca2+, and Fe2+ by restricting the flow of water. Leaves of M. charantia had lessen carotenoid and chlorophylls and enhanced activity of chlorophyllase in leaves. Biological content of antioxidants (CAT and POD), Protein and Proline were found maximum in NaHS treated plants while minimum in stressed plants. Nutrient content of root and shoot (K, Na and Ca) and morphological characteristics also behaved differently on different concentration of NaHS and Pb stress. Here, we concluded that the application of NaHS under the exposure to Pb stress significantly improved physiological and biochemical structures of M. charantia.

The effect of plant growth regulators on micropropagation of Melientha suavis Pierre. and assessment of genetic fidelity of regenerants based on iPBS and SRAP markers

Abstract. Siringam T, Vanijajiva O. 2023. The effect of plant growth regulators on micropropagation of Melientha suavis Pierre. and assessment of genetic fidelity of regenerants based on iPBS and SRAP markers. Biodiversitas 24: 4628-4634. Melientha suavis Pierre, a significant deciduous edible plant species with high nutritional value and belonging to the Opiliaceae family, holds paramount importance for both agricultural and conservation purposes in its native Southeast Asia. However, the species faces challenges due to habitat fragmentation and negative anthropogenic impacts, leading to a decline in its population. This study aimed to explore the effects of various concentrations of plant growth regulators (cytokinin and auxin) on the shoot and root development of M. suavis, and to assess the genetic stability of micropropagated plants obtained from nodal explants. In vitro shoot regeneration and proliferation were conducted on Murashige and Skoog (MS) semi-solid medium, supplemented with 6-benzylaminopurine (BAP) or kinetin (Kn) at different doses. The optimal shoot length, shoot number, and leaf volume were observed in the modified MS medium with 1.0 mg/L BAP after an 8-week incubation period. Efforts to enhance the rooting of micropropagated shoots included the addition of auxins, such as ?-naphthalene acetic acid (NAA) or indole-3-butyric acid (IBA). However, the species exhibited recalcitrant behavior during the reproduction and rooting stages, as the rooting percentage did not correlate with the increase in auxin concentration. Interestingly, the highest root number and length were achieved in the MS medium without plant growth regulators after 8 weeks of incubation. To ensure genetic fidelity, regenerants were subjected to inter-primer binding site (iPBS) and sequence-related amplified polymorphism (SRAP) marker analysis. The results revealed no genetic variation between the micropropagated plants and the mother trees, confirming the production of genetically stable progeny. Overall, this protocol presents a promising alternative for profitable propagation and establishment of genetically constant progeny of M. suavis, with important implications for sustainable applications and germplasm conservation in Southeast Asia. Addressing the challenges faced by this valuable species through micropropagation and genetic fidelity assessment can contribute significantly to its preservation and utilization for agricultural and conservation initiatives.

In vitro Propagation and Genetic Assessment by using Markers in Potato (Solanum tuberosum L.) cv Kufri Ganga

The purpose of this study was to determine the best types and concentrations of plant growth regulators to use during different stages of micropropagation, as well as to assess the genetic fidelity of tissue culture raised plants of a cv Kufri Ganga using reliable markers. According to the data on shoot bud initiation, a combination of 0.25 mgl-1 BAP and 0.5 mgl-1 Kinetin results in the greatest number of shoots per explant (3.4±0.11) in the shortest amount of time (3.2±0.09) days. The maximum number of in vitro shoots per shootlet (4.7±0.15) was recorded when auxins were combined with cytokinin (MS medium + 0.25 mgl-1 BAP + 0.01 mgl-1 IAA) for shoot proliferation. Root initiation was observed in (2.2±0.10) days on MS medium fortified with 1.5 mgl-1 NAA. The maximum number of in vitro roots per shoot (14.2±0.31) was observed when MS media was fortified with 2.0 mgl-1 IBA. Molecular markers were used to assess genetic fidelity in plants grown in vitro. Monomorphic DNA banding patterns were found in all tissue culture raised plants and mother plants, indicating true to type planting material. This protocol for tissue culture propagation, as well as testing its genetic fidelity, may be beneficial for the successful application of better conservation of germplasm.

Shoot Cultures of Vitis vinifera (Vine Grape) Different Cultivars as a Promising Innovative Cosmetic Raw Material-Phytochemical Profiling, Antioxidant Potential, and Whitening Activity.

The primary purpose of this work was the initiation and optimization of shoot cultures of different Vitis vinifera L. cultivars: cv. Chardonnay, cv. Hibernal, cv. Riesling, cv. Johanniter, cv. Solaris, cv. Cabernet Cortis, and cv. Regent. Cultures were maintained on 30-day growth cycles using two media, Murashige and Skoog (MS) and Schenk and Hildebrandt (SH), with various concentrations of plant growth regulators. Tested media ('W1'-'W4') contained varying concentrations of 6-benzylaminopurine (BA) in addition to indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA). High performance liquid chromatography coupled with mass spectrometry (UPLC-MS) was used for metabolomic profiling. In all tested extracts, 45 compounds were identified (6 amino acids, 4 phenolic acids, 13 flavan-3-ols, 3 flavonols, and 19 stilbenoids). Principal component analysis (PCA) was performed to assess the influence of the genotype and medium on metabolic content. PCA showed that metabolic content was mainly influenced by genotype and to a lesser extent by medium composition. MS media variants induced the amino acid, procyanidin, and flavan-3-ol production. In addition, the antioxidant potential and anti-tyrosinase activity was measured spectrophotometrically. The studies on antioxidant activity clearly reveal very high efficiency in reducing free radicals in the tested extracts. The strongest tyrosinase inhibition capacity was proved for shoots cv. Hibernal cultured in SH medium and supplemented with NAA, with an inhibition of 17.50%. These studies show that in vitro cultures of V. vinifera cvs. can be proposed as an alternative source of plant material that can be potentially used in cosmetic industry.

Optimization of an Efficient Micropropagation Protocol and the Assessment of Genetic Fidelity of Solanum tuberosum L. cv Kufri Neelkanth

The aim of our experiment was to determine the optimal types and concentrations of plant growth regulators used during different stages of micropropagation and assess the genetic fidelity of tissue culture raised plants of a purple colored cv Kufri Neelkanth by using reliable markers. According to the shoot bud initiation data, it was found that utilizing a combination of 0.25 mg l-1 BAP and 0.5 mg l-1 Kinetin resulted in the greatest number of shoots per explant (3.5±0.12) within a timeframe of (3.3±0.08) days. The maximum number of in vitro shoots per shootlet (10.4±0.59) were recorded when auxins were used in combination with NAA (0.01 mg l-1 NAA) and Kinetin (0.25 mg l-1) for shoot proliferation. In vitro root initiation was observed in (2.1±0.07) days on MS medium fortified with 2.5 mg l-1 NAA. The maximum number of in vitro roots per shoots (13.0±0.55) were observed when MS media fortified with 2.5 mg l-1 IBA. Maximum 100% rooting was observed in all MS media supplemented with different concentrations of auxins. In vitro raised plants were assessed for genetic fidelity by using twenty RAPD primers (genetic markers). Out of twenty primers used only four primers produced amplification. DNA banding patterns of all tissue culture raised plants and mother plants were monomorphic showing true to type planting material. This protocol for tissue culture propagation along with testing its genetic fidelity could be useful for better conservation of germplasm and genetic transformation studies in potato.

The effect of various natural growth regulatory substances and the origin of stem cuttings on the vegetative growth of white jasmine (Jasminum sambac L.)

Jasmine is a native plant of the archipelago in Indonesia. Jasmine (Jasminum sambac L.) is a plant that has many ingredients, including as an ornamental plant, the chemical content of jasmine flowers, namely indole, benzyl, livalyacetat, is believed to be able to treat diseases such as headaches, shortness of breath, fever, excess breast milk and eye pain. Shallot extract is one of the natural growth regulators, namely the auxin group which contains indole and naphthalene compounds which function to stimulate the development of adventitious root meristems. Bean sprouts are one type of vegetable that is commonly used for consumption, easy to obtain, has economic value, and does not have toxic compounds. Bean sprout extract contains growth regulators, namely auxins and cytokinins. Based on the research results, it was found that the concentrations of various natural growth regulators were significantly different with respect to the number of shoots, shoot length, and number of leaves. The best treatment of natural growth regulators was found in shallot extract 50% (Z2). The type of cutting material differed significantly with the number of shoots, shoot length, and number of leaves. The best cutting material treatment was found in the middle of the branch (B2).

In vitro regeneration, phytochemical profiling and antioxidant activity in Ruta chalepensis plants established from alginate encapsulated synthetic seeds

We developed a highly efficient protocol for the short-term conservation and in vitro regeneration of Ruta chalepensis L. This involved encapsulating nodal segments (NS) in a sodium-alginate solution (3% w/v) and calcium chloride (100 mM) to produce synthetic seeds. The effect of various concentrations of plant growth regulators (PGRs) on the conversion of synthetic seeds into plantlets was examined and the best results, with a conversion rate of 96.8%, were obtained on Murashige and Skoog (MS) medium supplemented with 5.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA) after 12 weeks of culture. When stored at 4 °C for different durations (2-12 weeks), encapsulated and non-encapsulated NS displayed distinct conversion responses. Encapsulated NS stored at 4 °C for 10 weeks exhibited a regeneration rate of 72.7%, while non-encapsulated NS did not survive under the same conditions. The proliferated shootlets derived from cold-stored NS were successfully rooted in agar-solidified MS medium containing 0.5 μM indole-3-butyric acid (IBA) and successfully acclimatized to ex vitro conditions. UV-Visible spectroscopy (UV), gas chromatography-mass spectrometry (GC–MS), and high-performance liquid chromatography (HPLC) were used to identify bioactive chemicals in the aqueous methanolic extracts of the acclimatized plants and compared with the donor plant. The bioactive chemicals were found considerably higher in the acclimatized plants (Phenols 95.0 mg GAE/g DW, flavonoids 99.2 mg QE/g DW, tannins 38.7 mg TAE/g DW, hesperidin 94.5 µg/mg, and quercetin 25.4 µg/mg). The antioxidant potential was also evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and demonstrated high scavenging activity at a concentration of 800 µg/mL in the acclimatized plants (83.0%). This study represents the first successful demonstration of in vitro regeneration, phytochemical profile analysis, and assessment of antioxidant activity in R. chalepensis plants established from synthetic seeds following short-term cold storage.

The Effect of NAA and BA on In Vitro Callus Induction of Biduri (Calotropis gigantea)

A family of medicinal plants known as biduri (Calotropis gigantea) has a number of useful traits. Alkaloids, glycosides, phenolics, tannins, flavonoids, terpenes, and sterols are just a few of the chemical substances found in biduri. One method used to boost the generation of secondary metabolites is biduri callus induction by tissue culture. The goal of the study was to find the ideal growth regulator concentration for callus biduri induction. The study employed a completely randomized design (CRD), which was carried out three times and was set up in a factorial fashion with two treatment components and 25 treatment combinations. The growth regulator NAA and the growth regulator BA were the first and second factors, respectively. ANOVA was used to examine the data with a 5% level test. The Duncan Multiple Range Test (DMRT) was conducted at a 5% level if there was a significant difference. A compact, green callus texture was generated by applying one ppm NAA and three ppm BA. At the same time, a callus on the biduri could not have been treated without an NAA application. Further research can be done by subculture callus biduri when the callus is still fresh and green to produce secondary metabolites.

Multifaceted Role of Nanomaterials in Modulating In Vitro Seed Germination, Plant Morphogenesis, Metabolism and Genetic Engineering.

The agricultural practices of breeding, farm management and cultivation have improved production, to a great extent, in order to meet the food demands of a growing population. However, the newer challenges of climate change, global warming, and nutritional quality improvement will have to be addressed under a new scenario. Plant biotechnology has emerged as a reliable tool for enhancing crop yields by protecting plants against insect pests and metabolic engineering through the addition of new genes and, to some extent, nutritional quality improvement. Plant tissue culture techniques have provided ways for the accelerated clonal multiplication of selected varieties with the enhanced production of value-added plant products to increase modern agriculture. The in vitro propagation method has appeared as a pre-eminent approach for the escalated production of healthy plants in relatively shorter durations, also circumventing seasonal effects. However, there are various kinds of factors that directly or indirectly affect the efficiency of in vitro regeneration like the concentration and combination of growth regulators, variety/genotype of the mother plant, explant type, age of seedlings and other nutritional factors, and elicitors. Nanotechnology as one of the latest and most advanced approaches in the material sciences, and can be considered to be very promising for the improvement of crop production. Nanomaterials have various kinds of properties because of their small size, such as an enhanced contact surface area, increased reactivity, stability, chemical composition, etc., which can be employed in plant sciences to alter the potential and performance of plants to improve tissue culture practices. Implementing nanomaterials with in vitro production procedures has been demonstrated to increase the shoot multiplication potential, stress adaptation and yield of plant-based products. However, nanotoxicity and biosafety issues are limitations, but there is evidence that implies the promotion and further exploration of nanoparticles in agriculture production. The incorporation of properly designed nanoparticles with tissue culture programs in a controlled manner can be assumed as a new pathway for sustainable agriculture development. The present review enlists different studies in which treatment with various nanoparticles influenced the growth and biochemical responses of seed germination, as well as the in vitro morphogenesis of many crop species. In addition, many studies suggest that nanoparticles can be useful as elicitors for elevating levels of important secondary metabolites in in vitro cultures. Recent advancements in this field also depict the suitability of nanoparticles as a promising carrier for gene transfer, which show better efficiency than traditional Agrobacterium-mediated delivery. This review comprehensively highlights different in vitro studies that will aid in identifying research gaps and provide future directions for unexplored areas of research in important crop species.

Increasing Amounts of Secondary Metabolites and Medicinal Compounds in Callus Culture of Moringa Oleifera (Lam.) Using Abiotic Elicitors

Moringa oleifera Lam. tree has often been considered a medicinal and economical plant for its nutritional and medicinal properties. This study is intended to use plant tissue culture and elicitation technologies to increase secondary metabolites in the callus cultures of the M. oleifera. Young nodal segments were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of plant growth regulators (PGRs). A combination of 1.5 mg/l NAA plus 0.5 mg/l TDZ resulted in a high callusing rate (70%) with the largest size. Various concentrations of Salicylic acid (SA) and Methyl jasmonate (MeJa) were individually added to the growth medium to evaluate their impact on the biomass of callus cultures, DPPH antioxidant activity, and accumulation of some flavonoids. Results indicated that all elicitors doses reduced weights and growth index of callus. While DPPH assay recorded high significant values at high concentrations of SA and MeJa compared to other treatments. HPLC analysis revealed that every concentration of SA and MeJa used led to rising flavonoids content, including Baicalin, Rutin, Hesperidin, Rosmarinic acid, Quercitrin, Quercetin, and Kaempferol. It should be noted that 20 mg/l MeJa gave the highest flavonoid content compared to the control and other treatments. In conclusion, the in vitro elicitation by SA or MeJa can improve beneficial biological substances like flavonoids and other antioxidant compounds in Moringa callus cultures.

Application of NAA and BA to Calotropis gigantea (L.) W.T. Aiton in vitro

Crown flower (Calotropis gigantea (L.)) is a weed plant potentially to be used as raw material for textile fibers yet the utilization is not optimal. Efforts to optimize the utilization of C. gigantea are needed. This research aimed to determine the appropriate concentration of plant growth regulators, including NAA (Naphthalene Acetic Acid) and BA (Benzyl Adenine) to stimulate optimal growth of C. gigantea to be used as the basis for in vitro propagation of C. gigantea. The research was arranged in a completely randomized design (CRD). Explants were planted on Murashige and Skoog (MS) medium with various concentrations of NAA (0; 0.5; 1; 1.5; and 2 ppm) and BA (0; 1; 2; 3; and 4 ppm). Based on the research result, the addition of 3 ppm BA followed by increasing the concentration of NAA to 1.5 ppm could increase the number of C. gigantea shoots, and giving 1 ppm BA followed by adding 1.5 to 2 ppm NAA increased shoot height. The best plantlet response was found in media with 0 ppm NAA and 4 ppm BA, which could produce an optimal shoot height of 5.9 cm and a large number of shoots of 5.67 shoots. Increasing NAA concentration retarded root formation and reduced the root length and number of leaves, while the medium without NAA gave the earliest root emergence of 11 days after planting, the number of leaves of 20 strands, and the longest root of 10.9 cm. Applying a single substance BA did not accelerate all the variables observed.

IN VITRO MORPHOGENESIS OF GYPSOPHILA: RESPONSE TO EXOGENOUS PHYTOHORMONES

IN VITRO MORPHOGENESIS OF GYPSOPHILA: RESPONSE TO EXOGENOUS PHYTOHORMONES

Shoot regeneration in Nepenthes mirabilis as affected by flurprimidol and GA3 application

Abstract. Dwiati M, Widodo P, Susanto AH. 2023. Shoot regeneration in Nepenthes mirabilis as affected by flurprimidol and GA3 application. Biodiversitas 24: 4168-4174. Nepenthes mirabilis (Lour.) Druce. is a pitcher plant species at a higher risk of extinction mainly due to overexploitation. Previous investigations have shown that in vitro micropropagation can be used as an approach for ex-situ conservation of N. mirabilis. Therefore, this study aimed to assess the new shoot formation of N. mirabilis under in vitro conditions stimulated with the application of flurprimidol and GA3. The three concentrations of flurprimidol and GA3 used as treatments were arranged factorially in a Randomized Complete Block Design (RCBD), with internodes 2, 3, 4, and 5 serving as blocks. Based on the results, F1G0 (flurprimidol of 1 mgL-1 without GA3) had the highest shoot formation, which was not accompanied by sufficiently high contents of chlorophyll, leading to improper shoot development. Despite the smaller number of newly formed shoots in F2G2 (flurprimidol of 2 mgL-1 and GA3 of 2 mgL-1), significantly higher chlorophyll contents were observed, enabling better development of shoot. Leaf length and shoot diameter also showed similar results under F2G2, indicating that higher concentrations of exogenous GA3 were necessary for better shoot development. This indicated that appropriate concentrations of both plant growth regulators should be applied for micropropagation of N. mirabilis in support of ex-situ conservation.