Shoot Multiplication Research Articles

Establishment of a Highly Efficient Micropropagation System of Aquilaria crassna Pierre ex Lecomte

Aquilaria crassna Pierre ex Lecomte is a principal species renowned for its production of agarwood. However, the active components of agarwood are not universally in compliance with the standards set by the Chinese Pharmacopoeia. We have identified an elite A. crassna tree with agarotetrol and alcohol extract levels that exceed these standards and have successfully established a stable in vitro micropropagation system using stem segments from this elite tree. The effects of auxins and minerals on axillary-bud induction, shoot multiplication, and rooting were investigated. The most effective medium for axillary-bud induction was a half-strength Murashige and Skoog (1/2MS) medium supplemented with 0.50 mg/L 6-benzylaminopurine (6-BA), achieving an induction rate of 53.33% with minimal hyperhydricity. The optimal shoot proliferation medium was an MS medium with 0.40 mg/L 6-BA, yielding a propagation coefficient of 2.96 without hyperhydricity. The best rooting medium comprised quarter-strength MS (1/4MS) macroelements and 1/2MS microelements with 0.10 mg/L naphthaleneacetic acid (NAA), resulting in an 82.54% rooting rate. Substrate effects on transplant survival and growth were also evaluated, and peat soil was identified as the best substrate, achieving a survival rate of 96.67%. This study introduces a straightforward and efficient in vitro micropropagation system utilizing mature A. crassna as explants. It holds significant importance for the consistent production of agarwood that complies with the standards of the Chinese Pharmacopoeia and provides a model for the targeted breeding of medicinal plants.

Micropropagation of Selected Hybrid Tomato (Solanum lycopersicum L.) Varieties Using Shoot Tip Culture

Tomato (Solanum lycopersicum L.) is one of the most important vegetable crops in the world. The use of hybrid varieties is a great option to increase its production but using F2 seed leads to lack of uniformity as a result of segregation. Therefore mass propagation using tissue culture could help to solve these problems. The objective of this study was to develop an optimum micropropagation protocol for Valouro, Uwezo and Shelter hybrid tomato varieties using shoot tip culture. Three successive experiments: shoot initiation; shoot multiplication and root inductions were conducted. Different concentrations of BAP (0.0, 0.25, 0.5, 0.75, 1.0 and 1.25 mg/l) and (0.0, 0.5, 1.0, 1.5, 2.0, 2.5 and 3 mg/l) were used for shoot initiation and shoot multiplication, respectively. While different concentrations of IBA (0.0, 0.25, 0.5, 0.75, 1.0, 1.25 and 1.5 mg/l) were used for root induction. The three separate experiments were laid out as factorial arranged in completely randomized design. The result of shoot initiation showed that the interaction of BAP*Varieties highly significant (P<0.01) effect. In the multiplication experiment, all parameters showed highly significant difference (P<0.01). In rooting experiment the interaction effect of IBA*Variety showed highly significant difference for the studied parameters. In conclusion MS medium without growth regulators was the optimum for shoot initiation. For shoot multiplication experiment 2mg/l, 0.5mg/l and 1.5mg/l of BAP was the optimum concentration for number of shoots per explant, shoot length, and leaf number respectively. For in vitro rooting MS medium containing 0.5 mg/l IBA was optimum for all varieties.

Development of efficient and scalable regeneration tissue culture method for Cannabis sativa

Large scale production of uniform disease-free plants is crucial for Cannabis sativa biotechnology. Existing micropropagation protocols rely heavily on shoot multiplication from existing meristems via direct organogenesis. Such protocols do not allow multiplication of plant material through continuous sub-culturing. Protocols that use indirect regeneration are usually not efficient enough and have very low multiplication rates. In the present study, an efficient protocol that uses a combination of direct organogenesis and callogenesis to induce multiple shoot development cultures is developed. Callogenesis was induced from various explants cultured on the media having various combinations of thidiazuron (TDZ) and naphthaleneacetic acid (NAA); best callogenesis and shoot regeneration was achieved from hypocotyl explants cultured on TDZ 0.4mgl-1 NAA 0.2mgl-1. Hypocotyls with cotyledonary node and shoot apical meristem were significantly better for shoot regeneration than explants without it. Shoots obtained from multiple shoot cultures were successfully rooted and then acclimatized under greenhouse conditions to develop into adult cannabis plants.

Silicon nanoparticle(s) induced morpho-anatomical traits and improved micropropagation of white fleshed dragon fruit [Selenicereus undatus (Haworth)]

Silicon nanoparticles (SiNPs) improve plant growth and yield by enhancing nutrient uptake and stress tolerance. These nanoparticles boost plant defense mechanisms against pathogens and environmental challenges. In this study, seedlings-derived explants of white-fleshed dragon fruit (Selenicereus undatus) were used for differentiation and high-frequency shoot regeneration on Murashige and Skoog (MS) medium with 6-benzylaminopurine (BAP, 0.5 mg L−1) and α-naphthalene acetic acid (NAA, 0.25 mg L−1). The complementation of 3.0 mg L−1 SiNPs in the optimized nutrient medium enhanced the shoot multiplication rate to 23.0 cladodes per explants. The regenerated cladodes were structurally well-suited with thick cuticles, well-developed epidermis, hypodermis, aquiferous cortex, and collateral vascular bundles compared to the control and other concentrations of SiNPs tested. SiNPs promoted the development of 3-5 aerial and adventitious roots per cladode enabling preferred direct hardening of cloned plants on soilrite ex vitro. The morphological transformations improved by SiNPs, including the formation of robust cladode and spines, played a pivotal role in achieving a high rate of acclimatization success (96%) in the field. The incorporation of SiNPs in vitro at the shoot amplification stage of micropropagation of S. undatus induced morphogenesis and structural changes, and promoted qualitative and quantitative improvements in the regenerated cladodes which helped in enhanced survival of plants in vivo.

Micropropagation, micromorphological evaluation and genetic homogeneity validation of Cucumis collosus (Rottl.) Cogn.: A wild progenitor of melon

Cucumis collosus (Rottl.) Cogn., a wild ancestor of melon cultivars, is an underutilized cucurbit of arid horticulture having nutraceutical and therapeutic significance. C. collosus has great potential for enhancing the genetic base of cultivated melon and other cucurbits. This investigation is the first report on a robust and reliable micropropagation system for C. collosus (var. AHK-119) using nodal explants, followed by micromorphological and genetic stability assessment. More than 95% explants showed axillary shoot-bud induction on MS (Murashige and Skoog, 1962) medium containing 0.5 mg L−1 BAP and developed a maximum of 3.2 ± 0.6 shoots measuring 3.2 ± 0.1 cm length. After evaluating the various parameters, the best shoot multiplication (Shoot Number 34.9 ± 1.7, Shoot Length 6.7 ± 0.9 cm) was achieved on MS medium having half-strength of nitrates (825 mg L−1 NH4NO3 and 950 mg L−1 KNO3) + 0.25 mg L−1 6-benzylaminopurine (BAP) + 0.1 mg L−1 Kinetin (Kin) + 0.1 mg L−1 Indole-3-acetic acid (IAA) + additives (50 mg L−1 ascorbic acid and 25 mg L−1 each of adenine sulfate, L-arginine and citric acid), after 4 weeks. Micro-shoots were rooted using three approaches: In Vitro Continuous Treatment (IVCT), In Vitro Pulse-Treatment (IVPT) and Ex Vitro Pulse Treatment (EVPT). Of these, EVPT approach proved to be most successful in terms of rooting response (80%), root number (6.4 ± 0.9) and root length (5.7 ± 0.7 cm). The foliar micro-morphology analysis of in vitro leaves (IVL), ex vitro leaves (EVL) and mother plant leaves (MPL) demonstrated notable adaptive alterations during the transitional stages and produced structurally stable plants similar to the mother plant. Furthermore, the validation of genetic homogeneity using Start Codon Targeted (SCoT) and Inter Simple Sequence Repeats (ISSR) markers revealed monomorphic banding patterns among the micropropagated and the mother plant. The discussed method can be applied for large-scale commercial production of C. collosus for arid horticulture, urban/vertical farming, breeding purposes and improvement of other cucurbits.

EFFECT OF THE CONCENTRATION OF THE PLANT GROWTH REGULATORS ON IN VITRO SHOOT GROWTH OF BLACKBERRY

Berry fruit species, such as Ribes nigrum, are highly esteemed in numerous countries for their biological and economic value, being regarded as important species of small fruits. Ribes nigrum, indigenous to northern Europe, and northern and central Asia. Conserving species like blackcurrant, as highlighted in the International Scientific reports, is crucial due to their potential health benefits and contributions to maintaining vascular health, especially in postmenopausal populations. The discovery that blackcurrant extract, rich in phytoestrogens, shows promise in preventing elastin degradation and pathological vascular remodeling underscores the importance of preserving this species. In Kazakhstan, the preservation of Ribes nigrum includes various factors, such as ecological significance, local economic value through agriculture, and conservation priorities since blackcurrant plays a significant role in the country's biodiversity and traditional uses. The potential of in vitro propagation for rapid mass production of valuable species is substantial. Our study addresses biodiversity conservation by employing this approach. Specifically, we aimed to investigate the impact of nutrient media and growth regulators on shoot multiplication of Ribes nigrum. Our findings reveal that the optimal concentration of plant growth regulators (PGR) significantly influences both the number and length of shoots. Notably, the highest multiplication rate for Ribes nigrum was achieved on MS with a combination of BAP at 0.5 μ/L and GA 0.5 μ/L. In conclusion, our study elucidates the effects of manipulating PGR concentrations in culture media on Ribes nigrum. We recommend optimizing these concentrations to enhance the growth of Ribes nigrum for potential reintroduction efforts and biodiversity conservation. The research is being conducted as part of program BR21882166, which focuses on the scientific and practical aspects of reproducing, conserving, and utilizing fruit and berry plants from the natural flora of Western, Eastern, Central, and Northern Kazakhstan to ensure food security. This program is financially supported by the Ministry of Science and Higher Education of the Republic of Kazakhstan for the period 2023-2025.

Influences of Thidiazuron Concentrations and Exposure Duration on Axillary Shoot Proliferation of Faba Bean [Vicia faba (L.)] Genotypes

Background: Faba bean is one of the most important feed grain legume crops in the world. However, this crop is a recalcitrant species for in vitro regeneration. Methods: In this study, in vitro shoot proliferation of ten faba bean genotypes, that are classified according to their seed size into small, medium and large, were investigated in response to different light regimes and thidiazuron (TDZ) treatments. Result: Incubation under different light regimes, explant type as well as genotypes influenced seedling growth and axillary shoot multiplication of faba bean. The explants with single cotyledon and incubated under continuous darkness for 6 days resulted higher number of shoot than nodal explants. Medium size seeded genotype (ILB4347) produced the highest shoot and root length and shoot proliferation as compared with large (Luz) or small (Triple White) seeded genotypes. The effects of TDZ concentrations (2-10 mM/L) and exposure duration (10 and 20 days) on axillary shoot multiplication were investigated for the ten faba bean using cotyledonary explants. The overall mean of TDZ time exposure showed that treatment of 20 days exposure had higher number of shoot compare to 10 days exposure treatment with 3.77 shoot per explant. On the basis of genotype used, the three most responsive genotype in tissue culture were medium sized seeds genotypes (ILB4347, Hassawi 2 and Misr 3 with 4.76, 4.40 and 4.15 shoot per explant, respectively). ILB4347 were the most responsive genotype in tissue culture. Treatment of 8 mM TDZ after 20 days followed by 2 weeks on MS medium without PGRs generated the highest proliferation with 6.63 shoot per explant. These findings contribute towards effective in vitro propagation of faba bean.

Effect of Different Concentrations and Combinations of Benzyl Aminopurine and Indole-3-Butyric Acid on Micropropagation of <i>Vanilla Planifolia</i>

Micropropagation of explants in vitro has potency to address propagule demands for promoting large-scale vanilla production. Plant growth regulators (i.e., cytokinin, auxin) are necessary for the plant micropropagation success. Objective of this study is to determine the shoot multiplication and root development of Vanilla planifolia under the influence of different concentrations and combinations of BA and IBA for micropropagation. Sixty stem nodal segments of Vanilla planifolia were cultured on MS medium supplemented with IBA (0, 0.5, or 2.0 mg/L) and combined with BA (0, 0.5, 1.0, or 2.0 mg/L). Shoot multiplication and root induction were measured after 60 days of culture. The results show that the MS medium with 1 mg/L IBA hindered shoot growth, while the medium containing 1 mg/L BA yielded the highest number or weight of shoots per explant. For the root development, supplementing the medium with 0.5 mg/L or 1 mg/L IBA improved root length or number of roots per explant, respectively. This research establishes a valuable approach for vanilla micropropagation by utilising low concentrations of plant growth regulators and a rapid protocol. This paves the way for significant advancements in large-scale commercial production.

In vitro micropropagation and mass multiplication of Stevia rebaudiana Bertoni

Stevia rebaudiana Bertoni, an ancient perennial herb produces steviol glycosides including stevioside and rebaudioside-A that are valuable as low calorie sweeteners, about 300-400 times sweeter than saccharose. Conventional propagation methods are not produce adequate planting material. Micro propagation is an imperative technology that could be extensively exploited to meet the growing demands of elite planting material for commercially cultivated. The prime objective of this study was to established standard protocol for in vitro regeneration and mass multiplication in Stevia. In present investigation, different concentrations of Auxin and Cytokinin were tried for optimization of shoot initiation, shoot multiplication, root initiation and callus induction of Stevia from different explant. BAP and IBA (1.0+0.5) mg/l, IBA (2.0) mg/l shows promising results of shoot multiplication and root initiation respectively.

Micropropagation of Zuccagnia punctata, an argentine medicinal plant, by means of “in vitro” technology to promove its conservation and the sustainable production of bioactive compounds

Zuccagnia punctata Cav. (“jarilla poposa”) is a monotypic argentine native medicinal plant used by indigenous communities for the treatment of several pathologies. The pharmacological properties have been scientifically validated and its potential use in phytocosmetic and pharmaceutic industry as antiaged, hypoglycemic, anti-inflammatory, antitumoral, antioxidant, antibacterial and antimicotic has been previously demonstrated. However, the species has a low propagation rate in natural conditions and has been included in the preliminary red list of endangered plants in category 3. Therefore, it is necessary to develop methodologies leading to its conservation and propagation to achieve its sustainable use. The objective of this work is to perform a micropropagation protocol for Z. punctata species for the sustainable use and conservation of this species. Furthermore, the study evaluated also the impact of “in vitro” propagation on the content of bioactive compounds of ethanolic extracts obtained from Z. punctata plants obtained “in vitro” with collected wild plants. Seeds have been used as mother material. The seed germination to obtain explants were performed on Murashige and Skoog medium (MS) without plant growth regulators (PGRs) containing sucrose, agar, and ascorbic acid. The same medium was employed for the multiplication of shoots. “In vitro” rooting was achieved in liquid and semisolid medium, and a higher yield was obtained in the latter. The plants were transferred into commercial substrate and acclimatized in a greenhouse. The micropropagated plants (MP) had a high content of total phenolic compounds similar to those obtained from wild adult plants (WAP) collected in the Monte Desert. The flavonoid content of MP was higher than that of WAP. The main chalcones identified in WAP, 2′,4′ dihydroxychalcone (DHC) and 2′,4′-dihydroxy-3′-methoxychalcone (DHMC) were also present in the MP with a rate 1/1, and 1/3 in WAP. The extracts obtained from the MP showed antioxidant, antiinflammatory and antimicrobial activity, similar to WAP. The “in vitro” propagation and acclimatization of Z. punctata constitutes a fundamental tool to achieve large-scale multiplication of this species in the future and the sustainable production of bioactive compounds with commercial applications. Reimplantation in the natural habitat could prevent the loss of this medicinal genetic resource.

High-Efficiency In Vitro Root Induction in Pear Microshoots (Pyrus spp.).

Extensive research has been conducted on the in vitro mass propagation of pear (Pyrus spp.) trees through vegetative propagation, demonstrating high efficiency in shoot multiplication across various pear species. However, the low in vitro rooting rates remain a significant barrier to the practical application and commercialization of mass propagation. This study aims to determine the favorable conditions for inducing root formation in the in vitro microshoots of Pyrus genotypes. The base of the microshoots was exposed to a high concentration (2 mg L-1) of auxins (a combination of IBA and NAA) for initial root induction at the moment when callus formation begins. The microshoots were then transferred to an R1 medium (1/2 MS with 30 g L-1 sucrose without PGRs) to promote root development. This method successfully induced rooting in three European pear varieties, one Asian pear variety, and a European-Asian hybrid, resulting in rooting rates of 66.7%, 87.2%, and 100% for the European pear (P. communis), 60% for the Asian pear (P. pyrifolia), and 83.3% for the hybrid pear (P. pyrifolia × P. communis) with an average of 25 days. In contrast, the control group (MS medium) exhibited rooting rates of 0-13.3% after 60 days of culture. These findings will enhance in vitro root induction for various pear varieties and support the mass propagation and acclimatization of pear. The in vitro root induction method developed in this study has the potential for global commercial application in pear cultivation.

Multiplication of apical and axillary shoots of coffee arabica using various types of cytokinins

Abstract In-vitro shoot multiplication presents a promising avenue for replicating the genetic characteristics of parent plants, commonly utilized in various plant biotechnological techniques such as mutation replication, protoplast fusion, anther culture, and genetic engineering. The primary aim of this research was to assess the impact of different cytokinins on the proliferation of apical and axillary shoots in Arabica coffee multiplication. A Completely Randomized Factorial Design was employed, considering two factors: explant source and cytokinin type. Explants were sourced from both apical and axillary branches of AS2K Arabica coffee plantlets. Three cytokinins, namely 6-Benzyl adenine purine (BAP), isopentenyl adenine (2-IP), and kinetin, were incorporated into the Murashige and Skoog (MS) 1962 basal medium at concentrations of 0, 1, 2, and 3 mg/l. The findings revealed significant effects of all three cytokinins on shoot number, plantlet height, and leaf growth, albeit without influencing root development. Notably, axillary shoot-derived explants demonstrated a more pronounced effect on shoot count compared to those from apical shoots. Among the cytokinin treatments assessed, BAP at a concentration of 2 mg/l in the MS medium yielded optimal results for propagating the AS2K variety of Arabica coffee, thus representing the recommended medium for this purpose.

Propagasi In Vitro Kaliandra Merah (Calliandra calothyrsus Meisn.) II: Induksi Perakaran Tunas dan Aklimatisasi Planlet

Red calliandra is one of woody plants that has been extensively developed as biofuel. Tissue culture techniques are employed to produce a large and uniform quantity of seedlings in a relatively short period. Previous stages of in vitro initiation and multiplication of shoot from calliandra explants have shown promising results. The shoots that were multiplied by in vitro technique need to be induced for rooting process to produce plantlets. These plantlets are then prepared for the subsequent acclimatization process. This research was conducted in two stages; root induction of shoots using a modification of MS half-strength media (MS ½) supplemented with 1-4 mg/L 1-naphthaleneacetic acid (NAA), and acclimatization of the plantlets into soil media. The results shown that MS ½ medium and media supplemented with 1 mg/L NAA were suitable for root induction, particularly to induce the root length. Furthermore, MS ½ media supplemented with 1-2 mg/L NAA stimulates the increment of plantlet height and leaves numbers. Plantlets which were transferred into soil exhibit robust growth during 3-weeks of acclimatization process, up to 100% plantlets survived during this stage. These results underscore the potential of tissue culture techniques in providing large scale calliandra seedlings production for future biofuel development.

Development of a Suitable in vitro Regeneration Protocol for Rahangala Pear (Pyrus pyrifolia L.) in Sri Lanka

The limitation of planting material is the major drawback to expanding pear cultivation in Sri Lanka. Hence, an in vitro protocol was developed to produce planting material for the pear variety of Rahangala. Pear shoots were used as explants and surface sterilization of explants was done using 0.5% and 1% (w/v) AgNO3 for 15 min, 5% and 10% (v/v) NaOCl for 5, 10 and 15 min. In vitro shoot multiplication and root induction were found to be suitable in full and half strengths of MS medium respectively supplemented with various concentrations of BAP (1, 1.5, 2, 2.5, 3 mg/l), IBA (0, 0.5, 1, 2, 3, 4 mg/l) and NAA (0.1 mg/l). Acclimatization was conducted in pots containing different proportions of topsoil, compost, coir dust and sand. Completely Randomized Design (CRD) was used as an experimental design for this study. During sterilization of explants, the survival rate of 86.6% was recorded after 3 weeks where the treatment was done with 1% AgNO3, and 10% NaOCl for 10 min. BAP (3.0 mg/l) produced the best shoot multiplication (1.46 ± 0.2). Significantly (p<0.05), the highest mean number of roots (1.93 ± 0.98), average length (60.6 mm), and rooting percentage (68%) were observed in ½ MS medium with 4.0 mg/l IBA. After five weeks of transplantation in pots, 100% of plantlets survived in topsoil, compost, coir dust, and sand in a ratio of 1:1:1:1 Plant Tissue Cult. & Biotech. 34(1): 25-33, 2024 (June)

The effect of zinc oxide nanoparticles on the growth and development of Stevia plants cultured in vitro

Stevia (Stevia rebaudiana Bertoni) is an essential herbal plant used as a sweetener. The demand for stevia is growing due to its low caloric and medicinal value, hence the need for a more thorough investigation of its nutritional and biological properties. Nanoparticles of metal oxides have been found to have broad applications in agriculture for the stimulation of plant growth and development. The study aimed to assess the effect of various zinc oxide nanoparticles (ZnONPs) concentrations on stevia plants’ quantitative and qualitative traits obtained in in vitro cultures. Micropropagation of two stevia varieties, Candy and Morita, was carried out using explants of shoot tips placed on MS medium supplemented with 1.0 mg dm–3 BA and 0.1 mg dm–3 IBA and with ZnONPs at concentrations of 0 (control), 10, 20, 30 and 40 mg dm–3. The obtained results indicated that high concentrations of ZnONPs stimulated the propagation of shoots. On the other hand, they negatively influenced shoot length, root number and length, and the fresh weight of the plantlets. The presence of zinc oxide nanoparticles in the medium increased the potassium, calcium, magnesium, and zinc content while decreasing the sodium and iron content in the regenerated stevia plantlets. The total phenolic content in the Candy variety was higher in the treatments with ZnONPs than in the control plants, while it was varied in the Morita variety. In both varieties, total antioxidant content measured by the ABTS method showed significantly higher in the treatments with 20–30 mg dm–3 ZnONPs than in the control. The content of chlorophyll a, chlorophyll b and chlorophyll a + b in the Morita variety was higher in the treatments with 10 and 20 mg dm–3 ZnONPs than in the control. On the other hand, high concentrations of ZnONPs negatively affected the content of carotenoids in both varieties. The study showed that stevia plants obtained in in vitro cultures on control media and media containing ZnONPs had a high content of valuable minerals, phytocompounds with antioxidant properties, and photosynthetic pigments.

ESTABLISHMENT OF AN EFFICIENT MICROPROPAGATION PROTOCOLS FOR THREE KENAF (HIBISCUS CANNABINUS L.) CULTIVARS

This study was aimed to establish an efficient micropropagation protocol for three popular kenaf (Hibiscus cannabinus L.) cultivars (V36, HF992 and KB6) using different types of plant growth regulators (PGR’s) on Murashige and Skoog medium (MS). The shoot tip, cotyledon and hypocotyl isolated from 2 weeks old seedlings were used as explants. MS medium fortified with different types and concentrations of cytokinins and auxin were evaluated for establishment callus formation, shoot multiplication and rooting. Among different PGR’s used, 5.0 mg/l 2iP + 0.1 mg/l NAA was the best concentration for direct shoot regeneration from cotyledon with shooting percentage of 55% in HF992 cultivar. In multiple shoot induction experiment, 1.0 mg/l TDZ + 0.1 mg/l NAA mg/l concentration has induced the highest (10.4) shoots/explant. Moreover, indirect shoot regeneration was established from callus which was induced from cotyledon, maximum callus induction percentage (88.75%) was recorded on 0.5 mg/l TDZ and 1.0 mg/l NAA in V36 variety. While, for shoot elongation and rooting, MS media half strength supplemented with 1.0 mg/l kinetin + 1.0 mg/l IBA was found to be an optimum concentration.

In vitro morphogenesis and micro-morpho-anatomical developments in Moringa concanensis Nimmo.: An endemic tree of Indian sub-continent

Moringa concanensis Nimmo. is a regionally important and underutilized medicinal tree of the Indian subcontinent. An in vitro morphogenesis protocol for Moringa concanensis has been developed using seedling-derived explants. The cotyledonary node explants were found to be the most suitable exhibiting cent-percent bud break with each regenerating 2.33±0.76 shoots of 1.32±0.69 cm length within 15 days on 1.0 mg/L 6-benzyl aminopurine (BAP) supplemented modified Murashige and Skoog Medium (MMS) media. The rate of shoot multiplication increased up to 10.17±2.54 shoots/inoculum by sub-culturing of regenerated shoots on 0.5 mg/L BAP along with 0.1 mg/L Indole acetic acid (IAA). The in vitro cloned shoots were rooted using both in vitro and ex-vitro methods by applications of root-inducing auxins, indole butyric acid (IBA), and naphthalene acetic acid (NAA). IBA proved to be more effective and induced roots in all in vitro raised shoots. It produced 17.72±4.83 roots/shoot with 2.12±0.88 cm length on 1/4th strength MMS media fortified with 1.0 mg/L of IBA. Ex vitro rooted plantlets with concurrent acclimatization were achieved by pre-treating shoot bases with IBA at 250 ppm for 5 min. Comparative micro-morpho-anatomical evaluation of leaf and stem revealed differentiation of non-glandular trichomes, anomocytic stomata, and vascular tissue development under applied in vitro conditions, which suggests the survivability of plantlets. Sixty percent of the in vitro regenerated plants survived in the field. It is the foremost report on in vitro morphogenesis of Moringa concanensis protocol using cotyledonary nodal explant and ex vitro rooting method. The tissue culture methods defined and developed can be used for large-scale multiplications of Moringa concanensis as non-conventional approaches.

Response of Shoots from Porang Leaf Bulbs to Cytokinins and IAA in Shoot Multiplication In Vitro

The growth regulators in media have the potential to significantly influence the process of in vitro plant regeneration. The response of explants in forming shoots also depends on the type and the combination of growth regulators. This research aimed to identify the most optimal type and concentration of cytokinin for the multiplication of in vitro porang shoots. Additionally, it determined whether the addition of cytokinins should be accompanied by auxin/IAA for porang shoot multiplication. To achieve these objectives, a completely randomized design with six replications was employed, followed by a 5% BNJ. The tested treatments consisted of various combinations of growth regulators. Several observations were recorded, including the speed of explants forming prospective shoots/shoots (days), the age of the explants forming prospective shoots/shoots, the number of prospective shoots and shoots produced per explant, and shoot height (cm). The results showed that 1) Tdz 2 and IAA 0.5 mg.l-1 formed the highest shoots candidates and shoots in porang multiplication in vitro, and 2) the addition of BAP and Tdz 2 mg.l-1 should be combined with IAA 0.5 mg.l-1, while BAP and Tdz 3 mg.l-1 did not require IAA.

In vitro shoot multiplication of Haplophyllum virgatum and flavonoid elicitation in proliferated shoots by methyl jasmonate

In vitro shoot multiplication of Haplophyllum virgatum and flavonoid elicitation in proliferated shoots by methyl jasmonate

EFFECT OF BAP ALONG WITH 2.4, D ON THE INDUCTION OF SHOOTS FORMATION ON POTATO CULTIVAR ARIZONA IN VITRO

This examination delves into the many-sided transaction between two key development controllers, BAP (6-benzylaminopurine) and 2,4-Dichlorophenoxyacetic acid (2,4-D), exploring their aggregate effect on shoot induction in the in vitro development of the esteemed potato cultivar Arizona. The study is fastidiously designed to unwind the nuanced effect of these controllers on shoot commencement inside a controlled lab setting.Potato micropropagation stands as a fundamental feature of horticultural advancement, and the feeling of proficient shoot development fills in as a crucial stage in this cycle. The exceptional development attributes of Cultivar Arizona provide an ideal stage for assessing the synergistic impacts of BAP and 2,4-D on the commencement and multiplication of shoots. By carefully noticing and measuring the reaction of potato explants to fluctuating focuses and mixes of BAP and 2,4-D, this study expects to discern ideal conditions conducive to enhanced shoot development. Through detailed analysis and observation, the research seeks to contribute valuable insights into refining in vitro cultivation methodologies for potato propagation, aiming at elevating efficiency and yield.