Showing papers on "Murashige and Skoog medium published in 2022"
TL;DR: In this article , a reliable protocol for direct in vitro propagation using nodal shoot meristem of Thunbergia coccinea as explant was standardized, and the best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 1.32) in cm.
Abstract: Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR's) and concentrations tested. The aforesaid PGR's combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542-1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants.
16 citations
TL;DR: In this paper , the applicability of a temporary immersion system (TIS) bioreactor for plantlet regeneration was studied for chrysanthemum, strawberry, and Cnidium officinale.
Abstract: A temporary immersion system (TIS) bioreactor has been used as an efficient and cost-effective method for the in vitro propagation of many plant species. In the current study, the applicability of a TIS bioreactor for plantlet regeneration Chrysanthemum morifolium Ramat., Fragaria × ananassa Duch., and Cnidium officinale Makino was studied. Shoot length, a number of leaves per regenerated shoot, fresh, and dry biomass of plantlets were optimal with the TIS compared to semi-solid and liquid immersion cultures. The leaf area in cryshanthmum, strawberry, and C. afficinale were 2.87 cm2, 3.51 cm2, and 1.43 cm2, respectively, in the plants regenerated by TIS. The photosynthetic pigments were highest in strawberry plants grown in TIS bioreactor culture, and there was no significant difference between semi-solid and liquid culture while the highest values were obtained in C. officinale maintained in semi-solid culture. The chrysanthemum and strawberry plants showed a 100% acclimatization rate in all culture systems. C. officinale plants showed the highest survival rate at 96.9%, which were regenerated in the TIS. TIS bioreactor culture, thus, provides a convenient method that could be adopted for commercial in vitro propagation of chrysanthemum, strawberry and C. officinale plants.
12 citations
10 citations
TL;DR: In this paper , the genome size estimation of in vitro regenerated Tylophora indica was made by flow cytometric method using start codon targeted (SCoT) molecular marker.
Abstract: Tylophora indica (Burm.f.) Merrill. is a pharmacologically important plant, popular for alkaloidal and non-alkaloidal richness. Large scale propagation of T. indica is difficult in the wild as the seeds are small and the frequency of germination is very poor. In the present study, the genome size estimation of in vitro regenerated (indirect, direct and somatic embryo mediated) T. indica was made by flow cytometric method. Clonal fidelity of the regenerants was assessed using a start codon targeted (SCoT) molecular marker. Initially, the explants were inoculated on Murashige and Skoog basal medium supplemented with various concentrations of plant growth regulators like 2,4-dichlorophenoxy acetic acid (2,4-D), Kinetin, 6-benzyl amino purine (BAP) and 1-naphthalene acetic acid either singly or in combinations. The highest callus induction frequency (87.75%) was obtained in 6.7 µM 2,4-D added MS medium which metamorphosed into progressive stages (globular, heart, torpedo, and cotyledonary) of embryos. Mature and healthy somatic embryos efficiently germinated into plantlets on 8.8 µM BAP + 1.4 µM GA3 enriched MS medium. Histological and scanning electron microscopic study confirmed the above developing stages. The regenerated shoots were rooted best in 2.45 µM Indole-3-butyric acid supplemented solid MS medium. The plants were hardened and acclimatized with 90% survivability. The flow cytometric 2C DNA content of indirect, direct and somatic embryo derived plants was 1.896 pg, 1.940 pg and 1.926 pg respectively, very similar to the mother plant (1.928 pg). SCoT marker generated a high percentage of monomorphic bands (94%) revealing similarity with the mother plant, thus ensuring genetic fidelity. To the best of our knowledge, this is perhaps the first ever report of 2C DNA content estimation and SCoT marker based genetic homogeneity study in T. indica. An efficient regeneration method has been developed in T. indica. Histology and SEM indicated somatic embryogenesis based morphogenesis. 2C genome size and SCoT marker revealed genetic homogeneity of the regenerated population.
9 citations
TL;DR: In this paper, the effects of MS basal media (half or full strength) and MS medium supplemented with 6-Benzylaminopurine (BAP) were studied for in vitro seed germination.
8 citations
TL;DR: In this article, shoot tips as explants were cultured in Murashige and Skoog medium with major salts at half strength supplemented with silver nanoparticles concentrations (0, 5, 10, 20, 40 µM AgNPs) which were then exposed to three different light intensities (40, 80, 160 µmol m−2 s−1).
7 citations
7 citations
TL;DR: In this article , the efficacy of aromatic and synthetic cytokinins, which influence the direct organogenesis, biochemical responses, and ex vitro and in vivo performance of the in vitro propagated plantlets of Dioscorea pentaphylla L., an underrated famine tuber crop, was described.
7 citations
TL;DR: In this article , the authors used liquid medium in temporary immersion bioreactors to propagate axillary shoots of Cannabis sativa L. using liquid medium for 1 min, 3 times per day in β-A medium supplemented with 2-μM metatopoline and 0.5% of sucrose and subcultured every 4 weeks.
Abstract: The aim of this study was to propagate axillary shoots of Cannabis sativa L. using liquid medium in temporary immersion bioreactors. The effect of immersion frequency (3 or 6 immersions per day), explant type (apical or basal sections), explant number (8, 10, and 16 explants), mineral medium (Murashige and Skoog half-strength nitrates, β-A and β-H, all supplemented with 2-μM metatopoline), sucrose supplementation (2, 0.5, and 0% sucrose), culture duration (4 and 6 weeks), and bioreactor type (RITA® and Plantform™) were investigated. As a result, we propose a protocol for the proliferation of cannabis apical segments in RITA® or Plantform™ bioreactors. The explants (8 per RITA® and 24 per Plantform™) are immersed for 1 min, 3 times per day in β-A medium supplemented with 2-μM metatopoline and 0.5% of sucrose and subcultured every 4 weeks. This is the first study using temporary immersion systems in C. sativa production, and our results provide new opportunities for the mass propagation of this species.
6 citations
6 citations
TL;DR: In this article , the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyn segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine).
Abstract: Dragon's head plant (Lallemantia iberica), is a flowering species belongs to the mint family (Lamiaceae). The species contains valuable essential oils, mucilage and oil which are used in pharmaceutical and food industries. Tissue culture is a feasible strategy to attain large-scale production of plantlets with a huge potential to produce plants with superior quality. The objective of this study was to develop a simple and efficient method for regeneration and transformation of L. iberica. To reach this goal, the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyledon segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine). According to the results, cotyledonary nodes showed the best regeneration response. The maximum rate of regeneration (and number of induced shoots was achieved in 1 mg l-1 BAP in combination with 0.05 mg l-1 NAA from the cotyledonary nodes. Additionally, through the optimized regeneration technique Agrobacterium-mediated transformation of L. iberica was successfully accomplished. Gene transfer was assessed on leaf samples from regenerated plantlets under a fluorescent microscope to detect the GFP signals. Moreover, transgene integration and its expression were confirmed by PCR and RT-PCR analysis, respectively. The establishment of these efficient regeneration and genetic transformation methods paved the way for further application such as plant improvement, functional analysis and gene editing.
TL;DR: In this paper , the authors reported successful androgenic plant development via embryogenesis from microspore calluses in anther cultures and estimation of bioactive metabolites in in vitro regenerants and parent plant (control) of Cambod tea, Camellia assamica ssp. lasiocalyx (Planch MS) cultivar TV19.
Abstract: This pioneering work reports successful androgenic plant development via embryogenesis from microspore calluses in anther cultures and estimation of bioactive metabolites in in vitro regenerants and parent plant (control) of Cambod tea, Camellia assamica ssp. lasiocalyx (Planch MS) cultivar TV19. Anthers bearing microspores at early-to-late uni-nucleate stage were selected to initiate androgenesis. A pre-treatment of 5 °C for 5 days in the dark was most effective to initiate profusely growing white callusing from microspores within 10 weeks of culture on MS medium (6% sucrose) supplemented with high cytokinin/auxin ratio maintained by 6-benzylaminopurine (BAP) and 2,4-dichlorophenxoyacetic acid. Nodular structures on the callus surface differentiated into embryos. Further developement of the embryos occurred on embryogenesis medium but, with ten times reduced concentration of growth regulators and additives. Germination of embryos into complete plantlets was achieved when major salts in medium were reduced to half MS (½ MS) and augmented with BAP, GA3 and IBA along with glutamine and serine. Cytological examination of the root-tip cells revealed that regenerated plantlets were haploids (2n = x = 15), which was further confirmed through flow cytometry. The hot-water extracts from in vitro haploid calluses, embryos and field-grown donor plant were utilized for quantification of (+)-catechin, (−)-epicatechin, (−)-epigallocatechin gallate, caffeine and theophylline. Our findings revealed that the metabolite profile of in vitro regenerated haploid cultures is comparable to that of the mother plant, thereby presenting them as potential source for genome duplication and development of genetically stable homozygous pure breeding lines. This is first report on haploids in out-breeding tree, Cambod tea. It’s a significant achievement towards generating homozygous lines, which is impossible using conventional methods. Haploids showed consistent metabolite production.
24 Feb 2022
TL;DR: In this article , a new viable biotechnological approach for the in vitro propagation and long-term conservation of endangered Agave germplasm is presented. But the results presented in this paper are limited to shoot-tips of A. peacockii.
Abstract: More than 50% out of 129 of Agave species are endemic to Mexico. Among them, Agave peacockii is among the list of threatened species that require special protection. In this work, we aimed at developing new supplementary strategies to achieve micropropagation and perform cryopreservation of in vitro grown shoot-tips of A. peacockii. For multiplication, the addition of two cytokinins, 6-benzylaminopurine (26.6 μM) and kinetin (27.84 μM) to MS semisolid medium significantly favoured the morphogenetic response and produced the highest (87.00 ± 17.18) shoot generation number after 60 days of culture. This interaction was more effective than using the same growth regulators separately. Propagated and rooted plantlets were acclimatized with 100% survival and normal morphological development. For cryopreservation, an optimized protocol following droplet-vitrification allowed obtaining 98% and 96% regrowth before and after cryopreservation, respectively. Shoot-tips (1 mm in length × 1 mm wide) were excised of in vitro-propagated plants, subjected to preculture on MS semisolid medium with 0.3 M sucrose for 1d, loaded in solution with 0.4 M sucrose and 1.6 M glycerol for 20 min, exposed to Plant Vitrification Solution 2 for 15 min, and then, immersed in liquid nitrogen in droplets of PVS2 placed on aluminium foil strips. The vegetative growth of cryo-derived plants and of the in vitro propagated plants was compared under greenhouse conditions. No significant differences were detected in most assessed characteristics after 120 days of culture. The results presented here constitute new viable biotechnological approaches for the in vitro propagation and long-term conservation of endangered Agave germplasm. Agave peacockii shoot micropropagation was induced combining 6-benzylaminopurine (BAP) and 6-furfuryl-aminopurine (kinetin). A droplet-vitrification protocol was optimized to cryopreserve shoot-tips. Greenhouse performance of in vitro and cryo-derived plants was similar.
TL;DR: Aconitum violaceum Jacq. ex Stapf is a threatened medicinal plant with restricted global distribution, and the highest frequency of seed germination was recorded on Murashige and Skoog's (MS) basal medium, supplemented with 0.5 mg L−1 kinetin with a germination rate of 77.32% and mean germination time of 27 days as discussed by the authors .
Abstract: Aconitum violaceum Jacq. ex Stapf is a threatened medicinal plant with restricted global distribution. The highest frequency of seed germination was recorded on Murashige and Skoog’s (MS) basal medium, supplemented with 0.5 mg L−1 kinetin with a germination rate of 77.32% and mean germination time of 27 days. Among the various plant growth regulators examined, 0.1 mg L−1 kinetin (Kn) + 0.5 mg L−1 indole-3-acetic acid (IAA) proved to be effective for maximum embryogenic callus production (51.0%) within 31 days of inoculation. The conversion rate of somatic embryos into complete plantlets was highest in the MS medium augmented with 0.1 mg L−1 Kn + 0.5 mg L−1 IAA (68.00%), with an average root initiation time of 25 days. The rooted plantlets were subsequently hardened into jiffy pots with a combination of loamy soil, coco-peat, and vermicompost (1:1:1 v/v), and then transplanted into a greenhouse with a 60% survival rate. To our knowledge, this is the first study on direct in vitro propagation and embryogenic callus induction from seeds. The established regeneration protocol could be employed to propagate A. violaceum on a large scale in a short time. This would contribute significantly to its rapid propagation and germplasm conservation, and establish a framework for the domestication of this highly valued threatened medicinal plant.
TL;DR: In this article , the in vitro-derived seedlings studied were used as the starting material for the establishment of various media, such as Murashige and Skoog (MS) media, hormone-free and containing 0.5 mg L−1 6-benzyladenine (BA), leading to the satisfactory (84.0-89.0%) establishment of plantlets.
Abstract: Over the past few decades, both wildfires and human-sparked fires have ravaged Mount Parnitha, destroying the mountain’s unique natural ecosystem, applying pressure to its flora, and subjecting the vulnerable populations of Sideritis raeseri subsp. attica to excessive stress. The present study aims to establish an efficient micropropagation method starting from in vitro-grown seedlings. The in vitro germination study carried out during the production of seedlings revealed a higher germination rate (34.0% and 37.0%, respectively) at 20.0 °C and 25.0 °C. The in vitro-derived seedlings studied were used as the starting material for the establishment of various media. Murashige and Skoog (MS) media, hormone-free and containing 0.5 mg L−1 6-benzyladenine (BA), led to the satisfactory (84.0–89.0%) establishment of plantlets. During the multiplication phase, the study used BA in conjunction with α-naphthaleneacetic acetic acid and four different cytocinins (BA; kinetin (KIN); 6-(γ-γ-dimethylallylamino) purine; zeatin) at low concentrations (0.5 mg L−1). During the second subculture, a high multiplication index (7.3 and 6.4, respectively) was found for the hormone-free MS medium and the MS medium containing KIN at 0.5 mg L−1. Hyperhydricity took place on the media supplemented with hormones. Rooting occurred on the half-strength MS medium (51.0%). After two months, the plants’ survival rate stood at 100.0%, as did their ex vitro acclimatisation rate, which also registered at 100.0%. The present results could encourage not only the introduction of S. raeseri subsp. attica into the industry of floriculture as a new ornamental plant but also its ex vitro conservation.
TL;DR: In this article , the effects of explant source and plant growth regulators (PGRs) including naphthaleneacetic acid, picloram, 2,4-dichlorophenoxyacetic acids, 6-benzylaminopurine, kinetin, and thidiazuron on the in vitro regeneration capacity of Macodes limii plantlets were examined.
Abstract: Habitat disturbance and excessive collection of wild orchids from their natural habitat have threatened many orchids species at risk of extinction. In this study, the in vitro regeneration protocol for Macodes limii, a jewel orchid endemic to Sabah was established. The effects of explant source and plant growth regulators (PGRs) including naphthaleneacetic acid, picloram, 2,4-dichlorophenoxyacetic acid, 6-benzylaminopurine, kinetin, and thidiazuron on the in vitro regeneration capacity of M. limii plantlets were examined. Both factors showed a significant interaction in promoting axillary shoot formation. Nodal explants from the third and fourth positions cultured with 1.0 mg/L TDZ, induced 95% of shoot regeneration, with an average of three shoots/explant (1.6–1.8 cm of shoot length) after 90 days of culture. The well-developed plantlets went through an acclimatization phase for 60 days with a 60% of survival rate. An inter simple sequence repeat (ISSR) marker analysis confirmed the genetic stability of the in vitro regenerated plants to the mother plant. The successfully acclimatized plantlets were finally transferred to Poring Orchid Conservation Centre for reintroduction. The established protocol provides the means for large-scale production of this endemic jewel orchid, as well as a basis for further research aimed at the conservation and genetic improvement of this plant.
TL;DR: In this article , the authors showed that MS medium + 1.0 g activated charcoal + 30 g sucrose + 8.0 mg/L kinetin + 0.5mg/L α-NAA + 30g sucrose+8.0g agar per liter, pH 5.7-5.8 was the optimal medium to increase shoot length.
Abstract: Hoa Binh province is one of the best places for orchids in Vietnam. The climate and environment of Hoa Binh province are favorable for the development of orchids, especially rare indigenous ones. Dendrobium anosmum Lindl., which stands out because of the unique fragrance and colors, is one of the most popular varieties in Hoa Binh province. To meet the increasing demands of the industrial market as well as to contribute to the preservation and development of genetic resources of Dendrobium sp. in Hoa Binh province, propagating D. anosmum Lindl. is a crucial step. Plant tissue culture, which has been applied to improve reproducibility of orchids for many years, is still an effective method, especially for large-scale propagation. Studies on in vitro propagation of D. anosmum Lindl. from Hoa Binh province showed that growth regulators (BA, kinetin, α-NAA) did not have a significant effect on protocorm initiation because D. anosmum Lind. from Hoa Binh province already has a high rate of regeneration. However, MS medium + 1.0 mg/L kinetin + 0.5 mg/L α-NAA + 30 g sucrose + 8.0 g agar per liter, pH 5.7–5.8 was the optimal medium to increase shoot length. The MS medium + 1.0 g activated charcoal + 30 g sucrose + 8.0 g agar per liter, pH 5.7–5.8 was the most suitable medium for shoot growth—after 6 weeks of culture, the average shoot length was 1.09 cm, the average number of leaves was 6.13, the average number of roots was 3.17, and the average root length was 1.11 cm—about 3.3, 4.17, 3.41, and 1.67 times higher, respectively, than in the control (without activated charcoal).
TL;DR: In this paper , the authors introduce and optimize a protocol for inducing callus and cell suspension cultures as well as estimating phenolic, flavonoid compounds, and antioxidant activity in the cultures of the studied species.
Abstract: Lycium schweinfurthii is a traditional medicinal plant grown in the Mediterranean region. As it is used in folk medicine to treat stomach ulcers, it took more attention as a source of valuable secondary metabolites. The in vitro cultures of L. schweinfurthii could be a great tool to produce secondary metabolites at low costs. The presented study aimed to introduce and optimize a protocol for inducing callus and cell suspension cultures as well as estimating phenolic, flavonoid compounds, and antioxidant activity in the cultures of the studied species. Three plant growth regulators (PGRs) were supplemented to MS medium solely or in combination to induce callus from leaf explants. The combination between 2,4-dichlorophenoxy acetic acid (2,4-D) and 1-naphthyl acetic acid (NAA) induced callus in all explants regardless of the concentration. The highest fresh weight of callus (3.92 g) was obtained on MS medium fortified with 1 mg L−1 of both 2,4-D and NAA (DN1) after 7 weeks of culture. DN1 was the best medium for callus multiplication regarding the increase in fresh weight and size of callus. Otherwise, the highest phenolics, flavonoids, and antioxidant activity against DPPH free radicals were of callus on MS fortified with 2 mg L−1 NAA (N2). The cell suspension cultures were cultivated on a liquid N2 medium with different sucrose concentrations of 5–30 g L−1 to observe the possible effects on cells’ multiplication and secondary metabolite production. The highest fresh and viable biomass of 12.01 g was obtained on N2 containing 30 g L−1 sucrose. On the other hand, the cell cultures on N2 medium of 5 and 30 g L−1 sucrose produced phenolics and flavonoids, and revealed antioxidant activity against DPPH and ABTS+ free radicals more than other sucrose concentrations. The presented protocol should be useful in the large-scale production of phenolic and flavonoid compounds from callus and cell cultures of L. schweinfurthii.
TL;DR: In this paper , the effect of 2,4-D and picloram herbicides on the development of palm Euterpe edulis embryos was evaluated. But, the results showed that the effect was limited to those embryos that were induced with 2, 4-D (159.42 μmol L−1) and PIC (300 μmol l−1).
TL;DR: In this paper , the effects of plant growth regulators (PGRs) on the callus induction, shoot organogenesis from callus and in vitro rooting were evaluated, and the parthenolide yield of the micro-propagated and conventionally grown plants was determined by using high-performance liquid chromatography (HPLC).
Abstract: Feverfew (Tanacetum parthenium) is a well-known multi-functional plant with anti-inflammatory, cardiotonic, antiangiogenic, and anticancer effects. The therapeutic value of this plant is due to its phytochemical constitutes, especially parthenolide. Tissue culture techniques have been applied to improve the bioactive components of many herbal plants. Hence, this study, was carried out to establish a protocol for micropropagation of the feverfew plant and to quantify parthenolide content in its micropropagated and conventionally grown plants. To establish an aseptic culture, different concentrations of sodium hypochlorite (NaOCl) were investigated for seed surface sterilization. Besides, the effects of plant growth regulators (PGRs) on the callus induction, shoot organogenesis from callus and in vitro rooting were evaluated. Additionally, the parthenolide yield of the micropropagated and conventionally grown plants was determined by using high-performance liquid chromatography (HPLC). The results showed that surface sterilization of feverfew seeds with 6% NaOCl for 15 min obtained 65.00 ± 2.69% aseptic seeds. Murashige and Skoog (MS) medium supplemented with 0.4 mg/L thidiazuron (TDZ) and 2 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) resulted in 86.00 ± 1.72% callus induction. The highest number of shoots (5.00 ± 0.15) per explant was obtained in the treatment of MS medium supplemented with 5 mg/L zeatin. MS medium fortified with 3 mg/L indole-3-butyric acid (IBA) produced the maximum number of roots per plantlet (8.90 ± 0.35). A total of 90% of the micropropagated plantlets survived when planted in perlite + peat moss (1:1 v/v); the micropropagated plantlets were successfully established in the ex vitro conditions. According to parthenolide analysis, its level was significantly higher in the micropropagated plants than conventionally grown plants. Among different solvents, ethanolic extraction obtained the highest parthenolide content of the feverfew plant. Hence, it can be concluded that micropropagation of feverfew could be applied to produce disease-free planting materials and to improve the parthenolide content of the feverfew plant.
TL;DR: In this paper , the authors developed an efficient protocol for in vitro propagation of a rare orchid species, Vanda brunnea (syn. Vanda henryi Schltr.).
TL;DR: In this article , a high-performance in vitro propagation system of Plectranthus amboinicus through direct shoot organogenesis was developed using axillary node explants cultured on MS (Murashige and Skoog) medium augmented with 0.5, 2.5 and 5.5 µM NAA.
Abstract: Plectranthus amboinicus (Lour.) Spreng is a medicinally important aromatic perennial herb used for the treatment of skin diseases, constipation, asthma, flu, fever, cough, and headache as well as a flavoring ingredient in traditional drinks, food, and meat stuffing. In this study, a high-performance in vitro propagation system of P. amboinicus through direct shoot organogenesis was developed using axillary node explants cultured on MS (Murashige and Skoog) medium augmented with 0.5, 2.5, 5.0, 7.5, and 10.0 µM of 6-benzyladenine (BA) or kinetin (Kin), alone or with 0.1, 0.5, 2.5, and 5.0 µM of indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA). To optimize the regeneration potential of node explants, the effects of basal media strength and pH were also investigated. After 8 weeks of culture, explants cultured in full strength MS basal medium (pH 5.7) with 5.0 µM BA and 2.5 µM NAA exhibited the highest percentage (97.1%) of regeneration and the maximum number (19.3) of shoots per explant. Individual elongated shoots were rooted on half strength MS basal medium containing 0.25 µM indole 3-butyric acid (IBA) after 4 weeks of culture, producing 5.3 roots/shootlets with a root induction frequency of 93.7%. First time genetic stability of in vitro raised P. amboinicus plants was determined using SPAR markers, such as DAMD and ISSR, as well as flow cytometric tests, assuring the availability of authenticated raw materials for commercial production of the plant and its bioactive components.
TL;DR: In this paper , the authors developed an effective micropropagation system of the economically important Musa acuminata Colla (AAA) having high structural fidelity and significant potential to multiply shoots under in vitro conditions using the non-conventional cytokinin 6-benzoyladenine (BOA, C12H9N5O).
TL;DR: In this paper , an efficient encapsulation and slow growth conservation protocol was developed for Nyctanthes arbor-tristis L. an antiviral medicinal plant of the family Oleaceae.
Abstract: An efficient encapsulation and slow growth conservation protocol was developed for Nyctanthes arbor-tristis L. an antiviral medicinal plant of the family Oleaceae. A gel matrix with 3% sodium alginate and 100 mM calcium chloride (CaCl2⋅2H2O) was found best for the encapsulation of nodal segments. The viability and shoot development potential of encapsulated nodal segments was optimized. Encapsulated nodal segments stored at 4 °C and 24 °C remained viable for up to 90 days and showed shoot development potential 42.89 ± 6.04% and 33.53 ± 7.15% respectively. Nodal segments maintained under slow growth conditions up to 180 days on one-eighth strength Murashige and Skoog (MS) medium supplemented with 0.5% sucrose was suitable for satisfactory viability (40.28 ± 2.04%), while further addition of 0.5 mg/l abscisic acid supported 40.36 ± 1.01% viability of the nodal segments. The best rooting response was achieved on half-strength MS medium supplemented with 4 mg/l indole-3-acetic acid. The field survival of rooted plants was 65%. The clonal fidelity of in-vitro derived plantlets was studied with start codon targeted primer profile, which showed the same banding mobility patterns as the source parent plant. The maximum banding profile was monomorphic and consistent, confirming the clonal stability of regenerated plants. The method developed will permit the in-vitro conservation of this species and facilitate an easy exchange of plant germplasm.