Production of in vitro strawberry (Fragaria × ananassa) plantlets in large-scale system supplemented with silver nanoparticles

The growth of strawberry plantlets in the rooting stage on culture medium supplemented with silver nanoparticles (AgNPs) and the ethylene gas accumulation in plantlet culture bottles were investigated. In addition, different culture systems were first used to produce large-scale Strawberry plantlets. The results showed that shoots (3 cm) were cultured on MS medium supplemented with 0.02 mg/L NAA, 1 g/L activated charcoal, 30 g/L sucrose, 8 g/L agar and 0.5 mg/L AgNPs showed about 4 days earlier rooting formation and the plantlet growth such as plantlet height (5.60 cm), fresh weight (242.67 mg), dry weight (34,67 mg), number of roots/plantlet (6.67), root length (3.40 cm), SPAD (39.30 nmol/cm2) were higher than those in the control after 15 days of culture. Besides, the ethylene gas content in the culture bottle (0.06 ppm) in the 0.5 mg/L AgNPs treatment was lower than as compared to that in the control (0.15 ppm) after 15 days of culture. A shoot density (10 shoots) in 250 mL culture bottle with 40 mL of medium gave optimal growth than those in other treatments after 15 days of culture. Square plastic box culture system (length × width × height: 19 cm × 19 cm × 7 cm; 2.5 L in volume) containing 250 mL MS medium added to 0.5 mg/L AgNPs produced 100 vigorous plantlets; meanwhile, rectangular plastic box system (34 cm × 23 cm × 13 cm; 10 L in volume; 10 L in volume) produced 200 vigorous plantlets. Plantlets derived from 0.5 mg/L AgNPs treatment in the plastic box systems exhibited well acclimatization after 30 and 60 days of culture in the greenhouse.

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In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

Acta Botanica Croatica ◽

10.1515/botcro-2017-0012 ◽

2018 ◽

Vol 77 (1) ◽

pp. 80-87 ◽

Cited By ~ 3

Author(s):

Mahipal S. Shekhawat ◽

M. Manokari

Keyword(s):

Medicinal Plant ◽

Large Scale ◽

Nodal Segments ◽

Ms Medium ◽

Ex Vitro ◽

Culture Bottle ◽

Hybanthus Enneaspermus ◽

Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

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Silver Nanoparticles Effects on In Vitro Germination, Growth, and Biochemical Activity of Tomato, Radish, and Kale Seedlings

Materials ◽

10.3390/ma14185340 ◽

2021 ◽

Vol 14 (18) ◽

pp. 5340

Author(s):

Alicja Tymoszuk

Keyword(s):

Silver Nanoparticles ◽

Practical Importance ◽

Plant Cells ◽

Dry Weight ◽

Guaiacol Peroxidase ◽

In Vitro Germination ◽

Vegetable Crops ◽

Fresh Weight ◽

Biochemical Activity

The interactions between nanoparticles and plant cells are still not sufficiently understood, and studies related to this subject are of scientific and practical importance. Silver nanoparticles (AgNPs) are one of the most commonly produced and used nanomaterials. This study aimed to investigate the influence of AgNPs applied at the concentrations of 0, 50, and 100 mg·L−1 during the process of in vitro germination as well as the biometric and biochemical parameters of developed seedlings in three vegetable species: Solanum lycopersicum L. ‘Poranek’, Raphanus sativus L. var. sativus ‘Ramona’, and Brassica oleracea var. sabellica ‘Nero di Toscana’. The application of AgNPs did not affect the germination efficiency; however, diverse results were reported for the growth and biochemical activity of the seedlings, depending on the species tested and the AgNPs concentration. Tomato seedlings treated with nanoparticles, particularly at 100 mg·L−1, had shorter shoots with lower fresh and dry weights and produced roots with lower fresh weight. Simultaneously, at the biochemical level, a decrease in the content of chlorophylls and carotenoids and an increase in the anthocyanins content and guaiacol peroxidase (GPOX) activity were reported. AgNPs-treated radish plants had shorter shoots of higher fresh and dry weight and longer roots with lower fresh weight. Treatment with 50 mg·L−1 and 100 mg·L−1 resulted in the highest and lowest accumulation of chlorophylls and carotenoids in the leaves, respectively; however, seedlings treated with 100 mg·L−1 produced less anthocyanins and polyphenols and exhibited lower GPOX activity. In kale, AgNPs-derived seedlings had a lower content of chlorophylls, carotenoids, and anthocyanins but higher GPOX activity of and were characterized by higher fresh and dry shoot weights and higher heterogeneous biometric parameters of the roots. The results of these experiments may be of great significance for broadening the scope of knowledge on the influence of AgNPs on plant cells and the micropropagation of the vegetable species. Future studies should be aimed at testing lower or even higher concentrations of AgNPs and other NPs and to evaluate the genetic stability of NPs-treated vegetable crops and their yielding efficiency.

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Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽

10.21273/hortsci.44.1.106 ◽

2009 ◽

Vol 44 (1) ◽

pp. 106-112 ◽

Cited By ~ 15

Author(s):

Alice Noemí Aranda-Peres ◽

Lázaro Eustáquio Pereira Peres ◽

Edson Namita Higashi ◽

Adriana Pinheiro Martinelli

Keyword(s):

New Media ◽

Mineral Composition ◽

Culture Media ◽

Dry Weight ◽

Defined Medium ◽

Ms Medium ◽

Media Formulation ◽

Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

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Biosynthesis and characterization of silver nanoparticles using Ochradenus arabicus and their physiological effect on Maerua oblongifolia raised in vitro

Scientific Reports ◽

10.1038/s41598-020-74675-9 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Hassan O. Shaikhaldein ◽

Fahad Al-Qurainy ◽

Mohammad Nadeem ◽

Salim Khan ◽

Mohamed Tarroum ◽

...

Keyword(s):

Silver Nanoparticles ◽

Chlorophyll Content ◽

Total Protein ◽

Dry Weight ◽

Shoot Length ◽

Antioxidant Enzyme Activities ◽

Fresh Weight ◽

Wide Range ◽

Shoot Number

Abstract Silver nanoparticles (AgNPs) are presently the most commonly generated engineered nanomaterials and are found in a wide range of agro-commercial products. The present study was designed to synthesize AgNPs biologically using Ochradenus arabicus leaves and investigate their effect on the morphophysiological properties of Maerua oblongifolia raised in vitro. Physicochemical methods (ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy were performed for characterization and for obtaining microphotographs of the AgNPs. Shoots of M. oblongifolia (2–3 cm) grown in Murashige and Skoog medium supplemented with different concentrations of AgNPs (0, 10, 20, 30, 40, or 50 mg L−1) were used. Following 6 weeks of in vitro shoot regeneration, the shoot number, shoot length, leaf number, fresh weight, dry weight, chlorophyll content, total protein, proline level, and antioxidant enzyme activities of the plants were quantified. We found that 20 mg L−1 AgNPs increased the shoot number, shoot length, fresh weight, dry weight, and chlorophyll content of the plants. The maximum total protein was recorded in plants that were administered the lowest dose of AgNPs (10 mg L−1), while high concentrations of AgNPs (40 and 50 mg L−1) increased the levels of proline and the enzymes superoxide dismutase and catalase. Our results indicate that green-synthesized AgNPs may be of agricultural and medicinal interest owing to their effects on plants in vitro.

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Callogenesis in leaves of Kalanchoe pinnata Lam. by 2,4-D and BA action

Revista Brasileira de Plantas Medicinais ◽

10.1590/1983-084x/13_031 ◽

2014 ◽

Vol 16 (3 suppl 1) ◽

pp. 760-764 ◽

Cited By ~ 3

Author(s):

M.R.A. Santos ◽

M.G.R. Ferreira ◽

M.C.M. Guimarães ◽

R.A. Lima ◽

C.L.L.G. Oliveira

Keyword(s):

Callus Induction ◽

Large Scale ◽

Leaf Explants ◽

Ms Medium ◽

Main Technique ◽

Kalanchoe Pinnata ◽

Medicinal Properties ◽

Industrial Level ◽

Inflammatory Effect

The Kalanchoe pinnata Lam. is a bush species of the Crassulaceae that is distinguished by its important medicinal properties. Its leaves are used as cataplasm to treat headaches and wounds. There is evidence for a hypotensive and anti-inflammatory effect. Techniques of plant tissue culture have been applied to plant species that produce substances likely to be explored in pharmacology, cell suspension being the main technique. At the industrial level, this method utilizes bioreactors in order to produce secondary metabolites on a large scale. The objective of this study was to evaluate the effects of in vitro combinations of 2,4-dichlorophenoxiacetic acid (2,4-D) and benzylaminopurine (BA) on callus induction in leaf explants of K. pinnata. Leaf fragments were inoculated in MS medium supplemented with 3.0% sucrose, 0.8% agar and factorial combinations of 2,4-D (0.00, 4.52, 9.06, 18.12 µM) and BA (0.00, 4.44, 8.88, 17.76 µM). The cultures were kept in the darkness at 24±2ºC for 50 days. The percentage of callus induction and the area of explants covered by callus cells were evaluated. In the absence of growth regulators, callus induction did not occur, with necrosis of all explants. The highest percentage of callus induction was 100%, obtained with the combination of 9.06 µM 2,4-D and 8.88 µM BA, but the calluses covered only 25% of the leaf area. The most efficient combination was 4.52 µM 2,4-D and 8.88 µM BA, resulting in 91% callus induction with 50 to 100% of the explants being covered by callus cells.

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In Vitro Propagation via Organogenesis and Formation of Globular Bodies of Salvia Plebeia: A Valuable Medicinal Plant

10.21203/rs.3.rs-39443/v1 ◽

2020 ◽

Author(s):

Qinggui Wu ◽

Honglin Yang ◽

Yuxi Sun ◽

Jinyao Hu ◽

Lijuan Zou

Keyword(s):

Medicinal Plant ◽

Large Scale ◽

Natural Habitat ◽

Shoot Tips ◽

Direct Organogenesis ◽

Ms Medium ◽

Hypocotyl Explants ◽

Cotyledonary Nodes ◽

Over Exploitation

Abstract Background: As a highly valued medicinal plant, Salvia plebeia R. Brown belongs to the Lamiaceae family that has been subjected to over exploitation in its natural habitat for phytochemical and pharmacological studies. Alternative collection methods need to be developed for the large-scale propagation of Salvia plebeian. Results: Here, efficient and simple, direct organogenesis (from shoot tips and cotyledonary nodes explants) and Globular bodies (GBs) induction (from hypocotyl explants) systems were developed for the in vitro propagation of Salvia plebeia. The highest and number of regenerated shoots (7.0±0.82) per shoot tips was obtained on Murashige and Skoog (MS) medium supplemented with a combination of 0.1 mg L-1 indole-3-acetic acid (IAA) and 1.0 mg L-1 6-benzyladenine (6-BA), the proliferation of shoots and shoots rooted were carried out on the same medium treatments almost synchronously. Similarly, MS medium supplemented with 0.1 mg L-1 IAA and 1.0 mg L-1 thidiazuron (TDZ) yielded the maximum number of shoots (37.5±1.34) with 100% shoot sprouting frequency. Simultaneously, a protocol was developed for GBs induction from hypocotyl explants, and it produced 17.4 GBs per explant with 82.7% response on MS medium supplemented with TDZ (1.0 mg L-1) and IAA (0.1 mg L-1), and produced GBs that were morphologically similar to globular embryos and successfully germinated on hormone-free MS medium. The acclimatized plantlets with well-developed root systems were successfully shifted to the natural soils with a 100% survival rate. Conclusions: Taken together, this protocol can be efficiently used for mass propagation, germplasm preservation and likely also for gene transfer of Salvia plebeia.

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IN VITRO CULTURE OF IMMATURE TAXUS EMBRYOS

HortScience ◽

10.21273/hortsci.27.6.698d ◽

1992 ◽

Vol 27 (6) ◽

pp. 698d-698 ◽

Cited By ~ 1

Author(s):

Ching-yeh Hu ◽

Lee Wang ◽

Bernard Wu

Keyword(s):

Embryo Culture ◽

Large Population ◽

Dry Weight ◽

Ms Medium ◽

Embryonic Growth ◽

Embryo Dormancy ◽

Taxol Production ◽

Initial Medium ◽

Mature Seeds

Embryo culture can by-pass yew (Taxus) seed dormancy and produce large population of seedlings to be screened for the anticancer drug, taxol, production. Immature linear embryos from seeds of T. baccata, T. brevifolia. T. cuspidata, and T. media were dissected and cultured. B5 medium supported the best embryonic growth during the initial two week's culture for T. cuspidata and T. baccata. T. brevifolia grew faster on MS medium. Weak embryo dormancy was encountered in T. brevifolia and T. cuspidata from the mature seeds but not from the immature ones. No embryonic growth had been observed in T. media dissected from mature seeds due to strong dormancy. Developing embryos were subsequently transferred to 1/2X B5 medium for germination. Rooting percentage in the mature seed derived T. brevifolia embryos increased from 12.5 to 63.6 when 30 μM GA3 was added to the initial medium. Several hundreds of seedlings of T. baccata. T. brevifolia and T. cuspidata had been acclimatized to the greenhouse conditions. The taxol content of resultant T. cuspidata seedlings was 0.027% (dry weight), while that of T. brevifolia obtained from the wild twig was 0.030%.

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Growth and Photosynthetic Capability of Momordica grosvenori Plantlets Grown Photoautotrophically in Response to Light Intensity

HortScience ◽

10.21273/hortsci.44.3.757 ◽

2009 ◽

Vol 44 (3) ◽

pp. 757-763 ◽

Cited By ~ 11

Author(s):

Meijun Zhang ◽

Duanduan Zhao ◽

Zengqiang Ma ◽

Xuedong Li ◽

Yulan Xiao

Keyword(s):

Callus Formation ◽

Dry Weight ◽

Control Treatment ◽

Photon Flux ◽

Photon Flux Density ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Light Intensities ◽

Four Levels

Momordica grosvenori plantlets were cultured in vitro for 26 d on sucrose- and hormone-free Murashige and Skoog (MS) medium with four levels of photosynthetic photon flux density (PPFD), namely 25, 50, 100, or 200 μmol·m−2·s−1, and a CO2 concentration of 1000 μmol·mol−1 in the culture room [i.e., photoautotrophic micropropagation (PA) treatments]. The control treatment was a photomixotrophic culture using MS medium containing sucrose and NAA with a CO2 concentration of 400 μmol·mol−1 in the culture room and a PPFD of 25 μmol·m−2·s−1. Based on the results, a second experiment was conducted to investigate the effects of α-naphthaleneacetic acid (NAA) and sucrose on callus formation. For this, plantlets were grown in the absence and presence of either NAA or sucrose. Compared with the control, the PA plantlet had a well-developed rooting system, better shoot, greater chlorophyll content, and higher electron transport rate and the ex vitro survival percentage was increased by 31%. Both sucrose and NAA stimulated callus formation on the shoot bases of control plantlets, whereas calluses did not form on the plantlets grown in sucrose- and hormone-free medium. The stronger light intensities increased the fresh and dry weight of plantlets. A PPFD of 100 μmol·m−2·s−1 was more suitable for the growth of M. grosvenori plantlets. Therefore, photoautotrophic plantlets grown at high light intensities would be better suited to the intense irradiance found in sunlight.

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INFLUENCE OF STRESS ON SECONDARY METABOLITES PRODUCTION FROM CALLUS OF MORINGA OLEIFERA IN VITRO

IRAQI JOURNAL OF AGRICULTURAL SCIENCES ◽

10.36103/ijas.v48i4.367 ◽

2017 ◽

Vol 48 (4) ◽

Author(s):

Ibrahim & Ameen

Keyword(s):

Tissue Culture ◽

Moringa Oleifera ◽

Active Material ◽

Dry Weight ◽

Ms Medium ◽

Poly Ethylene Glycol ◽

College Of Agriculture ◽

Poly Ethylene ◽

Metabolites Production

An experiment was conducted to study the effect of sucrose, poly ethylene glycol (PEG) on hypocotyl induced callus of Moringa oleifera at the plant tissue culture lab.- College of Agriculture– University of Baghdad from February 2015 to May 2016. Sucrose concentrations were 30, 60, and 90, 120 g .L -1 and PEG 0, 25, 50 and 100 g .L -1 added to MS medium supplemented with 2.0 mg .L -1 of 2, 4-D and 0.1 mg .L -1 of NAA. MS medium supplemented with120 g .L -1 of sucrose gave the best amount of Zeatin, Quercetin and Kaempferol reached to 103.4, 1324.6 and 966.5 µg. g dry weight of callus-1 respectively. The concentrations of active compound increasing with adding PEG, MS medium supplemented with 100 g .L -1 PEG gave the highest value of Zeatin, Quercetin and Kaempferol which recorded 92.01, 3528.0 and 931.0 µg. g dry weight of callus-1 respectively. We found that we could increase the production of active material from callus that induced from explant by exposure the callus to several stress and then could separate the pure active material and used it as a drug in medicine.

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(447) Response of Two Salt-stressed Cultivars of Sweetpotato to Changes in Calcium Levels under In Vitro Conditions

HortScience ◽

10.21273/hortsci.40.4.1035a ◽

2005 ◽

Vol 40 (4) ◽

pp. 1035A-1035

Author(s):

Devi Prasad V. Potluri ◽

Nkechiyere Nwami

Keyword(s):

Salt Stress ◽

Dry Weight ◽

Proline Accumulation ◽

Metabolic Changes ◽

Soluble Carbohydrate ◽

Ms Medium ◽

Shoot Height ◽

Weight Number ◽

Adverse Affects

Two cultivars of sweetpotato, `Commensal' and `Salyboro', were subjected to salt stress using axillary bud cultures. The salt levels ranged from 0 to 10 g·L-1. After the initial experiments, levels of calcium in the medium were changed from 3 mm in the MS medium to 1.5, 6, and 12 mm. After 10 weeks of growth, plantlet shoot height, dry weight, number of nodes, levels of proline, soluble carbohydrate, and protein were measured. `Commensal' was tolerant to salt levels up to 4 g·L-1, but `Salyboro' was sensitive to concentrations of salt even at lower concentrations as evidenced by the growth and dry weight. Proline accumulation was higher in the shoot than in the root. The protein: carbohydrate ratios did not change much in `Commensal', but levels of carbohydrates increased in `Salyboro'. Reduction in calcium levels had a synergistic affect on salt-stressed cultivars. Enhanced levels of calcium reduced the inhibitory affects of salt stress. This was more pronounced in `Salyboro', which was susceptible. Proline levels were higher in plants subjected to salt stress and higher levels of calcium than controls, but lower than the plants subjected to salt stress. These and other metabolic changes suggest that calcium can reduce the adverse affects of salt stress in these two sweetpotato cultivars.

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