Recent Articles

Below are the titles and abstracts from the most recent articles published in In Vitro Cellular and Developmental Biology – Plant. Click on the title to view the full article at Springerlink.


  • Enhancement of recombinant human keratinocyte growth factor 1 protein production in transgenic hazelnut (Corylus avellana L.) plant cell suspension cultures under RAmy3D inducible promoter
    on June 1, 2023 at 12:00 am

    Abstract Recombinant human KGF1 (rhKGF) is a fibroblast growth factor that promotes epidermal cell proliferation, differentiation, migration, and wound repair. In this study, we used the cell culture of a hazelnut expression system to produce rhKGF. These cells have a high growth rate. A cassette containing the rhKGF gene fused to hydrophobin tag was modified based on the hazelnut optimized codon usage and subsequently introduced into a hazelnut suspension culture under the control of a RAmy3D promoter, and secreted into the plant cell culture medium. The results showed that the highest transcription rate of the rhKGF gene was obtained under the RAmy3D promoter on the third day of transgenic cell placement in the promoter induction medium. To optimize the production of rhKGF secreted into the culture medium, the pH of the medium was adjusted during the production phase. After exposing the hazelnut cells to a pH of 7 for 12 d in the production phase, it was […]

  • Ex vitro rooting of minicutting of Carica papaya L. single shoot from tissue culture
    on May 31, 2023 at 12:00 am

    Abstract Papaya (Carica papaya L.) is an important economic crop. An efficient and low-cost propagation ex vitro process is required for commercial papaya plantlet production. In this study, a propagation process of papaya was established by minicutting and evaluating parameters affecting survival. Individual shoots (1.5 cm) were cut from papaya multiple shoots in vitro as the minicuttings. After dipping in 400.0 mg L−1 indole-3-butyric acid (IBA), they were planted into the nursery pro trays with 104 holes containing vermiculite, perlite, and King Root® substrate at 1:1:1 ratio, and then covered with a lid to keep the ratio of humidity (RH) at 100% for root development. After a 3-wk treatment in a growth chamber (28°C), the rooting rate, root number, root length, and stem length were promoted to 91.1%, 2.9, 3.71 cm, and 1.88 cm, respectively. Combining the treatment with 400 mg L−1 IBA and 1/2 Murashige and Skoog (MS) salt solution supplemented at the 14th day […]

  • In vitro plant regeneration of ‘Prelude’ red raspberry (Rubus idaeus L.)
    on May 30, 2023 at 12:00 am

    Abstract In this study, shoots were regenerated from in vitro leaf tissues of ‘Prelude,’ a red raspberry known for its hardiness, vigorous growth, and early fruit ripening. Experiments were conducted to discover the effect of plant growth regulators (PGRs) on plant regeneration of ‘Prelude’ red raspberry. Four cytokinins, thidiazuron (TDZ), 6-benzylaminopurine (BA), zeatin, and kinetin, at various levels were analyzed for shoot regeneration when in vitro leaf tissues were cultured on woody plant medium (WPM). The maximum shoot regeneration rate was observed in the medium containing 0.5 μM TDZ with an average of 65.7% of the explants forming shoots. BA at 5.0 μM resulted in a shoot regeneration rate of 11.6%, and increasing the BA concentration slightly reduced the regeneration rate. Neither zeatin nor kinetin exhibited a significant effect on plant regeneration of ‘Prelude.’ In vitro shoots were dipped in a commercial rooting powder, TakeRoot®, and […]

  • Micropropagation of Quercus spp., complications and solutions—an overview
    on May 25, 2023 at 12:00 am

    Abstract The Quercus genus provides ecological importance by improving the ecosystem in urban and forest settings. However, the massive exploitation of Quercus species without regulation causes deforestation in many areas. Reforestation of the affected areas with Quercus plantations derived from in vitro propagation is the best option. The Quercus micropropagation permits the cultivation of different parts of the plant in various medium compositions. Buds, meristems, and shoot tips can produce genetically stable clones, but meristems and embryos could be used to avoid contamination. Otherwise, the highest multiplication rate of Quercus is obtained by somatic embryogenesis. During in vitro multiplication, several problems can occur, such as basal calluses, vitrification, tissue necrosis, and phenol accumulation. Alternatively, adding supplements to the basal medium, like polyvinylpyrrolidone or ascorbic acid as well as casein hydrolysate, improves the Quercus […]

  • Conservation strategy, phytochemical diversity, and genetic consistency assessment of in vitro propagated perennial herb Vincetoxicum hirundinaria Medik
    on May 22, 2023 at 12:00 am

    Abstract This study has developed efficient protocols for callus induction and mass propagation as well as reports phytochemical and genetic profile of the wild and acclimatized V. hirundinaria plant. Friable green and soft off-white texture callus was observed from seeds and leaf explant with frequency of 85 to 92% and 75 to 90% respectively. Earliest shoot buds initiation 13 d was noticed on media fortified with 1.0 mg L−1 BAP and 1.0 mg L−1 NAA, while highest shoot proliferation (4.20) was observed with 0.5 mg L−1 kinetin and 0.5 mg L−1 NAA. Earliest rooting (15 d) was achieved with 1.0 mg L−1 BAP and 0.5 mg L−1 IBA, while maximum root length (6.1 cm) was recorded on medium supplemented with 1.0 mg L−1 BAP and 1.5 mg L−1 IBA. The in vitro raised plantlets were acclimatized with 76% survival rate. Moreover, in phytochemical investigation, maximum sugar contents (756.65 µM mL−1), proline (15.31 mg g−1), ascorbic acid […]

  • Transverse thin cell layer (tTCL) technology: a promising tool for micropropagation of Centratherum punctatum Cass.
    on May 18, 2023 at 12:00 am

    Abstract The present study describes an efficient protocol for the micropropagation of Centratherum punctatum Cass. through transverse thin cell layers (tTCLs). The effect of plant growth regulators (PGRs), thickness of tTCLs, and source of tTCLs were evaluated. The tTCLs of varying thickness (0.5 to 5.0 mm) were excised from 10-d-old leaf and 45-d-old node and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of PGRs (BAP, TDZ, KN; 0.1 to 2.0 mg L−1), alone or in combination with NAA (0.1 to 1.0 mg L−1) for shoot induction. The leaf and node tTCLs responded with direct shoot regeneration. A significant effect of thickness or width on shoot induction was observed. For leaf, 1.0-mm-wide and for node 2.0-mm-thick tTCLs showed a maximum response. MS medium supplemented with 1.5 mg L−1 TDZ and 1.0 mg L−1 BAP in combination with 0.2 mg L−1 NAA was found to be optimum for shoot induction from tTCLs of leaf (96% response […]

  • Improved cryopreservation protocol for tea (Camellia sinensis (L.) O. Kuntze) using preconditioning of shoot tip donor plants and V cryo-plate technique
    on May 17, 2023 at 12:00 am

    Abstract Indian tea germplasm is known for its diversity, which not only assists the development of superior planting material for tea breeding but also functions as a genetic resource for other tea-producing countries. In this study, the aim was to develop an improved cryo-protocol for long-term conservation of tea germplasm. Shoot tips (approximately 2 mm) of Camellia sinensis were cryopreserved using vitrification and V cryo-plate techniques. The recovery rate of cryopreserved shoot tips obtained using vitrification was 33.2% at 90 min of Plant Vitrification Solution 2 (PVS2) exposure. To improve recovery rates and tolerance to cryo-injury, shoot tip donor plants were preconditioned with cold hardening, high sucrose, abscisic acid (ABA), and proline prior to cryopreservation. The vitrification technique resulted in an increase of regrowth of shoot tips to 60.5% after preconditioning of shoot tip donor plants for 30 d at 25 °C with 10 mg L−1 ABA and 3 mg […]

  • Arbuscular mycorrhizal fungi increase the survival and growth of micropropagated coconut (Cocos nucifera L.) plantlets
    on May 17, 2023 at 12:00 am

    Abstract Coconut is a crop of economic importance, and protocols for the propagation in vitro of coconut are already in use. However, during acclimatization, some micropropagated plants or plantlets do not survive. There are reports that show that the application of arbuscular mycorrhizal fungi (AMF) improves the development of plants. So far, there are no reports of acclimatization of coconut plantlets with or without AMF. Therefore, this study reports the evaluation of survival and growth during acclimatization of coconut plantlets (obtained by somatic embryogenesis) testing inoculation with native or commercial AMF. Survival increased from 1.19 to 1.24-fold with native AMF, but no increase occurred with commercial AMF. Growth and photosynthetic parameters were evaluated, and there were no significant changes among treatments at 180 d. However, 6 mo later, there was a significant increase in height, leaf area, and stem diameter in plantlets inoculated with […]

  • An efficient protocol for production of rosmarinic acid in Salvia nemorosa L.
    on May 17, 2023 at 12:00 am

    Abstract Rosmarinic acid (RA), as the major secondary metabolite of Salvia species, is a valuable bioactive compound for pharmaceutical applications. Toward rosmarinic acid production, Murashige and Skoog (MS) medium supplemented with 0.5 mg L−1 2,4-D and 2 mg L−1 BA or 1 mg L−1 2,4-D and 1 mg L−1 BA showed the highest percentage of callus induction (100%). In this research, MS medium supplemented with various concentrations of sucrose, phenylalanine, salicylic acid (SA), and methyl jasmonate (MeJA) were evaluated using cell suspension cultures of Salvia nemorosa L. Sucrose treatment (4.5%) increased cell biomass (20.58 g L−1) and RA content (17.1 mg g−1 DW). Phenylalanine had no significant effect on cell biomass; however, phenylalanine led to a higher accumulation of RA (16.2 mg g−1 DW) compared to non-treated cells. MeJA caused a slight decrease in cell growth. Maximum RA content (14.2 mg g−1 DW) observed in cells treated with […]

  • A novel soybean transcription factor, DREB7, regulates RD29A and SODFe gene expression in transgenic tobacco plants
    on May 16, 2023 at 12:00 am

    Abstract Glycine max dehydration–responsive element binding (GmDREB) protein is a group of transcription factors that regulate the expression of several downstream genes that respond to abiotic stresses. Among the Glycine max DREB subfamily genes, the role and relationship of GmDREB7 to functional genes that respond to abiotic stresses are also unknown. In this study, the GmDREB7 gene of G. max was analyzed for expression, determining the relationship with two functional genes, NtRD29A and NtSODFe, in Nicotiana tabacum under normal (non-treated) and saline stress conditions daily for 3 wk (NaCl-treated). The 35S-GmDREB7-c-MYC construct was generated and transformed to produce transgenic N. tabacum plants. Three GmDREB7 transgenic N. tabacum lines L27, L42, and L50 were analyzed for their expression at the transcriptional level. Under non-treated and NaCl-treated conditions, the transcript level of the GmDREB7 gene was strongly expressed in all three transformed […]

  • Impact of different Agrobacterium rhizogenes strains on secondary metabolites accumulation in Nitraria schoberi L. hairy roots and antiviral activity of their extracts against influenza virus of subtypes A (H5N1) and A (H3N2)
    on April 13, 2023 at 12:00 am

    Abstract To optimize protocol for obtaining hairy roots of Nitraria schoberi L. with high antiviral activities, factors such as four strain types of Agrobacterium rhizogenes (A4, ATCC15834, R-1601, 8196), two explant types, namely cotyledonous and primary leaves of seedlings, and different cultivation durations (30 and 90 d) were studied. The formation of hairy roots was observed after 2 to 4 wk of incubation, depending on the type of explant and the strain of A. rhizogenes used. The maximum transformation frequency (85.7%) was observed in the cotyledons genetically modified with the strain ATCC15834. The transgenic nature of hairy roots was revealed by PCR with primers to the Agrobacterium oncogenes rolB and rolC. The absence of contamination of the culture by A. rhizogenes was confirmed by primers to the virC and virD1 genes. Phytochemical analysis showed that accumulation of individual metabolites in the line samples exceeded their levels in the native Nitraria […]

  • Optimization of sucrose concentration to promote root proliferation and secondary metabolite accumulation in adventitious root cultures of Ocimum basilicum
    on April 12, 2023 at 12:00 am

    Abstract Ocimum basilicum L. (sweet basil) is a medicinal herb that contains valuable secondary metabolites belonging to different groups. Therefore, the plant displays a wide range of pharmacological activities. In vitro cultures of sweet basil are a very promising approach to produce bioactive compounds, but studies on optimization of essential medium components are very limited. To the current author’s knowledge, there is no previous report on the determination of the optimal sucrose concentration for rosmarinic acid production in adventitious roots of sweet basil in airlift bioreactors. This study aimed to determine the effects of 2.0, 3.0, 4.0, and 5.0% sucrose on rosmarinic acid content, biomass accumulation, and antioxidant activities (DPPH, ABTS, and FRAP) in adventitious root culture of sweet basil under balloon-type bubble bioreactors. Also, phenylalanine ammonia lyase activities (PAL) and accumulation of total phenolic and flavonoid were measured. […]

  • Clonal fidelity and phytochemical analysis of in vitro propagated Kaempferia rotunda Linn.—an endangered medicinal plant
    on April 12, 2023 at 12:00 am

    Abstract Kaempferia rotunda Linn. (Zingiberaceae) is an important herb that has both therapeutic and cosmetic applications. An efficient protocol has been developed for in vitro propagation of K. rotunda using axillary bud explants from unsprouted rhizomes. Murashige and Skoog medium containing 3.0 mg L−1 6-Benzyladenine (BA) in combination with 1.0 mg L−1 indoleacetic acid (IAA) was found to be optimum for the regeneration, multiplication, and in vitro maintenance of plantlets. Two-yr-old ex vitro grown micropropagated plants were assessed for stable drug-yielding potential through the evaluation of essential oil contents, its phytoconstituents, and antioxidant activity. Gas chromatography and mass spectroscopy (GCMS) analysis of essential oil of rhizome showed the presence of benzoic acid (61.34% and 58.27%), bornyl ester (15.11% and 14.66%), zingiberene (5.15% and 5.74%), and camphor (3.72% and 3.82%) in both micropropagated and conventionally grown K. […]

  • In vitro establishment and micropropagation of mango (Mangifera indica L.) from cotyledonary nodes
    on April 1, 2023 at 12:00 am

    Abstract Mango is the fifth most important fruit crop in the world in terms of production with an increasing demand for high-quality plant material for new plantings. Compared to other fruit tree crops, vegetative propagation in mango is slow and allows only a relatively limited production of plant material. To date, efficient procedures for in vitro establishment and micropropagation are not yet available in mango. This work aims at filling this gap. Germination of mango seeds in vitro, compared with ex vitro conditions, significantly increased the germination rate for the monoembryonic genotype tested (‘Irwin’). In vitro germination also increased the number of developed embryos for the two polyembryonic genotypes analysed, ‘Ataulfo’ and ‘Gomera-4’. Regarding the use of shoot tips for in vitro establishment and micropropagation, our results confirmed that this explant is not adequate for this purpose. We report for the first time the use of […]

  • Metabolic changes in hairy root cultures of Centella asiatica treated with methyl-jasmonate and coronatine: a 1H-NMR-based metabolomics approach
    on April 1, 2023 at 12:00 am

    Abstract Hairy root cultures of Centella asiatica–based coupled with exogenous application of methyl-jasmonate and coronatine are suitable bioproduction platforms to overproduce this plant-occurring high-valuable metabolite. These two molecules orchestrate cellular responses in metabolism that have not been yet deeply explored in hairy roots of this plant. The exploration of metabolic changes is crucial since the biotechnological system productivity depends on the knowledge of the biosynthetic machinery of the culture. To understand how methyl-jasmonate and coronatine cause changes in the metabolism of hairy roots of C. asiatica, the metabolic responses to these molecules were monitored through a nuclear magnetic resonance time-series metabolomics approach. The acquired data was analyzed to perform untargeted and targeted metabolomics approaches. Multivariate data analysis statistical models revealed that coronatine caused an early and superior reprogramming […]

  • An efficient semi–in vivo zygotic embryogenesis system in Arabidopsis
    on April 1, 2023 at 12:00 am

    Abstract Embryogenesis is the critical developmental phase in the life cycle of flowering plants, which starts with the double fertilization. In the model plant Arabidopsis, since the single-celled zygote follows a defined cell division and differentiation pattern to form a mature embryo, Arabidopsis embryogenesis is an ideal system for research of the cell fate determination and cell–cell communication. Here, we present an efficient semi–in vivo zygotic embryogenesis system in Arabidopsis via ovule culture. To achieve highly efficient embryogenesis, some key factors were carefully investigated. B5 medium supplemented with 10% sucrose was suitable for embryogenesis. After 5 d of culture, about 43% of all ovules could develop from the zygote stage to the globular embryo stage. By applying the system, we found that exogenous spermidine was not required for embryo development in ovules, and dicyclohexylamine, an inhibitor of spermidine synthase, significantly […]

  • Salicylic acid and jasmonic acid induced enhanced production of total phenolics, flavonoids, and antioxidant metabolism in callus cultures of Givotia moluccana (L.) Sreem
    on April 1, 2023 at 12:00 am

    Abstract Givotia moluccana (L.) Sreem. is a highly valued forest tree species known for its medicinal properties and the presence of bioactive compounds. Elicitation is an effective strategy used for the biotechnological production of secondary metabolites. The study aimed at determining the effects of jasmonic acid (JA) and salicylic acid (SA) elicitation on accumulation of total phenolics, flavonoids, and regulation of antioxidant metabolism in callus cultures of G. moluccana L. Callus cultures treated with 150 µM JA for 20 d resulted in highest contents of total phenolics (81.83 mg GAE g−1 DW) and flavonoids (60.89 mg RE g−1 DW) as compared to SA-treated and control cultures. SA-treated callus cultures showed relatively higher contents of total phenolics (31.43 GAE g−1 DW) and flavonoids (38.49 RE g−1 DW) when treated with 50 µM for 5 d and 20 d, respectively, compared to control callus cultures. Treatment of callus cultures with 150 µM JA for 20 […]

  • Establishment of adventitious root culture system of Cynanchum wilfordii in air-lift bioreactors for the efficient production of bioactive compounds
    on April 1, 2023 at 12:00 am

    Abstract Cynanchum wilfordii is a traditional Chinese herb and has various pharmacological effects. However, its raw plant material is insufficient to meet the market demand. Adventitious roots (ARs) are a novel alternative material for producing plant products. Herein, C. wilfordii ARs were cultured in 5-L bioreactors to produce bioactive compounds and the effects of salt strength and sucrose concentration were investigated in this study. Furthermore, the kinetic study was implemented for confirming a suitable culture period. Finally, the culture efficiency of different size bioreactors was compared. The full salt strength of Murashige and Skoog medium supplemented with 30 g L−1 sucrose was most favorable for AR biomass and the production of flavonoids, ρ-hydroxyacetophenone, 2,5-dihydroxyacetophenone, and total acetophenone derivatives, whereas the media with 50 and 70 g L−1 sucrose were optimal for the production of 2,4-dihydroxyacetophenone and […]

  • A practical framework for the cryopreservation of palm species
    on March 6, 2023 at 12:00 am

    Abstract Palms (Arecaceae) contribute remarkable economic and environmental values to human life. However, many elite and commercial palm species are becoming critically endangered and demand immediate action to preserve their genetic resources. Cryopreservation has established itself as the definitive in vitro conservation method for recalcitrant-seeded species, such as those within the Arecaceae. Progress in this area has moved steadily forward over the last three decades with the development of various techniques for different explants and palm genotypes and for molecular testing methods to ensure genetic fidelity is maintained in the regenerants. There remains a key challenge to properly identify the components that will enable the long-term conservation of palms using cryopreservation. This review methodically analyzes the state-of-the-art cryopreservation techniques developed for palms and places them within a practical framework. This framework encompasses […]

  • Shoot proliferation and de novo shoot organogenesis induction in Pandanus tectorius Sol.—new insights for in vitro plant regeneration
    on February 27, 2023 at 12:00 am

    Abstract Pandanus tectorius Sol. is an economically important coastal and insular tree with various uses. One objective of this study was to multiply shoots from tiller explants. In addition, the effects of basal media, plant growth regulators (PGRs), and the concentration of coconut water (CW) in P. tectorius tissue culture, specifically shoot proliferation and root formation, were assessed. Woody Plant Medium (WPM) was a more suitable basal medium than Murashige and Skoog (MS) medium for shoot proliferation. The optimal shoot proliferation medium was WPM with 0.5 mg L−1 6-benzyladenine (BA), 0.1 mg L−1 α-naphthaleneacetic acid (NAA), and 5.0% CW, resulting in a shoot proliferation efficiency of 7.9 within 35 d. The optimal rooting medium was WPM with 0.5 mg L−1 indole-3-butyric acid (IBA), leading to 100% rooting without the formation of callus from the shoot base. Survival percentage of transplanted plantlets was 100% after 90 d of observation. […]

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