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.


  • Functional identification of apple Baby Boom in genetic transformation and somatic embryogenesis
    on January 23, 2023 at 12:00 am

    Abstract  Agrobacterium-mediated genetic transformation is inefficient in apple, which limits the development of apple-related research and industry. Using molecular methods to create a new and more efficient method of genetic transformation has become the focus of research, and improving the efficiency of the three key steps of transformation, regeneration, and rooting has become an effective means of achieving this goal. Baby Boom (BBM) is widely used in plant biotechnology as an important transcription factor regulating adventitious shoot regeneration and somatic embryogenesis. A homologous gene, MDP0000125317, with high amino acid sequence similarity to AtBBM was cloned in this study, and it was identified from the Royal Gala apple genome (Malus × domestica Borkh.), which was named as MdBBM. The qRT-PCR analysis revealed that MdBBM was expressed at higher levels in the ovary, roots, and seeds. Agrobacterium-mediated transformation of pRI101-MdBBM […]

  • Micropropagation of sweet acacia (Vachellia farnesiana), an underutilized ornamental tree
    on January 13, 2023 at 12:00 am

    Abstract Sweet acacia (Vachellia farnesiana (L.) Wight and Arn) is an attractive small- to medium-sized evergreen tree native to southern North America spanning from southern California, east to southern Florida. Despite its ornamental appeal, sweet acacia has limited commercial availability. A series of experiments were conducted to determine the effect of hydrothermal scarification on in vitro seed germination, and an efficient level of BA (6-benzylaminopurine) for in vitro multiplication, auxin rooting treatments, and acclimatization of sweet acacia. Results revealed that hydrothermal scarification and visual selection of imbibed seeds are the most effective for rapid in vitro seed germination. The source of seed utilized for the in vitro germination exhibited an unusual characteristic of producing seedlings with 2, 3, or 4 cotyledons. Regardless of the cotyledon number observed, sweet acacia can be efficiently micropropagated using multiplication media […]

  • An efficient semi–in vivo zygotic embryogenesis system in Arabidopsis
    on December 27, 2022 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 […]

  • Establishment of adventitious root culture system of Cynanchum wilfordii in air-lift bioreactors for the efficient production of bioactive compounds
    on December 27, 2022 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 […]

  • Morphophysiology of Ananas comosus during in vitro photomixotrophic growth and ex vitro acclimatization
    on December 19, 2022 at 12:00 am

    Abstract Plants in conventional in vitro culture have a mixotrophic-type metabolism and they can present morphophysiological alterations that may prevent adequate photosynthetic capacity. For this, the objective was to verify whether photomixotrophic conditions could optimize photosynthesis, growth, and development of Ananas comosus under in vitro and ex vitro conditions. The experiment was carried out using a completely randomized design in a 2 × 3 factorial arrangement of treatments evaluating types of sealing (flasks with sealed lids or with permeable membranes) and sucrose concentrations (10, 20, and 30 g L−1) with five replications and the experimental unit consisted of one flask containing three plants each. After 90 d of in vitro culture, the following parameters were assessed: shoot and longest root length; number of leaves; leaf area; specific leaf mass; shoot and root fresh and dry mass and the ratio of these parameters; total dry mass; […]

  • In vitro culture using nicotine and d-tubocurarine and in silico analysis depict the presence of acetylcholine receptor (AChR) in tomato (Solanum lycopersicum L.)
    on December 16, 2022 at 12:00 am

    Abstract  Acetylcholine, an established neurotransmitter, is now known to regulate several plant processes. The researchers of this current study have already reported that ACh promotes root and inhibits shoot and callus formation, demonstrating its role in differentiation and morphogenesis in tomato (Solanum lycopersicum) in vitro leaf cultures (Bamel et al. Life Sci 80:2393–2396, 2007; Plant Signal Behav 11:6, 2016). To explore the presence of nicotinic acetylcholine receptor in plant cells, the effects of nicotine and tubocurarine, the agonists and antagonists for this receptor, were also assessed and it was found that nicotine mimicked the action of acetylcholine (ACh) by promoting root formation (Bamel et al. Int Immunopharmacol 29: 231–234, 2015). In the present study, nicotine simulated ACh and reduced the shoot and callus formation. d-Tubocurarine showed antagonistic effect on shoot formation. These investigations demonstrate indirect evidence for the […]

  • Rapid and efficient CRISPR/Cas9-mediated genome editing in potato via hairy root induction
    on December 8, 2022 at 12:00 am

    Abstract Potato is one of the most important crops, and its trait improvement through genetic engineering is of utmost need. CRISPR/Cas9-mediated genome editing tool and its advancement revolutionized genetic and metabolic engineering. Cas9 endonuclease requires gRNA and PAM for targeted genome editing. Delivery of CRISPR/Cas9 component via Rhizobium is most commonly used. Different factors affecting genetic transformation, for example bacterial strains, bacterial density, co-cultivation duration, acetosyringone, explants type, and cultivars, were optimized for Indian potato. Wild-type Rhizobium rhizogenes strain A4 showed the highest (60.7 ± 6.9%) hairy root induction followed by ATCC15834 (53.4 ± 10.7%). The highest transformation efficiency (68.8 ± 6.8%) was achieved onto Murashige and Skoog medium (MS) supplemented with 0.5 mg L−1 naphthaleneacetic acid (NAA). The putative transgenic hairy roots were induced within 8 to 10 d on […]

  • Shoot organogenesis and plant regeneration from leaf and petiole explants of Corydalis saxicola Bunting
    on December 8, 2022 at 12:00 am

    Abstract Corydalis saxicola Bunting, a medicinal herb known for alkaloids, grows mainly in Guizhou province and northwest Guangxi province of China. In this study, for the first time for this plant, an efficient adventitious shoot proliferation and regeneration system via shoot organogenesis from leaf and petiole explants was established. Cytokinins, including 6-benzyladenine (BA) and thidiazuron (TDZ), induced callus and then shoot organogenesis. Most adventitious shoots were induced from leaf explants on Murashige and Skoog (MS) medium (pH 6.5) supplemented with 2.0 µM BA and 0.5 µM α-naphthaleneacetic acid (NAA). This medium was also optimal for shoot proliferation. The rooting ½MS medium was supplemented with 1.0 µM indole-3-butyric acid (IBA) and 0.2 µM NAA; 95.7% of shoots induced roots within 30 d and 91% of plantlets survived on a peat:vermiculite (1:1, v:v) substrate after 90 d.

  • Improvement of mineral nutrition and rooting efficiency of Cannabis sativa L. for in vitro large-scale propagation
    on December 6, 2022 at 12:00 am

    Abstract Cannabis micropropagation is an effective method for producing clean plants in the new and expanding cannabis industry. However, existing protocols are not efficient enough to mass propagate for industry demands and to preserve large numbers of elite cultivars. This research was designed to validate optimal mineral nutrition for cannabis micropropagation. Currently, Murashige and Skoog (MS) basal salts are used in commercially available cannabis medium. Three mineral nutrient factors were defined from MS salts: ammonium nitrate, potassium nitrate, and mesonutrients (CaCl2·2H2O, KH2PO4, MgSO4·7H2O). A total of 20 treatments (MS1-20) were designed, and each factor was varied over a range of concentrations. Leaf canopy area, shoot fresh weight, and multiplication number were measured to quantify the effects of mineral nutrients on the in vitro growth of three cannabis cultivars. All cultivars responded positively to increased nitrogen and mesonutrients […]

  • Indirect organogenesis of Sechium edule (Jacq.) Swartz
    on December 1, 2022 at 12:00 am

    Abstract This article reports an efficient protocol for the indirect organogenesis of Sechium edule, a Mesoamerican species with important nutritional and pharmaceutical characteristics. The in vitro establishment was carried out with hypocotyl segments; subsequently, the formation of calluses was induced from leaves and nodal stem segments using different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D; 0.5, 1.0, and 1.5 mg L−1). Then, shoot formation was induced using different concentrations of thidiazuron (TDZ; 0.5, 1.0, and 1.5 mg L−1) in medium supplemented with 1.0 mg L−1 benzylaminopurine (BAP). Different concentrations of 1-naphthaleneacetic acid (NAA; 1.0, 2.0, and 3.0 mg L−1) were assessed to stimulate shoot rooting; chlorophyll-a, chlorophyll-b, and total chlorophyll were quantified during this stage. The results showed that callus formation and weight increased in parallel with 2,4-D concentration, with nodal segments […]

  • Induction of multiple shoots in Oryza sativa: roles of thidiazuron, 6-benzylaminopurine, decapitation, flooding, and Ethrel® treatments
    on December 1, 2022 at 12:00 am

    Abstract The impact of different concentrations of thidiazuron (TDZ) and 6-benzylaminopurine (BAP) and the role of decapitation, flooding, and Ethrel® treatments were evaluated on direct in vitro caryopsis culture of rice. After 28 d of culture, multiple-shoot formation was observed from 50 to 90% of TDZ-treated seedlings. A similar but lower frequency response was observed using BAP-supplemented media. Furthermore, multiple shoots appeared to arise from the mesocotyl region of the seedlings. In subsequent experiments, isolation and transfer of mesocotyl segments to TDZ-supplemented media resulted in increased number of multiple shoots (10 to 12) as compared to intact seedlings (5 to 6). The formation of multiple shoots per seedling (13 to 15) and percent responding cultures with multiple shoots (70%) increased when decapitated rice seedlings were used instead of intact seedlings or mesocotyl segments. This demonstrated a possible role of physical stress in […]

  • Critical factors governing the efficient direct organogenesis in green-fleshed kiwifruit (Actinidia deliciosa) [A. Chev.] var. deliciosa
    on December 1, 2022 at 12:00 am

    Abstract The present study was focused towards determining the factors governing direct organogenesis in kiwifruit cultivar ‘Allison’. Using in vivo buds or nodal segments as explants, the maximum 75.5% uncontaminated explants were obtained upon consecutive individual disinfection treatment with 1% carbendazim (3 min), 0.1% HgCl2 (1 min), and 2% NaOCl (2 min), respectively. The maximum shoot regeneration response of 84.81% was recorded on Murashige and Skoog (MS) medium fortified with 0.45 μM BA, 0.1 μM IBA, and 0.13 μM GA3. The months of May displayed the highest in vitro explant regeneration response for two consecutive years. The highest shoot proliferation rate was 1:6 with a 5.34 mean number of shoots and a maximum mean shoot length of 6.32 cm on MS medium containing 0.22 μM BA, 0.1 μM IBA, and0.69 μM GA3. A two-step or sequential approach for in vitro root induction showed good quality rooting response of 91.03% producing a higher number of […]

  • Evaluating suitable low-cost agar substitutes, clarity, stability, and toxicity for resource-poor countries' tissue culture media
    on December 1, 2022 at 12:00 am

    Abstract Over 20% of plant species assessed are threatened with extinction. Most of these plants have food security implications, especially in developing countries. Traditional seeds and cutting propagation techniques cannot counter the loss rate, and tissue culture provides a fast alternative to conventional propagation techniques. However, tissue culture has been considered too expensive for developing countries negatively impacted by food insecurity. A gelling agent is the costliest media component in plant tissue culture. This study aims to assess different gelling agents to find suitable ones with low cost and acceptable gelling properties for developing countries, especially in rural areas. Plantain explants were propagated on 16 starch-based substrates to evaluate their suitability as tissue culture gelling agents. This study compared the cost of various substrates and their gelling properties, such as clarity, toxicity, and texture, with agar as a […]

  • Factors affecting in vitro regeneration in the model tree Populus trichocarpa I. Medium, environment, and hormone controls on organogenesis
    on December 1, 2022 at 12:00 am

    Abstract In preparation for a major GWAS (Genome Wide Association Study) of plant regeneration and transformation, a large number of factors were examined for their effects on indirect regeneration rate in diverse wild genotypes—seeking a high rate of regeneration, but also highly genetically variable and heritable treatments. Many of the factors examined have never before been reported on for their effects on callus, shoot, or root organogenesis in poplar (Populus). Stems had the highest regeneration potential, followed by petioles and leaves, with greenhouse grown explant sources superior to in vitro growth explant sources. Changes of ± 50% to Murashige and Skoog (MS) basal medium salts and micronutrients had a minor effect on regeneration. Many popular treatments that were evaluated also had little to no useful effect at the levels studied, including activated charcoal, ascorbic acid, silver nitrate, melatonin, serotonin, sucrose concentration, and lipoic […]

  • Microgram-grade concentration of chlorine dioxide induces one-step plant regeneration in chrysanthemum
    on December 1, 2022 at 12:00 am

    Abstract Efficient in vitro regeneration of chrysanthemum requires virus-free plants for production. Previous studies have employed various exogenous plant growth regulators (PGRs) to induce shoot and root growth. Here, this study reports an efficient one-step regeneration protocol for chrysanthemum through incorporating a 10 μg L−1 chlorine dioxide (ClO2) solution into a medium; this method was compared with the previously reported PGR-mediated regeneration and the PGR-free medium (control). The results showed that the use of 10 μg·L−1 ClO2 achieved one-step regeneration of chrysanthemum by simultaneously inducing the growth of roots and shoots within 3 wk. Transplanting survival reached 100%. The ClO2-induced regenerants showed greater axillary buds, biomass, root number, and root length compared with the control and PGR treatment (P < 0.05). Inter-simple sequence repeat (ISSR) analysis suggested a > 95% molecular fidelity in ClO2-induced […]

  • In vitro medium-term conservation of Garcinia indica: a tropical recalcitrant seeded fruit tree of India
    on December 1, 2022 at 12:00 am

    Abstract Garcinia indica (Thouars) Choisy is one of the important fruit species of the Clusiaceae family distributed in the Western Ghats of India in wild and semi-domesticated forms. This is an under-exploited, dioecious tree with high economic potential. Germplasm conservation of this species is limited to in situ or on-farm level only due to the highly recalcitrant nature of its apomictic seeds. As the seeds are sensitive to moisture as well as low-temperature stresses, conservation has so far not been reported from any seed genebank or cryogenebank. The present work aimed to develop a medium-term conservation strategy under in vitro culture conditions. Adventitious in vitro shoots were developed through direct organogenesis in one accession (IC638184) sourced from Konkan region of the Western Ghats of Maharashtra. These shoots were grown on different concentrations of plant growth regulators to increase the subculture duration of the in vitro plants. The […]

  • Effects of culture methods and plant growth regulators on in vitro propagation of Brassavola nodosa (L.) Lindl. hybrid
    on December 1, 2022 at 12:00 am

    Abstract Brassavola nodosa (L.) Lindl. is an epiphytic orchid with great potential for the ornamental plant industry. The lack of information on propagation and production techniques limits the development of large-scale commercial production. Furthermore, this species is experiencing a reduction in population due to habitat destruction, the impact of climate change, and over-collection from native habitats. This study aimed at developing an efficient protocol for micropropagation of B. nodosa, which could be valuable towards the large-scale commercial production as well as for conservation of this species. Six different concentrations of plant growth regulators (BA or IBA, with or without adenine sulfate) were evaluated in modified Murashige and Skoog (MS) medium for shoot multiplication. In addition, two concentrations of either NAA or IBA were evaluated for rooting. Explants were cultured under three different culture media conditions: semi-solid medium, liquid […]

  • The genetic fidelity study of different callus-derived plantlets of Ficus religiosa L. using ISSR markers
    on December 1, 2022 at 12:00 am

    Abstract Ficus religiosa contributes huge significance in medicinal and ornamental properties. Recently, some obstacles have arisen for their in vivo propagation. Therefore, the present investigation was undertaken to produce genetically stable plants from callus of this species using shoot tip explants. Here, morphologically different calluses were formed based on several plant growth regulator treatments on Murashige and Skoog (MS) medium. Maximum callus (98.89%) was achieved with 2 mg L−1 α-naphthaleneacetic acid (NAA) and 0.5 mg L−1 both for 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP) in combination. The green compact callus exhibited superior result of organogenesis (50%) whereas light brown fragile callus recorded the lowest results (16.17%). The highest organogenesis response was 94.44% with green compact callus cultured on MS medium containing 0.5 mg L−1 BAP and 0.1 mg L−1 NAA. The maximum number of buds per shoot and […]

  • Elicitor-induced production of aervine in adventitious shoot cultures of Aerva lanata (L.) Juss. Ex Schult. and its biological applications
    on December 1, 2022 at 12:00 am

    Abstract Aerva lanata L. with enhanced aervine content can be used as an alternative source of antioxidative and antibacterial compounds through in vitro cultures. This study discusses a new method of aervine production using elicitors in A. lanata adventitious shoot culture. The elicitation of aervine in suspension culture was tested using five elicitors at different concentrations, and the most effective enhancement was recorded on the 20th d of treatment. Multiple shoot development was recorded for nodal explants (5.33 ± 1.15) in the Murashige and Skoog (MS) medium containing 1.0 mg L−1 benzylaminopurine (BAP) and 0.5 mg L−1 kinetin (KIN). Among the elicitors, chitosan (50 mg L−1) and NaH2PO4 (25 mg L−1) produced a 1.26- and 0.79-fold increase in the aervine content than the control. The elicited extract was subjected to liquid chromatography-mass spectrometry analysis to confirm the presence of aervine and the amount was quantified using […]

  • Improving the accumulation of recombinant human serum albumin (HSA) in transgenic tobacco plants by fusion with the N-terminal proline-rich domain of γ-zein (Zera)
    on December 1, 2022 at 12:00 am

    Abstract Plants have long played a major role in human health as a source of herbal remedies. Recently, transgenic plants have become convenient bioreactors for the large-scale production of recombinant biopharmaceuticals at affordable prices. Manufacture of human serum albumin (HSA) in transgenic plant tissue is viable as a safe and economical alternative to the plasma fractionation method. Here, we investigated improvement of recombinant HSA accumulation in tobacco by inducing protein body formation via addition of the Zera (N-terminal proline-rich region of maize γ-zein protein) domain. Genetically modified (GM) tobacco plants were regenerated from leaf explants infected with Agrobacterium tumefaciens strains LBA4404 and GV3101 bearing pBI121-HSA or pBI121-Zera-HSA expression vectors. The T1 tobacco plantlets were examined using southern and western blotting to confirm the integration and expression of the transgenes, respectively. The confirmed T1 tobacco […]

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