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.

 

  • A stable method for Agrobacterium-mediated transformation of Mentha piperita and Mentha canadensis using internodal explants
    on October 4, 2022 at 12:00 am

    Abstract Mentha species are planted worldwide for extraction of their metabolites, including essential oils and phenolic acid compounds, which have therapeutic and commercial value. Mentha piperita L. and Mentha canadensis L. both belong to the menthol chemotype. M. piperita is widely used in North America, and M. canadensis is one of the most important mint species in Asia. Here, an improved Agrobacterium-mediated transformation method using internode segments instead of leaves as explants of M. piperita was reported. Histological examination showed that adventitious buds formed from cortical cells of internode. An efficient method for regeneration of M. canadensis was also established. Internode pieces were excised and cultured on a regeneration medium containing Murashige and Skoog (MS) salts plus vitamin B5, 5.0 mg L−1 TDZ, 0.2 mg L−1 IAA, and 25% coconut water. Explants were then moved to the basal MS medium without plant growth regulator after initiation […]

  • Enhancement of galegine production in cell suspension culture of Galega officinalis through elicitation
    on September 30, 2022 at 12:00 am

    Abstract Galega officinalis L. is known for its dominant secondary metabolite “galegine” a guanidine derivative used in the synthesis of biguanides, metformin, and phenformin as drugs for type 2 diabetes. This study aimed to evaluate the effect of type, concentration, and duration of elicitor application on the content of galegine in the cell suspension culture of G. officinalis. For this purpose, the best cell line in terms of growth rate was selected and propagated for treatment with elicitors. Then, at the end of the log phase of cell growth, chitosan (100, 200, and 300 mg L−1), salicylic acid (SA) (50, 75, and 100 mM), and ultrasound (1, 2, and 4 min) were applied as elicitors. Three and 6-d after elicitation, we harvested the cells and measured total phenol, total flavonoid, and galegine contents. Ultrasound treatment significantly decreased the cell biomass and increased medium electrical conductivity (EC) at 1, 2, and 4 min treatment as compared with […]

  • In vitro propagation for conservation of the rare date palm (Phoenix dactylifera L.) ‘Amri’ using immature inflorescence
    on September 28, 2022 at 12:00 am

    Abstract Immature female inflorescence plays a significant role in date palm micropropagation because inflorescences are available with no practical limit as the source of explants. Moreover, using floral buds for propagation helps in the conservation of date palm biodiversity and the enhancement of socioeconomically valuable landraces. With the goal of avoiding undesirable genetic variability, the optimal combinations and concentrations of plant growth regulators and other medium compositions were investigated to achieve direct organogenesis and multiplication from the immature female inflorescence of date palm (Phoenix dactylifera L.) cultivar Amri. For the initiation stage, the best response was achieved using Murashige and Skoog (MS) medium containing 1.0 mg L−1 zeatin and 1.0 mg L−1 thidiazuron (TDZ) after 16 wk of culturing. For the multiplication stage, the best culture medium contained 0.5 mg L−1 TDZ solidified with GelriteTM, without activated […]

  • Development of efficient Agrobacterium rhizogenes-mediated hairy root system in Curcuma longa L. and elicitation driven enhanced production of pharmaceutically important curcuminoids
    on September 20, 2022 at 12:00 am

    Abstract The Agrobacterium rhizogenes-mediated transformation of plants for hairy root induction epitomizes an attractive biotechnological strategy for enhancing the production of bioactive compounds from medicinal plants. The Curcuma longa L. (turmeric) belonging to Zingiberaceae family is a prevalent crop grown in India and other Southeast Asian countries. The three curcuminoids, viz. curcumin (CUR), demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC), are the key bioactive components attributing to its pharmacological activity. The Curcuma longa hairy root lines were established from shoot base explants of in vitro grown plants using the A. rhizogenes strains A4, A4T, and 8196. The acetosyringone augmentation in bacterial culture and co-cultivation media proved to be advantageous for effective induction of hairy roots. The hairy root lines induced with A4 strain displayed maximum biomass growth after 25 days of inoculation, followed by 8196 and A4T. […]

  • Morphological and transcriptome analyses reveal mechanism for efficient regeneration of adventitious buds from in vitro leaves of Rhododendron delavayi regulated by exogenous TDZ
    on September 19, 2022 at 12:00 am

    Abstract The efficient differentiation of adventitious buds from in vitro leaves has been of great significance for improving Rhododendron traits via genetic transformation. Yet, the actual mechanism of leaf differentiation into adventitious buds remains unclear. In this study, morphological and transcriptome analyses revealed the mechanism by which exogenous thidiazuron effectively regulates the buds’ regeneration from in vitro leaves of Rhododendron delavayi. The experimental treatment consisted of 0.36 µM thidiazuron combined with 0.05 µM 1-naphthaleneacetic acid; for the control, 0.05 µM 1-naphthaleneacetic acid was added to the medium. The results showed that s2 (15 to 22 d) was the key stage for leaf budding. At this stage, the content of endogenous zeatin riboside in leaves regulated by exogenous thidiazuron reached its peak value, and the leaves gained regeneration ability. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway […]

  • Microgram-grade concentration of chlorine dioxide induces one-step plant regeneration in chrysanthemum
    on September 14, 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 […]

  • Towards ex situ conservation of globally rare Turkish endemic Tripleurospermum fissurale (Asteraceae)
    on August 22, 2022 at 12:00 am

    Abstract A rapid and efficient in vitro micropropagation system was developed to conserve Tripleurospermum fissurale (Sosn.) E.Hossain (Asteraceae), a rare endemic species for Turkey. Murashige and Skoog basal medium (MS) supplemented with 4.7 µM KIN was found to be the most appropriate basal medium with a 60% germination rate. MS medium supplemented with 4.9 µM 2iP and 0.5 µM IBA gave the highest shoot length of 54.3 ± 3.53 mm. Furthermore, 4.4 µM 6-BA combined with 0.5 µM IBA was superior for the highest shoot number with 3.4 ± 0.49 after 4-wk culture. Cytogenetic analyses indicated that all propagated plants have the same DNA ploidy level (x) and chromosome number (2n = 18) compared with the mother plants. After 4 wk, the rooting percentage achieved 100% in all tested rooting media. MS medium supplemented with 2.7 µM NAA favored the highest root number, root length, and secondary root number with 3.44 ± 0.5, […]

  • Micropropagation using direct and indirect organogenesis in Artemisia maritima L.: scanning electron microscopy of somatic embryos and genome size analysis by flow cytometry
    on August 18, 2022 at 12:00 am

    Abstract In this study, an efficient, reproducible, and genetically stable regeneration protocol has been developed in Artemisia maritima L. The experiments were conducted for callus induction, plant regeneration, and somatic embryogenesis using stem and leaf of A. maritima as explants. The optimal callus induction (81.3%) was observed on 2.5 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg L−1 6-benzylaminopurine (BAP). The shoot regeneration was observed on different concentrations of BAP, α-naphthaleneacetic acid (NAA), and thidiazuron (TDZ), using nodal segments and microshoot tips as explants. The microshoot tips were more responsive compared to nodal segments with the highest induction frequency (90.33%) obtained on 1.5 mg L−1 BAP. Maximum root induction frequency (74.36%) was obtained on 1.5 mg L−1 NAA. The somatic embryogenesis was induced on Murashige and Skoog (MS) medium amended with TDZ and indole-3-butyric acid (IBA) with maximum […]

  • In vitro medium-term conservation of Garcinia indica: a tropical recalcitrant seeded fruit tree of India
    on August 16, 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 […]

  • High-level expression of sucrose inducible sweet potato sporamin gene promoter: β-glucuronidase fusion gene in transgenic Nicotiana plumbaginifolia hairy roots
    on August 12, 2022 at 12:00 am

    Abstract We developed transgenic Nicotiana plumbaginifolia hairy roots with sucrose-inducible minimal promoter (Spomin)-β-glucuronidase (GUS) gene–fused constructs with signal sequences for sorting to cytosol, apoplast, and ER, and we analyzed the GUS activities of hairy roots after sucrose treatment. Induced GUS activities by Spomin were about 10 times higher than those by the CaMV 35S promoter. GUS activities in hairy roots induced with a Spomin-UTR-GUS construct after 6% sucrose treatment were higher than those with either Spomin-UTR-GUS construct after 10% sucrose treatment, Spomin-apoplast or ER-GUS constructs after sucrose treatment. High GUS activities in hairy roots with Spomin constructs were induced during 20 wk by culturing hairy roots in LS liquid medium containing 6% sucrose in 300 ml conical flasks. Hairy roots in Linsmaier and Skoog (LS) liquid medium showed the increase of fresh weight (FW) for 16 wk. Total yield (μg) of GUS by the hairy roots […]

  • Shoot apex priming, antioxidants, and mechanical injuries modulating regeneration and transformation potential of grain legume crop ricebean (Vigna umbellata (Thunb.) Ohwi & Ohashi)
    on August 3, 2022 at 12:00 am

    Abstract Vigna umbellata is an unexplored and underutilized grain legume of high nutritional values. This crop can be improved for several agronomic attributes via genetic engineering. The presented study emphasizes the first-time standardization of parameters affecting regeneration and transformation using shoot apices. Various parameters comprise concentration of growth regulators, age of explants, explant’s priming with 6-benzylaminopurine (BAP), genotype effect, modulation of explant’s compatibility towards Agrobacterium infection via mechanical wounding, and assessment of the type and concentration of antioxidants (silver nitrate and thiourea). Out of various variables, the 2-d-old apices primed with 15 μM BAP formed an average of 10.4 shoots per explant with 100% regeneration frequency on shoot regeneration medium [SRM2: MSB5V (MS – Murashige and Skoog salts and B5 – Gamborg’s vitamins), 2.5 μM BAP and 5.0 mg L−1 silver nitrate]. Scanning […]

  • Enhanced productivity of atropine in cell suspension culture of Hyoscyamus muticus L.
    on August 1, 2022 at 12:00 am

    Abstract For the first time, suspension cultures produced tropane alkaloids and were obtained from root explants of Hyoscyamus muticus L. It was shown that to obtain a well-proliferating suspension culture from the plant under investigation, a root explant was used. It is advisable to add 6-benzyladenine (0.5 mg L-1) in addition to α-naphthaleneacetic acid (1.0 mg L-1) to the nutrient Murashige and Skoog medium. Histological studies have shown that suspension cultures of H. muticus L. have a complex structure and are characterized by the presence of nodular structures, which are apparently necessary for synthesis and the accumulation of tropane alkaloids. The use of methyl jasmonate (100 and 200 μМ) made it possible to increase the productivity of the suspension culture of H. muticus L., which forms atropine four times more than in plants and more than 5.5 times the root.

  • 2022 In Vitro Biology Meeting, June 4 – 7, San Diego, California
    on August 1, 2022 at 12:00 am
  • In vitro culture and biochemical and antioxidant potential of the critically endangered medicinal plant Atropa acuminata Royle ex Lindl of Kashmir Himalaya
    on August 1, 2022 at 12:00 am

    Abstract Atropa acuminata Royle is a critically endangered annual herb growing in Kashmir Himalaya and is used as a source of medicine for many ailments like arthritis, muscle pain, and joint pain. Due to its immense medicinal importance, the plant needs a tissue culture protocol for mass propagation. The present study is an attempt to develop a highly reliable and reproducible tissue culture protocol for this valuable herb and also an attempt to evaluate and estimate primary biochemical attributes and antioxidant activities. The tissue culture protocol was standardized using in vitro–raised nodal segments and leaves as explants followed by the studies on biochemical evaluation and antioxidant potential of both in vitro–regenerated and field-raised plant material. Best regeneration was observed on MS media augmented with BAP and IAA with the maximum number of shoots obtained as 17.33 ± 4.15. For biochemical studies, field-growing leaf explants, in […]

  • In vitro assessment of strawberry (Fragaria × ananassa Duch.) plant responses to water shortage stress under nano-iron application
    on August 1, 2022 at 12:00 am

    Abstract Strawberry is a susceptible plant to water stress, and its growth is severely declined under water shortage. Therefore, the aim of this study was to assess the impacts of iron, as a micro-nutrient, in the form of a nano-oxide on the tolerance of strawberry plants to water stress conditions under in vitro culture. Three levels of PEG-induced water stress (0, 5%, and 7% w v−1 PEG 6000) and three levels of iron nano-particles (0, 1, and 1.2 mg L−1) were supplemented to Murashige and Skoog (MS) culture medium. The results demonstrated that PEG-induced water stress negatively affected shoot fresh and dry weight, root fresh and dry weight, and physiological traits such as relative water content, membrane stability index, and pigment content. The biochemical traits were also affected by PEG-induced water stress; consequently, antioxidant enzyme activity, leaf proline content, total soluble sugar, and malondialdehyde (MDA) and H2O2 content were increased. The […]

  • A new economical storage technique for strawberry (Fragaria × ananassa Duch.) in vitro
    on August 1, 2022 at 12:00 am

    Abstract Strawberry plants grown in vitro are typically stored and maintained on agar containing Murashige and Skoog (MS) media and sucrose as a carbohydrate source. This method of storing strawberry plants in vitro is expensive and time consuming, requiring sub-culturing onto fresh media every 2 to 3 mo. This study aimed to establish the viability of using a substrate alternative as an economical replacement for MS media, for both long-term storage in vitro and ease of transfer (ex vitro). A protocol was developed for strawberry in vitro using commercially available sterilized peat pellets (Jiffy-7® pellets), to optimize culture conditions and tissue culture practices. Suitability of the peat substrate was measured by the plant’s overall response to culture in vitro and subsequent health and survival ex vitro following deflasking. Included in this study was a comparison of the use of vented vs non-vented tissue culture vessels for their effect on plant […]

  • Production and antioxidant activity of secondary metabolites in Hassawi rice (Oryza sativa L.) cell suspension under salicylic acid, yeast extract, and pectin elicitation
    on August 1, 2022 at 12:00 am

    Abstract Plants that produce bioactive chemicals provide a viable in vitro method for producing key nutraceutical substances, especially in the presence of appropriate elicitors. The goal of this study is to use elicitors in Hassawi rice cell suspension culture to enhance production of phenolic compounds. Salicylic acid (SA), yeast extract (YE), and pectin were utilized at 50, 100, and 200 mg L−1. Impacts of elicitors on cell culture were measured in terms of biomass packed cell volume (PCV), dry and fresh weight, total phenolic, total flavonoid, anthocyanin, tannins content, antioxidant activity, and phenolic compounds using ultra performance liquid chromatography mass spectrometry (UPLC-mass spec). Except for high doses of pectin (200 mg L−1), the 50, 100, and 200 mg L−1 SA–, YE–, and pectin–treated cell culture produced increased PCV, dry (DW), and fresh weight (FW), and all of the aforementioned contents. The cell suspension supplied with […]

  • Shoot organogenesis from leaf and stem explants of Heliotropium foertherianum Diane and Hilger
    on August 1, 2022 at 12:00 am

    Abstract Heliotropium foertherianum Diane and Hilger (Boraginaceae), an evergreen shrub or tree that grows naturally on tropical atoll sandbanks above the high tide line of beaches, is used for landscaping in tropical islands but also has medicinal and ecological value. In this study, shoot proliferation and regeneration protocols were established for H. foertherianum for the first time. Adventitious shoots were successfully induced from young leaves and internode explants on Murashige and Skoog (MS) medium supplemented with 1.0 mg L–1 thidiazuron, applied alone or in combination with 0.2 mg L–1 6-benzyladenine (BA). Shoot proliferation was optimal on MS medium with 2.0 mg L–1 BA and 0.2 mg L–1 indole-3-butyric acid (IBA), achieving a shoot proliferation coefficient of 4.7 within 45 d of culture. When BA concentration was too high (> 3.0 mg L–1), fasciated stems and abnormal leaves formed. Adventitious roots were induced from 100% of […]

  • Exogenous dehydrin NnRab18 improves the Arabidopsis cryopreservation by affecting ROS metabolism and protecting antioxidase activities
    on August 1, 2022 at 12:00 am

    Abstract Seeds usually dehydrate and reach a vitrification state during maturation, conferring stress tolerance and storage tolerance on mature seeds. Late embryogenesis abundant (LEA) proteins accumulate and play multiple protective roles in this process. In this study, recombinant dehydrin NnRab18 of LEA family from Nelumbo nucifera seeds was added into loading solution and plant vitrification solution (PVS2) to verify the optimization effect of NnRab18 on plant cryopreservation. The NnRab18-optimized method approximately doubled the survival rate of Arabidopsis thaliana L. seedlings, and the survival rates were linearly dependent on concentrations of exogenous NnRab18 protein. Evans blue staining revealed that NnRab18 alleviated the plasma membrane damages of seedling roots. In the NnRab18-optimized group, OH· scavenging activity was increased by 50.0%, and H2O2 and malondialdehyde contents were decreased by 9.3% and 29.7%, respectively. Quantitative gene […]

  • Correction to: Enhanced productivity of atropine in cell suspension culture of Hyoscyamus muticus L.
    on August 1, 2022 at 12:00 am
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