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.


  • Plant regeneration from root segments of Anthurium andraeanum and assessment of genetic fidelity of in vitro regenerates
    on April 8, 2021 at 12:00 am

    Abstract A highly efficient regeneration system of Anthurium andraeanum Linden was established using root segments as explants of four genotypes, Arizona, SWD, OYH, and Mississippi. The root explants were firstly incubated on callus induction media containing NH4NO3 level of 206 mg L−1 or 825 mg L−1, and the calluses were formed from both ends of the root segments after 1 mo of culture. Root-derived calluses were then transferred to shoot regeneration media with NH4NO3 level of 412 mg L−1 or 1650 mg L−1, and numbers of shoot were successfully produced from the surface of calluses after 2 mo of culture. Medium containing 206 mg L−1 NH4NO3 induced more callus compared to 825 mg L−1. In contrast, the number of shoots per callus was the same between the above two shoot regeneration media indicating that NH4NO3 level had little effect on shoot regeneration. Long-termin vitro maintenance of the callus-bearing shoot initials was possible without compromising […]

  • Melatonin alleviates NaCl-induced damage by regulating ionic homeostasis, antioxidant system, redox homeostasis, and expression of steviol glycosides-related biosynthetic genes in in vitro cultured Stevia rebaudiana Bertoni
    on April 1, 2021 at 12:00 am

    Abstract Melatonin has been shown to play a variety of roles in regulating plant growth and response to various abiotic stresses, but its main function remains unclear in response to NaCl toxicity. The aim of the current study was to evaluate the effect of melatonin (0, 5, 10, and 20 μM) on growth, photosynthetic pigments, compatible solutes, ionic homeostasis, enzymatic and non-enzymatic antioxidants, the content of steviol glycosides, and expression of kaurenoic acid hydroxylase (KAH) and uridine diphosphate glycosyltransferase (UGTs) genes in stevia under 25 mM NaCl in in vitro culture. The results showed that in the presence of 25 mM NaCl, melatonin at 5 and 10 μM improved growth parameters, photosynthetic pigments, proline, and soluble sugar contents and reduced electrolyte leakage in stevia leaves. Interestingly, melatonin (5 and 10 μM) reduced the accumulation of malondialdehyde and the level of ROS by improving the redox state of the […]

  • Optimizing micropropagation conditions for a recalcitrant ninebark ( Physocarpus opulifolius L. maxim.) cultivar
    on April 1, 2021 at 12:00 am

    Abstract Ninebark is a very popular ornamental shrub. Micropropagation is an efficient method for mass production of uniform plant material. This study was designed to develop and optimize conditions at all phases of ninebark micropropagation. For the multiplication stage, the Murashige and Skoog (MS) medium at full concentration and pH 5.8 was chosen as the basal medium. Sorbitol proved a more effective carbohydrate source than fructose, with no adverse effects on shoot vitrification or the medium itself. The best shoot production, both in number and length, was on the medium enriched with 2 and 3 mg·L−1 zeatin. High numbers of shoots were also obtained in treatments with 1 mg·L−1 6-benzyladenine (BA) or 2 mg·L−1 meta-Topolin (mT) in the basal medium. BA was the most cost-effective cytokinin. There was a positive effect of the gibberellic acid on proliferation: the highest shoot number per explant was produced in the presence of 1 mg·L−1 GA3. No […]

  • Meta- topolin and liquid medium enhanced in vitro regeneration in Scaevola taccada (Gaertn.) Roxb
    on April 1, 2021 at 12:00 am

    Abstract Scaevola taccada (Gaertn.) Roxb. is a hemi-sclerophyllous littoral shrub of the family Goodeniaceae. It is a salt-tolerant plant and used in soil reclamation and coastal landscaping programs to control beach erosion. The present article reports a reproducible in vitro regeneration system using meta-topolin (mT) through liquid medium employing nodal segment cultures. The highest number of axillary shoots (5.8 ± 0.29) was differentiated on Murashige and Skoog’s (MS) medium containing meta-topolin (1.5 mg L−1). The amplification of shoots (49.8 ± 0.37) and fresh growth was achieved by recurrent subcultures on liquid MS medium supplemented with 0.25 mg L−1 mT and 0.15 mg L−1 indole-3-acetic acid (IAA) in the presence of additives (50 mg L−1 ascorbic acid + 25 mg L−1 citric acid, adenine sulphate and l-arginine). Rooting and acclimatization was simultaneously achieved using concurrent ex vitro rooting and acclimatization (CEVRA) […]

  • Production of some benzylisoquinoline alkaloids in Papaver armeniacum L. hairy root cultures elicited with salicylic acid and methyl jasmonate
    on April 1, 2021 at 12:00 am

    Abstract Papaver armeniacum hairy roots were induced by four Rhizobium rhizogenes strains on three explants (shoot, root, and hypocotyl). Also, the effects of two concentrations (100 and 200 μM) of methyl jasmonate (MJ) and salicylic acid (SA) were assessed on productions of papaverine, noscapine, thebaine, morphine, and codeine and expression of some related genes (TYDC, DBOX, BBE, SalAT, T6ODM, and COR) in P. armeniacum L. hairy root culture at 24 and 48 h after elicitation. R. rhizogenes strain C58C1 induced the highest hairy root rate on hypocotyl explant. Application of 100 μM MJ resulted in the highest contents of thebaine, codeine, and morphine by enhancing the expression of SalAT, COR, and T6ODM genes, respectively, while application of 100 μM SA resulted in the highest contents of papaverine and noscapine by upregulating DBOX and BBE genes, respectively. 100 μM MJ can be used as an effective elicitor in P. armeniacum hairy root culture to increase […]

  • Anatomical changes and cytogenetic stability in bird of paradise plants after zygotic embryo cryopreservation by desiccation method
    on April 1, 2021 at 12:00 am

    Abstract Bird of paradise (Strelitzia reginae Aiton.) is an ornamental plant of great economic value; however, the floricultural market is shaped by trends that vary throughout seasons and years. Thus, conservation techniques, such as cryopreservation, may be used for species and cultivar maintenance. It is important to understand the effect of low temperature used for cooling on the genetic stability and anatomical aspects of conserved species. The objective of this study was to evaluate the occurrence of anatomical changes and the maintenance of the cytogenetic stability of plants recovered from cryopreserved bird of paradise zygotic embryos. Zygotic embryos were cryopreserved in liquid nitrogen (+LN) after being dehydrated for 0, 30, and 60 min using silica gel (moisture content 16.79, 11.38, and 11.49%, respectively). Considering the dehydration duration, seedling development was higher (66.13%) in embryos that were cryopreserved after 30-min dehydration […]

  • In vitro regeneration of the colorful fern Pteris aspericaulis var. tricolor via green globular bodies system
    on April 1, 2021 at 12:00 am

    Abstract Pteris aspericaulis var. tricolor T. Moore ex Lowe is a fern species with colorful fronds, potentially used as an ornamental plant. In the present study, we reported the establishment of in vitro regeneration of P. aspericaulis var. tricolor via green globular bodies (GGBs) system by using in vitro juvenile sporophytes as explants. GGB induction, multiplication and plantlet regeneration media, as well as the influence of light-emitting diodes (LEDs) on GGB multiplication and plantlet regeneration were investigated. The highest GGB induction frequency obtained on the medium supplemented with 2.73 μM thidiazuron (TDZ). The medium containing 1.32 μM 6-Benzyladenine (BA) was the optimum for GGB multiplication. GGB cultivated on 1/4 MS medium with the addition of 0.1% (w/v) activated charcoal (AC) behaved the highest differentiation frequency (94.44%), and 1/4 MS medium without AC was the most effective for plantlet growth. The light quality of LEDs (white, […]

  • Optimisation of gentiopicroside production in Gentiana kurroo Royle from adventitious root cultures in a liquid culture system
    on April 1, 2021 at 12:00 am

    Abstract Gentiana kurroo Royle is a critically endangered medicinal herb of the Indian Himalaya. It has bioactive seco-iridoid glycosides, notably gentiopicroside, in the rhizome and roots. In this study, we report on the in vitro production of gentiopicroside (GPD) from adventitious (AD) roots induced directly from the leaf, nodal, and suspension cell cultures of G. kurroo. Murashige and Skoog (MS) media fortified with Indole-3-butyric acid (IBA) 2 mg L−1 + Naphthaleneacetic acid (NAA) 0.5 mg L−1 and IBA 2 mg L−1 + Indole-3-acetic acid (IAA) 1 mg L−1 produced the maximum number of roots. Suspension cell cultures derived AD roots showed a 1.41-fold higher biomass production than adventitious roots induced from leaf and nodal explants. Among the various concentrations of MS media salts evaluated, half-strength MS suspension media favored the higher biomass and GPD accumulation. The maximum accumulation of GPD (2.58 mg g –l dry weight (DW)) and AD root […]

  • Phytosynthesis of poly (ethylene glycol) methacrylate-hybridized gold nanoparticles from C. tuberculata : their structural characterization and potential for in vitro growth in banana
    on April 1, 2021 at 12:00 am

    Abstract Here, we are analyzing the comparative study of green-synthesized biogenic Caralluma tuberculata gold nanoparticles (Ca-AuNPs) and poly (ethylene glycol) methacrylate coated Ca-AuNPs nanocomposites (PEGMA-AuNPs), formed by hybridization of Ca-AuNPs with PEGMA. Both the virgin and polymer-capped materials (polymer hybrid) were physico-chemically characterized, using Fourier transform infrared spectroscopy (FTIR) that revealed physical interactions during coating without new peak, as PEGMA did not disturb the crystal structure of the AuNPs. However, a red shift of AuNPs from 450 to 520 nm was observed and the overall particle size of AuNPs was increased after PEGMA coating (50 to 100 nm). Furthermore, an efficient protocol of biomass enhancement of in vitro micropropagated Musa acuminate was achieved upon supplementation with Ca-AuNPs and PEGMA-Ca-AuNPs, in the Murashige and Skoog (MS) media in a dose-dependent manner. The maximum fresh, dry weight, and […]

  • Improved in planta genetic transformation efficiency in bitter gourd ( Momordica charantia L.)
    on April 1, 2021 at 12:00 am

    Abstract Production of transformed bitter gourd plants through in vitro regeneration is a laborious practice, which may also result in somaclonal variations. Thereby, in the current study, we have established a less tedious in planta protocol for more efficient genetic transformation of bitter gourd var. NBGH-470 (Apurva) using the seed as a target material. Bitter gourd seeds were transformed by Agrobacterium tumefaciens strain LBA4404 bearing the pCAMBIA1301 binary vector, and the transformed plants were selected against hygromycin B. The putatively transformed bitter gourd (To) plants were examined by GUS assay. Two parameters, including vacuum infiltration and sonication improving the in planta transformation, have been investigated in the present work. Amongst several time durations examined, the highest transformation rate (37%) was achieved upon subjecting the pre-cultured bitter gourd seeds to sonication for 15 min, followed by vacuum infiltration for 6 min […]

  • Isolation and preliminary functional characterization of MxWRKY64 , a new WRKY transcription factor gene from Malus xiaojinensis Cheng et Jiang
    on April 1, 2021 at 12:00 am

    Abstract Malus xiaojinensis Cheng et Jiang is a special and significant germplasm resource of semi-dwarf apple in China. Abiotic stresses, such as Fe and high salinity, affect Malus xiaojinensis growth and development. WRKY transcription factors (TFs) are widely involved in the responses of plants to different stresses. In the present study, a new WRKY gene was isolated from Malus xiaojinensis and designated as MxWRKY64. Subcellular localization showed that MxWRKY64 was a nucleus localized protein. The expression level of MxWRKY64 was highly affected by Fe and salt stress in M. xiaojinensis seedlings. When MxWRKY64 was introduced into Arabidopsis thaliana, it greatly increased the Fe and salt tolerance in transgenic plant. When dealt with Fe stress, overexpression of MxWRKY64 in transgenic A. thaliana contributed to higher levels of fresh weight, root length, and contents of chlorophyll and Fe than wild type (WT). Increased expression of MxWRKY64 in transgenic A. […]

  • Comparative transcriptome analysis reveals molecular regulators underlying pluripotent cell induction and callus formation in Anthurium andraeanum “Alabama”
    on April 1, 2021 at 12:00 am

    Abstract Shoot regeneration from pluripotent cell masses is an important process for genetic improvement of Anthurium andraeanum through Agrobacterium-mediated transformation. However, the mechanisms underlying pluripotent cell induction are not well understood. Here, three half-leaf explant samples collected after 0-, 2-, and 30-d culture on callus-inducing medium were used for RNA isolation and transcriptome profiling. In total, 225,752 non-redundant unigenes were assembled with a mean length of 1299 bp. Differentially expressed genes among the three samples were identified by comparative transcriptome analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed the greatest number of differentially expressed genes in the comparison between 0- and 2-d cultures were related to the “ribosome.” Many differentially expressed genes in the comparison between 2- and 30-d cultures were involved in “plant hormone signal transduction,” “starch and […]

  • Establishment of a micropropagation supporting technology for the Fraxinus mandshurica × Fraxinus sogdiana
    on April 1, 2021 at 12:00 am

    Abstract The interspecific hybridization can take advantage of heterosis and combine the double parental traits most extensively, and is an effective strategy for improving species and stress tolerance breeding for cross-pollinated trees. Species of Fraxinus L. are important for landscaping, timber, and afforestation tree species. To solve problems of poor cold adaption associated with the introduction of Fraxinus sogdiana in Maoershan (Heilongjiang province, Northeast China), to improve pest resistance of Fraxinus mandshurica, and to take advantages of the characteristics of F. mandshurica and F. sogdiana, the interspecific hybridization of F. mandshurica × F. sogdiana was conducted. Tissue culture is the key technology for the promotion of excellent clones and is a powerful tool for germplasm conservation of endangered or valuable plant species. Basing on the embryo culture of hybridization seeds, a complete and efficient micropropagation technology of the […]

  • In vitro regeneration system of Halogeton glomeratus : an important halophyte
    on April 1, 2021 at 12:00 am

    Abstract Halogeton glomeratus (H. glomeratus) is one of the most important halophytes in Asia, and the research on the genes and salt-tolerant mechanisms of this species is limited because of the lack of an optimal and efficient in vitro regeneration system. Here, we developed an efficient plant regeneration protocol using H. glomeratus leaves as explants, which has not been previously reported. High-quality calli were successfully obtained from the H. glomeratus leaves at a frequency of 100% supplemented with 2.00 mg/L 2,4-dichlorophenoxyacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose; the optimum callus subculture medium is 1.00 mg/L 2,4-dichlorophenoxyacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose. Shoots were regenerated on shoot induction medium at a frequency of 100% added with 0.50 mg/L 6-benzylaminopurine, 2.00 mg/L kinetin, 0.20 mg/L naphthaleneacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose; then, shoots […]

  • Steviol glycoside accumulation and expression profiling of biosynthetic pathway genes in elicited in vitro cultures of Stevia rebaudiana
    on April 1, 2021 at 12:00 am

    Abstract In the present study, different elicitors, viz., polyethylene glycol (PEG), alginate (ALG), chitosan (CHI), salicylic acid (SA), and yeast extract (YE), were used in in vitro shoot cultures of Stevia rebaudiana for 4 wk to decipher their effect on growth, biomass yield, and accumulation of steviol glycosides (SG), especially rebaudiosides. The highest leaf number (16.33), root number (4.67), and shoot length (3.80 cm) were observed in media supplemented with 0.5 mg/L YE, whereas maximum shoot number (2.44) and root length (1.89 cm) were recorded in 1.5 mg/L CHI and 1.0 mg/L PEG respectively. Elicitation by 1.0 mg/L YE resulted in enhanced biomass production with a maximum fresh weight of leaves (114.01 mg/plantlet), stem (90.27 mg/plantlet), and shoots (204.28 mg/plantlet). SG content was quantified through ultra-high-performance liquid chromatography (UHPLC). Stevioside (ST) content was increased by 5-fold (0.77 mg/g leaf DW) in 2.0 mg/L ALG and […]

  • Establishment of an efficient shoot regeneration system in vitro in Brassica rapa
    on March 29, 2021 at 12:00 am

    Abstract To improve the genetic transformation system for Brassica rapa L., we established a high-efficiency shoot regeneration protocol. A double haploid line ‘CX001’ with strong regeneration ability was identified from among 25 genotypes. Cotyledons from 4-d-old ‘CX001’ exhibited high-frequency (77.14%) shoot regeneration when explants were cultured on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg L−1 6-benzylaminopurine (BA) and 0.5 mg L−1 α-naphthaleneacetic acid (NAA). The highest regeneration frequency (85.40%) and the maximum shoots number per explant (1.79) were achieved on medium containing 2.0 mg L−1 BA and 0.5 mg L−1 NAA. The addition of 2.5 mg L−1 AgNO3 was critical to shoot regeneration of the cotyledonal explants. The highest rooting rate was obtained on MS medium supplemented with 0.1 mg L−1 NAA. Flow cytometric analysis indicated that there was no ploidy variation between the mother plant and the regenerated […]

  • Cannabis sativa L. photoautotrophic micropropagation: a powerful tool for industrial scale in vitro propagation
    on March 22, 2021 at 12:00 am

    Abstract Global demands for an in vitro culture of cannabis have never been more sought after as countries shift their paradigm towards legalization. Cannabis conventional (photomixotrophic) micropropagation has not been suitable enough for large-scale propagation due to a high degree of plant hyperhydricity, low growth rate, poor rooting, and acclimation efficiency. In the present study, cannabis photoautotrophic micropropagation method is introduced with the purpose of overcoming the difficulties that conventional micropropagation entails when conducted at a large scale. The roles of rockwool medium pH and moisture content, cutting length, basal wounding methods, light intensity, and culture vessel gas exchange capacity were assessed with the intention of increasing productivity of micropropagation method. The results showed 300 mL per vessel of fertilizer solution containing 5-mM MES buffer stabilized medium pH and increased rooting success. Both 5- and 7-cm […]

  • Morpho-physiological investigations in transgenic tomato ( Solanum lycopersicum L.) over expressing OsRGLP1 gene
    on March 17, 2021 at 12:00 am

    Abstract The OsRGLP1 gene was overexpressed under the control of CaMV 35S promoter in tomato (Solanum lycopersicum L.) plants using Agrobacterium-mediated transformation. The transgene expression was validated using RT-qPCR (real-time quantitative polymerase chain reaction) in the transgenic plants. Noticeable morpho-physiological traits, higher SOD activity, and elevated histological H2O2 were manifested by transgenic tomato lines. An increase in total chlorophyll and relative water content (RWC) in the leaves of transgenic lines was noted. The electrolytic leakage/conductivity (EC) in leaves of transgenic lines was lower than the control. The transgenic tomato lines showed dwarf phenotypic features in vegetative and floral traits. Interestingly, the vascular cambium regions in cross sections of stem showed secondary growth in radial symmetry. The transgenic lines also produced higher density and diversity of trichomes. The differences observed in transgenic lines […]

  • In vitro somatic embryogenesis of Texas ebony ( Ebenopsis ebano [Berland.] Barneby & J.W. Grimes)
    on March 15, 2021 at 12:00 am

    Abstract Texas ebony (Ebenopsis ebano [Berland.] Barneby & J.W. Grimes) is a member of the Fabaceae that is native to Mexico. Its wood has multiple uses, and its natural propagation is limited and only by seed. The present investigation describes the first report of Texas ebony somatic embryogenesis with the objective of identifying medium, growth regulator components, and culture conditions. Immature cotyledons were cultured on semisolid and in liquid media containing Murashige and Skoog (MS) basal salts plus 0.0 to 9.06 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The most frequent generation of embryogenic callus occurred with cotyledons cultured in the dark in liquid medium containing 9.06 μM 2,4-D. For the maturation phase, the embryogenic calluses were subcultured on semisolid MS medium with 3.78 to 30.24 μM abscisic acid (ABA), 3% and 6% polyethylene glycol (PEG) 4000, or 3 to 5% sucrose. Sucrose at 4% and 5% induced the greatest total number of somatic […]

  • Silver nanoparticles as the sterilant in large-scale micropropagation of chrysanthemum
    on March 15, 2021 at 12:00 am

    Abstract Micropropagation has proven to be an effective method for large-scale plant production in a short time and a useful tool for plant breeding. Microbial contamination is one of the most difficult micropropagation challenges, resulting in reduced plant quality and loss of valuable stocks. Therefore, sterilization of culture media is a critical step in plant micropropagation. However, sterilized media might reduce the activity of plant growth regulators and nutritional components of culture media. The sterilization effects of silver nanoparticles (AgNP) on the growth of explants and culture media were examined. The treatment with 250 ppm AgNP for 15 to 20 min of 4-wk-old ex vitro leaves proved optimal for controlling the contamination. Furthermore, the Murashige and Skoog medium containing 4 ppm AgNP resulted in 100% medium disinfection (no contamination) after 4 wk of culture. The plantlets obtained from non-sterilized MS medium (NoM) containing 4 ppm AgNP and […]

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