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.


  • Physiological and morphological analyses of Thymus vulgaris L. in vitro cultures under polyethylene glycol (PEG)-induced osmotic stress
    on April 16, 2019 at 12:00 am

    Abstract Morphological and physiological parameters of polyethylene glycol (PEG)-induced drought-stressed Thymus vulgaris L. (thyme) seedlings were examined. To start in vitro selection for drought stress tolerance and to provide a more efficient method for the production of pharmaceutical substances, callus induction and production of essential oils were analyzed. Thyme seeds were germinated on medium containing 0, 2, 4, 6, and 8% (w/v) PEG; and, for callus induction, stem explants were cultured on medium supplemented with 1 mg L−1 2,4-dichlorophenoxyacetic acid with different concentrations of PEG. Four weeks after PEG-induced water stress, a significant decrease in seedling growth, chlorophyll and carotenoid contents, and callus weight and induction was observed. However, no significant difference was found in the germination rate of seeds. Organic osmolytes, e.g., proline and carbohydrates, were influenced by PEG-induced osmotic stress. Reducing […]

  • Evaluation of canavanine as an allelochemical in etiolated seedlings of Vicia villosa Roth: protoplast co-culture method with digital image analysis
    on April 16, 2019 at 12:00 am

    Abstract Strong allelopathic activities of etiolated seedlings of a leguminous plant, Vicia villosa Roth (hairy vetch), were confirmed by a protoplast co-culture method. The combination of 1% Cellulase RS and 1% Driselase 20 in 0.4 M mannitol solution was used for protoplast isolation. The inhibitory effects of V. villosa protoplasts on the growth of recipient Lactuca sativa L. (lettuce) protoplasts were determined using 96-well culture plates with 50 μL of liquid medium consisting of Murashige and Skoog’s basal medium containing 1 μM 2,4-dichlorophenoxyacetic acid, 0.1 μM 6-benzyladenine, 3% (w/v) sucrose, and 0.4 M mannitol in each well. The allelopathic activity of V. villosa epicotyl protoplasts (2 × 102 to 105 mL−1), on cell division of lettuce protoplasts (6 × 103 to 5 × 104 mL−1), was stronger than that of roots and was stronger than on […]

  • In vitro cloning via direct somatic embryogenesis and genetic stability assessment of Paphiopedilum niveum (Rchb.f.) Stein: the endangered Venus’s slipper orchid
    on April 16, 2019 at 12:00 am

    Abstract Wild populations of Paphiopedilum niveum (Rchb.f.) Stein, an endangered orchid species, have been threatened with extinction due to climate change and overcollection. Conventional methods of propagation are unable to meet market demands. The current study reports the production of genetically stable, protocorm-derived protocorm-like bodies (PLBs) of P. niveum that can be used for commercial production. These PLBs are referred to hereafter as somatic embryos (SEs). Factors affecting SE induction and SE proliferation were examined. The highest percentage of SE formation (68.33 ± 11.77%) and the maximum number of SEs per explant (5.19 ± 0.67) were obtained on modified Vacin and Went (MVW) medium containing 0.1 mg L−1 1-naphthaleneacetic acid (NAA). The highest increase in fresh weight (FW) of proliferated SEs (183.33 ± 28.93 mg per 100 mg initial FW) was gained on […]

  • Plant tissue culture media and practices: an overview
    on April 15, 2019 at 12:00 am

    Abstract This review presents an overview of the culture media and practices used in plant tissue culture and developmental biology. The compositions of the most commonly used basal media, especially Murashige and Skoog (MS) and modified MS (MMS), Gamborg’s B5 medium and B5 modifications, Woody Plant Medium (WPM), and Driver and Kuniyuki Woody plant medium (DKW) are discussed, along with typical basal medium manipulations to elicit and support various developmental responses. The most commonly used plant growth regulators and their applications to promote various developmental responses are examined, along with a presentation of the classical phytohormone developmental models for organogenesis and somatic embryogenesis. Elaborated developmental models for both organogenesis and somatic embryogenesis, with emphasis on discrete developmental steps, occasional need for multiple manipulations in culture to achieve a single developmental step, and identification of […]

  • The in vitro propagation system of Citrus × latifolia (Yu. Tanaka) Yu. Tanaka (Rutaceae) affects the growth and depletion of nutriments
    on April 15, 2019 at 12:00 am

    Abstract Among citrus fruits, Citrus × latifolia (Yu. Tanaka) Yu. Tanaka stands out given its diverse uses, and there is a great demand for plants of high phytosanitary and genetic quality. In the present study, the in vitro micropropagation process of C. latifolia was studied in RITA®-type bioreactor, semisolid medium, and partial immersion with liquid culture medium. Explants that were 2 mo of age were used and apices of 0.5 cm were extracted and placed in each culture system. In all the systems, MS (Murashige and Skoog) culture medium was used, supplemented with 6-benzylaminopurine (BAP) and kinetin (KIN). After 30 d, the number of shoots, the size of the shoots, the number of leaves, and the depletion of nutrients from the culture medium were analyzed. With the data obtained, ANOVA and Tukey’s mean comparison tests were performed. The RITA® bioreactors, with the addition of 40 mL of medium per explant in a 4-h-interval […]

  • Wounding and medium formulation affect de novo shoot organogenic responses in hypocotyl-derived explants of annatto ( Bixa orellana L.)
    on April 15, 2019 at 12:00 am

    Abstract Bixa orellana L. (annatto) is the unique exploited source for the production of bixin and norbixin natural dyes, which are widely used in the food, pharmaceutical, cosmetic, and textile industries. In tissue culture, adequate culture conditions are essential for the development of the plant. Among them, the saline composition of the culture medium and the type of explants are limiting factors. In this study, the effect of MS (Murashige and Skoog), Correia and colleagues (JADS) medium, Woody Plant Medium (WPM), and Driver Kuniyuki Walnut (DKW) culture media and wounding of hypocotyl explants on de novo shoot organogenesis (DNSO) of annatto were evaluated. It was demonstrated that JADS medium improved the nutritional status of shoots, which promoted the best development and quality of the plants generated after acclimatization, when compared to MS medium. The use of JADS medium for annatto DNSO during 45 d of culture was sufficient to regenerate and […]

  • Plant tissue culture and biotechnology: perspectives in the history and prospects of the International Association of Plant Biotechnology (IAPB)
    on April 12, 2019 at 12:00 am

    Abstract The evolutionary route from plant tissue culture (IAPTC) to plant biotechnology (IAPB). Plant biotechnology is an evolutionary scientific process, formulated and maintained by our accumulated cultural-societal knowledge and the invention of new technologies (Altman and Mesoudi submitted). It emerged thousands of years ago when wheat, rice, chickpeas, potatoes, and coffee (and other plants) were first domesticated; when grains were fermented by yeasts to produce bread; and when grape juice, barley, and tubers fermentation resulted in wine, alcohol, and beer. The modern era of plant biotechnology started in the beginning of the twentieth century and is associated with the ability to grow plant cells and tissues in vitro, to regenerate and clone new plants and, later, to modify their genetic characteristics by molecular breeding, including molecular marker-assisted selection (MAS), genetic modification (GM), and, more recently, genome editing. Additional novel […]

  • Histo-morphological analysis of rice callus cultures reveals differential regeneration response with varying media combinations
    on April 12, 2019 at 12:00 am

    Abstract Genetic transformation of most indica rice (Oryza sativa) cultivars is hampered by poor in vitro culture performance and low regeneration potential. Histological study of primary calli can provide substantial information on their regeneration potential and can be used for early grading of calli expected to develop plantlets on regeneration media. The study was aimed to undertake histological analysis of primary calli derived from mature seeds of five indica rice cultivars viz. KSK-133, KS-282, Shaheen Basmati, Super Basmati, and DilRosh in order to assess their regeneration potential on different media combinations supplemented with various hormone concentrations (N6 + 2 mg/L 2,4-Dichlorophenoxyacetic acid; N6 + 2 mg/L 2–4 D + 2 mg/L Benzylaminopurine and MS + 2 mg/L 2,4-D). Calli with regeneration capability were subjected to histological assays by examining toulidine blue […]

  • Increased freezing stress tolerance of Nicotiana tabacum L. cv. Bright Yellow-2 cell cultures with the medium addition of Ascophyllum nodosum (L.) Le Jolis extract
    on April 10, 2019 at 12:00 am

    Abstract The study of bioactive components of the brown alga Ascophyllum nodosum (L.) Le Jolis has shown that they promote growth and increase productivity of plants. However, a standardized model system providing consistent responses to such bioactive components has yet to be established. Given that freezing stress, especially in northern climates, reduces plant growth and productivity, a technique was developed to protect plant cells under freezing stress using the natural products made from the abundant A. nodosum as a cell culture supplement. In this study, a homogenous cell culture system of Nicotiana tabacum L. cultivar Bright Yellow-2 (BY-2) suspension cells was used to investigate the bioactivity and protection level of this alga extract under freezing temperatures, and BY-2 cell growth, physiology, and molecular changes were measured in the presence or absence of the extract. The results indicated that A. nodosum extract significantly improved BY-2 cell […]

  • Micropropagation of Nelumbo nucifera ‘Weishan Hong’ through germfree mature embryos
    on April 9, 2019 at 12:00 am

    Abstract Asian lotus (Nelumbo nucifera Gaertn.) is an economically important aquatic plant that is widely cultivated in Asia. It is mainly propagated by dividing rhizomes, which maintains genotype stability but is also laborious and time-consuming. An efficient in vitro method that ensures genetic stability is highly desirable. An optimized method was developed to produce intact and sterile mature embryos by soaking fruits in sterile water containing 2% (v/v) plant preservative mixture. Using mature embryos as explants, an improved micropropagation procedure was established in N. nucifera ‘Weishan Hong.’ The growth patterns of explants differed with the types and concentrations of cytokinins. Media containing 0.5 to 2.0 mg L−1 kinetin were suitable for elongation of unexpanded stems, while media with 1.0 to 2.0 mg L−1 6-benzylaminopurine (BAP) the stems were shorter. Higher concentrations of about 3.0 mg L−1 […]

  • Seed cryopreservation and micropropagation of the federally threatened species, Price’s potato-bean ( Apios priceana B.L. Robins.)
    on April 4, 2019 at 12:00 am

    Abstract Apios priceana, commonly known as Price’s potato-bean, is a perennial species native to the Southeastern US. Habitat destruction has caused A. priceana to become rare, and it is federally listed as threatened. Protocols developed for in vitro germination, shoot micropropagation, in vitro rooting, shoot establishment in soil, and seed cryopreservation will assist in the safeguarding and conservation of dwindling natural populations. Seeds were germinated in vitro on plant growth regulator (PGR)-free Murashige and Skoog (MS) medium after seed sterilization in H2O2 and seed nicking. Greatest shoot multiplication occurred on MS medium with BAP/IBA/GA3 at 2.22/0.49/1.44 μM with 2.0 g/l Phytagel and pH adjusted to 5.7. Shoots rooted in vitro in MS medium with 3.2 μM IBA. Shoots rooted in vitro rapidly established in greenhouse potting mix, usually showing new growth within 2 wk, with tuber formation by the end of the growing season. […]

  • In vitro storage of micropropagated grapevine rootstocks at low temperature
    on April 3, 2019 at 12:00 am

    Abstract A long-term in vitro conservation protocol for five micropropagated grapevine rootstocks (3309 Couderc, 110 Richter, Vitis riparia Gloire de Montpellier, 101-14 Millardet et De Grasset, and Teleki 5C) was developed and optimized by testing the effect of culture containers (polyethylene bags versus glass test tubes), explant types (rooted versus unrooted cuttings), low-temperature regimes (1.6, 4.0, and 7.2°C), and light intensity (low versus dark). Optimal storage conditions were achieved at 4.0°C and low light intensity (3 μmol m−2 s−1), with rooted plantlets established on woody plant medium (WPM) supplemented with 15 g L−1 sucrose and 37 mg L−1 cysteine in polyethylene bags. After 12 and 24 mo of optimal cold storage, the plant condition ratings (3.5 and 2.5 on a scale of 4) and survival rates (75% and 52%) were high, regardless of the rootstock genotype, as determined by visual […]

  • Goji berry ( Lycium barbarum L.) in vitro multiplication improved by light-emitting diodes (LEDs) and 6-benzylaminopurine
    on April 2, 2019 at 12:00 am

    Abstract In Brazil, Lycium barbarum L. (goji berry) is an agronomically valuable, imported, commercialized plant. The present study evaluated the effects of cytokinins and light sources on optimization of in vitro multiplication of goji berry by axillary bud proliferation. Nodal explants were excised from plants 90 d after in vitro germination and cultured on Murashige and Skoog (MS) medium containing 30 g L−1 sucrose; 7 g L−1 agar-agar; and 0, 2, 4, 8, or 16 μM 6-benzylaminopurine (BAP), 6-furfurylaminopurine (KIN), or thidiazuron (TDZ). After 60 days, regenerated lateral buds and shoot lengths were evaluated. The regenerated shoots were inoculated in MS supplemented with 0.0, 2.5, 5.0, 10.0, or 20.0 μM BAP under white fluorescent lamps or mixed treatment of red/blue (RB) light-emitting diode (LED) lamps. After 60 days in vitro cultivation, the number of formed shoots, leaf number, and shoot length were […]

  • Cell suspension culture of Rhizoma zedoariae in a two-stage perfusion bioreactor system for β-elemene production
    on April 1, 2019 at 12:00 am

    Abstract We presented a two-stage combined bioreactor system consisting of a stir-tank and an airlift column, and challenged with Rhizoma zedoariae cell suspensions for β-elemene production. Two-stage culture was initiated when the cell concentration in both vessels was maintained at an appropriate density. The cells were proliferated in stirred-tank with the maximal growth rate of 0.17 d−1 to present enough cells for β-elemene synthesis. In the airlift column, continuous cell separation from culture medium was achieved by using a cell retention device based on centrifugal and gravity settling when the system was performed in perfusion mode. The results indicated that additives can efficiently promote the accumulation of β-elemene in R. zedoariae cells. In addition, the β-elemene content showed higher levels in cell lines of overexpressing 3-hydroxy-3-methylglutaryl coenzyme-A reductase, Farnesyldi phosphate synthase, and ST02C genes. […]

  • Overexpression of cinnamyl alcohol dehydrogenase gene from sweetpotato enhances oxidative stress tolerance in transgenic Arabidopsis
    on April 1, 2019 at 12:00 am

    Abstract Cinnamyl alcohol dehydrogenase (CAD) is the enzyme in the last step of lignin biosynthetic pathway and is involved in the generation of lignin monomers. IbCAD1 gene in sweetpotato (Ipomoea batatas) was identified, and its expression was induced by abiotic stresses based on promoter analysis. In this study, transgenic Arabidopsis plants overexpressing IbCAD1 directed by CaMV 35S promoter were developed to determine the physiological function of IbCAD1. IbCAD1-overexpressing transgenic plants exhibited better plant growth and higher biomass compared to wild type (WT), under normal growth conditions. CAD activity was increased in leaves and roots of transgenic plants. Sinapyl alcohol dehydrogenase activity was induced to a high level in roots, which suggests that IbCAD1 may regulate biosynthesis of syringyl-type (S) lignin. Lignin content was increased in stems and roots of transgenic plants; this increase was in S lignin rather than guaiacyl (G) lignin. […]

  • RNAi targeting the gene for 1,8-cineole synthase induces recomposition of leaf essential oil in lavandin ( Lavandula  ×  intermedia Emeric)
    on April 1, 2019 at 12:00 am

    Abstract Recomposition of volatile compounds is an effective way to alter the fragrance of an essential oil. Using RNA interference (RNAi), we attempted to reduce the production of 1,8-cineole, a major monoterpene in essential oils, to alter the composition of the essential oil in lavandin. We obtained 12 transgenic regenerants via inoculation with Agrobacterium tumefaciens, including an RNAi inducing vector targeting five regions of the gene for 1,8-cineole synthase (CINS). Of these, two regenerants, targeting a coding region about 250 nt upstream with 5′-UTR and a coding region about 300 nt downstream of CINS mRNA (CINS I and V), respectively, showed a significant decrease of 1,8-cineole production in leaf essential oil, although the overall composition was barely altered because the production of other compounds decreased concurrently. By contrast, in two other regenerants, targeting a coding region about 1000 nt in the middle and a coding region […]

  • Improved plant regeneration in callus cultures of Sorghum bicolor (L.) Moench
    on April 1, 2019 at 12:00 am

    Abstract Sorghum bicolor is a recalcitrant species for tissue culture regeneration and genetic transformation. Browning of explants is one of the factors limiting organ and tissue cultures. To overcome this, callus tissue was initiated from the shoot tips of in vitro germinating seeds (S. bicolor cv. Róna 1), and then cultured on modified MS media (Murashige and Skoog in Physiol Plant 15:473–497, 1962). In the first experiment, we tested callus induction on several media supplemented with casein hydrolysate, polyvinylpyrrolidone, honey, and sucrose. The best callus induction was recorded for the medium with honey and sucrose (80.0%) and for control medium (79.8%). Shoot regeneration was tested on the MS medium with 6-benzylaminopurine (BAP) supplemented with honey and sucrose at a 1:1 ratio (by weight) or with sucrose only. The highest percentage of calluses regenerating shoots was noted for those induced on the medium with sucrose and […]

  • Phytostilbenoid production in white mulberry ( Morus alba L.) cell culture using bioreactors and simple deglycosylation by endogenous enzymatic hydrolysis
    on April 1, 2019 at 12:00 am

    Abstract Phytostilbenes are responsible for several biological activities of mulberry (Morus sp.), which has been widely used as a raw material in health products. This study aimed to investigate the capability of Morus alba L. cell in bioreactors to produce the major bioactive stilbenes. The cell obtained from air-driven bioreactors such as round bottom, flat bottom, and air-lift vessel shape bioreactors was collected and analyzed for the levels of mulberroside A and oxyresveratrol. The results showed that the cell culture in round bottom and air-lift vessel bioreactors had higher growth rate, as compared with the cell culture in shake flasks (1.38- and 1.41-fold, respectively). The optimized culture condition to produce mulberroside A was obtained from round bottom bioreactor culture (55.56 ± 11.41 μmol/L). Additionally, endogenous stilbenoid hydrolysis of cell from the bioreactor culture was examined. Under optimized hydrolytic […]

  • Thidiazuron induced efficient in vitro organogenesis and regeneration of Scutellaria bornmuelleri : an important medicinal plant
    on April 1, 2019 at 12:00 am


  • Data mining study of hormone biosynthesis gene expression reveals new aspects of somatic embryogenesis regulation
    on April 1, 2019 at 12:00 am

    Abstract Although the application of exogenous hormones and the status of endogenous hormones play a crucial factor in somatic embryogenesis (SE), our understanding of the status of endogenous hormones and the molecular framework for hormone biosynthesis during SE are still elusive. With a data mining approach, we analyzed public RNA-seq data of SE from two model plants, Arabidopsis thaliana and Oryza sativa var. Japonica, which showed that several plant hormone biosynthesis genes had distinct patterns during SE. The expressions of auxin, cytokinin, gibberellin, and ethylene biosynthesis genes were induced during SE in A. thaliana, while only jasmonic acid showed an expression increase during somatic embryogenesis in both plants. Furthermore, by analyzing public RNA sequencing and DNA affinity purification sequencing (DAP-seq), we revealed some possibilities of the role of ethylene during SE through the activity of EIN3, by increasing the expression level of major […]

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