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.


  • RNAi targeting the gene for 1,8-cineole synthase induces recomposition of leaf essential oil in lavandin ( Lavandula  ×  intermedia Emeric)
    on February 13, 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 […]

  • Peculiarities of calcium and iron effects on some wild terrestrial orchids in vitro compared to in vivo
    on January 31, 2019 at 12:00 am

    Abstract There are 32 species from the family Orchidaceae in Latvia, and 26 are rare and endangered, and their preservation in an in vitro bank is vital. The death of in vitro grown wild terrestrial orchids is mainly caused by the release of phenolic compounds from root tissues. As mineral nutrients form a significant component of culture media, a hypothesis was advanced that increased doses of both Ca and Fe could prevent phenol oxidation and improve micropropagation. Liparis loeselii (L.) Rich. and Gymnadenia conopsea (L.) R. Br. calcicole plants, and Dactylorhiza russowii (Klinge) Holub, a non-calcicole plant, were used as model species. Modification of the culture media with increased Ca gluconate monohydrate concentrations significantly improved the quality of L. loeselii and D. russowii plants, especially in the presence of elevated levels of ferric citrate. There was no benefit of Ca for G. conopsea, except in combination with the highest level of ferric […]

  • Programmed cell death induced by cadmium stress and/or boron deprivation in tobacco ( Nicotiana tabacum L.) cultivar Bright Yellow 2 cells
    on January 31, 2019 at 12:00 am

    Abstract Cadmium (Cd) is one of the most toxic and widespread heavy metal pollutants in soil. As an essential mineral nutrient, boron (B) plays critical roles in physiological processes of plants. In the present study, programmed cell death (PCD) induced by Cd stress and/or B deprivation was assessed and the underlying mechanisms were clarified in suspension-cultured Nicotiana tabacum L. cultivar Bright Yellow 2 (TBY-2) cells. The PCD in TBY-2 cells was analyzed by Hoechst 33258 staining and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and then, expression analysis of PCD-related genes was performed using quantitative real-time polymerase chain reaction (qPCR) assays. The production of reactive oxygen species (ROS) was determined using fluorescence microscopy of 2′,7′-dichlorofluorescein diacetate–labeled cells. The levels of lipid peroxides were quantified by the thiobarbituric acid–reactive substances […]

  • Correction to: Improving micropropagation of Mentha × piperita L. using a liquid culture system
    on January 31, 2019 at 12:00 am

    This correction reflects Sadanand A. Dhekney’s updated e-mail address and affiliation. […]

  • Improving callus regeneration of Miscanthus  ×  giganteus J.M.Greef, Deuter ex Hodk., Renvoize ‘M161’ callus by inhibition of the phenylpropanoid biosynthetic pathway
    on January 24, 2019 at 12:00 am

    Abstract In previous studies, the regeneration rates of Miscanthus × giganteus J.M.Greef, Deuter ex Hodk., Renvoize from callus tissue cultured on semi-solid media significantly declined after 4 mo of culture, which presents problems with germplasm conservation and use as an alternative propagation system. Due to the species’ lignocellulosic nature, it was hypothesized that the accumulation of phenolic compounds in the callus may be responsible for inhibiting regeneration. The current study aimed to optimize regeneration of M. × giganteus callus by culturing the callus tissue in the presence of 2-aminoindan-2-phosphonic acid (AIP), a competitive inhibitor of phenylalanine ammonia lyase (PAL), to reduce the biosynthesis of phenolics. Embryogenic callus was cultured on media supplemented with 9.0- or 11.3-μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0-, 1-, 10-, 100-, or 1000-μM AIP. Every 28 d for […]

  • An efficient callus-based in vitro regeneration protocol for Warburgia Ugandensi s Sprague, an important medicinal plant in Africa
    on January 22, 2019 at 12:00 am

    Abstract Warburgia ugandensis Sprague is a woody species in the family Canellaceae and an important source of medicines in Africa. Natural propagation of W. ugandensis is problematic due to its recalcitrant seeds and lack of an efficient in vitro regeneration system for this species. This study describes an efficient regeneration protocol. Petiole bases and shoot tips were used as explants. Callus tissue developed when the explants were cultured on Murashige and Skoog medium containing 30 g L−1 sucrose and 7 g L−1 agar (MS30 medium), supplemented with 1.0 mg L−1 indole-3-butyric acid (IBA), 1.6 mg L−1 6-benzylaminopurine (BA), and 0.1 mg L−1 thidiazuron (TDZ). Adventitious buds were efficiently induced from the callus when the MS30 medium was supplemented with 0.8 mg L−1 BA and 0.2 mg L−1 IBA. Root induction occurred within 7–10 d on […]

  • 2018 special reviewers thank you
    on January 22, 2019 at 12:00 am


  • Improving micropropagation of Mentha  ×  piperita L. using a liquid culture system
    on January 18, 2019 at 12:00 am

    Abstract In the current study, in vitro shoot proliferation and plant regeneration of Mentha × piperita L. (peppermint) cultivar ‘Black Mitcham’ was compared in semi-solid and liquid culture systems. Shoot tips from field-grown plants were used as explants to study shoot proliferation response on either Murashige and Skoog (MS) or Chee and Pool (C2D) medium containing varying levels of 6-benzylaminopurine (BAP), kinetin, and 6-γ,γ-dimethylallyl aminopurine (2iP). Differences in leaf ultrastructure and antioxidant capacity of greenhouse-grown and micropropagation-derived plants were studied to identify potential changes occurring during in vitro culture. Among the various media treatments tested, the maximum number of shoots was produced on the C2D medium with 4.0 μM BAP (40.7) followed by the MS medium with 4.0 μM BAP (32.2). Among the rooting treatments, shoots on the MS medium with 1.0 μM […]

  • Plantlet regeneration from primary callus cultures of Lilium brownii F.E.Br. ex Miellez var. giganteum G. Y. Li & Z. H. Chen, a rare bulbous germplasm
    on January 16, 2019 at 12:00 am

    Abstract Lilium brownii F.E.Br. ex Miellez var. giganteum G. Y. Li & Z. H. Chen, an endangered valuable genetic resource, was used to establish and optimize a callus propagation system and to investigate the effects of internal and external phytohormones for the purpose of germplasm conservation. Of the combinations and concentrations of auxins and cytokinins examined, Murashige and Skoog (MS) medium supplemented with 8 g L−1 agar, 30 g L−1 sucrose, 0.45 μM 2,4-dichlorophenoxyacetic acid, 2.69 to 5.37 μM α-naphthaleneacetic acid, and 0.44 μM 6-benzyladenine, 0.45 μM thidiazuron, and 0.28 μM zeatin riboside generated the best results, effectively promoting callus proliferation. Four callus types could be discriminated, of which type A (yellowish, granular) and type B (yellow, medium-granular) were dry, friable, and grew well. Periodic acid-Schiff staining revealed small and regular […]

  • Co-expressing Turnip Crinkle Virus -coat protein with the serine protease α-thrombin precursor (pFIIa) in Nicotiana benthamiana Domin
    on January 15, 2019 at 12:00 am

    Abstract The serine protease α-thrombin (FIIa) plays a fundamental role in blood clotting. In the present report, a FIIa precursor (pFIIa) was expressed in Nicotiana benthamiana Domin. The expression construct featured the Kozak consensus sequence and the 2S2 Arabidopsis thaliana (L.) Heynh. signal peptide to direct the protein into the secretory pathway (sec-pFIIa). A version carrying the KDEL endoplasmic reticulum (ER) retention signal (pFIIa-ER) was also constructed. Transient expression of pFIIa in N. benthamiana leaves was achieved by Agrobacterium tumefaciens infiltration. The influence of post-transcriptional gene silencing (PTGS) was analyzed by co-infiltrating with an A. tumefaciens strain carrying the construct for the Turnip Crinkle Virus-coat protein (TCV-CP) known for interfering with PTGS. Reverse transcription polymerase chain reaction and Western blot analyses confirmed the presence of the corresponding messenger RNA and the recombinant pFIIa […]

  • An efficient protocol for in vitro propagation of the wild legume Cicer microphyllum Benth., a crop wild relative of chickpea ( Cicer arietinum L.)
    on January 11, 2019 at 12:00 am

    Abstract Cicer microphyllum Benth. is a wild legume that is adapted to the extremely adverse climatic conditions of the cold Himalayan deserts. This rare species is a crop wild relative (CWR) that could be used for chickpea (Cicer arietinum L.) breeding programs to improve abiotic stress tolerance. However, the availability of C. microphyllum biological material is very limited due to its high altitude and remote location. These factors make in vitro plant tissue culture an excellent alternative for the conservation and propagation of this species. In this study, an efficient and reproducible in vitro regeneration protocol was established for C. microphyllum. This process enabled the regeneration of multiple shoots through indirect organogenesis from excised embryos. In addition, the effects of combinations of auxin and cytokinin concentrations in Murashige and Skoog (MS) medium were tested. The highest percentage of callus induction was observed on MS with […]

  • Overexpression of cinnamyl alcohol dehydrogenase gene from sweetpotato enhances oxidative stress tolerance in transgenic Arabidopsis
    on January 10, 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. […]

  • High-efficiency somatic embryogenesis of a broad range of Brazilian Saccharum spp. hybrids (sugarcane) varieties using explants from previously established in vitro plants
    on January 8, 2019 at 12:00 am

    Abstract The current study assessed the embryogenic potential of different Brazilian Saccharum spp. hybrid (sugarcane) varieties, using explants from previously established in vitro plants, and determined the morphogenic capacity of calluses to regenerate plants during consecutive subcultures. After examining various explant lengths (1 to 15 mm) for callus initiation, an optimal length of 12 mm was determined, and 14 sugarcane varieties were studied. Callus induction occurred on Murashige and Skoog medium with 3.0 mg L−1 2,4-dichlorophenoxyacetic acid, in the dark. After 40 d, calluses were divided according to their predominant type: the mucilaginous part remained in the dark on fresh induction medium, whereas the compact nodular callus fraction was transferred to a regeneration medium containing 1.86 mg L−1 1-naphthaleneacetic acid and 0.09 mg L−1 6-benzylaminopurine and was cultivated in light. This […]

  • Acid pretreatment improves microtuberization of potato plantlets
    on December 18, 2018 at 12:00 am

    Abstract To efficiently produce potato (Solanum tuberosum L.) microtubers, in vitro plantlets were pretreated with an acid solution before culturing in Murashige and Skoog medium supplemented with 60 g L−1 sucrose. The first tubers were formed at 36 h of acid pretreatment (AT), which was 6.2 d earlier than the untreated control. The maximum number of tubers (3.2), and highest fresh weight per tuber (282.5 mg) were also achieved with the AT. During microtuberization, the leaf dry weight of the control was higher than the AT from 1 to 5 d of culture. However, the dry weight of AT leaves increased rapidly after 5 d of culture and reached the same level as the control. The tuber dry weight in the AT was significantly higher (α = 0.05) than the control during the same culture period. Sucrose and starch content, and the enzyme activity of sucrose synthase (SS), ADP-glucose pyrophosphorylase (AGPase), and soluble […]

  • In vitro desiccation tolerance of the epiphytic Ghost Orchid, Dendrophylax lindenii (Lindl.) Benth x. Rolfe
    on December 7, 2018 at 12:00 am

    Abstract The Ghost Orchid, Dendrophylax lindenii (Lindl.) Benth x. Rolfe, is a rare and endangered epiphytic orchid native to south Florida and Cuba. The orchid is considered difficult to propagate under greenhouse conditions, requiring high humidity and low air movement. In contrast, the orchid’s native habitat seasonally dries out with decreased precipitation and humidity. This suggests some level of desiccation tolerance. Ghost Orchid plants were assessed for potential desiccation tolerance and ability to recover from desiccation stress under in vitro conditions. In vitro-derived plants were placed into sterile baby food jars and transferred to chambers maintained at 10% relative humidity, which is extremely low compared to relative humidity levels (40–100%) recorded under natural field conditions. Plants were removed every week for 4 wk and recovered on P723 medium supplemented with banana powder for 4 wk. Data were collected at the […]

  • Improved Agrobacterium tumefaciens- mediated transformation of soybean [ Glycine max (L.) Merr.] following optimization of culture conditions and mechanical techniques
    on December 1, 2018 at 12:00 am

    Abstract In the present study, Agrobacterium tumefaciens-mediated transformation of Glycine max (L.) Merr. (soybean) cv. DS-9712 using half-seed explants was optimized for eight different parameters, including seed imbibition, medium pH, infection mode (sonication and vacuum infiltration), co-cultivation conditions, concentrations of supplementary compounds, and selection. Using this improved protocol, maximum transformation of 14% and regeneration efficiencies of 45% were achieved by using explants prepared from mature seeds imbibed for 36 h, infected with A. tumefaciens strain EHA105 at an optical density (OD600) of 0.8, suspended in pH 5.4 medium containing 0.2 mM acetosyringone and 450 mg L−1 L-cysteine, followed by sonication for 10 s, vacuum infiltration for 2 min, and co-cultivated for 3 d on 35 mg L−1 kanamycin-containing medium. Independent transgenic lines were confirmed to be transgenic after […]

  • Quercetin and silver nitrate modulate organogenesis in Carissa carandas (L.)
    on December 1, 2018 at 12:00 am

    Abstract An in vitro organogenesis protocol for Carissa carandas L. was developed using an auxin transport inhibitor (quercetin) and silver nitrate (AgNO3), an inhibitor of ethylene action, in association with cytokinins in the culture medium. This protocol produced the maximum number of shoots from aseptic seedling-derived shoot apex explants of C. carandas. The highest rate of shoot multiplication was recorded on MS medium containing 2.0 mg L−1 6-benzylaminopurine; 0.5 mg L−1 kinetin, and 0.75 mg L−1 quercetin at after 4 wk of culture. Similar results were obtained when MS medium fortified with 2.0 mg L−1 BAP, 0.5 mg L−1 kinetin, and 1.5 mg L−1 AgNO3 was used. However, successful rooting was achieved on quarter strength MS medium with 0.5 mg L−1 indole-3-acetic acid. In this study, an inhibitor of auxin transport and ethylene action maximized shoot […]

  • Efficient callus-mediated regeneration and in vitro root tuberization in Trichosanthes kirilowii Maxim., a medicinal plant
    on December 1, 2018 at 12:00 am

    Abstract Trichosanthes kirilowii Maxim. is a climbing herb with considerable medicinal value. In this study, efficient protocols for callus-mediated regeneration and in vitro tuberization of this plant were developed. Sterilized stem and leaf tissues were cultured on Murashige and Skoog (MS) medium with plant growth regulators (PGRs), and additives that promoted callus induction and regeneration. Both stem and leaf tissues showed the best response (100%) for callus initiation on MS medium supplemented with 4.5-μM 2,4-dichlorophenoxyacetic acid (2,4-D). Efficient shoot organogenesis was obtained by exposing the callus tissue to 4.6-μM kinetin, 2.2-μM 6-benzylaminopurine, and 2.7-μM 1-naphthylacetic acid (NAA) along with 12.6-μM copper sulfate, which yielded a shoot regeneration rate of 85.5% and 28 shoots derived from each callus. In vitro shoots were best rooted on half-strength (1/2) MS medium with 2.7-μM NAA. Tuberous roots were efficiently […]

  • In vitro preservation and micropropagation of Oreocharis mileense (W.T. Wang) M. Möller & A. Weber (Gesneriaceae) through shoot organogenesis
    on December 1, 2018 at 12:00 am

    Abstract Oreocharis mileense (W.T. Wang) M. Möller & A. Weber is endemic to China and was considered to be extinct because it had not been seen in the wild since the first collection in 1906. In 2006, the species was rediscovered in Shilin County, Yunnan Province. Oreocharis mileense was considered critically endangered for its narrow geographic range and extremely small population. An efficient method to preserve plant germplasm by in vitro culturing of O. mileense has not been reported. In this study, an orthogonal array with three factors (6-benzyladenine, BA; α-naphthaleneacetic acid, NAA; and sucrose), at four levels was performed, and shoot induction as well as shoot proliferation were recorded. The results were analyzed to determine the most significant components and the optimum combination for micropropagation of O. mileense. The results showed that: (1) organogenesis was easily induced by different combinations of plant-growth regulators […]

  • Micropropagation of Glossonema varians (Stocks) Benth. ex Hook.f.—a rare Asclepiadeae of Indian Thar Desert
    on December 1, 2018 at 12:00 am

    Abstract In the present study, an in vitro regeneration protocol for Glossonema varians (Stocks) Benth. ex Hook.f. of family Asclepiadaceae was optimized. Cotyledonary nodes of in vitro cultured seedlings were used as explants for activation of axillary shoot buds. Axillary shoot buds were initially activated on 0.1 mg L−1 6-benzylaminopurine (BAP) and then multiplied on 0.05 mg L−1 BAP. Shoots were rooted in vitro on 1/4 strength Murashige and Skoog medium containing 0.1 mg L−1 2-naphthoxyacetic acid and 100 mg L−1 activated charcoal. The cultures were maintained in a 12 h photoperiod at 40–50 μmol m−2 s−1 spectral flux photon, 25–30 ± 2°C, and 60% relative humidity (RH). Up to 80% of in vitro regenerated plantlets were acclimatized on soilrite in cotton-plugged culture tubes in the greenhouse. This protocol can be a useful method […]

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