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.


  • Genetic homogeneity assessment of in vitro -regenerated plantlets of Nyctanthes arbor-tristis L. and comparative evaluation of bioactive metabolites and antioxidant activity
    on August 16, 2019 at 12:00 am

    Abstract In vitro propagation of Nyactanthes arbor-tristis L. was achieved by culturing N-phenyl-N′-benzothiazol-6-yl-urea (PBU)-pretreated nodal explants in Murashige and Skoog (MS) medium without any phytohormones. Pretreatment of nodal explants in liquid MS medium with 100 μM N-phenyl-N′-benzothiazol-6-yl-urea for 4 d showed the highest shoot proliferation by producing maximum number of shoots (17.40 ± 1.02) per explant, with average shoot length of 5.96 ± 0.08 cm at the end of 8 wk. Effective rooting was accomplished by preincubating the cut-end of shoots with half-strength MS medium containing 6 μM indole-3-butyric acid for 1 wk, followed by implantation into half-strength MS medium; an average of 6.20 ± 0.049 roots per shoot were produced. Seventy-eight percent of the plantlets regenerated in vitro were successfully acclimatized and transferred to […]

  • Micropropagation and molecular characterization of Thymus sibthorpii Benth. (Lamiaceae), an aromatic-medicinal thyme with ornamental value and conservation concern
    on August 13, 2019 at 12:00 am

    Abstract Thymus sibthorpii Benth. (Lamiaceae), with accession number 01,1796-22, is a biotype of native Greek thyme with ascending stems and potential use as a new medicinal-aromatic crop and ornamental plant. An efficient and reliable protocol for in vitro clonal propagation of T. sibthorpii from nodes and meristem tip explants was developed. Shoot proliferation succeeded on a new basal medium (BB) without plant growth regulators, as prior experiments with 6-benzyladenine generated hyperhydricity. Eight different basal media were compared; on two formulations using the new BB 5.9 and 5.6 shoots per explant were produced. Regenerated single shoots were rooted in the BB medium, supplemented with 5 μM of indole-3-butyric acid, and produced 3.1 roots along with 2.5 adventitious shoots. Three types of acclimatization were assessed: in vitro, using two different systems (no significant differences); ex vitro, using eight soil substrates under greenhouse and […]

  • Improved protocol for the transformation of adult Citrus sinensis Osbeck ‘Tarocco’ blood orange tissues
    on August 13, 2019 at 12:00 am

    Abstract The production of transgenic citrus plants from adult tissues is difficult because of low regeneration and transformation rates. To increase the transformation efficiency of adult citrus tissues, an improved protocol involving adult Citrus sinensis Osbeck ‘Tarocco’ blood orange tissues was developed. Explants were pre-incubated in a liquid medium prior to infection by Agrobacterium tumefaciens. Plant materials were also incubated on callus-induction medium supplemented with various combinations of cytokinin (Cyt) and kanamycin (Kan). An appropriate pre-incubation of the explants increased the transformation efficiency of adult tissues. During the callus-induction period, the Cyt type and Kan concentration had the largest and smallest effects on the transformation efficiency, respectively. The most effective combination of plant growth regulator and Kan for the transformation of ‘Tarocco’ blood orange tissues was […]

  • Development of a micropropagation protocol for Malus orientalis using axillary buds
    on August 9, 2019 at 12:00 am

    Abstract Caucasian apple trees (Malus orientalis) grow individually or in small groups with a scattered distribution pattern throughout the Hyrcanian forest. The wild gene pool in such trees has a high genetic diversity which is highly important for apple breeders. Micropropagation can be advantageous due to its quick propagation for apple breeders as well as its maintenance of in vitro and in vivo germplasm collection and exchange. Here, for the first time, we investigated the in vitro propagation of M. orientalis from three populations by collecting axillary buds and established a micropropagation protocol. Two strengths of Murashige and Skoog (MS) basal medium (full and half) in combination with different concentrations of 6-benzylaminopurine (BA) were tested for optimal multiplication. The interaction among the BA concentrations, MS strength, and populations was effective in the promotion of shoot development across the three populations. The average number of […]

  • Growth and development of carnation ‘Dreambyul’ plantlets in a temporary immersion system and comparisons with conventional solid culture methods
    on August 8, 2019 at 12:00 am

    Abstract The aim of the current study was to compare the effects of the culture method—conventional solid medium culture and temporary immersion system (TIS)—on the growth and development of carnation ‘Dreambyul’ plantlets. At the same time, different immersion intervals and immersion durations of TIS culture were also tested to find the optimal setting for mass production of high-quality carnation plantlets in vitro. In the first experiment, the results showed that the shoot length, root length, and number of nodes of plantlets cultured in the TIS were highest when the immersion interval was 8 h. Compared with that of plantlets cultured in the conventional solid medium culture, the fresh weight of plantlets cultured in the TIS was at least 3 times greater. The greatest total chlorophyll content, stomata with normal shapes was observed for plantlets grown in the TIS with an 8-h immersion interval. The lowest H2O2 level was recorded in […]

  • Use of DoE methodology to optimize the regeneration of high-quality, single-copy transgenic Zea mays L. (maize) plants
    on August 7, 2019 at 12:00 am

    Abstract The maize Agrobacterium tumefaciens-mediated transformation process normally takes about 10 to 15 wk from embryo infection and co-cultivation to sending “T0” plants to the greenhouse (GH). A new method was developed using the maize transcription factors Babyboom (BBM) and Wuschel2 (WUS2), to stimulate direct transgenic embryo formation and plant regeneration, that bypasses the need for prolonged tissue culture and regeneration from callus. In the present study, a design of experiment (DoE) method was used to test 10 factors to optimize the quality of somatic embryo maturation, root formation, and subsequent plantlet survival, without compromising the molecular event quality. The concentration of NO3− and the ratio of NH4+ to K+ had significant effects on the morphology of plantlets derived directly from germinated transgenic embryos. During early development, optimal tissue morphology required a NH4+/K+ ratio of 1:1 with 20 mM […]

  • Alkaloid synthesis is coupled to shoot morphogenesis in Argemone mexicana L. (Papaveraceae) in vitro cultures
    on August 5, 2019 at 12:00 am

    Abstract During the induction process of an in vitro callus culture of Argemone mexicana L. (Papaveraceae), the levels of two benzylisoquinoline alkaloids known as berberine and sanguinarine displayed opposing trends. While the berberine levels steadily decreased from the initial explant stage up to the early proliferation of unorganized parenchymatous cell masses, the sanguinarine content increased. Once the callus culture was established, sanguinarine was the primary alkaloid present and berberine could no longer be detected. However, upon shoot regeneration, the berberine accumulation recovered, but sanguinarine was found in the newly formed leafy tissue. After root formation, sanguinarine was relocated to this organ, whereas berberine was evenly distributed between both tissues. Explants from stem internodes did not form callus, and berberine—plus sanguinarine—containing axillary shoots emerged from lateral buds in the induction medium. In contrast to […]

  • Genetic homogeneity and high shoot proliferation in banana ( Musa acuminata Colla) by altering medium thiamine level and sugar type
    on August 5, 2019 at 12:00 am

    Abstract To enhance the multiplication rate in Musa acuminata Colla (banana; ‘Grand Nain’) organogenesis, higher amounts of thiamine along with different sugar types and concentrations were evaluated at the proliferation phase. Thiamine at 1, 10, 50, 100, and 200 mg L−1 was compared with 0.1 mg L−1 thiamine found in conventional Murashige and Skoog (MS) medium. Maximum proliferation of banana was induced with 100 mg L−1 thiamine. Additionally, 15, 30, and 45 g L−1 sucrose, glucose, fructose, and sorbitol combined with regular and optimal levels of thiamine were tested. Glucose at 30 g L−1 most improved shoot proliferation alone and enhanced shoot proliferation further, when combined with 100 mg L−1 thiamine, followed by sucrose and fructose, whereas sorbitol completely inhibited growth and caused tissue browning. All evaluated vegetative traits were […]

  • Sodium transport by endocytic vesicles in cultured Arabidopsis thaliana (L.) Heynh. cells
    on July 31, 2019 at 12:00 am

    Abstract The involvement of endocytosis in Na+ internalization by suspension-cultured Arabidopsis thaliana (L.) Heynh. cells under salt stress was investigated. Using epifluorescence and confocal laser-scanning microscopy in co-localization experiments with a marker of the endocytic structures, N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino) phenyl) hexatrienyl) pyridinium dibromide (FM 4-64), and a membrane-impermeable Na+ indicator, Asante Natrium Green-2 tetramethylammonium salt (ANG-2 TMA), intracellular Na+ ion uptake via endocytosis, but not through the plasma-membrane ion channels or transporters, was detected. The punctate fluorescence of the two markers was co-localized in the microvacuoles (MVs) with sizes up to 5 μm. The study of cell ultrastructure by transmission electron microscopy revealed the fusion of MVs into larger ones during an incubation of the cells in medium containing 100 mM NaCl. Wortmannin, an inhibitor of multivesicular […]

  • Morphophysiological in vitro performance of Brazilian ginseng ( Pfaffia glomerata (Spreng.) Pedersen) based on culture medium formulations
    on July 31, 2019 at 12:00 am

    Abstract Pfaffia glomerata has potential pharmacological and medicinal properties due to the production of a secondary metabolite known as the phytoecdysteroid 20-hydroxyecdysone (20E). There have been increasing efforts for massive in vitro propagation of Pfaffia plants due to high extractivism and overharvesting of this species. Research on the species has shown that photoautotrophic cultivation can improve the production of 20E. In addition, other abiotic factors such as the formulations of culture media can influence the morphophysiological behavior of the plants in vitro. Therefore, the objective of this study was to analyze the morphological and physiological performances of P. glomerata plants in different formulations of culture media, under photoautotrophic and photomixotrophic propagation conditions. Six medium formulations, the Driver and Kuniyuki medium (DKW), Correia et al. medium (JADS), Murashige and Skoog medium (MS), Quoirin and Lepoivre […]

  • A correlation between leaf shape and its related key genes in Viola albida complex
    on July 29, 2019 at 12:00 am

    Abstract Simple to compound leaves occur in the Viola albida complex, which comprises the simple, finely serrate leaves of V. albida Palib., the deeply lobed leaves of V. albida var. takahashii (Nakai) Nakai, and the compound leaves of Viola chaerophylloides (Regel) W. Becker. To identify a correlation between the different leaf forms and the expression of several key genes with roles in leaf morphogenesis, the distinct leaf forms occurring within these species were generated by tissue culture of the V. chaerophylloides petiole, for comparison with wild-type leaves. Compound leaves were generally formed from a petiole explant taken close to the leaf blade, whereas simple leaves resulted from petiole explants taken close to the petiole base. KNOTTED-1 (VaKN1), SHOOTMERISTEMLESS (VaSTM), CUP-SHAPED COTYLEDON-2 (VaCUC2), and ASYMMETRIC LEAVES 1 (VaAS1), which are known to play key roles during compound leaf patterning and morphogenesis, were isolated and multiple […]

  • Melatonin influence on in vitro callus induction and phenolic compound production in sweet basil ( Ocimum basilicum L.)
    on July 26, 2019 at 12:00 am

    Abstract The aim of the present study was to evaluate melatonin effects on the callus induction and phenolic compound production of Ocimum basilicum L. (sweet basil). Calluses, derived from leaf explants, were grown on Murashige and Skoog (MS) medium supplemented with 0, 100, or 200 μM melatonin, and subsequently extracted for determination of their phenolic contents. Melatonin decreased the callus induction in both concentrations. Based on the phytochemical analysis, the highest total phenolic acid contents (784.6 μg g−1 and 335.2 μg g−1, respectively) were recorded in calluses grown in 100 and 200 μM melatonin-supplemented medium, compared with the calluses induced with MS alone (192.0 μg g−1). Among the five phenolic acids confirmed in the callus samples, rosmarinic acid was the major constituent. The amount of rosmarinic acid increased significantly in callus grown on 100 μM […]

  • Somatic embryogenesis to overcome low seed viability and conserve wild banana ( Ensete superbum (Roxb.) Cheesman)
    on July 25, 2019 at 12:00 am

    Abstract The seed viability, ex vitro germination, and percentage of in vitro zygotic embryo germination were found to be very low in Ensete superbum (Roxb.) Cheesman. Only 33.33% of seeds were viable, and the ex vitro germination percentage was only 5%, while the percentage of in vitro zygotic embryo germination was 33%. Somatic embryogenesis experiments produced competent callus on Murashige and Skoog (MS) medium supplemented with 2.5 mg L−1 2,4-D and 3 mg L−1 BAP from inflorescence explants. The embryogenic callus produced the maximum number of somatic embryos on MS basal medium kept in a dark chamber for 15 wk. Half-strength MS medium supplemented with 500 mg L−1 glutamine was optimal for somatic embryo germination and development of plantlets. Regenerated plants had 80 to 90% survival rate. Therefore, somatic embryogenesis can be considered as an efficient method to overcome a drastic reduction in population and to […]

  • In vitro germination and seedling development of two European orchid species, Himantoglossum jankae Somlyay, Kreutz & Óvári and Spiranthes spiralis (L.) Chevall.
    on July 23, 2019 at 12:00 am

    Abstract New protocols for successful asymbiotic seed germination of two European orchid species, Himantoglossum jankae Somlyay, Kreutz & Óvári and Spiranthes spiralis (L.) Chevall., were established in this study. The influence of two basal media, organic supplements, illumination, and cytokinins on germination, protocorm formation, and seedling development was examined. A strong species-specific dependence on illumination conditions and nutrient medium composition was observed. Improved germination under continual darkness indicated that H. jankae had negatively photoblastic seeds, while S. spiralis seeds could germinate under both light conditions examined in this study, but slightly faster development was achieved under the 16-h photoperiod protocol. The H. jankae seeds cultured on Knudson C medium failed to germinate in any medium combination, irrespective of the organic supplement use. The highest germination rate of H. jankae was achieved on […]

  • The effect of slow-growth strategy on a production of Petunia × hybrida Vilm. microcuttings
    on July 23, 2019 at 12:00 am

    Abstract Petunia × hybrida Vilm. is a fast-growing ornamental plant that was cultured under varying storage conditions to address seasonal fluctuations in microcutting demand. The effects of storage period (16 to 32 wk), temperature (15 to 23°C), low and high light intensity, sucrose (1 to 5% w/v), and mannitol (0 to 4% w/v) in factorial arrangements were analyzed. Stored and non-stored shoots were compared for microcutting production, harvested twice at 3-wk intervals, and were subsequently transferred to the greenhouse for 17 d. Nearly all plants from 16-wk storage survived well at all treatment conditions and the quality and quantity of microcuttings were enhanced from shoots stored at 15°C and 3% (w/v) sucrose, without mannitol for all storage periods. Another experiment tested 11- to 25-wk storage period with Petunia hybrida ‘Ragtime’ and ‘Suncatcher’ at 12°C and low light intensity. Repeated cycles of […]

  • Root cryopreservation to biobank medicinal plants: a case study for Hypericum perforatum L.
    on July 18, 2019 at 12:00 am

    Abstract In this study, an effective root-based cryopreservation method was developed for Hypericum perforatum L., an important medicinal species, using in vitro plants. A systematic approach was applied to determine effective combinations of protocol steps such as preculture, osmoprotection, vitrification solution treatment, and unloading, followed by protocol optimization using a single-factor approach. The effects of root section type (root tips, middle sections, or basal sections), duration of root section culture after excision, and donor plant age were also investigated. In a wild genotype, middle and basal root sections excised from 8-wk-old plants and cryopreserved at the age of 10 d after excision showed the highest plant regrowth after cryopreservation. In the optimized protocol, root sections were precultured in 10% (w/v) sucrose for 17 h, osmoprotected with a solution composed of 17.5% (w/v) glycerol and 17.5% (w/v) sucrose for 20 min, […]

  • Development of an efficient micropropagation system for Tecoma stans (L.) Juss. ex Kunth using thidiazuron and effects on phytochemical constitution
    on July 16, 2019 at 12:00 am

    Abstract The present study was designed to investigate the stimulatory effects of different doses (0.1 to 2.5 μM) of thidiazuron (TDZ) on in vitro shoot induction and proliferation of mature nodal explants of Tecoma stans. Of the tested concentrations, 2.0 μM TDZ proved to be optimal for maximum regeneration (91%) with a mean shoot number of 5.6 ± 0.67, and length of 2.38 ± 0.08 cm, after 4 wk of incubation. To determine the negative effects of prolonged TDZ exposure, after 4 wk of incubation at optimized level of TDZ, the cultures were transferred to a secondary medium either lacking plant growth regulators or supplemented with benzyladenine (BA) alone, or in combination with different auxins (indole-3-acetic acid, indole-3-butyric acid, or α-naphthalene acetic acid; NAA). Among the tested concentrations, 2.5 μM BA in combination with 0.5 μM NAA yielded the maximum mean […]

  • Maize seedlings produced from dry seeds exposed to liquid nitrogen display altered levels of shikimate pathway compounds
    on July 10, 2019 at 12:00 am

    Abstract In light of climate change and risks of food insecurity, it is becoming increasingly important to preserve plant germplasm in genebanks. Storage of seeds, particularly via cryopreservation, is one of the most proficient methods for ex situ plant germplasm conservation. Whilst seed cryo-banking can have little, to no, or even beneficial effects on subsequent seedling vigor in some species, it can lead to a number of plant abnormalities (morphological and physiological). This study investigated the effects of maize seed cryopreservation on seedling growth (until 14 d) and levels of selected amino acids produced in the shikimate pathway, a major link between primary and secondary metabolism. Seed cryopreservation reduced FW in recovered seedlings, reduced caffeic acid (2.5-fold decrease), and increased levels of all other shikimate pathway–related compounds assessed: phenylalanine (2.9-fold increase), tyrosine (2.6-fold increase), and shikimic […]

  • 2019 In Vitro Biology Meeting, June 8 – 12, Tampa, Florida
    on July 10, 2019 at 12:00 am


  • An in vitro screening system to assess aluminum toxicity in sugarcane ( Saccharum spp.) cultivars
    on July 3, 2019 at 12:00 am

    Abstract Mutagenic breeding is an approach being developed especially for those characteristics that are not inherited through conventional genetic hybridization. Soil acidity and aluminum (Al3+) toxicity limit sugarcane production, particularly in leached soils in the coastal cultivation regions in South Africa. An in vitro screening procedure was developed based on plantlet re-rooting in Al3+-containing medium to select putative Al3+ tolerant variants produced by induced mutagenesis. A preliminary test did not show any difference among three cultivars in their ability to re-root in the presence of Al3+. Subsequent work focused on Saccharum sp. ‘NCo376’ and two Al3+ concentrations (15 and 45 mM), representing LC50 and LC90 re-rooting inhibition. In an attempt to increase the stringency of the selection conditions, the possible effects of inorganic nitrogen (N) forms in the culture media on Al3+ toxicity were investigated. The form of N affected root […]

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