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.


  • Phytochemical, biochemical, and growth changes in response to salinity in callus cultures of Nigella sativa L.
    on February 24, 2020 at 12:00 am

    Abstract Black cumin (Nigella sativa L.) is considered as a noteworthy herbal medicine. However, no study has been conducted on the physiological adaptive mechanism of it to salinity stress, especially under in vitro condition. To this aim, the callus cultures of ten different genotypes of N. sativa were applied to evaluate the changes occurring in biochemical traits under salinity stress. The calluses were exposed to the in vitro salt stress using different sodium chloride concentrations (0, 84, and 250 mM). A reduction occurred in the content of K+ and callus growth by enhancing the NaCl concentration. However, most of the content of Na+ (4 mgg− 1 DW), malondialdehyde (1.38 μmolg− 1 FW), total phenolic content (1.18 mg GAEg−1 FW), thymol (25.26 mgg− 1 DW), total flavonoids content (0.06 mg QEg− 1 FW), total flavonols (TFL) content (0.023 mg QEg− 1 FW), total […]

  • Micropropagation and essential oil characterization of Plectranthus amboinicus (Lour.) Sprengel, an aromatic medicinal plant
    on February 24, 2020 at 12:00 am

    Abstract Plectranthus amboinicus (Loureiro) Sprengel is a therapeutically valued herb highly sought by various industries. This has led to its uncontrolled harvest from the wild leading to the risk of extinction. As an alternative, commercial propagation of P. amboinicus is desirable to avoid further depletion of wild populations. This paper describes the establishment of an efficient protocol for the multiplication, rooting and acclimatization of P. amboinicus. The greatest frequency of shoot multiplication of P. amboinicus was on semi-solid Murashige and Skoog (MS) medium supplemented with 0.4 mg L−1 6-benzylaminopurine (BAP). Media supplemented with higher concentrations of BAP or used in combination with gibberellic acid (GA3) produced abnormal plantlets. Optimal rooting was observed on plantlets cultured in half strength MS medium. Acclimatization was achieved by transferring the rooted plants onto sterile peat-moss moistened with MS medium and […]

  • The effects of illumination, temperature and 6-benzylaminoprine on asymbiotic seed germination and protocorm development in vitro in the achlorophyllous orchid Gastrodia pubilabiata Sawa
    on February 20, 2020 at 12:00 am

    Abstract We examined the effects of culture conditions, namely, illumination, temperature and the addition of 6-benzylaminopurine (BAP) to the germination medium, on the asymbiotic germination of mature seeds of Gastrodia pubilabiata Sawa, an achlorophyllous orchid. Mature seeds were harvested from plants at two sites, one in Toyama Prefecture and one in Chiba Prefecture, Japan. We used gellan gum-solidified New Dogashima medium as the basal medium. The maximum frequency of germination of the seeds in darkness was approximately 65%. BAP had no stimulatory effect on seed germination, but accelerated protocorm development. Necessary conditions for successful germination of seeds and subsequent development of seedlings from mature seeds of Gastrodia pubilabiata clarified in this study are prerequisites not only for conservation of biodiversity but also for the elucidation of the life cycle of this achlorophyllous orchid.

  • Practical methodology for gametophyte proliferation and sporophyte production in green penny fern ( Lemmaphyllum microphyllum C. Presl) using mechanical fragmentation
    on February 10, 2020 at 12:00 am

    Abstract Propagation of gametophytes and sporophytes using mechanical fragmentation has been considered a suitable method for mass production of ferns. This study aimed to develop a practical propagation method for Lemmaphyllum microphyllum C. Presl, which is a fern of significant ornamental and medicinal value. Gametophytes were obtained through in vitro spore germination and used for propagation experiments. The gametophyte was mechanically fragmented using a scalpel into small fragments, which were then used to investigate gametophyte proliferation. In addition, the gametophyte was fragmented using a blender and then used to study sporophyte formation. Optimal proliferation conditions of the gametophyte were determined using Murashige and Skoog (MS) basal medium (double-, full-, half-, quarter-strength), Knop medium, and medium components (sucrose, nitrogen sources, activated charcoal), at various concentrations. The fresh weight of the gametophyte was 14-fold […]

  • Photomixotrophic growth response of sugarcane in vitro plantlets using different light intensities and culture vessel types
    on February 4, 2020 at 12:00 am

    Abstract Sugarcane is a very important crop in tropical areas around the world. Optimizing the light intensity and culture vessel for sugarcane in vitro propagation can improve protocols for large-scale in vitro mass clonal propagation. The objective of this study was to evaluate biometric and anatomical characteristics of in vitro propagated sugarcane plantlets using different light intensities and culture vessel types. The study also aimed to stimulate desired photoautotrophic and/or photomixotrophic traits and to study correlations between survival and ex vitro vigor with in vitro data. Fluorescent lights at 70 μmol m−2 s−1 and light-emitting diodes (LEDs) at 120 and 200 μmol m−2 s−1 were used combined with different culture vessel types, including microboxes without a filter, microboxes with a filter, and baby food jars. Propagation and rooting growth were observed in all in vitro culture conditions. […]

  • Reduced major minerals and increased minor nutrients improve micropropagation in three apple cultivars
    on February 4, 2020 at 12:00 am

    Abstract Mineral nutrient medium requirements for propagation of in vitro shoots of apple (Malus domestica Borkh) ‘Golden Delicious’, ‘Maksat’, and ‘Voskhod’ were studied using response surface methodology (RSM). The mineral nutritional factors evaluated were based on Murashige and Skoog (MS) mineral nutrients (NH4NO3, KNO3, CaCl2, KH2PO4, MgSO4, and minor nutrients), with concentrations ranging from 0.5 to 3.0× the MS concentrations. Nine plant growth qualities were evaluated. The most significant factors were NH4NO3 at 0.5 to 1.0× MS, and minor nutrients at 2.0× MS. Most of the other factors were optimal at 0.5×. The quality rating was highest when minor nutrients were 2.0× MS, and most other nutrients were standard concentrations or lower. Increased KH2PO4 and minor nutrients were the most significant for improved multiplication, and higher KNO3 for shoot length. Optimized media were developed for […]

  • An improved Agrobacterium -mediated transformation method for cotton ( Gossypium hirsutum L. ‘KC3’) assisted by microinjection and sonication
    on February 1, 2020 at 12:00 am

    Abstract An efficient and reproducible Agrobacterium tumefaciens-mediated transformation method was developed for Gossypium hirsutum L. ‘KC3’ using a shoot apex explant with the combination of microinjection and sonication. To optimize the transformation efficiency, preculture conditions, acetosyringone concentration, A. tumefaciens cell density, and precision microinjection with sonication were evaluated. A high-throughput A. tumefaciens-mediated genetic transformation method was developed using the uidA gene. The shoot apex explants were microinjected with an A. tumefaciens strain EHA105 carrying the uidA gene and co-cultivated for 3 d on mMS medium fortified with 100 mM acetosyringone. Among the different parameters and conditions tested, preculturing explants for 24 h, microinjecting three times with an A. tumefaciens cell suspension of OD600 nm = 0.6 containing 100 mM acetosyringone, and sonicating explants for 20 s produced the maximum […]

  • Genetic homogeneity assessment of in vitro -regenerated plantlets of Nyctanthes arbor-tristis L. and comparative evaluation of bioactive metabolites and antioxidant activity
    on February 1, 2020 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 […]

  • Plant growth-promoting rhizobacteria for in vitro and ex vitro performance enhancement of Apennines’ Genepì ( Artemisia umbelliformis subsp. eriantha ), an endangered phytotherapeutic plant
    on February 1, 2020 at 12:00 am

    Abstract Apennines genepì (Artemisia umbelliformis Lam. subsp. eriantha), is a high-altitude plant endangered by illegal and indiscriminate harvesting to produce the homonymous liqueur. Even if synthetic seeds represented a valid propagation technique to respond to this threat, this technology still requires methods to improve the low germination and plantlet development rates. The aim of the present study was to evaluate the suitability of the bacterial consortium formed by Azospirillum brasilense, Gluconacetobacter diazotrophicus, Burkholderia ambifaria, and Herbaspirillum seropedicae, as an enrichment agent of Apennines Genepì synthetic seeds. To evaluate the effectiveness as an improving agent, the consortium was incorporated in the synthetic seeds, compared to a control, and followed until ex vitro cultivations. Plants were evaluated for their growth and development parameters, total phenolic and flavonoid contents, antioxidant properties, […]

  • In vitro propagation of male and female Hippophae salicifolia D. Don
    on February 1, 2020 at 12:00 am

    Abstract Hippophae salicifolia D. Don. is a deciduous, temperate, and dioecious tree of great economic and ecological importance. In this study, an in vitro propagation protocol was established for male and female explants of H. salicifolia. The most suitable explants were selected by screening the various types of explants such as dormant and active axillary buds, apical buds, nodal parts, and leaves. The explants were selected every month throughout the year to test the response, and only dormant and active buds were able to generate a response. Accumulation of phenolics was one of the most significant problems in the in vitro propagation of H. salicifolia, which was overcome by using an antioxidant solution of 1500 mg L−1 each ascorbic acid and citric acid followed by a lower temperature treatment, and pre-culture in 1.2% (w/v) agar supplemented with 0.01% (w/v) myo-inositol and 3% (w/v) sucrose. Active buds proved to be the best explant type to […]

  • Optimization of a Liquid Culture System for Shoot Regeneration and Achieving an Enriched Level of Scopadulcic Acid B in the Leaf Organ Cultures of Scoparia dulcis L. by Response Surface Methodology
    on February 1, 2020 at 12:00 am

    Abstract Response surface methodology (RSM) approach was utilized in the present investigation to optimize the constituents of Murashige and Skoog’s (MS) liquid medium to enrich the scopadulcic acid B (SDB) content in Scoparia dulcis L. RSM approach using the central composite design (CCD) was employed to identify the precise concentration of growth regulators of medium and substantiated in shake-flask cultivation. CCD-RSM model revealed a strong agreement, with a predicted coefficient of determination (R2) values of 0.881 and 0.872 for the shoot regeneration percentage (Y1) and the average number of shoots per explant (Y2), respectively. RSM predicted augmented conditions of MS liquid medium considerably influence the shoot proliferation (Y1 and Y2) in the leaf explants of S. duclis. The medium was fortified with 3.59 μM kinetin (KN; X1), 6.00 μM 6-benzylaminopurine (BAP; X2), 3.93 μM indole-3-acetic acid (IAA; X3), and […]

  • cDNA-AFLP analysis of differentially expressed genes during microspore embryogenesis in non-heading Chinese cabbage
    on February 1, 2020 at 12:00 am

    Abstract Stress can induce microspores to change their developmental pathway from the gametophytic to the embryogenic pathway. To explore the molecular mechanisms of microspore embryogenesis, complement DNA-amplified fragment length polymorphism was used to isolate the transcript-derived fragments during microspore embryogenesis of the non-heading Brassica campestris L. ssp. chinensis ‘Wuyueman’. With 256 primer combinations screened, a total of 94 transcript-derived fragments were identified, and 15 were successfully sequenced. Based on a BLAST search in the Brassica database, 12 of the 15 sequenced transcript-derived fragments were homologous to genes with annotations; the remaining three transcript-derived fragments did not match any sequences. Transcript-derived fragments with annotations were involved in cell wall formation, hormones, and resistance. Analysis of cis-elements indicated that there were heat shock-related and stress-related […]

  • The proposed APHIS regulation modernization could enhance agriculture biotechnology research and development in the USA
    on February 1, 2020 at 12:00 am

    Abstract The USDA’s Animal and Plant Health Inspection Service (APHIS) is one of three agencies that govern the importation, interstate movement, or environmental release of certain genetically engineered (GE) organisms. APHIS regulations are in 7 CFR part 340, originally issued in 1987 ( and based on the premise that a GE crop could pose a plant pest risk. A major revision was proposed June 6, 2019 (, with public comments accepted for 60 d. Members of the Society for In Vitro Biology (SIVB)—especially within the plant section—are impacted by the current rules and will be impacted by changes. Therefore, the public policy committee studied the changes and prepared a response that was reviewed by the board of directors and submitted to APHIS. APHIS received 6186 comments overall. In general, the SIVB welcomes the proposed […]

  • Use of gamma radiation to induce mutations in rice ( Oryza sativa L.) and the selection of lines with tolerance to salinity and drought
    on February 1, 2020 at 12:00 am

    Abstract One of the strategies for genetic improvement of Oryza sativa L. (rice) is the use of radiation-induced mutagenesis to search for materials suitable for production in marginal conditions, such as drought and salinity. In this present study, indica rice ‘CR-5272’ embryogenic calluses and seeds were exposed to gamma radiation. Survival and growth of plants derived from embryogenic calluses and seeds were evaluated in vitro in the presence of selection factors for salinity and drought (NaCl and sorbitol). Plants regenerated from irradiated calluses and seeds were screened for tolerance to these conditions and promising lines were identified. Culture of plants regenerated from irradiated calluses (M1) allowed the selection of 60, 58 and 15 promising lines with tolerance to salinity, drought, and both factors, respectively. Six promising salt-tolerant lines and four drought-tolerant lines were selected from plants grown from irradiated seeds (M2). A […]

  • Identification and quantitative determination of feruloyl-glucoside from hairy root cultures of Turbinicarpus lophophoroides (Werderm.) Buxb. & Backeb. (Cactaceae)
    on February 1, 2020 at 12:00 am

    Abstract Transformed hairy root cultures have become an alternative for the biosynthesis of plant secondary metabolites with biological activities. In this present work, the effects of liquid Murashige and Skoog (MS) and Gamborg B5 (B5) medium on kinetic behavior, biomass and phenolic metabolite production were analyzed in Turbinicarpus lophophoroides (Werderm.) Buxb. & Backeb. hairy root cultures. Liquid MS medium showed the highest biomass production (13.67 g L−1 dry weight) after 77 d of culture. For B5 medium, highest biomass was achieved sooner, at day 56, but with lower total biomass (8.10 g L−1 dry weight). After isolation, structural elucidation of the major compound present in T. lophophoroides hairy roots was determined by nuclear magnetic resonance and mass spectral analysis. As a result, a ferulic acid derivative (feruloyl-glucoside) was isolated from T. lophophoroides hairy roots and reported for the first time. Quantitative analysis […]

  • Μicropropagation and cultivation of Salvia sclarea for essential oil and sclareol production in northern Greece
    on February 1, 2020 at 12:00 am

    Abstract Salvia sclarea L. is valuable for its essential oils and other perfumery products used in the food and pharmaceutical industries. The aim of this study was to develop a rapid micropropagation protocol and cultivate S. sclarea in northern Greece for essential oil and sclareol production. The initial plant material used was S. sclarea seeds (biotype SC1) collected from a local population in Bulgaria. Apical meristems from seedlings germinated in the greenhouse were disinfected and established in vitro. For shoot proliferation, six different modified basal culture media supplemented with plant growth regulators (benzyladenine, auxins, and gibberellic acid) at different concentrations and combinations were tested. After 30 d, the highest proliferation rate (1.78) was achieved with an MS medium supplemented with 1.5 μM benzyladenine (BA) and 0.15 μM IBA. The MS medium with 5 μM IBA was the most effective treatment for rooting (40% […]

  • Antioxidant enzyme activities during somatic embryogenesis in Musa acuminata Colla (AAA group) ‘Grand Naine’ and Musa spp. (AAB group) ‘Rasthali’
    on February 1, 2020 at 12:00 am

    Abstract Antioxidant enzymes play a major role in scavenging reactive oxygen species that arise during somatic embryogenesis (SE). Activity levels of antioxidant enzymes such as peroxidase (POX), catalase (CAT), and superoxide dismutase (SOD) were investigated during developmental stages of SE of Musa acuminata Colla (AAA group) ‘Grand Naine’ and Musa spp. (AAB group) ‘Rasthali’. Using native polyacrylamide gel electrophoresis, five POX isoforms were detected in ‘Rasthali’, and four POX isoforms were observed in ‘Grand Naine’. The POX1 isoform was highly expressed in non-embryogenic callus (NEC) of both cultivars, and the POX2 isoform was present in somatic embryos (se) at 0, 30, 45, and 60 d of development of ‘Rasthali’, but not ‘Grand Naine’. Among two CAT isoforms, the CAT2 isoform showed higher expression in embryonic stages than in vegetative stages in explants and NEC of both […]

  • Exogenous polyamines enhance somatic embryogenesis and Agrobacterium tumefaciens -mediated transformation efficiency in sugarcane ( Saccharum spp. hybrid)
    on February 1, 2020 at 12:00 am

    Abstract The influence of exogenous polyamines (PAs) on somatic embryogenesis from immature leaf roll explants and Agrobacterium tumefaciens-mediated transformation of embryogenic callus of Saccharum spp. (sugarcane) ‘Co 86032’ was examined. Immature leaf roll-derived embryogenic callus was obtained on Murashige and Skoog with Gamborg B5 vitamins (MSB5) medium containing 3 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D). Various concentrations of PAs along with 2 mg L−1 2,4-D and 0.5 mg L−1 kinetin (Kin) were tested for somatic embryo induction. A total of 106 somatic embryos per 250 mg of callus (96.3% responsive explants) were obtained on medium supplemented with 20 mg L−1 putrescine (PUT) and 92.0% of the somatic embryos matured and produced 98 shoots per 250 mg of callus. Somatic embryo induction and maturation was increased more than two- and threefold, respectively, on […]

  • Behavior of Opuntia ficus-indica (L.) Mill. Heat-Stressed Microspores Under In Vitro Culture Conditions as Evidenced by Microscopic Analysis
    on February 1, 2020 at 12:00 am

    Abstract When constrained by in vitro culture conditions, microspores from Opuntia ficus-indica (L.) Mill. (Barbary fig) anthers were forced to stall out their gametophytic pathway and switch towards androgenesis. Five microspore stages were characterized based on cellular architecture. A relationship was also established between anthers and bud flower features. Anthers were cultured on three culture media containing 2,4-dichlorophenoxyacetic acid, thidiazuron and gibberellic acid at 22°C for 4 wk, followed by heat shock treatment at 32°C or 42°C for 2 wk, and the stressed material was recultured at 22°C for 24 wk in the dark. When anthers were heat shocked at 32°C and 42°C, the cultivated anthers swelled and burst, followed by the microspores flowing onto the explant surface. Androgenesis occurred directly and indirectly at uni- and binucleate stages. The micromorphology of multicellular, proembryos, globular, torpedo and […]

  • Maximization of micropropagule production in banana cultivars Grand naine (AAA) and Elakki (AB)
    on January 31, 2020 at 12:00 am

    Abstract Shoot tips from two different cultivars of banana known as Grand Naine (AAA) and Elakki (AB), were evaluated for their in vitro response in different concentrations of cytokinin and auxin which included 6-benzylaminopurine (6-BAP; 2, 3, and 4 mg L−1) and α-naphthalene acetic acid (NAA; 1 mg L−1) which was added to Murashige and Skoog (MS) basal media. The mean survival rates for cultured Grand Naine and Elakki explants in the varied growth regulator treatments were 66.25% and 52.5%, respectively. Rooting was initiated in the seventh passage using half-strength MS medium supplemented with 200 mg L−1 activated charcoal and 20 mg L−1 adenine sulfate (AS). The highest mean number of shoots generated per explant at the end of the seventh subculture from Grand Naine was 20.10 ± 0.73, and in Elakki, it was 11.20 ± 0.53. MS media supplemented with […]

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