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.


  • The effect of endophytic bacteria on in vitro shoot growth of Prunus yedoensis and its identification and elimination
    on October 17, 2019 at 12:00 am

    Abstract Tissue cultures of Prunus yedoensis, a famous ornamental tree species, are important to mass-propagate true-to-type plants. Within in vitro cultures, bacterial contamination of explants hinders the propagation of sustainable cultures. Negative effects of endophytic bacteria vary from leaf yellowing to shoot-tip necrosis, ultimately resulting in shoot death. Explants with an overgrowth of endophytic bacteria on basal parts showed inferior growth than explants with no bacterial growth. Shoot growth and contamination became severe as the shoots were subcultured. Two endophytic bacteria, Pantoea spp. and Curtobacterium spp., were identified from the in vitro shoots using 16S rRNA gene sequences. Bacterial susceptibility to antibiotics was screened using different antibiotics. Two antibiotics, cefotaxime and tetracycline, effectively controlled bacteria growth. Although both antibiotics killed the bacteria, tetracycline adversely affected plant growth by reducing […]

  • Euclidean distance can recognize the Biojas ® concentration that produces the ideal physiological status of pineapple in vitro plantlets
    on October 17, 2019 at 12:00 am

    Abstract Biojas® is a fermented broth of Lasiodiplodia theobromae, a jasmonic acid–producing fungus characterized as a plant growth regulator and as biological control of phytopathogenic microorganisms and pests. The present work describes the use of Biojas® in in vitro culture of pineapple. On the other hand, plant scientists usually record multiple indicators in their experiments. The common statistical data evaluations involve univariate analyses such as t test, Mann-Whitney, and analysis of variance (ANOVA) followed by Tukey’s HSD. Such analyses do not evaluate integrally the effects of the experimental treatments because each indicator is analyzed independently. For this reason, we explored in this study the Euclidean distance combined with the data of the Biojas® treatment (0–2 mg l−1) on pineapple in vitro plantlets as an integrating indicator. Plant height; number of leaves; D leaf length, width, and area; […]

  • Modifying sugarcane mineral levels through sodium chloride and mannitol exposure in temporary immersion bioreactors
    on October 15, 2019 at 12:00 am

    Abstract Temporary immersion bioreactors (TIBs) have been shown to be useful for studying plant stress physiology and inducing chemical mutagenesis. This study describes the effects of exposure to NaCl (salt stress) and mannitol (osmotic stress) within TIB on sugarcane mineral levels in vitro. Shoots were exposed to concentration of NaCl (89.4 mM) and mannitol (123.1 mM) previously shown to result in a 50% reduction in multiplication rate for 30 d. Thereafter, shoot multiplication rate, shoot cluster fresh weight, and levels of selected minerals were measured. Using ICP-OES, the following minerals were quantified: Na, Ga, Mn, Cr, K, Zn, Ca, Li, Mg, Sr, Co, B, Fe, S, P, Al, Ba, and N. Both NaCl and mannitol decreased shoot multiplication by c. 53% and except for Al and Ba altered mineral levels significantly relative to the control: Na accumulation increased markedly (six-fold in NaCl treatment); levels of Ga, Mn, Cr, K, and Zn changed moderately; Ca, […]

  • Direct somatic embryogenesis and shoot regeneration from leaves and internodes of Pluchea lanceolata (DC.) C.B. Clarke
    on September 9, 2019 at 12:00 am

    Abstract Pluchea lanceolata (DC.) C.B. Clarke is a threatened native medicinal plant. Increasing the propagation of this plant will preserve the wild population and provide material for medicinal use. In vitro and field-collected shoots and leaves were tested for response to 2,4-dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ), for initiation of direct shoot regeneration (DSR), or direct somatic embryogenesis (DSE). Leaves and internodes collected from field-grown plants produced only callus, while in vitro–raised shoots exhibited DSR and DSE on Murashige and Skoog (MS) medium with 2,4-D and TDZ. Direct shoot regeneration occurred on medium with TDZ from internode and leaf segments obtained from in vitro–developed shoots. In vitro–grown shoots were rooted on half-strength MS medium with 2 mg L−1 indole-3-butyric acid and acclimatized. Survival in natural conditions was 62.5% for DSE and 79% for DSR plantlets. […]

  • In vitro propagation of medicinal and aromatic plants: the case of selected Greek species with conservation priority
    on September 9, 2019 at 12:00 am

    Abstract Worldwide, many medicinal and aromatic plants (MAPs) are still collected from the wild and only a small fraction of them are exclusively sourced from cultivation. This practice when performed non-sustainably threatens species and populations. Micropropagation of MAPs is a powerful tool to conserve rare, threatened, and valuable MAPs, and to massively produce high-value plant material for cultivation without seasonal constraints. In this study, the in vitro propagation protocols of 22 Greek native MAPs assigned with conservation priority were assessed (herbaceous perennials, bulbous, subshrubs, and trees), including 17 range-restricted plants and 5 taxa of Orchidaceae. For the latter, current micropropagation efforts include seed germination, callus induction, and protocorm formation for successful plantlet development; however, these propagation protocols are still fragmentary. For the rest (n = 17), a five-stage detailed procedure is outlined […]

  • In vitro propagation of Trichosanthes kirilowii Maxim. through nodal segment shoot proliferation
    on August 23, 2019 at 12:00 am

    Abstract Trichosanthes kirilowii Maxim. is a vital traditional herbal medicinal plant found in northeastern Asia. Its roots, fruits, and seeds are used as food and medicine. Roots harvested for medicinal use take over 3 yr to mature when the plant is grown in a traditional way through cultivation in the field. This coupled with uncertainty in identification of the plant when collected from the wild calls for a standard in vitro propagation system to meet the increasing demand for it. The purpose of this study was to develop a standard protocol for the in vitro micropropagation of T. kirilowii. Ten different media supplemented with different concentrations of plant growth regulators were evaluated. At 5 wk, De Greef and Jacobs medium supplemented with 0.1 mg L−1 kinetin led to optimal shoot growth, while the same medium supplemented with 0.5 mg L−1 indole 3-butyric acid induced optimal root growth, also at 5 wk. The […]

  • Effects of sodium alginate elicitation on secondary metabolites and antioxidant activity of safflower genotypes under in vitro salinity stress
    on August 20, 2019 at 12:00 am

    Abstract The present study was aimed at investigating the effects of different concentrations of sodium alginate (NaAlg) (0.075 and 0.15% (w/v)) on the production of secondary metabolites (SMs) and antioxidant activity of seven safflower genotypes under in vitro salinity stress. The results showed that total phenolic content (TPC), total flavonoids (TFD), total flavonols (TFL), anthocyanin (Ant), total antioxidant capacity (TAC), phenylalanine ammonia-lyase (PAL), catalase (CAT) activity, and lipid peroxidation significantly increased under salinity stress consisting of the concentration of 1.5% (w/v) of NaCl, but callus growth traits decreased. The highest amount of TPC, Ant, and callus growth traits was observed under the elicitation of the sample with the concentration of 0.075(%) NaAlg under salinity stress, but the highest amount for TFD, TFL, CAT, PAL, and TAC was observed under elicitation of the sample with the concentration of 0.15% of NaAlg under salinity […]

  • 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 […]

  • 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 […]

  • 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 […]

  • 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 […]

  • 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 […]

  • 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 […]

  • Morphophysiological in vitro performance of Brazilian ginseng ( Pfaffia glomerata (Spreng.) Pedersen) based on culture medium formulations
    on August 1, 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 […]

  • Somatic embryogenesis to overcome low seed viability and conserve wild banana ( Ensete superbum (Roxb.) Cheesman)
    on August 1, 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 […]

  • 2019 In Vitro Biology Meeting, June 8 – 12, Tampa, Florida
    on August 1, 2019 at 12:00 am


  • The effect of slow-growth strategy on a production of Petunia × hybrida Vilm. microcuttings
    on August 1, 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 […]

  • Development of an efficient micropropagation system for Tecoma stans (L.) Juss. ex Kunth using thidiazuron and effects on phytochemical constitution
    on August 1, 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 […]

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