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.

 

  • Spike morphology alternations in androgenic progeny of hexaploid triticale ( × Triticosecale Wittmack) caused by nullisomy of 2R and 5R chromosomes
    on December 14, 2019 at 12:00 am

    Abstract Induction of androgenesis, followed by chromosome doubling, is a crucial method to obtain complete homozygosity in one-generation route. However, in vitro androgenesis can result in various genetic and epigenetic changes in derived triticale plants. In this study, we evaluated chromosome alternations and we associated them with the changes of spike morphology in androgenic progeny of triticale. We karyotyped offspring plants that derived from double haploid plants using fluorescence in situ hybridization techniques. We distinguished four major groups of karyotypes: double ditelosomics, nullisomics N2R, nullisomics N5R, and triticale plants with a complete set of chromosomes. It is known that more than half of QTLs connected with androgenic response are located in R-genome of triticale but 2R, 5R, and 6R chromosomes are not included. We hypothesized that the reason why only aberrations of chromosomes 2R and 5R appear during androgenesis of triticale is that […]

  • Behavior of Opuntia ficus-indica (L.) Mill. Heat-Stressed Microspores Under In Vitro Culture Conditions as Evidenced by Microscopic Analysis
    on December 6, 2019 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 […]

  • Genome-wide analysis of ACO and ACS genes in pear ( Pyrus ussuriensis )
    on December 2, 2019 at 12:00 am

    Abstract The “Nanguo” pear is a typically climacteric fruit and ethylene is the main factor controlling the ripening process of climacteric fruit. Ethylene biosynthesis has been studied clearly and ACC synthase (ACS) is the rate-limited enzyme. ACO (ACC oxidase) is another important enzyme in ethylene biosynthesis. By exploring the pear genome, we identified 13 ACS genes and 11 ACO genes, respectively, and their expression patterns in fruit and other organs were investigated. Among these genes, 11 ACS and 8ACO genes were expressed in pear fruits. What’s more, 4 ACS and 3ACO genes could be induced by Ethephon and inhibited by 1-MCP treatment. This study is the first time to explore ACS and ACO genes at genome-wide level and will provide new data for research on pear fruit ripening.

  • Exogenous polyamines enhance somatic embryogenesis and Agrobacterium tumefaciens -mediated transformation efficiency in sugarcane ( Saccharum spp. hybrid)
    on December 2, 2019 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 […]

  • In vitro propagation of medicinal and aromatic plants: the case of selected Greek species with conservation priority
    on December 1, 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 […]

  • Alkaloid synthesis is coupled to shoot morphogenesis in Argemone mexicana L. (Papaveraceae) in vitro cultures
    on December 1, 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 […]

  • Use of DoE methodology to optimize the regeneration of high-quality, single-copy transgenic Zea mays L. (maize) plants
    on December 1, 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 […]

  • Direct somatic embryogenesis and shoot regeneration from leaves and internodes of Pluchea lanceolata (DC.) C.B. Clarke
    on December 1, 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.

  • Genetic homogeneity and high shoot proliferation in banana ( Musa acuminata Colla) by altering medium thiamine level and sugar type
    on December 1, 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 […]

  • In vitro propagation of Trichosanthes kirilowii Maxim. through nodal segment shoot proliferation
    on December 1, 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 […]

  • Thidiazuron-induced efficient biosynthesis of phenolic compounds in callus culture of Ipomoea turbinata Lagasca and Segura
    on December 1, 2019 at 12:00 am

    Abstract Ipomoea turbinata Lagasca and Segura (Purple Moonflower) belongs to the largest flowering genus Ipomoea in the Convolvulaceae family. Ipomoea turbinata has not been previously explored for its in vitro potential. This is the first study focused on thidiazuron-induced callus culture for efficient biosynthesis of commercially important phenolic compounds in this plant species. Among the two plant growth regulators tested on leaf, stem, and root explants, 5 mg L−1 thidiazuron (TDZ) induced the highest biomass accumulation (61.4 g L−1 fresh weight, 6.3 g L−1 dry weight) in leaf-derived callus cultures after 5 wk of culture. The highest total phenolic and flavonoid contents recorded were 9.04 mg g−1 and 1.16 mg g−1, respectively, in optimized callus cultures. High-performance liquid chromatography analysis indicated high levels of pharmacologically important anticancer compounds such as chlorogenic acid (13.48 mg g−1), […]

  • In vitro culture and micropropagation of the Baetic-Moroccan endemic plant Lapiedra martinezii Lag. (Amaryllidaceae)
    on December 1, 2019 at 12:00 am

    Abstract Lapiedra martinezii Lag. is a potential medicinal and ornamental plant facing conservation challenges. Thus, this study was focused on determining the conditions for culture initiation and propagation using in vitro techniques. The optimal sterilization procedure combined thermotherapy at 54°C for 60 min and immersion in 7% (w/v) Ca(ClO)2 solution for 20 min. The most suitable medium to initiate bulb scales cultures was Gamborg B5 medium containing 500 mg L−1 casein, 2 mg L−1 adenine, 10 mg L−1 glutathione and 10 g L−1 sucrose. The most productive multiplication medium tested was Murashige and Skoog medium containing 30 g L−1 sucrose, 4.0 mg L−1 6-benzylaminopurine, and 0.12 mg L−1 1-naphtaleneacetic acid. Most plants developed in vitro rooted spontaneously in the multiplication phase. The vast majority of the plants (89%) were successfully transferred to ex vitro conditions, and 100% survived over 1 yr of […]

  • Correction to: Genetic homogeneity assessment of in vitro -regenerated plantlets of Nyctanthes arbor-tristis L. and comparative evaluation of bioactive metabolites and antioxidant activity
    on December 1, 2019 at 12:00 am

    There was an error in this article as originally published. The surname of coauthor Sobha Chandra Rath was misspelled as “Ratha”. The original article has been corrected.

  • Micropropagation and molecular characterization of Thymus sibthorpii Benth. (Lamiaceae), an aromatic-medicinal thyme with ornamental value and conservation concern
    on December 1, 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 December 1, 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 […]

  • In vitro propagation of male and female Hippophae salicifolia D. Don
    on November 27, 2019 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 […]

  • Non-ionic surfactants improved microspore embryogenesis and plant regeneration of recalcitrant purple flowering stalk ( Brassica campestris ssp. chinensis var. purpurea Bailey)
    on October 30, 2019 at 12:00 am

    Abstract As Brassicaceae species are mostly cross-pollinated, breeding homozygous parental lines by traditional approaches is time-consuming and costly. Alternatively, microspore culture has been widely applied to produce double haploid lines in a short time. This study aimed to establish a highly efficient microspore culture protocol for purple flowering stalk. Among the five genotypes studied, the highest and lowest embryo induction rates were observed in J18 and J17 (13.5 and 7.67 embryos per bud, respectively). Microspores of genotypes J17 and J18 were successfully induced to produce embryos in NLN-13 medium, but the frequency of microspore embryogenesis was low. Three non-ionic surfactants (Pluronic F-68, Triton X-100, Tween-20) were evaluated independently for their effect on microspore embryogenesis of purple flowering stalk. Microspores of the two genotypes were cultivated in NLN-13 medium supplemented with different concentrations (0.0001%, 0.001%, 0.01%, […]

  • 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; […]

  • 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 […]

  • 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, […]


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