Plant regeneration from protoplasts of Gentiana straminea Maxim

Shi, Guomin; Yang, Lina; He, Tengbing

doi:10.1515/biol-2016-0007

Cited by 7 publications

(5 citation statements)

References 23 publications

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“…Thus, within the framework of our researches we observed a different ratio of cell population being developed by cytodifferentiation. The morphology of the microcolonies obtained in the result of our study was common with the early stages of development of protoplasts of both Gentiana kurroo (Niedziela & Rybczyński, 2007) and G. straminea (Shi et al, 2016), which developed through direct somatic embryogenesis in specially selected plant lines in the deficiency of exogenous gibberellin. Related morphology has been observed, also in the initial division of the zygote in planta (Zhao & Sun, 2015) as well as polarization (formation by root-like projection) in the brown algae (Bogaert et al, 2013).…”

Section: Discussionsupporting

confidence: 63%

Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

Kondratenko

Pasternak

Samovol

et al. 2020

Regul. Mech. Biosyst.

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The Major Histocompatability Complex (MHC) determines the immune response to pathogens, and its genes are promising candidates for the search of associations with diseases. A special role is played by BoLA-DRB3 gene, the product of which directly participates in the binding of alien antigens and conditions the specificity of the immune response. The second exon of this gene codes β1-domain of class II antigens, which is necessary for binding a broad spectrum of alien antigens. Exon 2 of BoLA-DRB3 gene is extremely polymorphic, giving the possibility to search the associations of its alleles with various diseases. The article provides the results of the study on polymorphism of alleles of BoLA-DRB3.2 gene for detection of its associations with sensitivity to fusobacteriosis (necrobacteriosis) of cows. The survey was performed using PCR-RFLP method with DNA of blood from 176 cows of two herds of Ukrainian black-and-white dairy breed. As a result of the studies, in the first herd, 25 BoLA-DRB3.2 alleles were found. In the selections of nectobacteriosis susceptible and resistant cows, we found 22 and 21 variants respectively. In the second herd, in the general selection and group of healthy animals, 27 alleles were typed, and 22 in the group of susceptible cows. BoLA-DRB3.2*22 allele was the commonest in both herds in both general selections and groups of nectobacteriosis-resistant cows. In the selection of susceptible animals, the commonest was the variant BoLA-DRB3.2*16. We determined statistically significant associations of BoLA-DRB3.2 alleles with sensitivity to nectobacteriosis of cattle. BoLA-DRB3.2*03 and *22 alleles associate with nectobacteriosis-resistant, while *16 and *23 – with nectobacteriosis-susceptible cows of the both studied groups. Also, in the first herd, another allele was found – *24, indicating close relationship with the disease. The studies of polymorphism of BoLA-DRB3 gene expand the knowledge about genetic peculiarities of the Ukrainian black-and-white dairy breed. The identified molecular-genetic markers could be useful for breeders whose work is oriented towards the formation of herds which are resistant to diseases of the limbs in cattle.

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Section: Discussionsupporting

confidence: 63%

Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

Kondratenko

Pasternak

Samovol

et al. 2020

Regul. Mech. Biosyst.

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“…In our work, the highest callus induction rate was obtained by using a combination of BAP and 2,4D. It has been demonstrated that the auxin 2,4-D is essential for the formation of the cell wall and the initial protoplast growth as previously observed in other species ( Shi et al., 2016 ; Tu et al., 2023 ). Although we failed in somatic embryo formation and plantlet regeneration under the procedure defined by us microcolonies and embryogenic callus formation were achieved and it provides a base for further optimization.…”

Section: Discussionsupporting

confidence: 82%

Advances in Quercus ilex L. breeding: the CRISPR/Cas9 technology via ribonucleoproteins

Pavese,

Moglia,

Milani

et al. 2024

Front. Plant Sci.

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The CRISPR/Cas9 ribonucleoprotein (RNP)-mediated technology represents a fascinating tool for modifying gene expression or mutagenesis as this system allows for obtaining transgene-free plants, avoiding exogenous DNA integration. Holm oak (Quercus ilex) has an important social, economic, and ecological role in the Mediterranean climate zones of Western Europe and North Africa and is severely affected by oak decline syndrome. Here we report the first example of the application of the CRISPR/Cas9-RNP technology in holm oak. Firstly, we evaluated the protoplast isolation from both in vitro leaves and proembryogenic masses. Proembryogenic masses represented the best material to get high protoplast yield (11 x 106 protoplasts/ml) and viability. Secondly, the protoplast transfection ability was evaluated through a vector expressing green fluorescence protein as marker gene of transfection, reaching a transfection percentage of 62% after 24 hours. CRISPR/Cas9 RNPs were successfully delivered into protoplasts resulting in 5.6% ± 0.5% editing efficiency at phytoene desaturase (pds) target genomic region. Protoplasts were then cultured in semisolid media and, after 45 days in culture, developed embryogenic calli were observed in a Murashige and Skoog media with half concentration of NH4NO3 and KNO3 supplemented with 0.1 mg/L benzylaminopurine and 0.1 mg/L 2,4-dichlorophenoxyacetic acid.

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“…Another protocol for plant regeneration via SE from callus-derived protoplasts was developed for G. straminea (Shi et al, 2016). The agar-pool culture system was the only method that allows the production of embryogenic microcallus.…”

Section: Embryogenesis In Protoplast Culturesmentioning

confidence: 99%

“…A very high yield of regenerants via SE in protoplast bead culture confirmed the usefulness of this method for gentian cell manipulation. An efficient protocol for plant regeneration through SE was also developed for embryogenic cell suspension-derived protoplasts of G. macrophylla with the use of very similar culture conditions to those applied for protoplasts of G. straminea (Hu et al, 2015; Shi et al, 2016).…”

Section: Embryogenesis In Protoplast Culturesmentioning

confidence: 99%

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

et al. 2019

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The family Gentianaceae consists of 1736 species, which play an important role in human being existence due to their pharmacological and horticultural values. Many species accumulate bitter iridoid substances used medicinally and in flavorings, while others are cultivated because of beauty of their flowers showing a wide range of colors and patterns. Out of 99 genera belonging to the gentian family, process of somatic embryogenesis (SE) was reported for 5. The first reports, aimed at micropropagation of ornamental cultivars and production of secondary metabolites, concerned Centaurium erythraea Rafn., Eustoma russellianum Grieseb. and Exacum affine Balf. Somatic embryos were induced on different explants cultured in the liquid Murashige and Skoog medium supplemented with auxins and cytokinins. In the 1990s of the last century, significant progress in the exploration of the phenomenon of SE and its biotechnological application was made for the genus Gentiana . The process was induced on various explants and studied at the structural and ultrastructural levels. Regenerated plants were screened for genetic stability using flow cytometry, chromosome counting, and molecular markers. Besides typical indirect SE, the use of leaf fragments enabled to obtain single-cell origin of somatic embryos. On the other hand, proliferation of embryogenic callus in liquid medium resulted in the establishment of long-term embryogenic cell suspension cultures, paving the way not only to study the formation of somatic embryos and the development of regenerants but also to preserve the morphogenic potential of cell aggregates by cryopreservation. Cell suspensions re-established after storage in liquid nitrogen maintained their embryogenic character and allowed to obtain somatic embryo-derived regenerants that were true-to-type at both genetic and epigenetic levels. Another application of SE was related to genetic manipulation purposes. Efficient protocols of plant regeneration from callus-, cell suspension-, or leaf mesophyll-derived protoplasts allowed engaging procedures of somatic hybridization or protoplast electroporation for gentian genome modifications. Also, high embryogenic potential existing in the numerous gentian species enabled successful Agrobacterium -mediated transformation of G. cruciata L. and G. dahurica Fisch.

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Plant regeneration from protoplasts of Gentiana straminea Maxim

Cited by 7 publications

References 23 publications

Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

Advances in Quercus ilex L. breeding: the CRISPR/Cas9 technology via ribonucleoproteins

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

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