In vitro response and pedigree analysis for somatic embryogenesis of long-day photoperiod adapted soybean

Our aim was to elaborate an efficient and reproducible protocol for medium-term culture of the threatened taxon Dianthus nardiformis. To reduce the growth, sucrose, mannitol, polyethylene glycol, Abscisic acid and Jasmonic acid were tested. For assessing the in vitro response, the growth and regeneration were registered after different time intervals. Mannitol is the most effective for medium-term preservation viable cultures which can be maintained unlimited time through transfer at every 3 months. In its presence, somatic embryogenesis was induced and in vitro growth in the minimal cultures was reduced between 9 and 12 times comparing to the control. Antioxidant enzymes assay revealed qualitative and quantitative differences among the experimental variants, and also between different concentrations of the same compound in correlation with the growth reduction and regeneration. POX was the most suitable to detect the efficiency of different treatments to induce medium-term cultures.

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Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

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Telomere Length in Norway Spruce during Somatic Embryogenesis and Cryopreservation

Plants ◽

10.3390/plants10020416 ◽

2021 ◽

Vol 10 (2) ◽

pp. 416

Author(s):

Tuija Aronen ◽

Susanna Virta ◽

Saila Varis

Keyword(s):

Somatic Embryogenesis ◽

Telomere Length ◽

Norway Spruce ◽

Genotypic Variation ◽

Production Capacity ◽

Stress Factors ◽

Embryo Production ◽

One Year ◽

Plant Telomeres

Telomeres i.e., termini of the eukaryotic chromosomes protect chromosomes during DNA replication. Shortening of telomeres, either due to stress or ageing is related to replicative cellular senescence. There is little information on the effect of biotechnological methods, such as tissue culture via somatic embryogenesis (SE) or cryopreservation on plant telomeres, even if these techniques are widely applied. The aim of the present study was to examine telomeres of Norway spruce (Picea abies (L.) Karst.) during SE initiation, proliferation, embryo maturation, and cryopreservation to reveal potential ageing or stress-related effects that could explain variation observed at SE process. Altogether, 33 genotypes from 25 families were studied. SE initiation containing several stress factors cause telomere shortening in Norway spruce. Following initiation, the telomere length of the embryogenic tissues (ETs) and embryos produced remains unchanged up to one year of culture, with remarkable genotypic variation. Being prolonged in vitro culture can, however, shorten the telomeres and should be avoided. This is achieved by successful cryopreservation treatment preserving telomere length. Somatic embryo production capacity of the ETs was observed to vary a lot not only among the genotypes, but also from one timepoint to another. No connection between embryo production and telomere length was found, so this variation remains unexplained.

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Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

Journal of Biological Research-Thessaloniki ◽

10.1186/s40709-021-00138-5 ◽

2021 ◽

Vol 28 (1) ◽

Author(s):

Renata Orłowska

Keyword(s):

Tissue Culture ◽

Somatic Embryogenesis ◽

Taguchi Method ◽

Culture Conditions ◽

Dna Demethylation ◽

Silver Ion ◽

Donor Plant ◽

In Vitro Tissue Culture ◽

Induced Variation

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.

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Microstructure and surface texture driven improvement in in-vitro response of laser surface processed AZ31B magnesium alloy

Journal of Magnesium and Alloys ◽

10.1016/j.jma.2020.11.002 ◽

2021 ◽

Author(s):

Tso-Chang Wu ◽

Sameehan S. Joshi ◽

Yee-Hsien Ho ◽

Mangesh V. Pantawane ◽

Subhasis Sinha ◽

...

Keyword(s):

Magnesium Alloy ◽

Surface Texture ◽

Laser Surface ◽

Az31b Magnesium Alloy ◽

In Vitro Response

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Loss of Nitric Oxide Induces Fibrogenic Response in Organotypic 3D Co-Culture of Mammary Epithelia and Fibroblasts—An Indicator for Breast Carcinogenesis

Cancers ◽

10.3390/cancers13112815 ◽

2021 ◽

Vol 13 (11) ◽

pp. 2815

Author(s):

Gang Ren ◽

Xunzhen Zheng ◽

Vandana Sharma ◽

Joshua Letson ◽

Andrea L. Nestor-Kalinoski ◽

...

Keyword(s):

Nitric Oxide ◽

Culture System ◽

Epithelial Mesenchymal Transition ◽

Precancerous Lesions ◽

Three Dimensional ◽

Epidemiological Studies ◽

Mesenchymal Transition ◽

In Vitro Response ◽

Mammary Epithelia

Excessive myofibroblast activation, which leads to dysregulated collagen deposition and the stiffening of the extracellular matrix (ECM), plays pivotal roles in cancer initiation and progression. Cumulative evidence attests to the cancer-causing effects of a number of fibrogenic factors found in the environment, diseases and drugs. While identifying such factors largely depends on epidemiological studies, it would be of great importance to develop a robust in vitro method to demonstrate the causal relationship between fibrosis and cancer. Here, we tested whether our recently developed organotypic three-dimensional (3D) co-culture would be suitable for that purpose. This co-culture system utilizes the discontinuous ECM to separately culture mammary epithelia and fibroblasts in the discrete matrices to model the complexity of the mammary gland. We observed that pharmaceutical deprivation of nitric oxide (NO) in 3D co-cultures induced myofibroblast differentiation of the stroma as well as the occurrence of epithelial–mesenchymal transition (EMT) of the parenchyma. Such in vitro response to NO deprivation was unique to co-cultures and closely mimicked the phenotype of NO-depleted mammary glands exhibiting stromal desmoplasia and precancerous lesions undergoing EMT. These results suggest that this novel 3D co-culture system could be utilized in the deep mechanistic studies of the linkage between fibrosis and cancer.

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