Development of an efficient tissue culture protocol for callus formation and plant regeneration of wetland species Juncus effusus L.

Tissue culture is an essential requirement in plant science to preserve genetic resources and to expand naturally occurring germplasm. A variety of naturally occurring and synthetic hormones are available to induce the processes of dedifferentiation and redifferentiation. Not all plant material is susceptible to tissue culture, and often complex media and hormone requirements are needed to achieve successful plant propagations. The availability of new hormones or chemicals acting as hormones are critical to the expansion of tissue culture potentials. Phloroglucinol has been shown to have certain hormone-like properties in a variety of studies. Ornithogalum dubium, an important geophyte species, was used to characterise the potential of phloroglucinol as the sole plant-like hormone in a tissue culture experiment. Tissue culture, plant regeneration, total phenolic and genetic variability were established by applying a variety of methods throughout long-term experiments. Phloroglucinol did induce callus formation and plant regeneration when used as the sole supplement in the media at a rate of 37%, thus demonstrating auxin/cytokines-like properties. Callus formation was of 3 types, friable and cellular, hard and compact, and a mixture of the two. The important finding was that direct somatogenesis did occur albeit more frequently on younger tissue, whereby rates of induction were up to 52%. It is concluded that phloroglucinol acts as a “hormone-like” molecule and can trigger direct embryogenesis without callus formation.

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Study on Tissue Culture of Sweet Broad Pea

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.644-650.5407 ◽

2014 ◽

Vol 644-650 ◽

pp. 5407-5410

Author(s):

Hui Fang Chi

Keyword(s):

Tissue Culture ◽

Plant Regeneration ◽

Culture Medium ◽

Callus Formation ◽

Formation Rate ◽

Adventitious Bud ◽

Experimental Basis ◽

Research Results

s. The cotyledons, Internodes, leaves and stems of sweet broad pea were studied on tissue culture. Research results show that: The ability of different explants for callus formation and adventitious bud differentiation in different culture medium is different. The callus formation rate and sprouting rate of Internodes is significantly higher than other explants, which is a ideal material for tissue culture. The callus formation rate of Internodes was 100% in MS +BA1.0 mg/L+NAA 1.0 mg/L and MS+ 2, 4-D 0.5 mg/L; The bud differentiation is best at the medium of MS+ 6-BA 2 mg/L, which reached 86.7%; the rooting rate was 83.3% at the medium of MS+ NAA 3mg/L. The study provides a experimental basis for further study on the plant regeneration in the sweet broad pea.

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Optimization of protoplast regeneration in the model plant Arabidopsis thaliana

Plant Methods ◽

10.1186/s13007-021-00720-x ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Yeong Yeop Jeong ◽

Hun-Young Lee ◽

Suk Weon Kim ◽

Yoo-Sun Noh ◽

Pil Joon Seo

Keyword(s):

Tissue Culture ◽

Arabidopsis Thaliana ◽

Plant Regeneration ◽

Tissue Regeneration ◽

De Novo ◽

Callus Formation ◽

Protoplast Regeneration ◽

Culture Methods ◽

Cell And Tissue Culture

Abstract Background Plants have a remarkable reprogramming potential, which facilitates plant regeneration, especially from a single cell. Protoplasts have the ability to form a cell wall and undergo cell division, allowing whole plant regeneration. With the growing need for protoplast regeneration in genetic engineering and genome editing, fundamental studies that enhance our understanding of cell cycle re-entry, pluripotency acquisition, and de novo tissue regeneration are essential. To conduct these studies, a reproducible and efficient protoplast regeneration method using model plants is necessary. Results Here, we optimized cell and tissue culture methods for improving protoplast regeneration efficiency in Arabidopsis thaliana. Protoplasts were isolated from whole seedlings of four different Arabidopsis ecotypes including Columbia (Col-0), Wassilewskija (Ws-2), Nossen (No-0), and HR (HR-10). Among these ecotypes, Ws-2 showed the highest potential for protoplast regeneration. A modified thin alginate layer was applied to the protoplast culture at an optimal density of 1 × 106 protoplasts/mL. Following callus formation and de novo shoot regeneration, the regenerated inflorescence stems were used for de novo root organogenesis. The entire protoplast regeneration process was completed within 15 weeks. The in vitro regenerated plants were fertile and produced morphologically normal progenies. Conclusion The cell and tissue culture system optimized in this study for protoplast regeneration is efficient and reproducible. This method of Arabidopsis protoplast regeneration can be used for fundamental studies on pluripotency establishment and de novo tissue regeneration.

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Studies on the Micropropagation of Anemonella thalictroides Spach by Tissue Culture. 1. Conditions of Callus Formation from Leaf, Stem and Petiole Explants, and Plant Regeneration from Stem-derived Callus.

Plant tissue culture letters ◽

10.5511/plantbiotechnology1984.11.96 ◽

1994 ◽

Vol 11 (2) ◽

pp. 96-102 ◽

Cited By ~ 1

Author(s):

Fumiaki KISHI ◽

Yuukichi KAGAMI ◽

Masao KOYAMA

Keyword(s):

Tissue Culture ◽

Plant Regeneration ◽

Callus Formation ◽

Petiole Explants

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Somatic Embryogenesis and Plant Regeneration in Cucumber

HortScience ◽

10.21273/hortsci.29.8.906 ◽

1994 ◽

Vol 29 (8) ◽

pp. 906-909 ◽

Cited By ~ 14

Author(s):

H. Lou ◽

S. Kako

Keyword(s):

Tissue Culture ◽

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryos ◽

Callus Formation ◽

Semisolid Medium ◽

Cotyledon Explants ◽

High Sucrose ◽

Embryogenic Callus Formation ◽

Internode Explants

The embryogenic capacity of seven cucumber (Cucumis sativus L.) cultivars was examined by tissue culture of cotyledon, young first-leaf, and internode explants. Somatic embryogenesis frequencies differed significantly among the tested cultivars, and `Fushinarimidori' produced the highest number of embryos from either cotyledons or young first leaves. Cotyledon- and first-leaf-derived calluses produced more embryos than calluses from internodes. Somatic embryos were induced from `Aonaga F1' internodes. With relatively high sucrose levels (6% and 9%) in the initiation medium, the frequency of embryogenic callus formation from `Fushinarimidori' cotyledon explants was >90%. The highest yield of somatic embryos occurred in cultures initiated with high sucrose levels (9% or 12%), although 12% sucrose inhibited callus formation and growth. Somatic embryos germinated in a basal liquid medium supplemented with 0.5% activated charcoal, and they developed into well-shaped, healthy plantlets on semisolid medium with 1% sucrose.

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