In Vitro Regeneration Potential of Thin Cell Layer Explants of Lentisk (Pistacia lentiscus var. Chia) Plant

A basic factor underlying the success of largeâ€scale micropropagation and genetic transformation of any plant species is regeneration. In order to regenerate propagules of Rosa damascena Mill. on a large scale, an efficient and improved in vitro propagation system has been established using transverse thin cell layer culture (tTCL). By optimizing the position of the tissue and applying an improved selection procedure, in vitro shoots were elongated in 8 weeks of culture. Modified Murashige and Skoog (1962)(MS) medium fortified with 4.0 mg lâ€1 6â€benzylaminopurine (BAP) and 0.4mg lâ€1 anaphthalene acetic acid (NAA) gave optimal shoot regeneration. The explant was inoculated on this medium in the upright position and exhibited a high frequency of shoot regeneration (~96.66%), and it also gave the highest number of shoots (22.33/explant). The horizontally placed explant on an average 7.66 shoots/explant. Our experiments indicate that explant orientation strongly influences the organogenesis response. The frequency of shoot initiation and the number of multiple shoots produced from each explant were significantly dependent on the plant source, concentration of plant growth regulators and the orientation of the explants and contributed significantly to in vitro regeneration. Rooting of well developed shoots was achieved on hormone free ¼ strength MS medium with 4% sucrose.

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Effects of Variations in Hormonal Treatments Upon In vitro Regeneration Potential in Embryo Explants of Arenga pinnata

Malaysian Journal of Fundamental and Applied Sciences ◽

10.11113/mjfas.v17n5.2120 ◽

2021 ◽

Vol 17 (5) ◽

pp. 495-503

Author(s):

Shamsiah Abdullah ◽

Siti Nurain Roslan

Keyword(s):

Acetic Acid ◽

In Vitro Regeneration ◽

Hormonal Treatment ◽

Height Increment ◽

Regeneration Potential ◽

In Vitro Method ◽

Indole 3 Acetic Acid ◽

Hormonal Treatments

One of the challenges related to propagation of Arenga pinnata is its lengthy period of seed dormancy. In this study, in vitro regeneration was carried out to determine the effect of hormonal treatment on the embryo explant of Arenga pinnata. Embryos were surface sterilized and cultured into different media supplemented with various hormones concentrations and combinations. Each treatment contained of Kinetin (KN) hormone (1.0, 2.0, and 3.0 mg/l) and in combination with indole-3-acetic acid (IAA) of 0.1, 0.2, 0.3 mg/l. The height of plumule and length of radical was observed and recorded. Treatment 8 (3 mg/ml KN + 0.1 mg/ml IAA) showed 59.09% in plumule height increment while treatment 4 (1 mg/ml KN + 0.3 mg/ml IAA) showed the highest radical increments with 93.62%. The knowledge gained in this study consequently helps us to better understand the role of KN and IAA in the in vitro regeneration protocol. Since in vitro method able to produce higher number of in vitro seedlings at one time, it is important to establish the in vitro regeneration protocol for this plant.

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Regeneration of Plants from Apical Meristem Tips and Nodal Segments of Arachis pintoi

Peanut Science ◽

10.3146/pnut.30.2.0001 ◽

2003 ◽

Vol 30 (2) ◽

pp. 75-79 ◽

Cited By ~ 4

Author(s):

H. Y. Rey ◽

L. A. Mroginski

Keyword(s):

Apical Meristem ◽

In Vitro Regeneration ◽

Nodal Segments ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Regeneration Potential ◽

Arachis Pintoi ◽

Solid Media ◽

One Step

Abstract The in vitro regeneration potential of shoot apical tips (2 to 3 mm in length), meristems (0.3 to 0.5 mm in length), and nodal segments (4 to 7 mm long with an axillary bud) of diploid (2n = 2x = 20) and triploid (2n = 3x = 30) cytotypes of Arachis pintoi was evaluated. Explants were cultured on MS medium supplemented with different concentrations and combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA). In one experiment the effect of gibberellic acid was tested. The cultures were done in liquid and solid media. Plant regeneration can be readily achieved from all explants in one step of 30 d culture on MS + 0.01 mg/L each of NAA and BA or two steps consisting of 1) shoots regeneration through culture of explants on MS + 0.01 mg/L each of NAA and BA, and 2) induction of rooting in regenerated shoots by reculture on MS + 0.01 mg/L NAA. The plantlets were successfully transferred to pots in a greenhouse.

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Silver nanoparticles enhanced efficiency of explant surface disinfection and somatic embryogenesis in Begonia tuberous via thin cell layer culture

Vietnam Journal of Biotechnology ◽

10.15625/1811-4989/15872 ◽

2021 ◽

Vol 19 (2) ◽

pp. 337-347

Author(s):

Hoang Thanh Tung ◽

Hoang Thi Van ◽

Huynh Gia Bao ◽

Le The Bien ◽

Hoang Dac Khai ◽

...

Keyword(s):

Silver Nanoparticles ◽

Somatic Embryogenesis ◽

Cell Layer ◽

Initial Study ◽

Thin Cell Layer ◽

Flower Stalk ◽

Time Treatment ◽

Induction Rate ◽

Thin Cell Layer Culture

In vitro culture establishment is one of the most important stages in micropropagation. The disinfectant effectiveness depends on the type of surface disinfectant, concentration and the time treatment. In this initial study, silver nanoparticles (AgNPs) were used as a disinfectant for petioles, flower stalks and stems of Begonia tuberous. In addition, thin cell layer culture (TCL) technique has been applied for the purpose of somatic embryogenesis. The results showed that AgNPs were effective in eliminating infectious microorganisms on B. tuberous explants; which were identified included 4 species of fungi (Fusarium sp., Aspergillus aculeatus, Trichoderma sp. and Penicillium sp.) and 1 species of bacteria (Pseudomonas sp.). At concentrations of 200 ppm and 300 ppm, AgNPs were not only effective in disinfection but also increased the induction rate of somatic embryogenesis in flower stalk TCL explants (approximately 40.00%); a similar effect was observed in stem TCL explants at the same concentration. Meanwhile, for petiole TCL explants, the induction rate of somatic embryogenesis was optimal when using AgNPs at a concentration of 100 - 300 ppm to disinfected the explant. In contrast, at high (400 ppm) or low (50 ppm) concentrations of AgNPs did not play a disinfecting role and stimulated somatic embryogenesis. In addition, explants derived from AgNPs sterilization did not show any abnormalities in somatic embryogenesis with shapes such as globular, heart, torpedo, and cotyledon. AgNPs showed double efficacy in sterilization of explants and improved efficiency of somatic embryogenesis from TCL petioles, flower stalks and stems explants; thus increasing the efficiency micropropagation of B. tuberous.

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High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

Notulae Scientia Biologicae ◽

10.15835/nsb.6.1.9225 ◽

2014 ◽

Vol 6 (1) ◽

Cited By ~ 1

Author(s):

Dikash Singh THINGBAIJAM ◽

Devi Sunitibala HUIDROM

Keyword(s):

Plant Regeneration ◽

High Frequency ◽

Cell Layer ◽

Thin Cell Layer ◽

Regeneration System ◽

Transverse Thin Cell Layer

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An improved thin cell layer culture system for efficient clonal propagation and in vitro withanolide production in a medicinal plant Withania coagulans Dunal

Industrial Crops and Products ◽

10.1016/j.indcrop.2018.04.012 ◽

2018 ◽

Vol 119 ◽

pp. 172-182 ◽

Cited By ~ 9

Author(s):

Deepika Tripathi ◽

Krishna Kumar Rai ◽

Sanjay Kumar Rai ◽

Shashi Pandey Rai

Keyword(s):

Medicinal Plant ◽

Culture System ◽

Clonal Propagation ◽

Cell Layer ◽

Thin Cell Layer ◽

Withania Coagulans ◽

Thin Cell Layer Culture

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Investigating the In Vitro Regeneration Potential of Commercial Cultivars of Brassica

Plants ◽

10.3390/plants8120558 ◽

2019 ◽

Vol 8 (12) ◽

pp. 558 ◽

Cited By ~ 1

Author(s):

Nisma Farooq ◽

Muhammad Asif Nawaz ◽

Zahid Mukhtar ◽

Iftikhar Ali ◽

Penny Hundleby ◽

...

Keyword(s):

In Vitro Regeneration ◽

Commercial Variety ◽

Regeneration Potential ◽

Regeneration Frequency ◽

Commercial Cultivars ◽

Normal Seed ◽

Regeneration Response ◽

Almost All ◽

Seed Yields

In vitro regeneration is a pre-requisite for developing transgenic plants through tissue culture-based genetic engineering approaches. Huge variations among different genotypes of the genus Brassica necessitate the identification of a set of regeneration conditions for a genotype, which can be reliably used in transformation experiments. In this study, we evaluated the morphogenesis potential of four commercial cultivars (Faisal canola, Punjab canola, Aari canola, Nifa Gold) and one model, Westar, from four different explants namely cotyledons, hypocotyls, petioles and roots on three different Brassica regeneration protocols, BRP-I, -II and -III. The regeneration efficiency was observed in the range of 6–73%, 4–79.3%, 0–50.6%, and 0–42.6% from cotyledons, petioles, hypocotyls and roots, respectively, whereas, the regeneration response in terms of average shoots per explant was found to be 0.76–10.9, 0.2–3.2, 0–3.4 and 0–2.7 from these explants. Of the commercial varieties tested, almost all varieties showed poorer regeneration than Westar except Aari canola. In comparison to Westar, its regeneration frequency from cotyledons was up to 7.5-fold higher on BRP-I, while it produced up to 21.9-fold more shoots per explant. Our data show that the explant has strong influence on the regeneration response, ranging from 24% to 92%. While the growth of commercial cultivars was least affected by the regeneration conditions provided, the effect on Westar was twice that of the commercial cultivars. After determining the optimal explant type and regeneration conditions, we also determined the minimum kanamycin concentration levels required to selectively inhibit the growth of untransformed cells for these cultivars. Regenerated shoots of Aari canola could be successfully grown to maturity within 16–18 weeks, with no altered phenotype noted and normal seed yields obtained. Therefore, the commercial variety, Aari canola, could be a good candidate for future genetic transformation studies.

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High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

Notulae Scientia Biologicae ◽

10.15835/nsb619225 ◽

2014 ◽

Vol 6 (1) ◽

pp. 85-91

Author(s):

Dikash Singh THINGBAIJAM ◽

Devi Sunitibala HUIDROM

Keyword(s):

High Frequency ◽

Somatic Embryos ◽

Cell Layer ◽

Sucrose Concentration ◽

Dichlorophenoxyacetic Acid ◽

Thin Cell Layer ◽

Regeneration System ◽

Transverse Thin Cell Layer ◽

2,4 Dichlorophenoxyacetic Acid

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

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