Introduction and Methods Employed in Plant Tissue Culture

Street, H. E.; Henshaw, G. G.

doi:10.1016/b978-1-4832-3146-4.50016-x

Cited by 27 publications

(15 citation statements)

References 183 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

Section: Measurement Of Root Growthmentioning

confidence: 99%

“…For the fresh weight determination, the roots were gently pressed on filter paper to remove excess water, and weighed. The growth rate (GR) was calculated using the formula [56]: GR = final weight (g) − initial weight (g)/initial weight (g) × 100%.The dry weight was determined after lyophilization. The conductivity (expressed in miliSiemens, mS) was determined using a conductivity meter (model CC-317, Elmetron, Poland; sensitivitity 0.5%).…”

mentioning

confidence: 99%

See 1 more Smart Citation

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

et al. 2014

View full text Add to dashboard Cite

IntroductionHairy, or transgenic roots are characterized by a rapid and stable growth, which importantly can be initiated and maintained in a hormone-free medium. They often synthetize approximately the same and in some cases even greater amounts of secondary metabolites compared to their mother plants [1,2] and the rate of production remains stable. This makes hairy roots of medicinal plants a good source of valuable compounds and obviously their large-scale culture in bioreactors holds immense potential for the pharmaceutical industry [3].Platycodon grandiflorum (Jacq.) A. DC. (Campanulaceae) is an ornamental plant growing wild in northern Asia, in China, Korea, Japan and east Siberia. For culture it needs light, medium moisture, organically rich, well-drained loams and full sun to part shade.Platycodi radix, the root of P. grandiflorum, has been used as food and in traditional oriental medicine to treat bronchitis, asthma and other pulmonary diseases. Triterpene saponins, called platycosides, are responsible for the medicinal value of this plant material. They have demonstrated a broad spectrum of therapeutic effects, such as antitumor [4][5][6][7], anti-inflammatory [8,9], hepatoprotective [10], antiatherosclerotic [11] and antinociceptive [12]. Additionally, these compounds were found to be a potent adjuvant of specific cellular and humoral immune responses, with the potential use in vaccine production [13][14][15]. Finally, saponins from Platycodi radix stimulated osteoblast differentiation via RUNX2, a principal osteogenic master gene for bone formation [16].Although the extensive pharmacological studies of these valuable compounds were undertaken, to date the sum of saponins, not the chosen, in hairy root cultures of P. grandiflorum was determined only by our research team [17]. Besides, this is the first report of the growth of P. grandiflorum transgenic roots cultured in a mist bioreactor and their yield of saponins. The method of Platycodi radix saponins determination published in "Chinese pharmacopeia" [18] exhibited as no suitable for quantitative analysis of saponins from P. grandiflorum transgenic roots. Therefore, the authors elaborated their own method [17].The compounds mentioned above are the main chemicals found in Platycodi radix. They are typical composed of oleaene backbones with two side chains: one is a 3-O-glucose linked by glycosidic bond, and the other is a 28-O-arabinoserhamnose-xylose-apiose linked by an ester bond [19]. To date, more than 55 triterpenoid saponins have been isolated from Platycodi radix [20]. Examples are platycodins A-I and polygalacins D and D2. According to WHO monographs AbstractThe growth and saponin accumulation were measured in two lines of transgenic hairy roots of Platycodon grandiflorum, Pl 6 and Pl 17, cultured for 8 weeks in 250-ml shake flasks containing 50 ml of hormone-free woody plant medium supplemented with 40 g/l sucrose and in the Pl 17 line cultured for 12 weeks in a 5-l mist bioreactor containing 1.5 l of the same medium. With both methods, the...

show abstract

Section: Measurement Of Root Growthmentioning

confidence: 99%

mentioning

confidence: 99%

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Indian Maid and Crysanthemunt mnorifoliurn cv. Indianapolis were performed using the technique and experimental conditions described previously (11), only this time with the root culture nutrient solution described by Street and Henshaw (20) (2).…”

Section: Methodsmentioning

confidence: 99%

Presence and Possible Mode of Action of a Proteinaceous Gonadotropin-like Growth Regulating Factor in Plant Systems

et al. 1969

View full text Add to dashboard Cite

Abstract. Antiserum to human chorionic gonadotropin (HCG) caused marked inhibition of adventitious rooting of Begonia semperflorens and Chrysanthemum morifolium stem cuttings. Immuno-absorption of crude protein extract from chrvsanthemum foliage through a column of polymerized and unsolubilized HCG antibodies resulted in a significant reduction in adventitious root promoting activity of the extract. These results are discussed in the light of a hypothesis that an endogenous protein growth regulating substance which immunologically resembles HCG exists in plant systems. Further experimentation with HCG suggests that its mode of action is possibly via the regulation of peroxidase enzymatic control of auxin levels.In previous studies Leshem (10) and Leshem and Lunenfeld (11) have shown that HCG, a proteinaceous human sex hormone, has growth stimulating effects on certain morphogenetic processes in plants, especially upon adventitious rooting of Vitis vinifera, Begonia semperflorens and Brassica oleracea. It was shown that HCG possibly acts via regulation of endogenous gibberellin levels, the latter being markedly repressed by the gonadotropin, and a hypothesis was suggested that HCG triggers mevalonic acid conversion to steroids thereby diminishing precursor for gibberellins.In the present study bv use of immunological techniques an attempt has been made to indicate the existence of an endogenous gonadotropin-like proteinaceous growth regulating factor in plants. Fur-ther experimentation was carried out in order to elucidate the mode of HCG action in plant systems, especially in the light of hormone-enzyme interactions which are prevalent both in plant (3,7,13,14,15,21) One ml of the antiserum neutralized 600 International Units (IU) of HCG as determined by the procedure of Loraine and Brown (12). As control active rabbit serum lacking HCG antibodies was tised.The approach used in the immuno-absorption trials was as follows: If a protein extract of plant material were to be passed through a medium conitaining HCG antibodies, the effluent would be depleted of any existent endogenous HCG or any immunologically similar antigenic substances which could "cross react" with the antibodies (1) and resultinglv evoke a weaker growth stimulating response. This system was obtained by insolubilizing HCG antibodies by covalent binding of the molecules to one another with ethyl-chloroformate using the technique described by Avramaes and Ternynck (2).The immunoabsorbent properties of such insolubilized proteins were found by these workers to be specific, stable and effective to isolate specific antigens from mixtures and to remove contaminating antigen proteins from solutions. The protein obtained in this manner was packed in a 50 cm X 3 cm 4 glass column through which protein extract was subsequently passed. In order to determine whether this procedure does actually work in the present trials, various concentrations of HCG solutions were run through test columns and resulting effluents were assayed for presence of HCG by immuno-...

show abstract

“…At 3 and 6 months, the index of sporophyte biomass growth (WP) was calculated for 30 plants and the calculation was repeated in duplicate. The WP value was calculated according to Street and Henshaw [43] where WP = final plant mass (g) − initial plant mass (g)/initial plant mass (g) × 100. The mean (n = 10) fresh mass of bulbils was 0.196 ±0.007 g. Statistical analysis was performed after 8 weeks with Graph Pad Prism version 6.0 (Graph Pad Software, San Diego, CA) using one-way analysis of variance (ANOVA) with Tukey-Kramer post-hoc test.…”

Section: Plant Materialsmentioning

confidence: 99%

A new and fast method to obtain in vitro cultures of Huperzia selago (Huperziaceae) sporophytes, a club moss which is a source of huperzine A

2013

View full text Add to dashboard Cite

This study presents a protocol for a fast and effective in vitro axenic culture of <em>Huperzia selago</em> (Huperziaceae Rothm.) sporophytes, a club moss which is a source of huperzine A, an alkaloid of a considerable therapeutic potential extensively investigated for its uses as treatment for some neurodegenerative diseases. The proposed procedure allowed approximately tenfold shortening of the species developmental stages with the omission of the gametophyte stage while the sporophyte mass could be increased tenfold within a 6-month period. The cultures were established using vegetative propagules (bulbils) procured from sporophytes growing in the wild without degrading the habitats of this endangered plant species. Explants underwent surface and internal disinfection to eliminate the epiphytic and endophytic bacteria and fungi. In in vitro cultures, the optimum results were achieved using Moore (Mr) medium without growth regulators or supplemented with 0.015 mg/l IBA and 0.3 mg/l kinetin. These media ensured both viability of the propagules and their further development. The biomass growth index for <em>H. selago</em> sporophytes grown from propagules, determined at 3 months of culture (1 passage) on Mr medium with IBA and kinetin was 650%. At 6 months, the biomass growth index increased to 1114%. Vigorous growth of adventitious roots, especially on Mr medium with the addition of 0.25 mg/l NAA, and callus formation on shoot apices were observed. At 6 months of culture, some sporophytes obtained from the bulbils were used as the initiating material for shoot subcultures, which developed best on Mr medium with IBA and kinetin.

show abstract

Introduction and Methods Employed in Plant Tissue Culture

Cited by 27 publications

References 183 publications

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

The growth and saponin production of Platycodon grandiflorum (Jacq.) A. DC. (Chinese bellflower) hairy roots cultures maintained in shake flasks and mist bioreactor

Presence and Possible Mode of Action of a Proteinaceous Gonadotropin-like Growth Regulating Factor in Plant Systems

A new and fast method to obtain in vitro cultures of Huperzia selago (Huperziaceae) sporophytes, a club moss which is a source of huperzine A

Contact Info

Product

Resources

About