Mass multiplication of protocorm-like bodies using bioreactor system and subsequent plant regeneration in Phalaenopsis
01 Jan 2000-Plant Cell Tissue and Organ Culture (Kluwer Academic Publishers)-Vol. 63, Iss: 1, pp 67-72
TL;DR: The feasibility of using PLBs for large-scale micropropagation was evaluated for scaled-up liquid cultures in bioreactors, rate of proliferation, and regeneration, and Hyponex medium was found to be suitable for conversion of PLBs into plantlets.
Abstract: Protocorm-like bodies (PLBs) formed on leaf segmentsin vitro were used as explants for bioreactor cultures. Continuous immersion cultures (air lift column and air lift-balloon bioreactor), and temporary immersion cultures (with or without charcoal filter attached) were used for the culture of PLB sections. A temporary immersion culture with charcoal filter attached was most suitable for PLB culture. About 18,000 PLBs were harvested from 20 g of inoculum (∼1000 PLB sections) in 2 l Hyponex medium after 8 weeks of incubation. Aeration in a bioreactor at 0.5 or 2.0 volume of air per volume of medium min−1 (vvm) yielded similar levels of biomass production. PLBs grown in bioreactors were cultured on solid Murashige and Skoog, Vacin and Went, Knudson C, Lindemann and Hyponex media. Hyponex medium was found to be suitable for conversion of PLBs into plantlets and 83% of PLBs transformed into plantlets on this medium. The feasibility of using PLBs for large-scale micropropagation was evaluated for scaled-up liquid cultures in bioreactors, rate of proliferation, and regeneration.
Citations
More filters
TL;DR: The effect of AC on growth regulator uptake is still unclear but some workers believe that AC may gradually release certain adsorbed products, such as nutrients and growth regulators which become available to plants.
403 citations
TL;DR: Photautotrophic micropropagation has the potential for development as a routine method for the in vitro conservation of endangered plants and the potential applications of cryopreservation are significant in this area.
Abstract: In vitro techniques have found increasing use in the conservation of threatened plants in recent years and this trend is likely to continue as more species face risk of extinction. The Micropropagation Unit at Royal Botanic Gardens, Kew, UK (RBG Kew) has an extensive collection of in vitro plants including many threatened species from throughout the world. The long history of the unit and the range of plants cultured have enabled considerable expertise to be amassed in identifying the problems and developing experimental strategies for propagation and conservation of threatened plants. While a large body of knowledge is available on the in vitro culture of plants, there are limited publications relating to threatened plant conservation. This review highlights the progress in in vitro culture and conservation of threatened plants in the past decade (1995–2005) and suggests future research directions. Works on non-threatened plants are also included wherever methods have applications in rare plant conservation. Recalcitrant plant materials collected from the wild or ex situ collections are difficult to grow in culture. Different methods of sterilization and other treatments to establish clean material for culture initiation are reviewed. Application of different culture methods for multiplication, and use of unconventional materials for rooting and transplantation are reviewed. As the available plant material for culture initiation is scarce and in many cases associated with inherent problems such as low viability and endogenous contamination, reliable protocols on multiplication, rooting, and storage methods are very important. In this context, photoautotrophic micropropagation has the potential for development as a routine method for the in vitro conservation of endangered plants. Long-term storage of material in culture is challenging and the potential applications of cryopreservation are significant in this area. Future conservation biotechnology research and its applications must be aimed at conserving highly threatened, mainly endemic, plants from conservation hotspots.
334 citations
Cites background from "Mass multiplication of protocorm-li..."
...Maize extract has been shown to induce high frequency plantlet regeneration from PLBs in Doritaenopsis sp. (Rahman et al., 2004)....
[...]
...PLBs have the advantage that they can be produced from almost any part of the plant and are usually formed directly on the tissue rather than through a callus stage....
[...]
...Progress made in orchids on production of PLBs in horticulturally important genera such as Dendrobium, Cymbidium, and Phalaenopsis have wider applicability to threatened taxa....
[...]
...Bioreactor technology may prove to be a useful tool for mass production of orchid PLBs (Young et al., 2000)....
[...]
...The majority of orchid micropropagation at RBG Kew is based on in vitro seed germination to maintain genetic diversity, with a few investigations producing protocorm-like bodies (PLBs) from somatic or meristematic regions to provide material for molecular studies....
[...]
TL;DR: This work endeavours to include the major investigations on explant-based orchid tissue culture starting from the pioneering works of Rotor and Wimber to date to date.
263 citations
TL;DR: Low cost and less labour-intensive clonal propagation through the use of modified air-lift, bubble column, bioreactors, together with temporary immersion systems for the propagation of shoots, bud-clusters and somatic embryos is described.
Abstract: Automation of micropropagation via organogenesis or somatic embryogenesis in a bioreactor has been advanced as a possible way of reducing costs. Micropropagation by conventional techniques is typically a labour-intensive means of clonal propagation. The paper describes lower cost and less labour-intensive clonal propagation through the use of modified air-lift, bubble column, bioreactors (a balloon-type bubble bioreactor), together with temporary immersion systems for the propagation of shoots, bud-clusters and somatic embryos. Propagation of Anoectochilus, apple, Chrysanthemum, garlic, ginseng, grape, Lilium, Phalaenopsis and potato is described. In this chapter, features of bioreactors and bioreactor process design specifically for automated mass propagation of several plant crops are described, and recent research aimed at maximizing automation of the bioreactor production process is highlighted.
233 citations
Cites background from "Mass multiplication of protocorm-li..."
...We have now reported the mass multiplication of Phalaenopsis in bioreactors (Figure 4a and b) (Park et al., 2000)....
[...]
TL;DR: In tuber-, bulb- and corm-producing plants, growth retardants and elevated sucrose concentrations in the media were found to enhance storage organ formation, providing a better propagule for transplanting or storage and better control of the contact of the plant tissue with the culture medium.
Abstract: Bioreactors provide a rapid and efficient plant propagation system for many agricultural and forestry species, utilizing liquid media to avoid intensive manual handling. Large-scale liquid cultures have been used for micropropagation through organogenesis or somatic embryogenesis pathways. Various types of bioreactors with gas-sparged mixing are suitable for the production of clusters of buds, meristems or protocorms. A simple glass bubble-column bioreactor for the proliferation of ornamental and vegetable crop species resulted in biomass increase of 3 to 6-fold in 3–4 weeks. An internal loop bioreactor was used for asparagus, celery and cucumber embryogenic cultures. However, as the biomass increased, the mixing and circulation were not optimal and growth was reduced. A disposable pre-sterilized plastic bioreactor (2–5-l volume) was used for the proliferation of meristematic clusters of several ornamental, vegetable and woody plant species. The plastic bioreactor induced minimal shearing and foaming, resulting in an increase in biomass as compared to the glass bubble-column bioreactor. A major issue related to the use of liquid media in bioreactors is hyperhydricity, that is, morphogenic malformation. Liquid cultures impose stress signals that are expressed in developmental aberrations. Submerged tissues exhibit oxidative stress, with elevated concentrations of reactive oxygen species associated with a change in antioxidant enzyme activity. These changes affect the anatomy and physiology of the plants and their survival. Malformation was controlled by adding growth retardants to decrease rapid proliferation. Growth retardants ancymidol or paclobutrazol reduced water uptake during cell proliferation, decreased vacuolation and intercellular spaces, shortened the stems and inhibited leaf expansion, inducing the formation of clusters. Using a two-stage bioreactor process, the medium was changed in the second stage to a medium lacking growth retardants to induce development of the meristematic clusters into buds or somatic embryos. Cluster biomass increased 10–15-fold during a period of 25–30 days depending on the species. Potato bud clusters cultured in 1.5 1 of medium in a 2-l capacity bioreactor, increased during 10–30 days. Poplar in vitro roots regenerated buds in the presence of thidiazuron (TDZ); the biomass increased 12-fold in 30 days. Bioreactor-regenerated clusters were separated with a manual cutter, producing small propagule units that formed shoots and initiated roots. Clusters of buds or meristematic nodules with reduced shoots, as well as arrested leaf growth, had less distortion and were optimal for automated cutting and dispensing. In tuber-, bulb- and corm-producing plants, growth retardants and elevated sucrose concentrations in the media were found to enhance storage organ formation, providing a better propagule for transplanting or storage. Bioreactor-cultures have several advantages compared with agar-based cultures, with a better control of the contact of the plant tissue with the culture medium, and optimal nutrient and growth regulator supply, as well as aeration and medium circulation, the filtration of the medium and the scaling-up of the cultures. Micropropagation in bioreactors for optimal plant production will depend on a better understanding of plant responses to signals from the microenvironment and on specific culture manipulation to control the morphogenesis of plants in liquid cultures.
131 citations
Cites background from "Mass multiplication of protocorm-li..."
...In Phalaenopsis, protocorm-like bodies were proliferated in bioreactors and regenerated on agar-based medium ( Young et al., 2000 )....
[...]
...Embryogenic expression was achieved, however, only after a short exposure to 2-isopentenyladenine (2iP) and further subculture to a growth regulator-free medium ( Lilien-Kipnis et al., 1994 )....
[...]
References
More filters
TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Abstract: In experiments with tobacco tissue cultured on White's modified medium (basal meditmi hi Tnhles 1 and 2) supplemenk'd with kiticthi and hidoleacctic acid, a slrikin^' fourlo (ive-told intTease iu yield was ohtaitu-d within a three to Tour week j^rowth period on addition of an aqtteotis exlrarl of tobacco leaves (Fi^'ures 1 and 2). Subse(iueutly it was found Ihiit this jnoniotiou oi' f^rowih was due mainly though nol entirely to inorj^auic rather than organic con.stitttenls in the extract. In the isolation of Rrowth factors from plant tissues and other sources inorj '̂anic salts are fre(|uently carried along with fhe organic fraclioits. When tissue cultures are used for bioassays, therefore, il is necessary lo lake into account increases in growth which may result from nutrient elements or other known constituents of the medium which may he present in the te.st materials. To minimize interference trom rontaminaitis of this type, an altempt has heen made to de\\eh)p a nieditmi with such adequate supplies of all re(iuired tnineral nutrients and cotntnott orgattic cottslitueitls that no apprecial»le change in growth rate or yield will result from the inlroduclion of additional amounts in the range ordinarily expected to be present in tnaterials to be assayed. As a point of referetice for this work some of the culture media in mc)st common current use will he cotisidered briefly. For ease of comparis4)n Iheir mineral compositions are listed in Tables 1 and 2. White's nutrient .solution, designed originally for excised root cultures, was based on Uspeuski and Uspetiskaia's medium for algae and Trelease and Trelease's micronutrieni solution. This medium also was employed successfully in the original cttltivation of callus from the tobacco Iiybrid Nicotiana gtauca x A', tanijadorffii, atitl as further modified by White in 194̂ ^ and by others it has been used for the
63,098 citations
01 Jan 1962
16,251 citations
"Mass multiplication of protocorm-li..." refers methods in this paper
...They were cultured on Murashige and Skoog (1962) solid medium supplemented with sucrose (45 g l−1) and BA (3 mg l−1)....
[...]
10 May 1946
485 citations
"Mass multiplication of protocorm-li..." refers methods in this paper
...PLBs which were harvested from bioreactor cultures were inoculated individually on MS, VW (Vacin and Went, 1949), KC (Knudson, 1946), LM (Lindemann et al....
[...]
...Plantlet regeneration from PLBs PLBs which were harvested from bioreactor cultures were inoculated individually on MS, VW (Vacin and Went, 1949), KC (Knudson, 1946), LM (Lindemann et al., 1970) and Hyponex (6.5N – 4.5P – 19K 1 g l−1 + 20N – 20P – 20K 1 g l−1) media....
[...]
TL;DR: An investigation indicated that amino acids are excellent buffers and a new nutrient solution is proposed, based on the results of studies on weakly buffered culture media used for asymbiotic germination of orchid seeds.
Abstract: 1. The culture media used generally for the asymbiotic germination of orchid seeds are so weakly buffered that, as the seedlings start to grow, the pH value drops below the level for optimum growth. Even when the phosphate buffer was increased twenty-four fold, the buffering action was still weak in the optimal growth range of pH 4.5-5.5, indicating that phosphate buffers are unsatisfactory. An investigation indicated that amino acids are excellent buffers. This work is being continued to determine what amino acids will be satisfactory not only as buffers but also as growth factors. 2. Autoclaving the culture media without agar produced significant changes in pH value. Similar changes in pH value were observed, only at a slower rate, when the solutions stood for some time. These results are equally applicable to ordinary nonsterilized culture media. The changes when the original pH value was 3 to 6 were due to the presence of iron sulfate, which should be replaced by one of the organic iron salts; when it...
465 citations
"Mass multiplication of protocorm-li..." refers methods in this paper
...Plantlet regeneration from PLBs PLBs which were harvested from bioreactor cultures were inoculated individually on MS, VW (Vacin and Went, 1949), KC (Knudson, 1946), LM (Lindemann et al., 1970) and Hyponex (6.5N – 4.5P – 19K 1 g l−1 + 20N – 20P – 20K 1 g l−1) media....
[...]
Book•
01 Jan 1993
TL;DR: This book discusses the origins of Orchid Micropropagation, its applications, and some of the sources of Supplies, Equipment, Terms, and Reagents, as well as methods for specific Genera for Specific Genera.
Abstract: History General Outline of Techniques and Procedures Methods for Specific Genera Summary References Indexes.
444 citations
"Mass multiplication of protocorm-li..." refers background in this paper
...Researchers (see Arditti and Ernst, 1993), tested many antioxidants like polyphenoloxidase inhibitors, polyvinylpyrrolidone, and activated charcoal to overcome phenolic compound accumulation....
[...]