The origin of plant cell and tissue culture can be found in a treatise published during the mid-18th century, entitled La Physique des Arbes, that describes the formation of callus tissue following the for mation of a ring of cortex from elm trees. Over the next two centuries, the discovery of plant growth hormones, in particular auxins and cytokinins, and detailed analyses on the nutritional requirements of plants, led to the formulation of media that could maintain actively dividing cultures derived from gymnosperms, and both dicotyledon ous and monocotyledonous angiosperms. However, much of the prog ress and technological development in the in vitro propagation of plant cells, tissues, and organs has occurred during the last 25 years. Recently, plant tissue culture techniques have been used as basic tools in the rapidly expanding field of plant biotechnology for the development and clonal propagation of new and/or improved plant varieties. Plant tissue culture is used for the micropropagation of commercially valuable cultivars that include ornamentals, oil palm, Glycyrrhiza, Pyrethrum, pine, Eucalyptus, sugar cane, and potatoes. Cultured plant tissue is also used for the selection of cells and, ul timately, the regeneration of plants that are tolerant to physical stresses such as pathogens, drought, and temperature extremes, and to chemical stress agents such as salinity, herbicides, proteins, and pyrethrins. In addition, new plants have been produced by the fusion of protoplasts prepared from cultured cells of different species in cluding sunflower and french bean, tomato and potato, and various cultivars of Datura. Finally, bacterial vectors and various mechanical methods have been used to introduce foreign genes into cultured plant tissues. Genetic transformation can result in profound changes in the phenotype and/or biochemical profile of the regenerated trans genic plants that are not characteristic of the wild type. An impressive variety of technologies in tissue culture, genetic manipulation, and molecular biology have been developed for nu merous plant species. Many of these techniques, sometimes referred to as plant biotechnology, have been extensively summarized and compiled in a well-balanced collection of easy-to-follow protocols presented in three separate, but complimentary, volumes. Plant Cell, Tissue and Organ Culture consists of 22 chapters (with 86 figures) and 5 appendices. The chapters cover critical aspects of (a) the es sential requirements for the operation of a plant tissue culture lab oratory; (b) the basic procedures of micropropagation, regeneration, and somatic embryogenesis; (c) some specific applications of organ culture systems such as embryo rescue and culture, and anther and microspore culture for haploid and double haploid production; (d) elementary transformation technology; and (e) useful microtechnique and analytical protocols specifically adapted to cultured tissues and cells. The appendices provide a convenient summary of media for mulations and commercial suppliers for the materials described in the text.

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Plant Cell, Tissue and Organ Culture

The role of activated charcoal in plant tissue culture.

Haploids are plants with a gametophytic chromosome number and doubled haploids are haploids that have undergone chromosome duplication. The production of haploids and doubled haploids (DHs) through gametic embryogenesis allows a single-step development of complete homozygous lines from heterozygous parents, shortening the time required to produce homozygous plants in comparison with the conventional breeding methods that employ several generations of selfing. The production of haploids and DHs provides a particularly attractive biotechnological tool, and the development of haploidy technology and protocols to produce homozygous plants has had a significant impact on agricultural systems. Nowadays, these biotechnologies represent an integral part of the breeding programmes of many agronomically important crops. There are several available methods to obtain haploids and DHs, of which in vitro anther or isolated microspore culture are the most effective and widely used. This review article deals with the current status of knowledge on the production of haploids and DHs through pollen embryogenesis and, in particular, anther culture.

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Anther culture for haploid and doubled haploid production

Micropropagation of orchids: A review on the potential of different explants

Chitinases of filamentous fungi: a large group of diverse proteins with multiple physiological functions

Micropropagation of the endangered terrestrial orchid, Geodorum densiflorum (Lam.) Schltr. was achieved through rhizome section culture from in vitro seed- derived rhizomes. Rhizome sections were cultured on Murashige and Skoog (MS) and Knudson C(KC) media supplemented with various growth regulators and 0.1% activated charcoal. The rhizome sections responded on MS medium. Naphthaleneacetic acid (NAA) at 2.0 μM stimulated rhizome growth. However, benzyladenine (BA) at 5.0 μM induced multiple shoots within four weeks of culture and inhibited rhizome growth. The regenerated shoots rooted on MS only or with NAA at 1.0 μM. Well-developed plantlets were transferred to community pots and then to a greenhouse where the plants have been acclimatised.

http://images.wikia.com/orchids/en/images/d/d0/In_vitro_propagation_of_the_endangered_orchid,_Geodorum_densiflorum_(Lam.)_Schltr._through_rhizome_section_culture.pdf

In vitro propagation of the endangered orchid, Geodorum densiflorum (Lam.) Schltr. through rhizome section culture

Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L.

Somatic embryogenesis and whole plant regeneration were achieved in callus cultures derived from hypocotyl, cotyledon and leaf explants excised from seedlings of Gymnema sylvestre. Embryogenic callus was induced on Murashige and Skoog (MS) medium containing 2,4-D (0.5–5.0 μM) +BA (0.5–2.0 μM) and 2% (w/v) sucrose in 6–8 weeks of culture. Globular/heart stage embryos developed on induction medium. These embryos produced torpedo and cotyledon stage embryos upon sub-culturing on embryo maturation medium EM8 (medium containing MS salts, B5 vitamins, 0.5 μM BA and 2% sucrose). Embryo germination and plantlet formation was achieved by sub-culturing mature embryos on fresh EM8 medium. The plantlets were acclimatized in the greenhouse.

Somatic embryogenesis and plant regeneration in Gymnema sylvestre

The effects of sugars (sucrose, maltose, glucose and fructose) and amino acids (glutamine, glycine, arginine, asparagine and cysteine) on embryogenesis and plantlet regeneration from cultured anthers of Cucumis sativus L. cv. Calypso and Green Long were studied. Type and concentration of sugar and amino acid influenced embryogenesis. Among the different sugars tested, sucrose was the best for embryo induction with an optimal concentration of 0.25 M. Maximum of 72 and 80 embryos per 60 anthers of Calypso and Green Long, respectively, were induced on embryo induction medium [B5 (Gamborg, Miller and Ojima (1968) Exp. Cell Res. 50: 151–158) supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D), 1.0 µM 6-benzyladenine (BA)] containing 0.25 M sucrose. The addition of amino acids to the embryo induction medium improved embryo yield with a combination of amino acids (glutamine, glycine, arginine, asparagine and cysteine of 1.0 mM each) giving the best response. Embryo differentiation was achieved on B5 medium supplemented with 0.25 µM of α-naphthaleneacetic acid (NAA), 0.25 µM kinetin (KN) and 0.09 M sucrose. Embryos were converted on B5 medium supplemented with abscisic acid (ABA) (10 µM) and 0.09 M sucrose. Embryos that developed on B5 medium supplemented with a combination of amino acids (glutamine, glycine, arginine, asparagine and cysteine of 1.0 mM each) exhibited the highest plantlet regeneration frequency.

Effect of sugars and amino acids on androgenesis of Cucumis sativus

The hypocotyl and internodal segments from in vitro grown seedlings of Feronia limonia (L.) Swingle (wood apple) were cultivated on Murashige and Skoog’s (1962, MS) medium supplemented with N6-benzyladenine (BA) or adenine (ADE) or kinetin (KN) at 0.5 to 5 µM. The optimum response was recorded on the medium containing 2 µM BA. An average of 12 and 8 shoots were developed from hypocotyl and internodal explants, respectively, after eight weeks of culture. The shoots were excised, and the residual explants were transferred to fresh medium where again they developed shoots. Up to three such passages resulted in the production of shoots from repeatedly subcultured explants and an average of 24 – 36 shoots per explant was obtained. The in vitro developed shoots produced roots when transferred to half strength MS medium supplemented with 1 µM 1-naphthaleneacetic acid (NAA). The developed plantlets were successfully transferred to mixture of soil, sand and coco-peat (1:1:1) and hardened in controlled environment. Hardened plants were transplanted to soil in greenhouse.

H. G. Ashok Kumar

Papers

In vitro propagation of the endangered orchid, Geodorum densiflorum (Lam.) Schltr. through rhizome section culture

Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L.

Somatic embryogenesis and plant regeneration in Gymnema sylvestre

Effect of sugars and amino acids on androgenesis of Cucumis sativus

In vitro culture of Feronia limonia (L.) Swingle from hypocotyl and internodal explants