Tissue culture company begins selling plants directly to growers

When Yongjian Chang built North American Plants in 1998, the company had 800 square feet of lab space to propagate plants through a process known as tissue culture — essentially cloning them to meet nursery demand for ornamental trees and shrubs.

It’s come a long way in the years since, with five expansions bringing lab space to some 23,000 square feet and a switch in 2006 to focus on berries and rootstocks for tree fruit and nuts.

Today, North American Plants produces 3 million blueberry, blackberry and raspberry plants and 10 million rootstocks for tree fruit and nuts annually.

Increasing demand for disease-resistant rootstocks, particularly in the apple industry, has the company poised for another change: selling directly to

In vitro Propagation of Critically Endangered Endemic Rhaponticoides mykalea by Axillary Shoot Proliferation

Turkey is one of the richest countries in variability of flora. It has nearly 9000 plant species about 3000 of which are endemic [1]. Asteraceae, is represented by 50 species in Turkey with an endemism of nearly 54% [2]. Rhaponticoides mykalea (Hub.-Mor.) M.V. Agab. & Greuter which belongs to the Asteraceae family, falls within the CR (Critically Endangered) category in the Red Data Book of Turkey [1]. While R. mykalea (Hub.-Mor.) was classified under the section Centaurea as Centaurea mykalea (Hub.-Mor.) before now. Today it has been separated from the section Centaurea [3]. It spreads very scarce in Kuşadası (Aydın), Muğla and Isparta, and faces with the danger of extinction. R. mykalea that has very limited number of individuals is under strong anthropogenic pressure such as the gradually increase in ongoing urbanization due to rapid developments of tourism sector, the conversion of natural habitats into human dominated lands, the over-grazing and collecting capitula of R. mykalea by local people for food. The species has already been under the threat of extinction and the situation above will increase the risk of extinction of this species even more [4]. For this reason, local protection measures and global conservation strategies are necessary [5].

Nowadays, the conservation of wild plant genetic resources is very important for preventing a decrease in genetic variability. Conservation of the endemic or threatened plants is carried out using different strategies. In vitro culture is an

In vitro Regeneration, Acclimatization and Antimicrobial Studies of Selected Ornamental Plants

Tissue culture has been applied to diverse research techniques such as viral elimination, clonal propagation, gene conservation, in vitro fertilization, mutation, induction for genetic diversity, genetic transformation, protoplast isolation and somatic hybridization, secondary metabolite production and other related techniques. The commercial production of ornamental plants is growing worldwide. Its monetary value has significantly increased over the last two decades and there is a great potential for continued further growth in both domestic and international markets. About 156 ornamental genera are propagated through tissue culture in different commercial laboratories worldwide. About 212.5 million plants including 157 million ornamental plants amounting to 78% of the total production were reported [1]. These plants are over exploited due to their high medicinal value and hence, propagation of the plants by tissue culture may be mandatory, which offers a greater potential to deliver large quantities of disease-free, true-to-type healthy stock within a short span of time. Biotechnological interventions for in vitro regeneration, mass micropropagation and gene transfer methods in forest tree species have been practiced with success, especially in the last decade. Against the background of the limitations of long juvenile phases and lifespan, developments of plant regeneration protocols of ornamental species are gaining importance. Ornamental industry has applied immensely in vitro propagation approach for large-scale plant multiplication of elite superior varieties. During in vitro condition, plantlets are grown under fixed and controlled environment in sterile formulated medium which contained macronutrients, micronutrients, vitamins and plant growth regulators. After the plantlets reached optimum growth in the culture containers after

Micropropagation of Anthurium spp.

Micropropagation as an alternative method to conventional propagation, the culture of somatic cells, tissues and organs of plants under controlled conditions is a suitable way to produce a large number of progeny plants which are genetically identical to the stock plant in a short time. The important property of the plant cells is totipotency which is a capacity to produce the whole plant from different plant parts. Micropropagation has some features to be chosen in commercial production such as multiplicative capacity in a relatively short time, healthy and disease-free production capacity and ability to generate population during a year [1-5].

The genetic pattern of the plant is key element to select the propagation method. Using micropropagation techniques in plant biotechnology applications are costlier than conventional propagation methods. Propagation by using in vitro techniques instead of conventional methods offer some advantages like utilizing small pieces of plants called as explants to maintain the whole plant and increase their number. The main point is to evolve new strategies to

Aseptic Techniques in Plant Tissue Culture

Aseptic techniques in plant tissue culture from kumarkanika

Production of Useful Secondary Metabolites Through Regulation of Biosynthetic Pathway in Cell and Tissue Suspension Culture of Medicinal Plants

Medicinal herbs played important roles in human history, from ancient times to now. They have been used for thousands of years to cure diseases, colorize clothes, adjust food taste and keep healthy. It was recorded that, 61% of currently used small molecular drugs are derived from or inspired by natural products from medicinal herbs [1]. However, to cure disease, we need to harvest medicinal herbs, and use part of their tissue for extraction, such as root, leaves, seeds, flowers and so on. That will directly cause problems for the reproduction of these herbs. It has also been noted that the natural habitats have been destroyed due to human activities. With the modern city under construction, the enlarging need for natural resources and the serious pollution, the environment has been never so tough for the growth of medicinal herbs. In another word, the naturally growing medicinal herbs can’t fulfill the need of increasing market. Furthermore, the naturally grown medicinal herbs are also different from before, they are carrying more and more herbicide, insecticide and heavy metals, which will cause contamination to the extract and finally cause side effects. Besides, due to the complex structures of secondary metabolites, the chemical synthesis is proved to be cost-inefficient in most cases. Thus, how to produce enough medicinal herbal material in an appropriate manner becomes more and more urgent for the development of pharmaceutical industry all over the world.

In 1934, White proposed the theory of totipotency, and Steward proved the theory in 1952~1953 using carrot cells cultured in liquid media to regenerate whole plant. From then, the cell and tissue culture techniques are developed. As an alternative choice to produce active secondary metabolites, cell and tissue culture of medicinal herbs has obvious advantages:

1. The culture system doesn’t need much field which can be used for crop growing;
2. The system is not limited by whether and season changes.
3. The secondary metabolism can be

Plant tissue culture Medium

Plant tissue culture medium from Rinkesh Joshi

Effect of Additives on Micropropagation of an Endangered Medicinal Tree Oroxylum indicum L. Vent

Sonpatha (Oroxylum indicum (L.) Vent.) is a threatened medicinal tree species [1,2] belonging to family Bignoniaceae. It is valued for its antimicrobial, antiarthritic, anticancerous and antihepatitic properties possessed by its various parts. Root extract of this tree has been used for long in ayurvedic preparations like Dashmularisht and Chyawanprash [3,4].This tree possesses a flavonoid viz. Baicalein used to check proliferation of human breast cancer cell line MDA - MB - 435 [5]. Sonpatha grows in India, Sri Lanka, South China, Celebes, Philippines and Malaysia[6,7]. In India, it is distributed throughout the country up to an altitude of 1200 m and found mainly in ravine and moist places in the forests [8].Owing to indiscriminate collection, over exploitation and uprooting of whole plants with roots, this valuable tree has become vulnerable in different states of India like Karnataka, Andhra Pradesh, Kerala, Maharastra, M.P. and Chhatisgarh [9,10]. Hence research towards mass multiplication, conservation and higher production of the active compound under in vitro culture conditions is essential [11]. Few reports are available on the in vitro regeneration of the species [12,13]. Optimum factors influencing growth and morphogenesis vary with the genotype and types of explants used for micropropagation. [14]. Murashige and Skoog (MS) medium with a high content of nitrate, ammonium and potassium is of widespread use in the successful culture of a wide variety of plants. Sometimes it requires supplementation of additional substances in the medium.

Application of additives is adapted to the cultural needs[15] i.e. objectives of the experimental studies like micropropagation, regeneration, cytodifferentiation, androgenesis, biosynthesis of secondary metabolites and biotransformation of cells as well as

Polyamines, Gelling Agents in Tissue Culture, Micropropagation of Medicinal Plants and Bioreactors

Currently, tissue cultures of species of agricultural importance have wide applicability in industrial production processes. Tissue culture is a name given to a set of techniques that allow the regeneration of cells, tissues and organs of plants, from segments of plant organs or tissues, using nutrient solutions in aseptic and controlled environment. This regeneration is based on the totipotency of plant cells. Totipotency is a capability indicating that plant cells, in different times, may express the potential to form a new multicellular individual. Tissue culture appears to be a good alternative to conventional propagation, requiring less physical space, with high multiplication rate, without incidence of pests and diseases during cultivation, and enabling higher control of the variables involved. Thus, in the in vitro environment, with the required stimuli and appropriate conditions, different cell types express different behaviors, possibly leading to cell multiplication and differentiation into a specific tissue, characterized by a form and a function, which may lead to the regeneration of a new individual.

The discovery of this feature in plant cells is indistinguishable from the first studies on tissue culture in the early twentieth century by Heberlandt in 1902, which were followed by

Tissue Culture Techniques for Native Amazonian Fruit Trees

The fruits of the Amazon have attracted great interest in recent years, both nationally and internationally, according to its exotic flavors and pleasant and varied ways to use its pulp by agribusiness [1], pharmaceutical industry [2], high vitamin and antioxidant content[3].

In recent decades, the production of native fruits of the Amazon showed significant growth, mainly due to expansion of area for fruit production. It is noteworthy that this activity has had little impact on native vegetation, since most of the orchards were planted in areas previously occupied by other crops for market problems or environmental issues and pressure for sustainable agriculture, ceased to be interesting for farmers [4].

The Amazon forest has large number of non-domesticated fruit species and a minority being exploited through crop in place of natural occurrence [5]. According to the

Detection and control of bacterial contaminants of plant tissue cultures.

Preventing or avoiding microbial contamination of plant tissue cultures is critical to
successful micro propagation. Epiphytic and endophytic organisms can cause severe
losses to micro propagated plants at each stage of growth (Cassells. 1991; Debergh arxi
Vanderschaeghe. 1988; Leifert eot at. 1991). Bacterial contaminants are often diffiCult to
detect because they remain mostly within the plant tissue (D:ebergh and Vanderschaeghe.
1988; De Fossard and De Fossard. 1988; Viss eot al.. 1991). Contaminated plants may
have no visible symptoms. reduced multiplication and rooting rates. or may die (Leifert
et ai., 1989; 1992). Introduction of microorganisms due to poor aseptic technique or
improperly sterilized equipment can be corrected with improvements in training or
equipment handling. but eliminating internal contaminants is more problematic (Buckley et al.. 1995).

Procedures for producing aseptic cultures require attention to some or all of the
following: 1) indexing explants and cultures for

Tissue Cultured Versus Traditionally Grown Pineapples

Ananas comosus, commonly known as pineapple, is an herbaceous perennial belonging to the order Bromeliales, family Bromelaceace and sub-family Bromelioideae [1-3]. According to Carlier et al. [4] there are 56 genera of the pineapple which include 2921 species. According to Gene Technology Regulation [5] and Carr [6], pineapples originated in South America and through travels and migration of different peoples, its cultivation spread to other parts of the world. The Jamaican pineapple is said to have been discovered by Christopher Columbus on his voyage to the New World in the 15th century.

Bartholomew et al. [1], in describing the pineapple, states that the plant grows to 1-2 m high and wide with a club shaped stem having dimensions of 25-50 cm in length, width of 2-5 cm at the base and 5-8 cm at the top. The leaves are sessile and enclose the stem on two thirds of its circumference. They tend to be sword like with sharply dentate edges often variegated or

Plant Tissue Culture Technique and its applications

Plant Tissue Culture Technique and its applications from Komal Jalan

Caulogenic response of in vitro raised nodal explants of Orthosiphon stamineus to selected auxins

Auxins play an important role in stem initiation and elongation which involves in different features of growth and developments in higher plants. The auxins 2, 4-D are strong promoters of callus induction and growth of cell suspensions. There are only a few examples of the shoot and root induction by phenoxy auxins in tree tissue cultures (Zaerr and Mapes, 1982). For shoot induction generally requires the combination of auxin and cytokinin. However, auxin should be used carefully since too much auxin favors callus growth. Moreover, for shoot initiation in some explants, the production of endogenous auxin is sufficient for induction of shoots in larix decidue (Bondga and Von-Aderkas, 1992). The number of shoots induced on MS medium (Murashige and Skoog, 1962). The auxin signal is predictable by plant cells and rapidly converted to a wide variety of responses in the growth and development of plant organs. These comprise alters in the direction of growth, shoot and root branching, and vascular differentiation (Leyser, 2001).

The plant cell division and growth of tissue, cells cultured in vitro require an external source of auxin (Petrasek et al., 2002). The proportion of external to

Characterization of the Embryogenic Tissue of the Norway Spruce Including a Transition Layer between the Tissue and the Culture Medium by Magnetic Resonance Imaging

Magnetic resonance imaging (MRI) is a non-invasive method widely applied in the study of molecules. The MRI approach is frequently employed not only in medicine but also in biological, biochemical, and chemical research. Most of the papers describing the use of MRI to investigate plant physiology have emphasized certain aspects of anatomy or functional morphology [1]. In plant biology, MRI supports several major activities, namely, the research of the water and mineral compounds transported within a plant [2], [3], the determination of plant metabolites [4], [5], the investigation of cellular processes [6], and the examination of the growth and development of plants [7]. MRI is also instrumental towards monitoring water changes in early somatic embryos (ESEs). Šupálková et al. [8] examine the influence of the ESEs of spruce treated with cadmium and/or lead ions for twelve days using multi-instrumental analytical procedures; these authors employ image analysis to estimate the growth, a fluorimetric sensor to detect the viability of enzyme-treated ESEs, and the MRI technique to facilitate non-destructive measurement of the volume of the ESEs.

 The interaction between the cells plays a fundamental role in the growth and development of multicellular organisms. In this context, let us note that

Somaclonal Variation in Tissue Culture: A Case Study with Olive

Micro propagation of woody plants and fruit crops constitutes a major success in the commercial application of in vitro cultures. An important aspect to be considered when deriving perennial plants from micro propagation is the maintenance of genetic integrity with regard to the mother plant. In this regard, soma clonal variation has been reported at different levels (morphological, cytological, cytochemical, biochemical, and molecular) in micro propagated plants [1]. The economic consequence of soma clonal variation among regenerated plants is enormous in fruit crops and woody plants, because they have long life cycles. In consequence, the behavior of micro propagated plants should be assessed after their long juvenile stage in field conditions. The occurrence of soma clonal variation is a matter of great concern for any micro propagation system. In order to evaluate its presence several strategies were used to detect soma clonal variants, based on one or more determinants from among morphological traits, cytogenetic analysis (numerical and structural variation in the chromosomes), and molecular and biochemical markers [2]. In addition, studies on soma clonal variation are important for its control and possible suppression with the aim of producing genetically identical plants, and for its use as a tool to produce genetic variability, which will enable breeders the genetic improvement. Soma clonal variation has been studied extensively in herbaceous plants, whereas few studies have focused on

Flow Cytometry Applied in Tissue Culture

Flow cytometry is a powerful technology that allows for the simultaneous analysis of multiple attributes of cells or particles in a liquid medium. The first cytometer used was built during World War II, when [1] developed an equipment where particles flowed through the system to diffuse light through a lens, producing electrical signals sensed by a photodetector. The instrument could detect objects in the order of ~ 0.5 µm in diameter, and is recognized as the first flow cytometer used for observation of biological cells [2]. This would be possible to identify aerosols, bacteria that would possibly biological warfare agents as well as check the efficiency of gas mask filters against particles. In 1950, the same principle was applied to the detection and enumeration of blood cells. As hematology and cellular immunology, two biological areas, that drove the development of flow cytometry [3]. Later, with improved equipment and methods, this technique was adapted to other areas of biology, including the plant kingdom [4]. Already in 1973 the German botanist Friedrich Otto Heller used the Impulszytophotometrie (pulse cytophotometry in German). This scientist did not imagine that it has launched a new field of scientific research, which would later be called flow cytometry in plants.

In reference to [5] that developed a rapid and convenient method for the isolation of plant nuclei by cutting the same tissue in a lysis buffer consisting of a buffer to destroy the

Modeling the Effects of Light and Sucrose on In Vitro Propagated Plants: A Multiscale System Analysis Using Artificial Intelligence Technology

Since the beginning of in vitro culture in 1902 when the Austrian botanist Gottlieb Haberlandt attempted to grow isolated plant cells and tissues (leaf mesophyll and hair cells) in nutritive solutions, a large body of work has emerged describing the optimization of different culture conditions to supply explants with all the components required for successful in vitro plant tissue propagation. During the past 70–80 years, more than 3000 scientific articles have described the use of over 2000 different culture media in plant tissue culture [1]. In vitro tissue propagation, however, is still a stressful procedure for plants, which can limit the successful establishment of plants upon transfer to ex vitro conditions [2]–[5]. In many cases, the best in vitro conditions do not lead to optimal ex vitro results. Therefore, a better understanding of the complex effects of the variables involved during the in vitro plant tissue growth on the in vitro culture and the ex vitro acclimatization results should lead to an improvement of the process. The effect of carbon in the media, light conditions and their interaction appear to be particularly important [6]–[8].

Sucrose is the most common carbon source used in plant cell, tissue and organ culture. Media with 3% sucrose have been the staple since Murashige and Skoog [9] described their MS medium. Sucrose acts during plant tissue culture as a fuel source for sustaining photomixotrophic metabolism, ensuring optimal development, although other important roles such as

Effects of Red and Blue LED on the in vitro Growth of Rosa Kordesii in Multiplication Phase

Light source such as fluorescent lamps, metal halide lamps and high-pressure sodium lamps are generally used for in vitro cultures. However, these lights contains unnecessary wavelength that are low in quality for stimulating growth [8] [9]. Furthermore, the use of these lights consumed a lot of electricity and it was reported that tissue cultured lab consumed 65% of total electricity [10]. Light Emitting Diode also known as LED light has been utilized globally in agriculture as an alternative light source for plant growth and photosynthesis. LEDs have attracted considerable interest because of their wavelength specificity and narrow bandwidth, small mass and volume, long-life and minimum heating [11][12].

Morphology and physiology of in vitro grown plants are regulated by various micro-environmental
factors such as light, temperature, humidity and carbon dioxide [12][13]. Inadequate or insufficient light may cause harmful for plant growth or led to

In vitro Tissue Culture, a Tool for the Study and Breeding of Plants Subjected to Abiotic Stress Conditions

Abiotic stress factors are the main limitation to plant growth and yield in agriculture. Among them, drought stress caused by water deficit, is probably the most impacting adverse condition and the most widely encountered by plants, not only in crop fields but also in wild environments. According to published statistics, the percentage of drought-affected land area in the world in 2000 was double that of 1970 [1].

Another major environmental factor that limits crop productivity, mainly in arid and semi-arid regions is high salinity. Approximately 19.5% of the irrigated soils in the world have elevated concentrations of salts either in the soil or in the irrigation water [2], damaging both the economy and the environment [3, 4]. The deleterious effects of salinity on plant growth are associated with low osmotic potential of soil solution (water stress), nutritional imbalance, specific ion effect (salt stress), or a combination of these factors [5].

Abiotic stress leads to a series of

The Prerequisite of the Success in Plant Tissue Culture: High Frequency Shoot Regeneration

Plant tissue culture is a term containing techniques used to propagate plants vegetatively by using small parts of living tissues (explants) on artificial growth mediums under sterile conditions. Explants regenerate shoots and roots, and consequently whole fertile plants under certain cultural conditions. Micropropagation is the production of whole plants through tissue culture from small parts such as shoot and root tips, leaf tissues, anthers, nodes, meristems and embryos. Micropropagation is the vegetative (asexual) propagation of plants under in vitro conditions and is widely used for commercial purposes worldwide [1-3].

Plant tissue culture techniques have certain advantages over traditional ones of propagation. These are:

• Thousands of mature plants can be produced in a short time that allows fast propagation of new cultivars,
• Endangered species can be cloned safely,
• Large quantities of genetically identical plants can

The Science of Plant Tissue Culture as a Catalyst for Agricultural and Industrial Development in an Emerging Economy

In the last few decades, the flow of biological discovery has swelled from a trickle into a torrent, driven by a number of new methodologies developed in plant tissue culture, recombinant DNA technology, monoclonal antibodies and micro chemical instrumentation [1]Biological research has been transformed from a collection of single discipline endeavors into an interactive science with bridges between numbers of traditional disciplines. This synergy has made biology the “sunrise field” of the new millennium. The whole gamut of new discoveries in biology and allied sciences can be grouped together under a single umbrella term of “Biotechnology”.

Biotechnology has been defined as “any technique that uses living organisms, or substances from these organisms, to make or modify a product, to improve plants or animals, or to develop microorganisms for specific uses” [2]. No society has advanced without deploying appropriate technology in place to set the pace for addressing its major problems. Public investment in relevant technology, the application to industries and capturing of the benefit accrue to it is what sets developed nations apart. Previous reports have shown that there is no National economic growth without proper investment in a right technology which is applied in a Nation. Real solutions to priority on national problems like job creation and poverty alleviation is investment in appropriate technology. This is evident in countries that embraced and adopted biotechnology in past technological revolutions and are practicing on an unprecedented scale. Such countries like

Plant Tissue Culture Media

Optimal growth and morphogenesis of tissues may vary for different plants according to their nutritional requirements. Moreover, tissues from different parts of plants may also have different requirements for satisfactory growth [1]. Tissue culture media were first developed from nutrient solutions used for culturing whole plants e.g. root culture medium of White and callus culture medium of Gautheret. White’s medium was based on Uspenski and Uspenska’s medium for algae, Gautheret’s medium was based on Knop’s salt solution [2]. Basic media that are frequently used include Murashige and Skoog (MS) medium [1], Linsmaier and Skoog (LS) medium [3], Gamborg (B5) medium [4] and Nitsch and Nitsch (NN) medium [5]

Media Composition

Plant Tissue Culture: Current Status and Opportunities

Tissue culture is the in vitro aseptic culture of cells, tissues, organs or whole plant under controlled nutritional and environmental conditions [1] often to produce the clones of plants. The resultant clones are true-to type of the selected genotype. The controlled conditions provide the culture an environment conducive for their growth and multiplication. These conditions include proper supply of nutrients, pH medium, adequate temperature and proper gaseous and liquid environment.

Plant tissue culture technology is being widely used for large scale plant multiplication. Apart from their use as a tool of research, plant tissue culture techniques have in recent years, become of major industrial importance in the area of plant propagation, disease elimination, plant improvement and production of secondary metabolites.Small pieces of

Regeneration of in Vitro Shoot and Root Structure through Hormone Manipulation of Coconut Zygotic Embryos

Zygotic embryos tissues have found to be more responsive explants for clonal propagation of coconut. In the present study, the feasibility of using zygotic embryos explants for clonal propagation of a local coconut variety MATAG F2 was assessed. Callus was induced by incorporating of cytokinin and auxin into the medium. The sliced embryos explants were immersed in 1 M maltose for 60 mins, then with 0.05 M maltose for 1 min and followed by 0.01 M maltose for 5 mins was the best for prevention of phenolic compounds excretion. The callus formation depended on the concentration of 2,4-D in the media and the best effect was observed with the high level (2,4-D and BAP) tested. Attempts at inducing multiple shoot from the zygotic embryos callus were unsuccessful. No multiple shoots was present; most of the callus became root structure.

In vitro propagation of a medicinal important plant Bacopa monnieri from nodal explants

Bacopa monnieri, a traditional Indian medicinal plant with high commercial
importance, is used as a neurotonic, immuno modulator, adaptogen transquilizing, memory
and learning enhancing, cerebral activator, anti-ulcer, antispasmodic, anti-asthmatic
ayurvedic herb. The study was undertaken in order to elaborate the tissue culture technique
for Brahmi (Bacopa monnieri). In the present study nodal explants were taken for in vitro
propagation. For sterilization, 0.01% HgCl2 was used as the sterilant. The explants gave best
results when treated with the sterilant for 5 minutes. To induce bud-break, nodal segments
were inoculated onto MS media supplemented with 1mg/l BAP. Basal MS medium without
any plant growth hormone was used as control. Further multiplication was obtained on MS
medium + BAP (1.0 mg/l) +IAA (0.5 mg/l). The shoots were rooted and best rooting was
observed by IBA when incorporated in MS at different concentrations (0.1 - 0.3 mg/l) and
0.2 mg/l gave good results. After 30 days about 2.2 cm length was observed. Almost 70%
of the rooted shoots survived during hardening.

In vitro callus induction and isolation of volatile compounds in callus culture of Lallemantia iberica

A modern biotechnological technique to obtain useful natural products from plants is to isolate them from their callus cultures. Lallemantia iberica is an annual herb of the Lamiaceae family known for its stimulant, diuretic, and expectorant effects in Iranian folk medicine. The present study investigated the induction of plant callus tissue and identification of its volatile compounds. For this purpose, plant seeds of Lallemantia iberica were sterilized and cultured in petri dishes lined with an MS medium. After the emergence of seedlings, cotyledon segments were transferred to another MS medium supplemented with different combinations of the plant hormones BAP and 2,4-D. The petri dishes were incubated in a growth chamber at 25 °C for a given photoperiod. The fresh weights of the calli thus produced in the hormonal treatments were measured. In a second stage of the study, the essential oil of the fresh calli was obtained using a Clevenger type apparatus and subjected to analysis by gas chromatography-mass spectrometry (GC-MS). Results showed that callus induction from the otyledon
segments of the seedlings was better accomplished in the MS medium containing phytohormones and in a dose-dependent manner. Maximum callus production was induced in the MS medium supplemented with 2, 4-D (4 mg/L) and BAP (1.5 mg/L) as 3.5g. GC/MS analysis showed that the dominant compounds in the essential oil were Thymol (53.03%), Octane (19.90%), Decane (5.73% ), Carvacrol (5.63%), and Octadecane (3.73%).

Tissue Culture of Gerbera

Gerbera (Gerbera jamesonii) somatic tissues and seeds were tried for raising tissue cultures. The explants of shoot tips, immature inflorescences, leaf sections, capitulum explants, axillary buds and receptacles explants from field grown plants had contamination problems. Another trouble was slow growth of explant cultures as they were treated with sterilizing chemicals which damage their growing regions. However, callusing was in all explants and shoot regenerations were obtained from shoot tip and axillary buds. The calluses were obtained from clean seeds which were pre-soaked (20mg/liter for six hours) in colchicine for induction of polyploidy. The germinating seedlings were crushed and tissue mass was put on callusing medium containing MS with BA+2,4-D (each 3 mg/liter). After six weeks, the calli were transferred to MS+BA (4mg/liter) +IBA (1mg/liter). The developed plantlets were separated and cultured on MS containing BA and IBA (each @ 0.5mg/liter). The plantlets were transferred to pots under 100% humidity during initial weaning period, acclimatized and grown as normal plants.

A Low Cost Novel Medium for Plant Tissue Culture

Plants with its innumerable benefits have been exploited by mankind for their own well being. Furthermore rapid increase in population has led to an increase in food and energy demand many folds. In comparison to the developed countries food situation in most of the developing countries is precarious. To meet the food demands, production has to be increased and the increased production has to be obtained from a declining cultivable land area.

Plant tissue culture (PTC) holds the key to accelerated food production and medicines by continuous production and supply of plantlets of desired variety/genotype and thereby reducing the use of land area. Micropropagation technology via plant tissue culture provides better approach to raise clones of plants in a large number in which rapid proliferation is achieved and has been widely applied for the production of a large number of economically important plants including valuable medicinal plants, trees, staple food crops, horticultural plants.

However commercialization of such technologies has been

The Many Dimensions of Plant Tissue Culture Research

The practice of plant tissue culture has changed the way some nurserymen approach plant propagation. In the recent past, the applicability of this technology to the propagation of trees and shrubs has been documented. Some firms have established tissue culture facilities and commercial scale operations are presently in operation for the mass propagation of apples, crabapples, rhododendrons, and a few other selected woody species. The intent of this research update is to briefly examine "what is being done" and to explore "what can be done" with regard to the tissue culture of ornamental plants. Such a consideration necessarily includes an overview of tissue culture as a propagation tool. The major impact of plant tissue culture will not be felt in the area of micropropagation, however, but in the area of controlled manipulations of plants at the cellular level in ways which have not been possible prior to the introduction of tissue culture techniques.

THE ART AND SCIENCE OF MICROPROPAGATION

Of all the terms which have been applied to

Somaclonal Variations

Somaclonal variation from Rakesh Kumar

Oxidative Stress Studies in Plant Tissue Culture

Higher plants are sessile therefore are continuously exposed to different environmental stress factors, such as drought, salinity, heavy metals, nutritional disorders, radiation without any protection. Most of these stresses produce certain common effects on plants, like induced oxidative stress by overproduction of reactive oxygen species (ROS), besides their own specific effects (Rao, 2006). Thus, plants have developed their own specific response(s)against each of these stresses as well as cross-stress response(s). Investigating these responses is difficult under field conditions, but plant tissue culture techniques are performed under aseptic and controlled environmental conditions. These advantages of plant tissue culture allow various opportunities for researcher to study the unique and complex responses of plants against environmental stresses (Sakthivelu et al., 2008, Lokhande et al., 2011).

History of Plant Tissue Culture

The theoretical basis for plant tissue culture was proposed by Gottlieb Haberlandt in his address to the German Academy of Science in 1902 on his experiments on the culture of single cells (2). He opioned that, to my knowledge, no systematically organized attempts to culture isolated vegetative cells from higher plants have been made. Yet the results of such culture experiments should give some interesting insight to the properties and potentialities that the cell, as an elementary organism, possesses. Moreover, it would provide information about the interrelationships and complementary influences to which cells within a multi cellular whole organism are exposed (from the English translation, [3]). He experimented with isolated photosynthetic leaf cells and other functionally differentiated cells and was unsuccessful, but nevertheless he predicted that one could successfully cultivate artificial embryos from vegetative cells. He, thus, clearly established the concept of totipotency, and further indicated that the technique of cultivating isolated plant cells in nutrient solution permits the investigation of important problems from a new experimental approach. On the basis of that 1902 address and

Plant Tissue Culture: A Review

Plant tissue culture refers to growing and multiplication of cells, tissues and organs of plants on defined solid or liquid media under aseptic and controlled environment. The commercial technology is primarily based on micro propagation, in which rapid proliferation is achieved from tiny stem cuttings, axillary buds, and to a limited extent from somatic embryos, cell clumps in suspension cultures and bioreactors. The cultured cells and tissue can take several pathways. The pathways that lead to the production of true-to-type plants in large numbers are the preferred ones for commercial multiplication. The process of micro propagation is usually divided into several stages i.e., pre-propagation, initiation of explants, subculture of explants for proliferation, shooting and rooting, and hardening. These stages are universally applicable in large-scale multiplication of plants. The delivery of hardened small micro propagated plants to growers and market also requires extra care.

1. Preparation of nutrient medium:
A semi-solid medium is prepared in double distilled water containing macro elements, micro elements, amino acids, vitamins, iron source, carbon source like

Role of Tissue Culture (in vitro) Techniques in the Conservation of Rare and Endangered Species

The conservation and maintenance of plant  biodiversity is an important issue relating to the global human population. The anthropogenic  pressure, the introduction of foreign species, as well as native species and chronic weed invasion has striking special effects on plant diversity. As a result there is an increase in number of threatened species. For the food and medicine industry, plant  biodiversity serves as a natural source of raw material. It supplies a variety of necessary raw materials and serves to provide new genetic information valuable for breeding programs furthermore for developing high yielding crops and resistant plants to environmental stresses (Rao, 2000). A huge amount of funds are spent annually to restock the lost biodiversity and many protocols are accessible recently. Unluckily, we are not seeing any enhancement in the condition of these plant species naturally and the number of endangered plant species is raising slowly but surely (Tripathi, 2008).

Public interest is getting more towards the preservation of  plant genetic resources because

The Prerequisite of the Success in Plant Tissue Culture: High Frequency Shoot Regeneration

Plant tissue culture is a term containing techniques used to propagate plants vegetatively by using small parts of living tissues (explants) on artificial growth mediums under sterile conditions. Explants regenerate shoots and roots, and consequently whole fertile plants under certain cultural conditions. Micropropagation is the production of whole plants through tissue culture from small parts such as shoot and root tips, leaf tissues, anthers, nodes, meristems and embryos. Micropropagation is the vegetative (asexual) propagation of plants under in vitro conditions and is widely used for commercial purposes worldwide [1-3].

Plant tissue culture techniques have certain advantages over traditional ones of propagation.
These are:

Tragacanth, a Novel and Cheap Gelling Agent in Carnation and Miniature Rose Tissue Culture Media

During the last three decades, there have been increased efforts to explore suitable substitutes for agar namely, carrageenan (Lines, 1977), alginates (Scheurich et al., 1980), ficol (Kao, 1981), gelrite (Pasqualetto et al., 1988), starch (Henderson and Kinnersley, 1988; Nene et al., 1996), isubgol (Babbar and Jain, 1998), and katira gum (Jain and Babbar, 2002). Consequently, a number of substances have been used with reasonable success as a substitute for agar. These agents are not expected to reach universal acceptance due to various reasons. Starch, the cheapest of the gelling agents used, is not expected to find universal acceptance because of its inferior gelling ability and poor clarity than agar; and it metabolizes too readily (Kuria et al., 2008). Carrageenan and alginates gel only in the presence of specific ions, and agarose and ficoll are cost prohibitive (Jain and Babbar, 2002). Gelrite, another gelling agent, though not a perfect replacement for agar, has found wide acceptance for plant tissue culture media (Pasqualetto et al., 1988). Isubgol, a highly cost-effective gelling agent, has all the desirable properties, however, its higher melting point (~70 °C) necessitates adjustments of pH and quick

Morphology of callus, shoots, roots and leaves of Withania somnifera (Cultivated and Wild) in vitro tissue culture conditions with different hormones concentration

Modern plant tissue culture is practiced under aseptic or favourable conditions under filtered air. Living plant materials which was taken from the environment or free space are being naturally contaminated or will show contamination on their external surfaces (and sometimes interiors) with pathogenic microorganisms, so surface sterilization of initial materials (explants) in chemical solutions (usually Sodium or calcium hypochlorite or mercuric chloride) is done. Mercuric chloride is mostly used as a plant sterilant now a days, unless other sterilizing agents which require for removal of contamination are dispose found to be least effective, as it is very dangerous and harmful to use, and is very difficult to dispose of (Aniel, et al., 2011) [1]. 

Explants are then generally placed or maintained on the upper surface of a solidified culture medium (normally MS) (Pawar & Maheshwari, 2004) [11], but during some cases it will be inoculate directly into a liquid nutrient medium, normally when cell suspension cultures are required. Solid nutrient and liquid nutrient media usually consists of hormones. Solidified media are prepared or produced from the liquid media with the use of a gelling or solidified agent, generally a purified agar. The constituents of the

Use of Plant Auxins Produced by Bacteria in Plant Tissue Culture and Seed Pre-treatment; A Possibility of Replacement of Synthetic Auxins

Phyto-hormones or plant growth regulators are organic substances synthesized in defined organs of the plant that can be translocated to the other sites, where it triggers specific biochemical, physiological and morphological responses (Vamil, 2011). Today, the synthetic plant growth regulators are available commercially. If these chemicals are applied in specific quantity, enhance the plant growth (Naeem, 2004). There are five different types of plant growth promoting substances produced in plants. Out of this IAA, IBA are important as having major growth promoting activity. Many investigations showed that presowing treatment of growth regulators to the seeds could lead to increase in tissue hydration, redistribution of nutrient reserves, higher respiratory activities and enhancement of seedling growth, dry matter production, early flowering and yield. (Vamil, 2011). IAA exerts influence on plant growth by enlarging leaves and increasing photosynthetic activities in plants (Naeem, 2004). IAA also activates the translocation of carbohydrates during their

The Rise of 3D Cell Culture Systems

The development of three-dimensional (3D) cell culture systems has greatly expanded over the past few decades. 3D cell culture models – which are used to better illustrate the cellular structure of human biological diseases in the laboratory–are now used in a variety of research and therapeutic studies. These include, but are not limited to: developmental biology, tissue morphogenesis and engineering, gene and protein expression, disease pathology, drug discovery and other therapeutic developments, and regenerative medicine. This technology is leading to biological and biochemical discoveries and will be the focus of many conversations around innovation at this year’s Society for Laboratory Automation and Screening (SLAS) Conference and Exhibition. It is helping scientists transform research breakthroughs into real-world outcomes.

The need for more physiologically relevant cell culture models grew out of the observed limitations of the two-dimensional (2D) cell culture techniques that have been used since the early

In vitro Tissue Culture, a Tool for the Study and Breeding of Plants Subjected to Abiotic Stress Conditions

Abiotic stress factors are the main limitation to plant growth and yield in agriculture. Among them, drought stress caused by water deficit, is probably the most impacting adverse condition and the most widely encountered by plants, not only in crop fields but also in wild environments. According to published statistics, the percentage of droughtaffected land area in the world in 2000 was double that of 1970 [1].

Another major environmental factor that limits crop productivity, mainly in arid and semiarid regions is high salinity. Approximately 19.5% of the irrigated soils in the world have elevated concentrations of salts either in the soil or in the irrigation water [2], damaging both the economy and the environment [3, 4]. The deleterious effects of salinity on plant growth are associated with low osmotic potential of

Potential Applications of Wastes from Energy and Forestry Industry in Plant Tissue Culture

     Plant tissue culture refers to the growth and multiplication of cells, tissues and organs on defined solid or liquid media in aseptic and controlled environment. Plant tissue culture technology is being widely used for large-scale plant multiplication.1,2 Regeneration and development of callus depend on the initiation and growth conditions, but also on the species or vegetative organ from which it arose. The induction of callus cultures, the growth rate, as well as the morphogenetic processes, depend on the type and concentration of the regulators present in the culture medium.3

     In agreement with literature data,4 once induced, the callus has the ability to synthesize phytohormones by itself. For example, the callus derived from vegetative fragments of Daucus carota was induced on culture media containing polyphenolic extracts separated from

Great Overview Video of the Tissue Culture Process

Tissue Culture Terminology

Plant tissue culture - a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition

Callus - Undifferentiated, swollen cell mass forming under the influence of elevated plant hormone levels.

Etiolation - Yellow and stretched plant; parts elongate until light is intercepted.

Explant - Part of an organism used in "in vitro" culture.

IAA - Indoleacetic acid; a plant hormone increasing cell elongation and, under certain circumstances, implicated in stimulating cell division and root formation. IAA moves in a polar manner in plants forming an IAA gradient in tissues. Orientation of plant organs, then, influence callus formation and morphogenesis.

"in vitro" - "In glass"; as in tissue culture methods

Morphogenesis - Change in shape

Polarity - Orientation in gravitational field.

Primordia - The earliest detectable stage of an organ, such as a leaf, root or root branch.

Root Hairs - Epidermal cell extensions of

What is Tissue Culture?

Tissue Culture is the process of replicating cells in a sterile environment. Typically this is done in glass containers. The plant cells are placed in or on a nutrient rich media: liquid, semi-liquid or solid (agar). Many times this process is called 'in vitro' culture.

• Agar is a jelly like substance with nutrients for the plant cells to pull from as they multiply.   

In the picture above you can see the plants are growing in agar, the translucent media at the base of the container.

There are several parts of the plant that can be considered a culture or starting point. This includes, shoots, nodes, roots, callus, single cells, seeds and embryo. Many factors go into whether or not the plant cells will replicate, including chemicals, viability of the plant parts, maintained sterile environment and the growing media. 

Check out this biology discussion to see more.