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DAPI Staining and Fluorescence Microscopy Techniques for Phytoplasmas

The 4′,6-diamidino-2-phenylindole (DAPI) stain technique is a simple method that was developed for confirming the presence of phytoplasmas in hand-cut or freezing microtome sections of infected tissues. DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized amo ...

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Cryopreservation of Embryogenic Cell Suspensions by EncapsulationVitrification and EncapsulationDehydration

Encapsulation–vitrification and encapsulation–dehydration are two newly developed techniques for cryopreservation of embryogenic cell suspensions. Here, we describe the two protocols using grapevine (Vitis) as a model plant. Cell suspensions at the exponential growth s ...

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Pathogen and Biological Contamination Management in Plant Tissue Culture: Phytopathogens, Vitro Pathogens, and Vitro Pests

The ability to establish and grow plant cell, organ, and tissue cultures has been widely exploited for basic and applied research, and for the commercial production of plants (micro-propagation). Regardless of whether the application is for research or commerce, it is essential that the cul ...

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Measurement of Cell Viability

An overview of the methods for assessing cell viability is presented. Different protocols of the most commonly used assays are described in detail so that the readers may be able to determine which assay is suitable for their own projects in plant biotechnology.

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Callus, Suspension Culture, and Hairy Roots. Induction, Maintenance and Characterization

The growth is a characteristic of each culture and it is determinate by the origin of the species, culture conditions, and type of culture. In this chapter, we make a comparison of the different growth parameters among three different species and three different types of cultures.

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Appendix B: Plant Biotechnology and Tissue Culture Resources in the Internet

This appendix compiles a list of useful Internet sites for cell culture scientists. A total of more than 100 sites have been selected, based on the quality of the information offered in them, as well as on their users’ friendliness. We anticipate that some of these sites will be included among the reader’s f ...

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Appendix A: The Components of the Culture Media

The success in the technology and application of plant tissue culture is greatly influenced by the nature of the culture medium used. A better understanding of the nutritional requirements of cultured cells and tissues can help to choose the most appropriate culture medium for the explant us ...

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Organelle Transformation

The source of genetic information in a plant cell is contained in nucleus, plastids, and mitochondria. Organelle transformation is getting a lot of attention nowadays because of its superior performance over the conventional and most commonly used nuclear transformation for obtain ...

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Improved Genetic Transformation of Cork Oak (Quercus suber L.)

An Agrobacterium-mediated transformation system for selected mature Quercus suber L. trees has been established. Leaf-derived somatic embryos in an early stage of development are inoculated with an AGL1 strain harboring a kanamycin-selectable plasmid carrying the gene of inte ...

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Biolistic- and Agrobacterium-Mediated Transformation Protocols for Wheat

After rice, wheat is considered to be the most important world food crop, and the demand for high-quality wheat flour is increasing. Although there are no GM varieties currently grown, wheat is an important target for biotechnology, and we anticipate that GM wheat will be commercially available in ...

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Plant Tissue Culture and Molecular Markers

Tissue culture can be used to propagate elite material or to generate new variability by employing somaclonal variation. Genetic stability of the process must be evaluated analyzing DNA profiles by the use of molecular markers. Several techniques have been reported for the screening of ge ...

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Basic Procedures for Epigenetic Analysis in Plant Cell and Tissue Culture

In vitro culture is one of the most studied techniques, and it is used to study many developmental processes, especially in forestry species, because of growth timing and easy manipulation. Epigenetics has been shown as an important influence on many research analyses such as cancer in mammals a ...

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Determination of Histone Methylation in Mono- and Dicotyledonous Plants

Epigenetics includes DNA methylation and histones posttranslational modifications such as methylation, acetylation, phosphorylation among others. One of the most abundant modifications in histone tail is the methylation. It has been found that the methylation pattern in the h ...

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The Potential Roles of microRNAs in Molecular Breeding

microRNAs (miRNAs), a class of small, endogenous, noncoding RNAs, are uncovered to play greatly expanded roles in a variety of plant developmental processes by gene silencing through inhibiting translation or promoting the degradation of target mRNAs. In virtue of their ability to inact ...

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Epigenetics, the Role of DNA Methylation in Tree Development

During development of multicellular organisms, cells become differentiated by modulating different programs of gene expression. Cells have their own epigenetic signature which reflects genotype, developmental history, and environmental influences, and it is ultimate ...

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Establishment of a Sanguinarine-Producing Cell Suspension Culture of Argemone mexicana L (Papaveraceae): Induction of Alkaloid Accumulation

A protocol for the induction of a cell suspension culture of Argemone mexicana is described. This suspension has been kept for over 3 years producing sanguinarine, a benzophenanthridine-type alkaloid. Sanguinarine levels can be increased by exposing these cultures to yeast or fungal el ...

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Removing Heavy Metals by In Vitro Cultures

In vitro roots cultures of Typha latifolia and Scirpus americanus aquatic plants have the capacity to remove Pb (II), Mn (II), and Cr (III) from the culture medium. Both species remove Cr and Pb by an absorption process, while Mn is mainly adsorbed to the root surface. This chapter describes a protocol for the ...

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An Introduction to Plant Cell Culture: The Future Ahead

Plant cell, tissue, and organ culture (PTC) techniques were developed and established as an experimental necessity for solving important fundamental questions in plant biology, but they currently represent very useful biotechnological tools for a series of important applicati ...

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Plant Tissue Culture of Fast-Growing Trees for Phytoremediation Research

The ability of plants to remove pollutants from the environment is currently used in a simple and low-cost cleaning technology known as phytoremediation. Unfortunately, little is known about the metabolic pathways involved in the transformation of xenobiotic compounds and the abil ...

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Production of Interspecific Hybrids in Ornamental Plants

In breeding of ornamental plants, interspecific hybridization and polyploidization have successfully been used to produce novel cultivars with blended traits of both parents and to introgress useful traits of one species to another. Embryo rescue techniques and molecular cytog ...

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