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Anther Culture of Chili Pepper (Capsicum spp.)

Chili pepper (Capsicum spp.) is a very important horticultural crop around the world and is especially important for Mexicans because of its impact in the culture and the cuisine. Biotechnological tools such as tissue culture techniques and specifically anther culture may be applied suc ...

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Cryopreservation of Shoot Tips and Meristems: An Overview of Contemporary Methodologies

Cryopreservation is the storage of viable bioresources at ultra-low temperatures in liquid nitrogen (LN). This chapter provides an overview of those protocols most commonly used to cryopreserve in vitro derived shoot tips and meristems; they are described generically, as sequenti ...

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Improved Shoot Regeneration from Root Explants Using an Abscisic Acid-Containing Medium

Arabidopsis thaliana is an excellent experimental system to study various aspects of plant biology. An efficient regeneration protocol of the plant is important to produce in vitro fertile plants, as well as a fundamental step to study the cellular, physiological, and molecular mechani ...

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Maize Somatic Embryogenesis: Recent Features to Improve Plant Regeneration

Plant regeneration capacity is maintained through the life of a plant by the stem cell niche present in the meristems. Stem cells are capable of differentiating into any plant organ, allowing propagation of new plants by different techniques. Among them, somatic embryogenesis is a widely used ...

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Production of Haploids and Doubled Haploids in Maize

The in vivo haploid induction approach offers several advantages compared to the in vitro induction approach and recurrent self-pollination. It is currently the method of choice for inbred line development in many commercial maize breeding programs. Here, we describe the in vivo approa ...

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Liquid In Vitro Culture for the Propagation of Arundo donax

We describe a simple and inexpensive plant micropropagation system for giant reed (Arundo donax L.) that uses axillary buds from the lateral stems of elite plants selected from field- or nursery-grown plants. The buds, attached to the stems are cultured in stationary liquid MS culture medium, s ...

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Micropropagation of Banana

Banana (Musa spp. AAA) is propagated vegetatively and can be rapidly and efficiently propagated by micropropagation. Conventional micropropagation techniques, however, may be too costly for commercial purposes. Our laboratory has found that depending on the combination of cult ...

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Tissue Culture Methods for the Clonal Propagation and Genetic Improvement of Spanish Red Cedar (Cedrela odorata)

The choice of a method to culture red cedar tissues depends on the final objectives pursued. If homogeneous clonal material is required for experimental purposes, the easiest way is to generate the lines through adventitious shoot induction from seedlings germinated from seeds. If the obj ...

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A Chemical Genetics Method to Uncover Small Molecules for Dissecting the Mechanism of ABA Responses in Arabidopsis Seed Germination

Functional redundancy widely exists in genes encoding receptors and signaling components of plant hormones, particularly the stress hormone abscisic acid (ABA). The redundancy hinders the use of conventional genetic approach to dissecting molecular mechanisms for ABA signal ...

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Activation Tagging

Insertional mutagenesis is one of the most effective approaches to determine the function of plant genes. However, due to genetic redundancy, loss-of-function mutations often fail to reveal the function of a member of gene families. Activation tagging is a powerful gain-of-function app ...

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Small-Molecule Dissection of Brassinosteroid Signaling

The growth-promoting hormones, the brassinosteroids (BRs), are perceived at the plant cell surface by receptor kinases that transduce the signal to the nucleus by an intracellular cascade of phosphorylation-mediated protein–protein interactions. BR signaling is also regula ...

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Identification and Verification of Redox-Sensitive Proteins in Arabidopsis thaliana

Environmental stresses often trigger rapid generation of reactive oxygen species (ROS) in plants. Excessive amount of ROS can cause damage to plant cells and thus need to be counteracted by cellular antioxidant systems. On the other hand, ROS also serve as signaling molecules that modulate v ...

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Quantitative Analysis of Plasma Membrane Proteome Using Two-Dimensional Difference Gel Electrophoresis

The plasma membrane (PM) controls cell’s exchange of both material and information with the outside environment, and PM-associated proteins play key roles in cellular regulation. Numerous cell surface receptors allow cells to perceive and respond to various signals from neighbor ce ...

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Quantitative Analysis of Protein Phosphorylation Using Two-Dimensional Difference Gel Electrophoresis

Posttranslational modifications of proteins, especially phosphorylation and dephosphorylation, play an important role in signal transduction and cellular regulation in plants. Both 2-DE gel-based and non-gel-based proteomic technologies can monitor the changes in pho ...

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Identification of O-linked -d-N-acetylglucosamine-Modified Proteins from Arabidopsis

The posttranslational modification of proteins with O-linked β-d-N-acetylglucosamine (O-GlcNAc) on serine and threonine residues occurs in all animals and plants. This modification is dynamic and ubiquitous, and regulates many cellular processes, including transcripti ...

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Quantitative Measurement of Phosphopeptides and Proteins via Stable Isotope Labeling in Arabidopsis and Functional Phosphoproteomic Strategies

Protein phosphorylation is one type of posttranslational modification, which regulates a large number of cellular processes in plant cells. As an emerging powerful biotechnology that integrates all aspects of advantages from mass spectrometry, bioinformatics, and genomics, ...

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Experimental Analysis of Receptor Kinase Phosphorylation

Ligand binding by the extracellular domain of receptor kinases leads to phosphorylation and activation of the cytoplasmic domain of these important membrane-bound signaling proteins. To thoroughly characterize receptor kinase function, it is essential to identify specific ...

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Genome-Wide Profiling of Uncapped mRNA

Gene transcripts are under extensive posttranscriptional regulation, including the regulation of their stability. A major route for mRNA degradation produces uncapped mRNAs, which can be generated by decapping enzymes, endonucleases, and small RNAs. Profiling uncapped mRNA m ...

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Computational Tools for Quantitative Analysis of Cell Growth Patterns and Morphogenesis in Actively Developing Plant Stem Cell Niches

Pattern formation in developmental fields involves precise spatial arrangement of different cell types in a dynamic landscape wherein cells exhibit a variety of behaviors, such as cell division, cell expansion, and cell migration . The information is exchanged between multiple cell ...

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Transient Expression Assays for Quantifying Signaling Output

The protoplast transient expression system has become a powerful and popular tool for studying molecular mechanisms underlying various plant signal transduction pathways. Arabidopsis mesophyll protoplasts display intact and active physiological responses and are easy ...

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