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Plant Growth and Cultivation

There is a variety of methods used for growing plants indoor for laboratory research. In most cases plant research requires germination and growth of plants. Often, people have adapted plant cultivation protocols to the conditions and materials at hand in their own laboratory and growth fac ...

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Roles and Functions of Plant Mineral Nutrients

Plants require macro- and micronutrients, each of which is essential for a plant to complete its life cycle. Adequate provision of nutrients impacts greatly on plant growth and as such is of crucial importance in the context of agriculture. Minerals are taken up by plant roots from the soil solution in ...

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Applications of High-Throughput Plant Phenotyping to Study Nutrient Use Efficiency

Remote sensing and spectral reflectance measurements of plants has long been used to assess the growth and nutrient status of plants in a noninvasive manner. With improved imaging and computer technologies, these approaches can now be used at high-throughput for more extensive physiol ...

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Large-Scale Plant Ionomics

Large-scale phenotyping methods are at the heart of efficiently deciphering the functions of genes and gene networks in the postgenomic era. In order to obtain meaningful results when comparing natural variants, and mutants and wild-types during large-scale quantitative analyses, ...

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Real Time Measurement of Cytoplasmic Ions with Ion-Selective Microelectrodes

Ion-selective microelectrodes can be used to report intracellular ion concentrations. The ion-selective barrels of microelectrodes are filled with a sensor cocktail containing several different components including an ion-selective molecule, sensor or exchanger, a solv ...

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Measurements of Cytosolic Ion Concentrations in Live Cells

Here, we describe a series of methods suitable for the measurement of cytosolic ion concentrations in living plant cells using ion selective dyes. We describe procedures for the use of SBFI for the measurement of Na+ in live cells. The resulting material is suitable for most standard cell biology pr ...

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Plant Single Cell Sampling

Plant single cell sampling and analysis allows the determination of solute concentrations in individual cells and tissues. This is particularly important when studying mineral nutrition, where the cell- and tissue-specific distribution of individual mineral nutrients incr ...

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Methods for Xylem Sap Collection

Xylem and phloem are essential for the exchange of solutes and signals among organs of land plants. The synergy of both enables the transport and ultimately the partitioning of water, nutrients, metabolic products and signals among the organs of plants. The collection and analysis of xylem sap a ...

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Sampling and Analysis of Phloem Sap

The transport tubes of the phloem are essential for higher plants. They not only provide the route for the distribution of assimilates produced during photosynthesis from source to sink organs but also (re-) distribute mineral nutrients. Additionally, the phloem is essential for sending ...

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Ion Flux Measurements Using the MIFE Technique

Noninvasive microelectrode ion flux measurements (the MIFE™ technique) allow the concurrent quantification of net fluxes of several ions with high spatial (several μm) and temporal (ca 5 s) resolution. The MIFE technique has become a popular tool for studying the adaptive responses of pl ...

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Flux Measurements of Cations Using Radioactive Tracers

Standard procedures for the tracing of ion fluxes into roots of plants are described here, with emphasis on cations, especially potassium (K+). We focus in particular on the measurement of unidirectional influx by use of radiotracers and provide a brief introduction to compartmental anal ...

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Herbicide Resistance Screening Assay

Herbicide resistance screening is a method that can be used not only to determine presence of the enzyme, phosphinothricin acetyltransferase, encoded by either the Bar or the Pat gene in transgenic maize, but also to assess the inheritance ratio of those genes in a segregating population. Herb ...

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Characterizing Transgene Inheritance

The analysis of transgene inheritance is an important step in the molecular and genetic characterization of transgenes. In this manuscript, two approaches to characterize the inheritance of transgenes are described. The first approach is based on the expression of the transgene phen ...

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Backcross Breeding

Backcross breeding enables breeders to transfer a desired trait such as a transgene from one variety (donor parent, DP) into the favored genetic background of another (recurrent parent, RP). If the trait of interest is produced by a dominant gene, this process involves four rounds of backcrossi ...

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Agrobacterium-Mediated Maize Transformation: Immature Embryos Versus Callus

Transformation with Agrobacterium tumefaciens is the preferred method for delivery of transgenes into a wide range of plant species including maize. Optimized protocols for the Agrobacterium-mediated transformation of freshly isolated immature embryos and embryogenic T ...

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Biolistic Gun-Mediated Maize Genetic Transformation

Biolistic gun-mediated transformation is one of the two most effective and popular methods for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype, Hi II. The protocol uses 0.6-μm gold particles as microcarriers a ...

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Transient Expression of GFP in Immature Seed Tissues

Transient expression is the nonstable expression of a transgene whereby the transgene does not integrate into the host’s genomic DNA. Transient expression assays have 20 years of history in plant molecular biology research, being used to answer a variety of questions. The method described ...

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Design of Gene Constructs for Transgenic Maize

The first step of any maize transformation project is to select gene expression elements that will make up an effective construct. When designing a gene construct, one must have a full understanding of the different expression elements that are currently available and of the strategies that h ...

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Methods for Generation and Analysis of Fluorescent Protein-Tagged Maize Lines

The use of fluorescent proteins to localize gene products in living cells has revolutionized cell biology. Although maize has excellent genetics resources, the use of fluorescent proteins in maize cell biology has not been well developed. To date, protein localization in this species has m ...

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Whiskers-Mediated Maize Transformation

There has been rapid progress in recent years in extending gene transfer capabilities to include plant species that fall outside the normal host range of Agrobacterium. Methods that allow direct DNA delivery into plant cells have contributed significantly to this expanded capability. ...

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