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Misexpression Approaches for the Manipulation of Flower Development
Time of Update: 2021-01-13
The generation of dominant gain-of-function mutants through activation tagging is a forward genetic approach that complements the screening of loss-of-function mutants and that has been successfully applied to studying the mechanisms of flower development.
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Quantitative Imaging Approaches for Small-Molecule Measurements Using FRET Sensors in Plants
Time of Update: 2021-01-13
Genetically encoded F�ster resonance energy transfer sensors afford a powerful method of measuring these dynamics in situ and hence are now widely used in order to decode information communicated through the dynamics of cellular metabolites and ions.
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Saponins
Time of Update: 2021-01-13
Saponins comprise a large family of structurally related compounds containing a steroid or triterpenoid aglycone (sapogenin) linked to one or more oligosaccharide moieties by glycosidic linkage(Fig. 1) .
The primary biological effect of saponins is the interactions with cellular and membrane components.
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Mechanical Measurements on Living Plant Cells by Micro-indentation with Cellular Force Microscopy
Time of Update: 2021-01-13
Indentation methods on the micro- and nanoscale are increasingly used to assess mechanical properties of living plant tissues.
Cellular force microscopy is a micro-indentation method that was designed to precisely measure and apply forces on living plant cells.
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Geminivirus Isolation and DNA Extraction
Time of Update: 2021-01-13
Curtoviruses, such as beet curly top and tomato pseudo curly top viruses (BCTV and TPCTV), occupy an intermediate position between Mastreviruses and Begomoviruses, in that these viruses have monopartite genomes and are transmitted by leafhopper species, but only infect dicotyledonous hosts.
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Suspension-Cultured Plant Cells as a Tool to Analyze the Extracellular Proteome
Time of Update: 2021-01-13
Despite the central role played by extracellular proteins in most processes that control growth and development, the secretome has been less well characterized than other subcellular compartments, meaning that our understanding of the cell wall physiology is still very limited.
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Production Systems
Time of Update: 2021-01-13
This chapter ties together ideas introduced in previous articles of this volume in the search for ways to use plant cell cultures as industrial production systems.
The economics of a process must always be carefully considered when examining the suitability of plant cell culture for the manufacture of target compounds.
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Forward Chemical Screening of Small RNA Pathways
Time of Update: 2021-01-13
Forward genetic screens have led to the identification of many components in small RNA biogenesis and functional pathways.
Forward chemical screens can overcome these limitations by targeting multiple redundant proteins within a family and by giving the user temporal control of the application of the chemical.
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Genetic Transformation of Conifers Utilizing Somatic Embryogenesis
Time of Update: 2021-01-13
Over the last 5 yr, the production of transgenic conifers has been greatly facilitated by the ability to transform somatic embryonal tissues (somatic embryos) via cocultivation withAgrobacterium tumefaciens .
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RNA Silencing and Antiviral Defense in Plants
Time of Update: 2021-01-13
This process may in fact reflect the ancient origins of RNA silencing: plants may have evolved RNA silencing pathways as a defense mechanism against foreign nucleic acid species in the absence of an immune system (Wang and Metzlaff, Curr Opin Plant Biol 8:216–222, 2005).
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High-Throughput Data Pipelines for Metabolic Flux Analysis in Plants
Time of Update: 2021-01-13
Central to this is developing an end to end data pipeline, crucial for integrating the wet lab experiments and analytics, combining hardware and software automation, and standardizing data representation providing importers and exporters to support third party tools.
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Yeast Two-Hybrid Approaches to Dissecting the Plant Defense Response
Time of Update: 2021-01-13
We describe a reliable GAL4-based yeast two-hybrid system for identifying and isolating clones encoding proteins interacting with a protein of interest. This two-hybrid system gives extremely low bac
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commercial Experience with Neem Products
Time of Update: 2021-01-13
Governmental policies committed to the institution of integrated pest management (IPM) programs, the increasing resistance developed by insects and pathogens to chemical pesticides, and the public concern about chemicals in general has initiated a re-evaluation of pesticide use.
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In Situ Detection of Plant Viruses and Virus-Specific Products
Time of Update: 2021-01-13
The ability to combine nucleic acid hybridisation or immunospecific reactions with structural and ultrastructural analysis of virus-infected tissues has provided the opportunity to resolve the spatia
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Development of Highly Efficient Genetic Transformation Protocols for Table Grape Sugraone and Crimson Seedless
Time of Update: 2021-01-13
This chapter describes highly efficient genetic transformation protocols for seedless table grapevine cultivars Sugraone and Crimson Seedless by co-culturing embryogenic calli withAgrobacterium tumefaciens .
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Targeting DNA to a Previously Integrated Transgenic Locus Using Zinc Finger Nucleases
Time of Update: 2021-01-13
Retransformation with a corresponding ZFN-expressing cassette and a donor DNA with sequences homologous to the integrated construct and capable of functionalizing the herbicide resistance gene following site-specific integration results in targeted DNA addition.
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Reverse Genetics in Medicago truncatula Using a TILLING Mutant Collection
Time of Update: 2021-01-13
In the last decade different types of mutant populations have been created in this species that can be screened by forward and reverse-genetic approaches to identify and functionally characterize genes of interest.
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Cytogenetic Analyses of Arabidopsis
Time of Update: 2021-01-13
Recently, the very small genome, the low content, and strong clustering of repetitive sequences even turned out to be advantageous for the first establishment of chromosome painting in a euploid plant.
Chromosome painting together with the high-resolution fluorescencein situ hybridization (FISH) mapping onArabidopsis pachytene chromosomes has opened new perspectives for plant cytogenetics.
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Using a Reverse Genetics Approach to Investigate Small-Molecule Activity
Time of Update: 2021-01-13
Here we present an approach to characterize a chemical of interest, as well as examples of studies demonstrating an effective combination of chemical genomics with reverse genetics strategies, drawn from recent literature on phytohormone signalling and auxin transport.
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Practical Applications of Metabolomics in Plant Biology
Time of Update: 2021-01-13
A range of different separation and detection techniques are being refined and expanded and their combination with advanced data management and data analysis approaches is already giving plant scientists far deeper insights into the complexity of plant metabolism and plant metabolic composition than was imaginable just a few years ago.