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Plant Methods

Amanda Hopes, Vladimir Nekrasov, Sophien Kamoun, Thomas Mock
BACKGROUND: CRISPR-Cas is a recent and powerful addition to the molecular toolbox which allows programmable genome editing. It has been used to modify genes in a wide variety of organisms, but only two alga to date. Here we present a methodology to edit the genome of Thalassiosira pseudonana, a model centric diatom with both ecological significance and high biotechnological potential, using CRISPR-Cas. RESULTS: A single construct was assembled using Golden Gate cloning...
2016: Plant Methods
Annegret Ross, Imre E Somssich
BACKGROUND: The interaction of Pseudomonas syringae with Arabidopsis is one of the most commonly used systems to study various bacterial-host interrelationships. Currently, most studies are based on the growth quantification of the pathogen to characterize resistance or virulence targets. However, the standard available method for determining bacterial proliferation in planta is laborious and has several limitations. RESULTS: Here we present an alternative robust approach, which is based on the quantification of bacterial DNA by real-time PCR...
2016: Plant Methods
Wen-Jing Cai, Tian-Tian Ye, Qing Wang, Bao-Dong Cai, Yu-Qi Feng
BACKGROUND: Phytohormones play crucial roles in almost all stages of plant growth and development. Accurate and simultaneous determination of multiple phytohormones enabled us to better understand the physiological functions and the regulatory networks of phytohormones. However, simultaneous determination of multiple phytohormones in plant is still a challenge due to their low concentrations, structural and chemical diversity, and complex matrix of plant tissues. Therefore, development of a simple and selective method for the simultaneous determination of multiple phytohormones is highly needed...
2016: Plant Methods
Kumud B Mishra, Anamika Mishra, Kateřina Novotná, Barbora Rapantová, Petra Hodaňová, Otmar Urban, Karel Klem
BACKGROUND: Non-invasive and high-throughput monitoring of drought in plants from its initiation to visible symptoms is essential to quest drought tolerant varieties. Among the existing methods, chlorophyll a fluorescence (ChlF) imaging has the potential to probe systematic changes in photosynthetic reactions; however, prerequisite of dark-adaptation limits its use for high-throughput screening. RESULTS: To improve the throughput monitoring of plants, we have exploited their light-adaptive strategy, and investigated possibilities of measuring ChlF transients under low ambient irradiance...
2016: Plant Methods
Hanna Ćwiek-Kupczyńska, Thomas Altmann, Daniel Arend, Elizabeth Arnaud, Dijun Chen, Guillaume Cornut, Fabio Fiorani, Wojciech Frohmberg, Astrid Junker, Christian Klukas, Matthias Lange, Cezary Mazurek, Anahita Nafissi, Pascal Neveu, Jan van Oeveren, Cyril Pommier, Hendrik Poorter, Philippe Rocca-Serra, Susanna-Assunta Sansone, Uwe Scholz, Marco van Schriek, Ümit Seren, Björn Usadel, Stephan Weise, Paul Kersey, Paweł Krajewski
BACKGROUND: Plant phenotypic data shrouds a wealth of information which, when accurately analysed and linked to other data types, brings to light the knowledge about the mechanisms of life. As phenotyping is a field of research comprising manifold, diverse and time-consuming experiments, the findings can be fostered by reusing and combining existing datasets. Their correct interpretation, and thus replicability, comparability and interoperability, is possible provided that the collected observations are equipped with an adequate set of metadata...
2016: Plant Methods
Mohamed A Salem, Jessica Jüppner, Krzysztof Bajdzienko, Patrick Giavalisco
BACKGROUND: The elucidation of complex biological systems requires integration of multiple molecular parameters. Accordingly, high throughput methods like transcriptomics, proteomics, metabolomics and lipidomics have emerged to provide the tools for successful system-wide investigations. Unfortunately, optimized analysis of different compounds requires specific extraction procedures in combination with specific analytical instrumentation. However, the most efficient extraction protocols often only cover a restricted number of compounds due to the different physico-chemical properties of these biological compounds...
2016: Plant Methods
Xiao Jin, Jin-Ying Gou
BACKGROUND: Protein phosphorylation is one of the most important post-translational modifications catalyzed by protein kinases in living organisms. The advance of genome sequencing provided the information of protein kinase families in many organisms, including both model and non-model plants. The development of proteomics technologies also enabled scientists to efficiently reveal a large number of protein phosphorylations of an organism. However, kinases and phosphorylation targets are still to be connected to illustrate the complicated network in life...
2016: Plant Methods
Zhicheng Zhang, Kurt Boonen, Piero Ferrari, Liliane Schoofs, Ewald Janssens, Vera van Noort, Filip Rolland, Koen Geuten
BACKGROUND: The complexity of RNA regulation is one of the current frontiers in animal and plant molecular biology research. RNA-binding proteins (RBPs) are characteristically involved in post-transcriptional gene regulation through interaction with RNA. Recently, the mRNA-bound proteome of mammalian cell lines has been successfully cataloged using a new method called interactome capture. This method relies on UV crosslinking of proteins to RNA, purifying the mRNA using complementary oligo-dT beads and identifying the crosslinked proteins using mass spectrometry...
2016: Plant Methods
Donna M Bond, Nick W Albert, Robyn H Lee, Gareth B Gillard, Chris M Brown, Roger P Hellens, Richard C Macknight
BACKGROUND: Transcription factors (TFs) coordinate precise gene expression patterns that give rise to distinct phenotypic outputs. The identification of genes and transcriptional networks regulated by a TF often requires stable transformation and expression changes in plant cells. However, the production of stable transformants can be slow and laborious with no guarantee of success. Furthermore, transgenic plants overexpressing a TF of interest can present pleiotropic phenotypes and/or result in a high number of indirect gene expression changes...
2016: Plant Methods
Chantal Le Marié, Norbert Kirchgessner, Patrick Flütsch, Johannes Pfeifer, Achim Walter, Andreas Hund
BACKGROUND: Phenotyping of genotype-by-environment interactions in the root-zone is of major importance for crop improvement as the spatial distribution of a plant's root system is crucial for a plant to access water and nutrient resources of the soil. However, so far it is unclear to what extent genetic variations in root system responses to spatially varying soil resources can be utilized for breeding applications. Among others, one limiting factor is the absence of phenotyping platforms allowing the analysis of such interactions...
2016: Plant Methods
Guo-Qian Yang, Yun-Mei Chen, Jin-Peng Wang, Cen Guo, Lei Zhao, Xiao-Yan Wang, Ying Guo, Li Li, De-Zhu Li, Zhen-Hua Guo
BACKGROUND: The double digest restriction-site associated DNA sequencing technology (ddRAD-seq) is a reduced representation sequencing technology by sampling genome-wide enzyme loci developed on the basis of next-generation sequencing. ddRAD-seq has been widely applied to SNP marker development and genotyping on animals, especially on marine animals as the original ddRAD protocol is mainly built and trained based on animal data. However, wide application of ddRAD-seq technology in plant species has not been achieved so far...
2016: Plant Methods
Patrick C Boyle, Simon Schwizer, Sarah R Hind, Christine M Kraus, Susana De la Torre Diaz, Bin He, Gregory B Martin
BACKGROUND: The plant plasma membrane is a key battleground in the war between plants and their pathogens. Plants detect the presence of pathogens at the plasma membrane using sensor proteins, many of which are targeted to this lipophilic locale by way of fatty acid modifications. Pathogens secrete effector proteins into the plant cell to suppress the plant's defense mechanisms. These effectors are able to access and interfere with the surveillance machinery at the plant plasma membrane by hijacking the host's fatty acylation apparatus...
2016: Plant Methods
Mitchell A Ellison, Michael B McMahon, Morris R Bonde, Cristi L Palmer, Douglas G Luster
BACKGROUND: Rust fungi are obligate pathogens with multiple life stages often including different spore types and multiple plant hosts. While individual rust pathogens are often associated with specific plants, a wide range of plant species are infected with rust fungi. To study the interactions between these important pathogenic fungi and their host plants, one must be able to differentiate fungal tissue from plant tissue. This can be accomplished using the In situ hybridization (ISH) protocol described here...
2016: Plant Methods
Filippo Biscarini, Nelson Nazzicari, Chiara Broccanello, Piergiorgio Stevanato, Simone Marini
BACKGROUND: Noise (errors) in scientific data is endemic and may have a detrimental effect on statistical analyses and experimental results. The effects of noisy data have been assessed in genome-wide association studies for case-control experiments in human medicine. Little is known, however, on the impact of noisy data on genomic predictions, a widely used statistical application in plant and animal breeding. RESULTS: In this study, the sensitivity to noise in the data of five classification methods (K-nearest neighbours-KNN, random forest-RF, ridge logistic regression-LR, and support vector machines with linear or radial basis function kernels) was investigated...
2016: Plant Methods
Atena Haghighattalab, Lorena González Pérez, Suchismita Mondal, Daljit Singh, Dale Schinstock, Jessica Rutkoski, Ivan Ortiz-Monasterio, Ravi Prakash Singh, Douglas Goodin, Jesse Poland
BACKGROUND: Low cost unmanned aerial systems (UAS) have great potential for rapid proximal measurements of plants in agriculture. In the context of plant breeding and genetics, current approaches for phenotyping a large number of breeding lines under field conditions require substantial investments in time, cost, and labor. For field-based high-throughput phenotyping (HTP), UAS platforms can provide high-resolution measurements for small plot research, while enabling the rapid assessment of tens-of-thousands of field plots...
2016: Plant Methods
Michal Slota, Miroslaw Maluszynski, Iwona Szarejko
BACKGROUND: Genetic studies on the molecular mechanisms of the regulation of root growth require the characterisation of a specific root phenotype to be linked with a certain genotype. Such studies using classical labour-intensive methods are severely hindered due to the technical limitations that are associated with the impeded observation of the root system of a plant during its growth. The aim of the research presented here was to develop a reliable, cost-effective method for the analysis of a plant root phenotype that would enable the precise characterisation of the root system architecture of cereals...
2016: Plant Methods
François Postic, Claude Doussan
BACKGROUND: To face climate change and subsequent rainfall instabilities, crop breeding strategies now include root traits phenotyping. Rapid estimation of root traits in controlled conditions can be achieved by using parallel electrical capacitance and its linear correlation with root dry mass. The aim of the present study was to improve robustness and efficiency of methods based on capacitance and other electrical variables, such as serial/parallel resistance, conductance, impedance or reactance...
2016: Plant Methods
Peter Lootens, Tom Ruttink, Antje Rohde, Didier Combes, Philippe Barre, Isabel Roldán-Ruiz
BACKGROUND: Genetic studies and breeding of agricultural crops frequently involve phenotypic characterization of large collections of genotypes grown in field conditions. These evaluations are typically based on visual observations and manual (destructive) measurements. Robust image capture and analysis procedures that allow phenotyping large collections of genotypes in time series during developmental phases represent a clear advantage as they allow non-destructive monitoring of plant growth and performance...
2016: Plant Methods
Christian Jeudy, Marielle Adrian, Christophe Baussard, Céline Bernard, Eric Bernaud, Virginie Bourion, Hughes Busset, Llorenç Cabrera-Bosquet, Frédéric Cointault, Simeng Han, Mickael Lamboeuf, Delphine Moreau, Barbara Pivato, Marion Prudent, Sophie Trouvelot, Hoai Nam Truong, Vanessa Vernoud, Anne-Sophie Voisin, Daniel Wipf, Christophe Salon
BACKGROUND: In order to maintain high yields while saving water and preserving non-renewable resources and thus limiting the use of chemical fertilizer, it is crucial to select plants with more efficient root systems. This could be achieved through an optimization of both root architecture and root uptake ability and/or through the improvement of positive plant interactions with microorganisms in the rhizosphere. The development of devices suitable for high-throughput phenotyping of root structures remains a major bottleneck...
2016: Plant Methods
Martin Schäfer, Christoph Brütting, Ian T Baldwin, Mario Kallenbach
BACKGROUND: Plant metabolites are commonly functionally classified, as defense- or growth-related phytohormones, primary and specialized metabolites, and so forth. Analytical procedures for the quantifications of these metabolites are challenging because the metabolites can vary over several orders of magnitude in concentrations in the same tissues and have very different chemical characteristics. Plants clearly adjust their metabolism to respond to their prevailing circumstances in very sophisticated ways that blur the boundaries among these functional or chemically defined classifications...
2016: Plant Methods
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