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Plant Methods

Samir Alahmad, Eric Dinglasan, Kung Ming Leung, Adnan Riaz, Nora Derbal, Kai P Voss-Fels, Jason A Able, Filippo M Bassi, Jack Christopher, Lee T Hickey
Background: Plant breeding requires numerous generations to be cycled and evaluated before an improved cultivar is released. This lengthy process is required to introduce and test multiple traits of interest. However, a technology for rapid generation advance named 'speed breeding' was successfully deployed in bread wheat ( Triticum aestivum L.) to achieve six generations per year while imposing phenotypic selection for foliar disease resistance and grain dormancy. Here, for the first time the deployment of this methodology is presented in durum wheat ( Triticum durum Desf...
2018: Plant Methods
Sruti Das Choudhury, Srinidhi Bashyam, Yumou Qiu, Ashok Samal, Tala Awada
Background: Image-based plant phenotyping facilitates the extraction of traits noninvasively by analyzing large number of plants in a relatively short period of time. It has the potential to compute advanced phenotypes by considering the whole plant as a single object (holistic phenotypes) or as individual components, i.e., leaves and the stem (component phenotypes), to investigate the biophysical characteristics of the plants. The emergence timing, total number of leaves present at any point of time and the growth of individual leaves during vegetative stage life cycle of the maize plants are significant phenotypic expressions that best contribute to assess the plant vigor...
2018: Plant Methods
Tania Ho-Plágaro, Raúl Huertas, María I Tamayo-Navarrete, Juan A Ocampo, José M García-Garrido
Background: Solanum lycopersicum , an economically important crop grown worldwide, has been used as a model for the study of arbuscular mycorrhizal (AM) symbiosis in non-legume plants for several years and several cDNA array hybridization studies have revealed specific transcriptomic profiles of mycorrhizal tomato roots. However, a method to easily screen candidate genes which could play an important role during tomato mycorrhization is required. Results: We have developed an optimized procedure for composite tomato plant obtaining achieved through Agrobacterium rhizogenes -mediated transformation...
2018: Plant Methods
Victoria Pastor, Paloma Sánchez-Bel, Jordi Gamir, María J Pozo, Víctor Flors
Background: Systemin has been extensively studied since it was discovered and is described as a peptidic hormone in tomato plants and other Solanaceae . Jasmonic acid and systemin are proposed to act through a positive feed-back loop with jasmonic acid, playing synergistic roles in response to both wounding and insect attack. Despite its biological relevance, most studies regarding the function of systemin in defence have been studied via PROSYSTEMIN (PROSYS) gene expression, which encodes the propeptide prosystemin that is later cleaved to systemin (SYS)...
2018: Plant Methods
Niels van Tol, Martijn Rolloos, Peter van Loon, Bert J van der Zaal
Background: The formation of crossovers during meiosis is pivotal for the redistribution of traits among the progeny of sexually reproducing organisms. In plants the molecular mechanisms underlying the formation of crossovers have been well established, but relatively little is known about the factors that determine the exact location and the frequency of crossover events in the genome. In the model plant species Arabidopsis , research on these factors has been greatly facilitated by reporter lines containing linked fluorescence marker genes under control of promoters active in seeds or pollen, allowing for the visualization of crossover events by fluorescence microscopy...
2018: Plant Methods
Janet M Paper, Thiya Mukherjee, Kathrin Schrick
Background: Phospholipids are important structural and signaling molecules in plant membranes. Some fluorescent dyes can stain general lipids of membranes, but labeling and visualization of specific lipid classes have yet to be developed for most components of the membrane. New techniques for visualizing membrane lipids are needed to further delineate their dynamic structural and signaling roles in plant cells. In this study we examined whether propargylcholine, a bioortholog of choline, can be used to label the major membrane lipid, phosphatidylcholine, and other choline phospholipids in plants...
2018: Plant Methods
Carolina Camacho-Fernández, David Hervás, Alba Rivas-Sendra, Mª Pilar Marín, Jose M Seguí-Simarro
Background: For in vitro culture of plant and animal cells, one of the critical steps is to adjust the initial cell density. A typical example of this is isolated microspore culture, where specific cell densities have been determined for different species. Out of these ranges, microspore growth is not induced, or is severely reduced. A similar situation occurs in many other plant and animal cell culture systems. Traditionally, researchers have used counting chambers (hemacytometers) to calculate cell densities, but little is still known about their technical advantages...
2018: Plant Methods
Stefan Böhmdorfer, Josua Timotheus Oberlerchner, Christina Fuchs, Thomas Rosenau, Heinrich Grausgruber
Background: Anthocyanins are abundant secondary metabolites responsible for most blue to blue-black, and red to purple colors of various plant organs. In wheat grains, anthocyanins are accumulated in the pericarp and/or aleurone layer. Anthocyanin pigmented wheat grains can be processed into functional foods with potential health benefits due to the antioxidant properties of the anthocyanins. The grain anthocyanin content can be increased by pyramidizing the different genes responsible for the accumulation of anthocyanins in the different grain layers...
2018: Plant Methods
Chiara Broccanello, Claudia Chiodi, Andrew Funk, J Mitchell McGrath, Lee Panella, Piergiorgio Stevanato
Background: PCR allelic discrimination technologies have broad applications in the detection of single nucleotide polymorphisms (SNPs) in genetics and genomics. The use of fluorescence-tagged probes is the leading method for targeted SNP detection, but assay costs and error rates could be improved to increase genotyping efficiency. A new assay, rhAmp, based on RNase H2-dependent PCR (rhPCR) combined with a universal reporter system attempts to reduce error rates from primer/primer and primer/probe dimers while lowering costs compared to existing technologies...
2018: Plant Methods
Henrik Failmezger, Janne Lempe, Nasim Khadem, Maria Cartolano, Miltos Tsiantis, Achim Tresch
Background: Accurate and automated phenotyping of leaf images is necessary for high throughput studies of leaf form like genome-wide association analysis and other forms of quantitative trait locus mapping. Dissected leaves (also referred to as compound) that are subdivided into individual units are an attractive system to study diversification of form. However, there are only few software tools for their automated analysis. Thus, high-throughput image processing algorithms are needed that can partition these leaves in their phenotypically relevant units and calculate morphological features based on these units...
2018: Plant Methods
Xuehui Yao, Wenchao Zhao, Rui Yang, Jianli Wang, Fukuan Zhao, Shaohui Wang
Background: Guard cell protoplasts (GCPs) isolated from various plants have proven to be especially useful for studies of signal transduction pathways and plant development. But it is not easy to isolate highly purified preparations of large numbers of GCPs from plants. In this research, our focus is on a method to isolate large numbers of guard cells from tomato leaves. The protocols described yield millions of highly purified, viable GCPs, which are also suitable for studies on guard cell physiology...
2018: Plant Methods
Prabu Ravindran, Adriana Costa, Richard Soares, Alex C Wiedenhoeft
Background: The current state-of-the-art for field wood identification to combat illegal logging relies on experienced practitioners using hand lenses, specialized identification keys, atlases of woods, and field manuals. Accumulation of this expertise is time-consuming and access to training is relatively rare compared to the international demand for field wood identification. A reliable, consistent and cost effective field screening method is necessary for effective global scale enforcement of international treaties such as the Convention on the International Trade in Endagered Species (CITES) or national laws (e...
2018: Plant Methods
Duc Quan Nguyen, Andrew L Eamens, Christopher P L Grof
Background: Quantitative real-time polymerase chain reaction (RT-qPCR) is the key platform for the quantitative analysis of gene expression in a wide range of experimental systems and conditions. However, the accuracy and reproducibility of gene expression quantification via RT-qPCR is entirely dependent on the identification of reliable reference genes for data normalisation. Green foxtail ( Setaria viridis ) has recently been proposed as a potential experimental model for the study of C4 photosynthesis and is closely related to many economically important crop species of the Panicoideae subfamily of grasses, including Zea mays (maize), Sorghum bicolor (sorghum) and Sacchurum officinarum (sugarcane)...
2018: Plant Methods
Jingyi Jiang, Alexis Comar, Philippe Burger, Pierre Bancal, Marie Weiss, Frédéric Baret
Background: Leaf biochemical composition corresponds to traits related to the plant state and its functioning. This study puts the emphasis on the main leaf absorbers: chlorophyll a and b ([Formula: see text]), carotenoids ([Formula: see text]), water ([Formula: see text]) and dry mater ([Formula: see text]) contents. Two main approaches were used to estimate [[Formula: see text] [Formula: see text], [Formula: see text], [Formula: see text]] in a non-destructive way using spectral measurements...
2018: Plant Methods
Jose A Fernandez-Gallego, Shawn C Kefauver, Nieves Aparicio Gutiérrez, María Teresa Nieto-Taladriz, José Luis Araus
Background: The number of ears per unit ground area (ear density) is one of the main agronomic yield components in determining grain yield in wheat. A fast evaluation of this attribute may contribute to monitoring the efficiency of crop management practices, to an early prediction of grain yield or as a phenotyping trait in breeding programs. Currently the number of ears is counted manually, which is time consuming. Moreover, there is no single standardized protocol for counting the ears...
2018: Plant Methods
A Fortineau, P Bancal
Background: In plants, three-dimensional (3-D) organs such as inflorescences or fruits carry out photosynthesis and thus play a significant role in carbon assimilation and yield. However, this contribution has been poorly characterized because there is no reliable method for measuring photosynthesis by 3-D organs. One of the major challenges is ensuring the uniform irradiation of samples that are placed within a sealed chamber. Results: In this study, we developed an innovative chamber with homogeneous lighting that can be used to measure photosynthesis by large 3-D organs...
2018: Plant Methods
Zohaib Khan, Vahid Rahimi-Eichi, Stephan Haefele, Trevor Garnett, Stanley J Miklavcic
Background: Unmanned aerial vehicles offer the opportunity for precision agriculture to efficiently monitor agricultural land. A vegetation index (VI) derived from an aerially observed multispectral image (MSI) can quantify crop health, moisture and nutrient content. However, due to the high cost of multispectral sensors, alternate, low-cost solutions have lately received great interest. We present a novel method for model-based estimation of a VI using RGB color images. The non-linear spatio-spectral relationship between the RGB image of vegetation and the index computed by its corresponding MSI is learned through deep neural networks...
2018: Plant Methods
C R Gedling, E M Ali, A Gunadi, J J Finer, K Xie, Y Liu, N Yoshikawa, F Qu, A E Dorrance
Background: Virus induced gene silencing (VIGS) is a powerful genomics tool for interrogating the function of plant genes. Unfortunately, VIGS vectors often produce disease symptoms that interfere with the silencing phenotypes of target genes, or are frequently ineffective in certain plant genotypes or tissue types. This is especially true in crop plants like soybean [ Glycine max (L.) Merr]. To address these shortcomings, we modified the inoculation procedure of a VIGS vector based on Apple latent spherical virus (ALSV)...
2018: Plant Methods
Karthikeyan Alagarsamy, Lubobi Ferdinand Shamala, Shu Wei
Background: Camellia sinensis var. sinensis is widely grown for tea beverages that possess significant health promoting effects. Studies on tea plant genetics and breeding are hindered due to its recalcitrance to Agrobacterium -mediated genetic transformation. Among the possible reasons, oxidation of phenolics released from explant tissues and bactericidal effects of tea polyphenols during the process of transformation play a role in the plant recalcitrance. The aim of the present study was to alleviate the harmful effects of phenolic compounds using in - planta transformation...
2018: Plant Methods
Lingxiao Zhao, Ting Pan, Dongwei Guo, Cunxu Wei
Background: Storage starch in starchy seed influences the seed weight and texture, and determines its applications in food and nonfood industries. Starch granules from different plant sources have significantly different shapes and sizes, and even more the difference exists in the different regions of the same tissue. Therefore, it is very important to in situ investigate the morphology and distribution of starch in the whole seed. However, a simple and rapid method is deficient to prepare the whole section of starchy seed for investigating the morphology and distribution of starch in the whole seeds for a large number of samples...
2018: Plant Methods
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