SA
Reviewer
Shahin S. Ali
  • Research scientist, USDA/ARS Sustainable Perennial Crops Lab
Research fields
  • Plant science
Design and Construction of Repacked Soil Columns for Measuring Solute Transport, Plant Growth and Soil Biota
Authors:  Israel Ikoyi and Achim Schmalenberger, date: 01/20/2021, view: 2828, Q&A: 0

Researchers face a number of challenges in the construction of soil columns which can affect the outcome of their experiments. The use of intact soil cores closely mimics actual field conditions. However, the excavation of large intact soil cores is a time-consuming, labor-intensive process and may lead to soil compaction that would influence the solute transport behavior of the soil column. Repacked soil columns are used as an option to circumvent these challenges of intact soil cores. However, repacked soil columns also have their limitations and introduce other challenges. Here, we present a step by step procedure for the design of repacked soil columns to achieve a realistic bulk density, prevent preferential flow paths, and ensure hydraulic connectivity between soil layers. This protocol will be beneficial to Soil Scientists, Hydrologists and other Environmental Scientists utilizing repacked soil columns.

Single and Multiplexed Gene Editing in Ustilago maydis Using CRISPR-Cas9
The smut fungus Ustilago maydis is an established model organism for elucidating how biotrophic pathogens colonize plants and how gene families contribute to virulence. Here we describe a step by step protocol for the generation of CRISPR plasmids for single and multiplexed gene editing in U. maydis. Furthermore, we describe the necessary steps required for generating edited clonal populations, losing the Cas9 containing plasmid, and for selecting the desired clones.
A Reliable Assay to Evaluate the Virulence of Aspergillus nidulans Using the Alternative Animal Model Galleria mellonella (Lepidoptera)
The greater wax moth Galleria mellonella has emerged as an effective heterologous host to study fungal pathogenesis and the efficacy of promising antifungal drugs (Mylonakis et al., 2005; Li et al., 2013). Here, a methodology describing the Aspergillus nidulans infection in G. mellonella larvae, along with insect survival analysis, is reported. This protocol allowed the distinction between virulent A. nidulans strains (such as TNO2A3), which induced high larval mortality rates, to those in which gene deletion was accompanied by reduced pathogenicity such as ∆gcsA and ∆sdeA (Fernandes et al., 2016).
Automated Tracking of Root for Confocal Time-lapse Imaging of Cellular Processes
Authors:  Mehdi Doumane, Claire Lionnet, Vincent Bayle, Yvon Jaillais and Marie-Cécile Caillaud, date: 04/20/2017, view: 10131, Q&A: 0
Here we describe a protocol that enables to automatically perform time-lapse imaging of growing root tips for several hours. Plants roots expressing fluorescent proteins or stained with dyes are imaged while they grow using automatic movement of the microscope stage that compensates for root growth and allows to follow a given region of the root over time. The protocol makes possible the image acquisition of multiple growing root tips, therefore increasing the number of recorded mitotic events in a given experiment. The protocol also allows the visualization of more than one fluorescent protein or dye simultaneously, using multiple channel acquisition. We particularly focus on imaging of cytokinesis in Arabidopsis root tip meristem, but this protocol is also suitable to follow root hair growth, pollen tube growth, and other regions of root over time, in various plant species. It may as well be amenable to automatically track non-plant structures with an apical growth.
Bioassay of Xanthomonas albilineans Attachment on Sugarcane Leaves
Authors:  Imène Mensi, Jean-Heinrich Daugrois and Philippe Rott, date: 01/20/2017, view: 8921, Q&A: 0
Sugarcane (interspecific hybrids of Saccharum species) is an economically important crop that provides 70% of raw table sugar production worldwide and contributes, in some countries, to bioethanol and electricity production. Leaf scald, caused by the bacterial plant pathogen Xanthomonas albilineans, is one of the major diseases of sugarcane. Dissemination of X. albilineans is mainly ensured by contaminated harvesting tools and infected stalk cuttings. However, some strains of this pathogen are transmitted by aerial means and are able to survive as epiphytes on the sugarcane phyllosphere before entering the leaves and causing disease. Here we present a protocol to estimate the capacity of attachment of X. albilineans to sugarcane leaves. Tissue-cultured sugarcane plantlets were immersed in a bacterial suspension of X. albilineans and leaf attachment of X. albilineans was determined by two methods: leaf imprinting (semi-quantitative method) and leaf washing/homogenization (quantitative method). These methods are important tools for evaluating pathogenicity of strains/mutants of the sugarcane leaf scald pathogen.
Determination of Intracellular ATP Levels in Mycelium of Fusarium oxysporum
Authors:  Carmen Ruiz-Roldan and M. Isabel G. Roncero, date: 07/20/2016, view: 9137, Q&A: 0
Glycolysis provides metabolites for energy production via oxidative phosphorylation during vegetative growth of Fusarium oxysporum. Therefore, determination of intracellular ATP levels might be of valuable help to analyze regulation of glycolysis/gluconeogenesis pathways. The protocol described here can be applied to other filamentous fungi.
Determination of Intra- and Extracellular Glucose in Mycelium of Fusarium oxysporum
Authors:  Carmen Ruiz-Roldan and M. Isabel G. Roncero, date: 07/20/2016, view: 10039, Q&A: 0
To study alterations in the metabolism and/or in the transport of glucose during Fusarium oxysporum vegetative growth, we determined intracellular glucose levels in different fungal strains, as well as the amount of glucose remaining in the supernatants after growth in synthetic medium (SM) supplemented with either 0.05 or 2.5% glucose. We used the Glucose (GO) Assay Kit (Sigma-Aldrich) following the instructions of the manufacturer with some modifications. The protocol described here can be applied to other filamentous fungi.
A Reliable Method for Phytophthora cajani Isolation, Sporangia, Zoospore Production and in Planta Infection of Pigeonpea
Authors:  Mamta Sharma and Raju Ghosh, date: 01/20/2016, view: 11842, Q&A: 0
Pigeonpea (Cajanus cajan L.) is an important legume crop of rainfed agriculture. High levels of protein in pigeonpea make it a valuable protein source for developing countries. Phytophthora blight caused by Phytophthora cajani (P. cajani) is a potential threat to pigeonpea (Cajanus cajan L.) production, affecting the crop irrespective of cropping system, cultivar grown and soil types (Pande et al., 2011; Sharma et al., 2006). The primary mode of infection of P. cajani is sporangium and zoospore. Therefore, sensitive and reliable methods for zoospore production and estimating infection severity are desirable in case of Phytophthora blight of pigeonpea (Sharma et al., 2015). Here we present a protocol for isolation of P. cajani from infected plants, sporangia and zoospore production and in planta infection technique of pigeonpea seedlings. These methods will be important tool to devise a platform for rapid and reliable screening against Phytophthora blight disease of pigeonpea as well as for host x pathogen x environment interaction studies.
We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.