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Hypochlorous Acid Staining with R19-S in the Drosophila Intestine upon Ingestion of Opportunistic Bacteria

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I Plate-based Assay for Studying How Fungal Volatile Compounds (VCs) Affect Plant Growth and Development and the Identification of VCs via SPME-GC-MS

Authors: Wenzhao Wang, Ningxiao Li, Xingzhong Liu and Seogchan Kang, date: 02/20/2019, view: 903, Q&A: 0

[Abstract] Biogenic volatile compounds (VCs) mediate various types of crucial intra- and inter-species interactions in plants, animals, and microorganisms owing to their ability to travel through air, liquid, and porous soils. To study how VCs produced by Verticillium dahliae, a soilborne fungal pathogen, affect plant growth and development, we ...

Soluble and Solid Iron Reduction Assays with Desulfitobacterium hafniense

Authors: Lucrezia Comensoli, Julien Maillard, Wafa M. Kooli, Pilar Junier and Edith Joseph, date: 09/05/2018, view: 1033, Q&A: 0

[Abstract] There is a pressing need to develop sustainable and efficient methods to protect and stabilize iron objects. To develop a conservation-restoration method for corroded iron objects, this bio-protocol presents the steps to investigate reductive dissolution of ferric iron and biogenic production of stabilizing ferrous iron minerals in the strict ...

Enzymatic Activity Assay for Invertase in Synechocystis Cells

Authors: Xiaoming Tan, Kuo Song and Xuefeng Lu, date: 05/20/2018, view: 1874, Q&A: 0

[Abstract] Invertase can catalyze the hydrolysis of sucrose, and is widely distributed in cells of cyanobacteria and plants. Being responsible for the first step for sucrose metabolism, invertase plays important physiological roles and its enzymatic activity is frequently needed to be determined. All the methods for determination of the invertase activity ...

Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells

Authors: Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 1865, Q&A: 0

[Abstract] Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...

Determination of Intracellular Osmolytes in Cyanobacterial Cells

Authors: Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 1667, Q&A: 0

[Abstract] Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...

Flow Cytometric Quantification of Fatty Acid Uptake by Mycobacterium tuberculosis in Macrophages

Authors: Evgeniya V. Nazarova, Maria Podinovskaia, David G. Russell and Brian C. VanderVen, date: 02/20/2018, view: 3095, Q&A: 0

[Abstract] Mycobacterium tuberculosis (Mtb) has evolved to assimilate fatty acids from its host. However, until recently, there was no reliable way to quantify fatty acid uptake by the bacteria during host cell infection. Here we describe a new method to quantify fatty acid uptake by intracellular bacilli. We infect macrophages with Mtb ...

Determination of Mn Concentrations in Synechocystis sp. PCC6803 Using ICP-MS

Authors: Fabian Brandenburg, Hanan Schoffman, Nir Keren and Marion Eisenhut, date: 12/05/2017, view: 2844, Q&A: 0

[Abstract] Manganese (Mn) is an essential micronutrient for all photoautotrophic organisms. Two distinct pools of Mn have been identified in the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), with 80% of the Mn residing in the periplasm and 20% in cytoplasm and thylakoid lumen (Keren et al., 2002). In this protocol, we ...

Quantification of Bacterial Polyhydroxybutyrate Content by Flow Cytometry

Authors: Antonio Lagares (Jr.) and Claudio Valverde, date: 12/05/2017, view: 2640, Q&A: 0

[Abstract] We describe here a detailed protocol for the quantification of the intracellular content of polyhydroxybutyrate (PHB) in a population of bacterial cells by flow cytometry, which is based on a consensus of several previously reported works.

Sterol Analysis in Kluyveromyces lactis

Authors: Yvetta Gbelska, Nora Toth Hervay, Marcela Morvova and Alexandra Konecna, date: 09/05/2017, view: 2870, Q&A: 0

[Abstract] Sterols are essential lipids of most eukaryotic cells with multiple functions (structural, regulatory and developmental). Sterol profile of yeast cells is often determined during the studies of ergosterol synthesis mutants used to uncover a number of functions for various sterols in yeast cells. Molecular studies of ergosterol biosynthesis have ...

A Protocol of Using White/Red Color Assay to Measure Amyloid-induced Oxidative Stress in Saccharomyces cerevisiae

Authors: Vidhya Bharathi, Amandeep Girdhar and Basant K Patel, date: 08/05/2017, view: 4259, Q&A: 0

[Abstract] The yeast Saccharomyces cerevisiae (S. cerevisiae) harboring ade1 or ade2 mutations manifest red colony color phenotype on rich yeast medium YPD. In these mutants, intermediate metabolites of adenine biosynthesis pathway are accumulated. Accumulated intermediates, in the presence of reduced glutathione, are ...

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