Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience

In this investigation, the students’ goal was to isolate and characterize Agrobacterium strains from soil. Following selection and enrichment on 1A-t medium, putative Agrobacterium isolates were characterized by Gram stain reaction and biochemical tests. Isolates were further evaluated using polymer...

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Autores principales:	Kim R. Finer, Lee Fox, John J. Finer
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/51417f374a254d8180c6c5ecff5552a5
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spelling	oai:doaj.org-article:51417f374a254d8180c6c5ecff5552a52021-11-15T15:13:57ZIsolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience10.1128/jmbe.v17i3.11241935-78851935-7877https://doaj.org/article/51417f374a254d8180c6c5ecff5552a52016-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.v17i3.1124https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885In this investigation, the students’ goal was to isolate and characterize Agrobacterium strains from soil. Following selection and enrichment on 1A-t medium, putative Agrobacterium isolates were characterized by Gram stain reaction and biochemical tests. Isolates were further evaluated using polymerase chain reaction (PCR) with different primer sets designed to amplify specific regions of bacterial deoxyribonucleic acid (DNA). Primer sets included AGRH to identify isolates that were members of the Rhizobiaceae, BIOVAR1 primers to identify members of Agrobacterium biovar group I, and a third set, VIRG, to determine presence of virG (only present in pathogenic Agrobacterium strains). During the investigation, students applied previously learned techniques including serial dilution, use of selective/differential media, staining protocols, biochemical analysis, molecular analysis via PCR, and electrophoresis. Students also gained practical experience using photo documentation to record data for an eventual mock journal publication of the capstone laboratory experience. Pre- and post-evaluation of class content knowledge related to the techniques, protocols, and learning objectives of these laboratories revealed significant learning gains in the content areas of Agrobacterium–plant interactions (p ≤ 0.001) and molecular biology (p ≤ 0.01). The capstone journal assignment served as the assessment tool to evaluate mastery and application of laboratory technique, the ability to accurately collect and evaluate data, and critical thinking skills associated with experimental troubleshooting and extrapolation. Analysis of journal reports following the capstone experience showed significant improvement in assignment scores (p ≤ 0.0001) and attainment of capstone experience learning outcomes.Kim R. FinerLee FoxJohn J. FinerAmerican Society for MicrobiologyarticleSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 17, Iss 3, Pp 444-450 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	Special aspects of education LC8-6691 Biology (General) QH301-705.5 Kim R. Finer Lee Fox John J. Finer Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
description	In this investigation, the students’ goal was to isolate and characterize Agrobacterium strains from soil. Following selection and enrichment on 1A-t medium, putative Agrobacterium isolates were characterized by Gram stain reaction and biochemical tests. Isolates were further evaluated using polymerase chain reaction (PCR) with different primer sets designed to amplify specific regions of bacterial deoxyribonucleic acid (DNA). Primer sets included AGRH to identify isolates that were members of the Rhizobiaceae, BIOVAR1 primers to identify members of Agrobacterium biovar group I, and a third set, VIRG, to determine presence of virG (only present in pathogenic Agrobacterium strains). During the investigation, students applied previously learned techniques including serial dilution, use of selective/differential media, staining protocols, biochemical analysis, molecular analysis via PCR, and electrophoresis. Students also gained practical experience using photo documentation to record data for an eventual mock journal publication of the capstone laboratory experience. Pre- and post-evaluation of class content knowledge related to the techniques, protocols, and learning objectives of these laboratories revealed significant learning gains in the content areas of Agrobacterium–plant interactions (p ≤ 0.001) and molecular biology (p ≤ 0.01). The capstone journal assignment served as the assessment tool to evaluate mastery and application of laboratory technique, the ability to accurately collect and evaluate data, and critical thinking skills associated with experimental troubleshooting and extrapolation. Analysis of journal reports following the capstone experience showed significant improvement in assignment scores (p ≤ 0.0001) and attainment of capstone experience learning outcomes.
format	article
author	Kim R. Finer Lee Fox John J. Finer
author_facet	Kim R. Finer Lee Fox John J. Finer
author_sort	Kim R. Finer
title	Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
title_short	Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
title_full	Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
title_fullStr	Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
title_full_unstemmed	Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience
title_sort	isolation and characterization of <italic toggle="yes">agrobacterium</italic> strains from soil: a laboratory capstone experience
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/51417f374a254d8180c6c5ecff5552a5
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Isolation and Characterization of <italic toggle="yes">Agrobacterium</italic> Strains from Soil: A Laboratory Capstone Experience

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