1887

Battle of the Bacteria: Characterizing the Evolutionary Advantage of Stationary Phase Growth

    Authors: Karin E. Kram1,*, Kristina M. Yim2, Aaron B. Coleman3, Brian K. Sato4,*
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    Affiliations: 1: Department of Biology, California State University, Dominguez Hills, CA 90747; 2: Department of Genetics, Yale University, New Haven, CT 06510; 3: Division of Biological Sciences, University of California, San Diego, CA 92093; 4: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 04 May 2016
    • ©2016 Author(s). Published by the American Society for Microbiology.
    • [open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

    • Supplemental materials available at http://asmscience.org/jmbe
    • *Corresponding authors. Mailing addresses: Brian Sato: 2238 McGaugh Hall MC3900, Irvine, CA 92697. Phone: 949-824-0661. Fax: 949-824-8551. E-mail: [email protected]; Karin Kram: 1000 E. Victoria St. NSM A-137, Carson, CA 90747. Phone: 310-243-1090. Fax: 301-243-2350. E-mail: [email protected].
    Source: J. Microbiol. Biol. Educ. May 2016 vol. 17 no. 2 261-268. doi:10.1128/jmbe.v17i2.981
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    Abstract:

    Providing students with authentic research opportunities has been shown to enhance learning and increase retention in STEM majors. Accordingly, we have developed a novel microbiology lab module, which focuses on the molecular mechanisms of evolution in , by examining the growth advantage in stationary phase (GASP) phenotype. The GASP phenotype is demonstrated by growing cells into long-term stationary phase (LTSP) and then competing them against un-aged cells in a fresh culture. This module includes learning goals related to strengthening practical laboratory skills and improving student understanding of evolution. In addition, the students generate novel data regarding the effects of different environmental stresses on GASP and the relationship between evolution, genotypic change, mutation frequency, and cell stress. Pairs of students are provided with the experimental background, select a specific aspect of the growth medium to modify, and generate a hypothesis regarding how this alteration will impact the GASP phenotype. From this module, we have demonstrated that students are able to achieve the established learning goals and have produced data that has furthered our understanding of the GASP phenotype. Journal of Microbiology & Biology Education

Key Concept Ranking

Bacterial Growth Phases
0.54399544
Stationary Phase
0.4794331
0.54399544

References & Citations

1. Alters BJ, Nelson CE, Mitton J 2002 Perspective: teaching evolution in higher education Evolution 56 1891 1901 10.1111/j.0014-3820.2002.tb00115.x 12449476 http://dx.doi.org/10.1111/j.0014-3820.2002.tb00115.x
2. Crowe A, Dirks C, Wenderoth MP 2008 Biology in bloom: implementing Bloom’s taxonomy to enhance student learning in biology CBE Life Sci Educ 7 368 381 10.1187/cbe.08-05-0024 19047424 2592046 http://dx.doi.org/10.1187/cbe.08-05-0024
3. Emmert EAB ASM Task Committee on Laboratory Biosafety 2013 Biosafety guidelines for handling microorganisms in the teaching laboratory: development and rationale J Microbiol Biol Educ 14 78 83 10.1128/jmbe.v14i1.531 23858356 3706168 http://dx.doi.org/10.1128/jmbe.v14i1.531
4. Farrell MJ, Finkel SE 2003 The growth advantage in stationary-phase phenotype conferred by rpos mutations is dependent on the ph and nutrient environment J Bacteriol 185 7044 7052 10.1128/JB.185.24.7044-7052.2003 14645263 296246 http://dx.doi.org/10.1128/JB.185.24.7044-7052.2003
5. Finkel SE 2006 Long-term survival during stationary phase: evolution and the GASP phenotype Nat Rev Microbiol 4 113 120 10.1038/nrmicro1340 16415927 http://dx.doi.org/10.1038/nrmicro1340
6. Howard GS 1980 Response-shift bias: a problem in evaluating interventions with pre/post self-reports Eval Rev 4 93 106 10.1177/0193841X8000400105 http://dx.doi.org/10.1177/0193841X8000400105
7. Krathwohl DR 2002 A revision of Bloom’s taxonomy: an overview Theory Pract 41 212 218 10.1207/s15430421tip4104_2 http://dx.doi.org/10.1207/s15430421tip4104_2
8. Luria SE, Delbrück M 1943 Mutations of bacteria from virus sensitivity to virus resistance Genetics 28 491 511 17247100 1209226
9. Miller JD, Scott EC, Okamoto S 2006 Public acceptance of evolution Science 313 765 766 10.1126/science.1126746 16902112 http://dx.doi.org/10.1126/science.1126746
10. Sato BK 2013 Attack of the killer fungus: a hypothesisdriven lab module J Microbiol Biol Educ 14 230 237 10.1128/jmbe.v14i2.612 http://dx.doi.org/10.1128/jmbe.v14i2.612
11. Sato BK, Alam U, Dacanay SJ, Lee AK, Shaffer JF 2015 Brewing for students: an inquiry-based microbiology lab J Microbiol Biol Educ 16 2 223 229 10.1128/jmbe.v16i2.914 http://dx.doi.org/10.1128/jmbe.v16i2.914
12. Smith M 2010 Current status of research in teaching and learning evolution: II. pedagogical issues Sci Educ 19 539 571 10.1007/s11191-009-9216-4 http://dx.doi.org/10.1007/s11191-009-9216-4

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2016-05-04
2019-01-18

Abstract:

Providing students with authentic research opportunities has been shown to enhance learning and increase retention in STEM majors. Accordingly, we have developed a novel microbiology lab module, which focuses on the molecular mechanisms of evolution in , by examining the growth advantage in stationary phase (GASP) phenotype. The GASP phenotype is demonstrated by growing cells into long-term stationary phase (LTSP) and then competing them against un-aged cells in a fresh culture. This module includes learning goals related to strengthening practical laboratory skills and improving student understanding of evolution. In addition, the students generate novel data regarding the effects of different environmental stresses on GASP and the relationship between evolution, genotypic change, mutation frequency, and cell stress. Pairs of students are provided with the experimental background, select a specific aspect of the growth medium to modify, and generate a hypothesis regarding how this alteration will impact the GASP phenotype. From this module, we have demonstrated that students are able to achieve the established learning goals and have produced data that has furthered our understanding of the GASP phenotype. Journal of Microbiology & Biology Education

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Figures

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FIGURE 1

Timeline for the four experiments in the GASP module. The module was implemented in a lab course that met twice a week (Lab Period A and B). Lab Period A was three hours in duration while Period B was one hour. Each activity during lab is associated with one of the four specific experiments. (1) Growth of cells into long-term stationary phase and GASP assay, (2) Examination of RpoS activity, (3) Cell stress measurement, (4) Examination of mutation frequency. Specific times required for each activity are indicated on the figure in minutes.

Source: J. Microbiol. Biol. Educ. May 2016 vol. 17 no. 2 261-268. doi:10.1128/jmbe.v17i2.981
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Image of FIGURE 2

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FIGURE 2

Examples of student data illustrating the GASP phenotype. (A) Dilution plating on selective media. Cells are from the co-culture experiment (started in week three of the module). Day 0 refers to the initial culture inoculation and day 7 refers to one week of co-culture growth. Aged cells are distinguished from un-aged cells by the presence of distinct antibiotic resistance markers present in each strain. Cultures are plated with ten-fold serial dilutions. (B) Colony forming units (CFU) per ml of culture are calculated from the dilution plates and graphed on a plot of CFU/ml versus time. The GASP phenotype is illustrated by greater survival values of aged cells over time compared to the un-aged population in the culture. Growth in different types of media can impact the GASP phenotype.

Source: J. Microbiol. Biol. Educ. May 2016 vol. 17 no. 2 261-268. doi:10.1128/jmbe.v17i2.981
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FIGURE 3

Assessments confirm that students achieved the module learning objectives. (A) A 12-question pre-/post-test was administered before and after the GASP module in laboratory sections during Fall quarter 2014 and Winter quarter 2015 ( 197 students combined). Performance on each question (Q) and the corresponding learning objective (LO) it assessed is indicated. All post-test gains are statistically significant ( 0.001 by -test) with the exception of question 4. (B) Students noted their agreement with the indicated statements on a 5 -point Likert scale (5 = strongly agree, 1 = strongly disagree) upon completion of the module. They were asked to state their current agreement and provide a retroactive agreement (the pre-module data). Post-test gains are statistically significant ( 0.001 by chi-square test). Questions were asked using the iClicker response system ( 190 students). (C) Student performance on the pipetting test before and after module completion ( 198 students). Passing refers to whether a student’s dilution fell within 0.050 OD units from a standard value obtained by the course instructor who performed the same dilutions multiple times. The number of students who passed post-module was significantly greater than pre-module ( 0.001 by -test). (D) Student performance on exam questions categorized by whether or not they were GASP module-specific and by Bloom’s level (Bloom’s 2 versus 3, 4, and 5) ( 201 students). The difference between GASP and Other questions of Bloom’s level 3, 4, and 5 was statistically significant ( 0.001 by -test). Question numbers in each category were GASP (BL2) = 5, Other (BL2) = 10, GASP (BL3, 4, 5) = 19, Other (BL3, 4, 5) = 38.

Source: J. Microbiol. Biol. Educ. May 2016 vol. 17 no. 2 261-268. doi:10.1128/jmbe.v17i2.981
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