The Effect of Silver Nanoparticles on Plant Growth and Herbivory

Author Profile

Erin Rehrig, Fitchburg State University

Location: Massachusetts

Abstract

In this CURE, during labratories, students will conduct experiments to determine the effects of silver nanoparticles on plant growth and insect herbivory. Students will synthesize their own nanoparticles and use them to treat Arabidopsis thaliana plants. Plants growth will be monitored weekly using digital photography. After 5 weeks, insects (Pieris rapae, caterpillars) will be placed on plants for 48 hours and insect herbivory levels will be assessed across different treatments. Insects will be weighed before and after feeding assays. Insect herbivory will also be monitored using digital photography. Image analysis for both growth rate calculations and herbivory measurements will be done using MatLab.

Student Goals

  1. Conduct a well-controlled study that builds on prior knowledge and makes progress toward a research goal
  2. Interpret and evaluate collected data and make decisions about whether experiments need to be modified or redone to draw conclusions.
  3. Communicate about research results and progress orally and in writing

Research Goals

  1. Assess whether the addition of silver nanoparticles affects Arabidopsis growth rates
  2. Assess whether the addition of silver nanoparticles affects Arabidopsis resistance to herbivores.

Context

This CURE is part of a senior-level Plant Biology capstone course. The CURE will be conducted in the laboratory portion of the course over 15 week semester in the fall of 2019. Approximately 12-16 students will be enrolled in the course. Students will have taken several core biology courses, including Genetics, Ecology, and General Biology I and II before being able to register for this course.

Target Audience:Major, Upper Division
CURE Duration:A full term

CURE Design

The research theme at the heart of the CURE is silver nanoparticles and plant physiology. Little is known about the effects of silver nanoparticles on plant growth and physiology. Silver nanoparticles are used in many medical products and can contaminate waterways and holding tanks. Students are responsible for synthesizing their own nanoparticles as part of a group. Students will also be using their own phones to take digital photos of plants as they grow and before and after insect treatment and will be using digital image analysis to analyze data. We will be collaborating with Dr. Emma Downs from the Dept. of Chemistry and Dr. Catherine Buelle from the Dept. of Mathematics.

The plant biology scientific community would be interested in this work. It is important to understand how potential environmental pollutants affect crop/plant growth and their susceptibility to insects. Additionally, it is important to develop non-destructive methods for measuring plant growth, phenotypes, and herbivory rates that students can use for collecting and analyzing data.

Core Competencies:
Nature of Research:Basic Research

Tasks that Align Student and Research Goals

Research Goals →
Student Goals ↓
Research Goal 1: Assess whether the addition of silver nanoparticles affects Arabidopsis growth rates
Research Goal 2: Assess whether the addition of silver nanoparticles affects Arabidopsis resistance to herbivores.

Student Goal 1: Design a well-controlled study that builds on prior knowledge and makes progress toward a research goal

By the end of this CURE, students will be able to:

-Summarize current, general knowledge about the role of nanoparticles on plant physiology including growth and plant herbivory
-Use Google Scholar to find at least 8 papers published within the past five years on the study of nanoparticles and growth in plants
-Compare destructive vs. non-destructive phenotyping
-Identify appropriate control conditions
-Create a timeline for experiments, including how and when plants will be grown, and when pictures will be taken
-Synthesize nanoparticles according to established protocol.
-Collect data using high-quality digital photography (ie. Cell phones and photobox)
-Design and keep a detailed, neat lab notebook

-Use Google Scholar to find ANY papers published within the past five years on the study of nanoparticles and herbivory in plants
-Compare destructive vs. non-destructive phenotyping
-Identify appropriate control conditions
-Create a timeline for their experiment, including how and when herbivory assays will take place and how data will be collected.
-Collect data using high-quality digital photography (ie. Cell phones and photobox) of plants before and after insect feeding.
-Explain how to rear and handle insect herbivores
-Measure insect weight before and after feeding.


Student Goal 2: Interpret and evaluate data collected and make decisions about whether experiments need to be modified or redone to draw conclusions.

-Compare data with other groups
-Propose and defend the next steps for their experiments based on initial data collected.
-Use Matlab software to analyze digital photographs of plants during the experiments using a manual
-Determine the appropriate statistical tests that should be run based on the data collected (ie. Understand differences between categorical vs. continuous data sets, etc.)
-Calculate growth rates for each plant treatment groups
-Create graphs using Microsoft excel and show the sources of error/variation
-Work in collaborative groups (with help from the instructor on delegating tasks, sharing information, and resolving student conflicts)
-Propose additional experiments that can be done to continue this area of research.

-Propose and defend the next steps for their experiments based on initial data collected from the insect trials.
-Use Matlab software to analyze digital photographs of plants during the experiments using a manual
-Determine the appropriate statistical tests that should be run based on the data collected (ie. Understand differences between categorical vs. continuous data sets, etc.)
-Create graphs using Microsoft excel and show the sources of error/variation
-Work in collaborative groups (with help from the instructor on delegating tasks, sharing information, and resolving student conflicts)
-Propose additional experiments that can be done to continue this area of research.


Student Goal 3: Communicate about research progress orally and in writing

-Create an short literature review (introductory section) using of 5 primary papers most relevant to our research goal (can use "Article summary sheets" as templates)
-Summarize the current status of an experiment in weekly meetings about the research and issues or problems and how they were addressed
-Write a final lab report (individual) using data collected by the entire class.
-Prepare a final poster presentation about the research to audiences outside class (for your portfolios)

-Create an short literature review (introductory section) using 5 primary papers most relevant to our research goal (can use "Article summary sheets" as templates)
-Write a final lab report (individual) using data collected by the entire class.
-Prepare a final poster presentation about the research to audiences outside class (for your portfolios)
-Work in collaborative groups to solve problems


Instructional Materials

Plant Biology CURE Syllabus (Microsoft Word 2007 (.docx) 79kB Jun5 19)
Silver Nanoparticle Handout (Microsoft Word 2007 (.docx) 129kB Jun5 19)
Measuring Damage in Plants (Microsoft Word 2007 (.docx) 594kB Jun5 19)

Assessment

Lab Report Prompt & Rubric (Microsoft Word 2007 (.docx) 1.8MB Jun5 19)
Lab Report Rubric Score Calculator (Excel 2007 (.xlsx) 10kB Jun5 19)
Lab Notebook Prompt & Rubric (Microsoft Word 2007 (.docx) 234kB Jun5 19)

Instructional Staffing

My colleagues and collaborators will come to my lab and instruct the students how to synthesize the nanoparticles and use the digital image software. I will work with our lab coordinators and give them a supplies list before the semester starts so all the supplies will be ready. The rest of the equipment is already in my lab.

Author Experience

Erin Rehrig, Fitchburg State University

This CURE is designed to teach students in an Advanced Plant Biology course the tools and skills need to conduct novel research based on a Silver Nanoparticle Project that is important to my collaborators and myself.


Advice for Implementation

Give students more time to do data analysis and make mistakes. Don't try to fill up a 3-hour lab with "stuff" as it takes a long time to do things.

Iteration

Every week for 5 weeks, students will be using the same recipe to make nanoparticles, treat plants, take digital photos and analyze data. If something looks "funky", they can redo it and then compare their data to other groups. Other cognate lab activities include hands-on lessons in experimental design, data analysis, statistics, and creating damage rubrics for plants.

Using CURE Data

This research question and overall nanoparticle project including nanoparticle synthesis, plant experiments, and image analysis are the intellectual property of my colleagues and me. Students will be helping us collect data. Because of the sheer number of plants and insects that need to be reared for this type of study, it requires lots of replication. Student groups will meet with me to discuss their data during weekly lab periods. If any publications are written, students will be acknowledged, but their contributions do not warrant authorship.

Resources

1. Anjum, N. A., Gill, S. S., Duarte, A. C., Pereira, E., & Ahmad, I. (2013). Silver nanoparticles in soil–plant systems. Journal of Nanoparticle Research, 15(9), 1896.

2. Green, J. M., Appel, H., Rehrig, E. M., Harnsomburana, J., Chang, J. F., Balint-Kurti, P., & Shyu, C. R. (2012). PhenoPhyte: a flexible affordable method to quantify 2D phenotypes from imagery. Plant Methods, 8(1), 45.

3. Harris, A. T., & Bali, R. (2008). On the formation and extent of uptake of silver nanoparticles by live plants. Journal of Nanoparticle Research, 10(4), 691-695.

4. Mulfinger, L., Solomon, S. D., Bahadory, M., Jeyarajasingam, A. V., Rutkowsky, S. A., & Boritz, C. (2007). Synthesis and study of silver nanoparticles. Journal of chemical education, 84(2), 322.

5. Spalding, E. P., & Miller, N. D. (2013). Image analysis is driving a renaissance in growth measurement. Current Opinion in Plant Biology, 16(1), 100-104.

6. Tackenberg, O. (2007). A new method for non-destructive measurement of biomass, growth rates, vertical biomass distribution and dry matter content based on digital image analysis. Annals of botany, 99(4), 777-783.

7. Wang, J., Koo, Y., Alexander, A., Yang, Y., Westerhof, S., Zhang, Q., ... & Alvarez, P. J. (2013). Phytostimulation of poplars and Arabidopsis exposed to silver nanoparticles and Ag+ at sublethal concentrations. Environmental science & technology, 47(10), 5442-5449.

8. Zuverza-Mena, N., Armendariz, R., Peralta-Videa, J. R., & Gardea-Torresdey, J. L. (2016). Effects of silver nanoparticles on radish sprouts: root growth reduction and modifications in the nutritional value. Frontiers in plant science, 7, 90.