Overexpressed GLK Roots
Light ExperimentPurpose: Find out how light interacts with the transcription
factor. Once the roots were sub-cultured in the different light
settings and sub-cultured once again after 3 weeks, less than a
week later the roots started to turn green.
Cloning ProcessPurpose: Move the gene of interest into the agrobacterium to perform
the Virus Induced Gene Silencing experiment.
Research Experiences for Undergraduates Pathways Opening World Energy Resources" (REU - POWER)
Bradley Lehman, Principal Investigator
This work was supported by the National Science Foundation
Grant # 1757650
Enhancing Photosynthetic Efficiency of Plants
for Biomass and Biofuel ApplicationsJessica Figueroa, REU student, Northern Essex Community College
Lauren Cole, Ph.D. Student, Northeastern University
Carolyn Lee-Parsons, Chemical Engineering Department, Northeastern University
IntroductionThis research was conducted in order to enhance
photosynthetic efficiency of plants for biomass and biofuel
production, and reduces the need for fossil fuel. The reason
plant are being looked into for biomass and biofuel
applications is because the more chlorophyll a plant has the
more sugar it makes. By increasing the sugar content it will
provide additional energy to produce additional biomass for
a higher yield content of biofuel products without increasing
the amount of plants needed to do so, hence making the
plant more efficient.
Project Overview
Virus Induced Gene Silencing
(VIGS) ExperimentPurpose: Silence the gene of interest and learn if it takes part in the
chloroplast development.
ConclusionI was able to determine that GLK is a transcription factor in
chloroplast development. I successfully accomplished:
Cloning transcription factors (TF)
Demonstrated decreased chlorophyll content in GLK-silenced
plants
Found increased photosynthetic efficiency in GLK- overexpressed
roots
Determined the influence of light on GLK-induced chlorophyll
development
Steps ForwardDetermine the role of GNC and Hy5 in chlorophyll development:
Measure the phenotype of the Hy-5 VIGS plants
Perform VIGS experiment with GNC
Overexpress the transcription factors in roots to see if
chloroplast development increases
Develop transgenic plants
Ultimately, grow higher efficiency plants
AcknowledgementsI would like to give special thanks to Professor Carolyn Lee-
Parsons, especially to my mentor Lauren Cole, who really made
this a truly enjoyable experience. Also, I would like to thank Brad
Lehman, Principal investigator of the REU – Power, and Claire
Duggan, Director of The Center for STEM Education.
Amplify whole gene from cDNA
BP reaction to move PCR product into pDONR221 plasmid
Transform E. Coli
Colony PCR (3 colonies per construct)
Start liquid cultures
Miniprep kit
PCR amplify VIGS fragment from pDONR221 (GNC, Hy5, GLK)
One pot BP/LR reaction to form pTRV2 plasmid w/ VIGS fragment
Transform E. Coli
Colony PCR 3 colonies per construct (pTRV2 seq. & Fwd.
primer)
Liquid cultures of E. Coli get started
Miniprep E. Coli
Prepare for sequencing with difficult template
Analyze sequencing results
Electroporate into agro
Colony PCR
Start liquid cultures
Perform VIGS experiment
Only Hy5-V1 worked
for me
High
Lo
w
Medium
Overexpressed GLK in LowNegative Control in Dark
Clone plasmid for
VIGS
Overexpress gene in
roots to learn if
chloroplast
development increases
Higher efficiency solar
panel plants!
Amplify candidate
gene sequence
VIGS phenotype:
Discover gene
function to find if
regulates
chloroplast
development and
photosynthesis
Develop transgenic
plant
