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