Metabolic Engineering of Physcomitrium patens (moss) for Oleic Acid-rich Vegetable Oil Production

Authors' Affiliations

Jyoti Behera Ranjan, Department of Biological Sciences, East Tennessee State University, TN.

Location

D.P. Culp Center Ballroom

Start Date

4-5-2024 9:00 AM

End Date

4-5-2024 11:30 AM

Poster Number

150

Name of Project's Faculty Sponsor

Aruna Kilaru

Faculty Sponsor's Department

Biological Sciences

Classification of First Author

Graduate Student-Master’s

Competition Type

Competitive

Type

Poster Presentation

Presentation Category

Science, Technology and Engineering

Abstract or Artist's Statement

ABSTRACT Triacylglycerols (TAGs), the predominant storage lipids in plants, are integral to vegetable oil composition. The anticipated surge in vegetable oil demand by 2030 necessitates sustainable production strategies. Oleic acid, a vital omega-9 monounsaturated fatty acid with diverse industrial applications, faces challenges in current production methods using high oleic plant oils. Extended growth cycles and climatic limitations highlight the need for sustainable alternatives. Preliminary results in Nicotiana benthamiana leaves show a significant increase in TAG content, particularly oleic acid, through co-expression of avocado (Persea americana) WRINKLED genes (PaWRI1 and PaWRI2) and enzymes diacylglycerol acyltransferase 1 (PaDGAT1) and phospholipid: diacylglycerol acyltransferase 1(PaPDAT1). This study aims to metabolically engineer Physcomitrium patens (moss) to improve oil production in moss, mostly TAG, which has a higher proportion of oleic acid through stable expression of avocado WRINKLED transcription factors and enzymes PaWRI1, PaWRI2, PaDGAT1, and PaPDAT1. P. patens, a non-seed plant model organism, offers advantages like efficient transgene integration, and suitability for large-scale bioreactor cultivation. We hypothesize that expressing avocado genes in moss will enhance oleic acid production, providing a sustainable alternative for the oleochemical industry. The experimental approach involves cloning the avocado gene cDNA into an expression vector, Polyethylene glycol (PEG)-mediated protoplast transformation, transformant screening, and subsequent lipid analysis via TLC followed by gas chromatography for TAG quantification, and fatty acid composition analysis. Nile Red staining followed by confocal fluorescence microscopy will be done to visualize and quantify the lipid droplets. We expect to demonstrate a substantial increase in oleic acid production in P. patens. The transgenic P. patens will provide an eco-friendly solution for the oleochemical industry, addressing the growing demand for oleic acid in an environmentally conscious manner.

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Apr 5th, 9:00 AM Apr 5th, 11:30 AM

Metabolic Engineering of Physcomitrium patens (moss) for Oleic Acid-rich Vegetable Oil Production

D.P. Culp Center Ballroom

ABSTRACT Triacylglycerols (TAGs), the predominant storage lipids in plants, are integral to vegetable oil composition. The anticipated surge in vegetable oil demand by 2030 necessitates sustainable production strategies. Oleic acid, a vital omega-9 monounsaturated fatty acid with diverse industrial applications, faces challenges in current production methods using high oleic plant oils. Extended growth cycles and climatic limitations highlight the need for sustainable alternatives. Preliminary results in Nicotiana benthamiana leaves show a significant increase in TAG content, particularly oleic acid, through co-expression of avocado (Persea americana) WRINKLED genes (PaWRI1 and PaWRI2) and enzymes diacylglycerol acyltransferase 1 (PaDGAT1) and phospholipid: diacylglycerol acyltransferase 1(PaPDAT1). This study aims to metabolically engineer Physcomitrium patens (moss) to improve oil production in moss, mostly TAG, which has a higher proportion of oleic acid through stable expression of avocado WRINKLED transcription factors and enzymes PaWRI1, PaWRI2, PaDGAT1, and PaPDAT1. P. patens, a non-seed plant model organism, offers advantages like efficient transgene integration, and suitability for large-scale bioreactor cultivation. We hypothesize that expressing avocado genes in moss will enhance oleic acid production, providing a sustainable alternative for the oleochemical industry. The experimental approach involves cloning the avocado gene cDNA into an expression vector, Polyethylene glycol (PEG)-mediated protoplast transformation, transformant screening, and subsequent lipid analysis via TLC followed by gas chromatography for TAG quantification, and fatty acid composition analysis. Nile Red staining followed by confocal fluorescence microscopy will be done to visualize and quantify the lipid droplets. We expect to demonstrate a substantial increase in oleic acid production in P. patens. The transgenic P. patens will provide an eco-friendly solution for the oleochemical industry, addressing the growing demand for oleic acid in an environmentally conscious manner.