Transcriptional Regulation of Oil Biosynthesis genes in Arabidopsis Seed tissue Induced by Avocado WRINKLED2

Authors' Affiliations

Jyoti Ranjan Behera, Department of Biomedical Science, College of Arts and Sciences, East Tennessee State University, Johnson City, TN. Aruna Kilaru, Department of Biological Science, College of Arts and Sciences, East Tennessee State University, Johnson City, 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

147

Name of Project's Faculty Sponsor

Aruna Kilaru

Faculty Sponsor's Department

Biological Sciences

Classification of First Author

Graduate Student-Doctoral

Competition Type

Competitive

Type

Poster Presentation

Presentation Category

Science, Technology and Engineering

Abstract or Artist's Statement

Triacylglycerol (TAG), a primary storage lipid found in plants' seed and non-seed tissues, is the primary component of vegetable oils. The transcription factor WRINKLED1 (WRI1) directly or indirectly controls its synthesis. AtWRI2 is not functional. In avocado (Persea americana) mesocarp, which constitutes 60-70% oil by dry weight, three paralogs of the WRI1 are highly expressed. We previously demonstrated that PaWRI1 and PaWRI2 can interact with AW-box present in the promoters of target genes and transactivate them. Recognizing the importance of enhancing seed oil content to meet the global demand for vegetable oil, we hypothesize that PaWRI1 and PaWRI2 can induce oil accumulation in seed tissue through trans-activating the genes associated with fatty acid (FA) biosynthesis. We propose performing functional complementation of Arabidopsis mutant seeds, wri1-1 (CS69538) and wri2 (SALK_111105C) with PaWRI1 and PaWRI2. The strategy involves introducing the cDNA sequence of PaWRI2 and PaWRI1, cloned under a seed-specific promoter (Napin), into Arabidopsis using a floral dip method. Transformed T0 seeds will be selected, grown in selective kanamycin media, and further cultivated for T1, and T2 generations to harvest homozygous transformants. Further, Seeds will be collected at maturation stages from T3 plants to analyze total lipid, FA, and TAG using Gas Chromatography-Flame Ionization Detection (GC-FID). A comparison will be made between the T3 plants and the control group. Quantitative Reverse Transcription PCR (qRT-PCR) will be done using mRNA extracted from the seeds to measure the relative expression of oil biosynthesis-related genes. The outcome of this study will provide insights into the possible functional role of PaWRI2 in seed tissue, which is otherwise considered non-functional in other oilseed crops, and how it can be harnessed to increase oil production in other oil-rich plants.

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

Transcriptional Regulation of Oil Biosynthesis genes in Arabidopsis Seed tissue Induced by Avocado WRINKLED2

D.P. Culp Center Ballroom

Triacylglycerol (TAG), a primary storage lipid found in plants' seed and non-seed tissues, is the primary component of vegetable oils. The transcription factor WRINKLED1 (WRI1) directly or indirectly controls its synthesis. AtWRI2 is not functional. In avocado (Persea americana) mesocarp, which constitutes 60-70% oil by dry weight, three paralogs of the WRI1 are highly expressed. We previously demonstrated that PaWRI1 and PaWRI2 can interact with AW-box present in the promoters of target genes and transactivate them. Recognizing the importance of enhancing seed oil content to meet the global demand for vegetable oil, we hypothesize that PaWRI1 and PaWRI2 can induce oil accumulation in seed tissue through trans-activating the genes associated with fatty acid (FA) biosynthesis. We propose performing functional complementation of Arabidopsis mutant seeds, wri1-1 (CS69538) and wri2 (SALK_111105C) with PaWRI1 and PaWRI2. The strategy involves introducing the cDNA sequence of PaWRI2 and PaWRI1, cloned under a seed-specific promoter (Napin), into Arabidopsis using a floral dip method. Transformed T0 seeds will be selected, grown in selective kanamycin media, and further cultivated for T1, and T2 generations to harvest homozygous transformants. Further, Seeds will be collected at maturation stages from T3 plants to analyze total lipid, FA, and TAG using Gas Chromatography-Flame Ionization Detection (GC-FID). A comparison will be made between the T3 plants and the control group. Quantitative Reverse Transcription PCR (qRT-PCR) will be done using mRNA extracted from the seeds to measure the relative expression of oil biosynthesis-related genes. The outcome of this study will provide insights into the possible functional role of PaWRI2 in seed tissue, which is otherwise considered non-functional in other oilseed crops, and how it can be harnessed to increase oil production in other oil-rich plants.