AUTHOR=Yeom Wan Woo , Kim Hye Jeong , Lee Kyeong-Ryeol , Cho Hyun Suk , Kim Jin-Young , Jung Ho Won , Oh Seon-Woo , Jun Sang Eun , Kim Hyun Uk , Chung Young-Soo 

TITLE=Increased Production of α-Linolenic Acid in Soybean Seeds by Overexpression of Lesquerella FAD3-1

JOURNAL=Frontiers in Plant Science

VOLUME=10

YEAR=2020

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2019.01812

DOI=10.3389/fpls.2019.01812

ISSN=1664-462X

ABSTRACT=<p>Soybean is a major crop that is used as a source of vegetable oil for human use. To develop transgenic soybean with high α-linolenic acid (ALA; 18:3) content, the <italic>FAD3-1</italic> gene isolated from lesquerella (<italic>Physaria fendleri</italic>) was used to construct vectors with two different seed-specific promoters, soybean β-conglycinin (Pβ-con) and kidney bean phaseolin (Pphas), and one constitutive cauliflower mosaic virus 35S promoter (P35S). The corresponding vectors were used for <italic>Agrobacterium</italic>-mediated transformation of imbibed mature half seeds. The transformation efficiency was approximately 2%, 1%, and 3% and 21, 7, and 17 transgenic plants were produced, respectively. T-DNA insertion and expression of the transgene were confirmed from most of the transgenic plants by polymerase chain reaction (PCR), quantitative real-time PCR (qPCR), reverse transcription PCR (RT-PCR), and Southern blot analysis. The fatty acid composition of soybean seeds was analyzed by gas chromatography. The 18:3 content in the transgenic generation T<sub>1</sub> seeds was increased 7-fold in Pβ-con:<italic>PfFAD3-1</italic>, 4-fold in Pphas : <italic>PfFAD3-1</italic>, and 1.6-fold in P35S:<italic>PfFAD3-1</italic> compared to the 18:3 content in soybean “Kwangankong”. The increased content of 18:3 in the Pβ-con:<italic>PfFAD3-1</italic> soybean (T<sub>1</sub>) resulted in a 52.6% increase in total fatty acids, with a larger decrease in 18:1 content than 18:2 content. The increase in 18:3 content was also maintained and reached 42% in the Pphas : <italic>PfFAD3-1</italic> transgenic generation T<sub>2</sub>. Investigations of the agronomic traits of 12 Pβ-con:<italic>PfFAD3-1</italic> transgenic lines (T<sub>1</sub>) revealed that plant height, number of branches, nodes, pods, total seeds, and total seed weight were significantly higher in several transgenic lines than those in non-transgenic soybean. Especially, an increase in seed size was observed upon expression of the <italic>PfFAD3-1</italic> gene with the β-conglycinin promoter, and 6%–14% higher seed lengths were measured from the transgenic lines.</p>