PDAT1 genome editing reduces hydroxy fatty acid production in transgenic Arabidopsis
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The fatty acids content of castor (Ricinus communis L.) seedoil is 80-90% ricinoleic acid, which is a hydroxy fatty acid(HFA). The structures and functional groups of HFAs are different from those of common fatty acids and are useful forvarious industrial applications. However, castor seeds containthe toxin ricin and an allergenic protein, which limit theircultivation. Accordingly, many researchers are conducting studiesto enhance the production of HFAs in Arabidopsis thaliana, amodel plant for oil crops. Oleate 12-hydroxylase from castor(RcFAH12), which synthesizes HFA (18:1-OH), was transformedinto an Arabidopsis fae1 mutant, resulting in the CL37 lineproducing a maximum of 17% HFA content. In addition, castorphospholipid:diacylglycerol acyltransferase 1-2 (RcPDAT1-2),which catalyzes the production of triacylglycerol by transferringHFA from phosphatidylcholine to diacylglycerol, was transformed into the CL37 line to develop a P327 line that produces 25% HFA. In this study, we investigated changes in HFAcontent when endogenous Arabidopsis PDAT1 (AtPDAT1) ofthe P327 line was edited using the CRISPR/Cas9 technique. The successful mutation resulted in three independent lineswith different mutation patterns, which were transmitted untilthe T4 generation. Fatty acid analysis of the seeds showed thatHFA content decreased in all three mutant lines. These findings indicate that AtPDAT1 as well as RcPDAT1-2 in the P327line are involved in transferring and increasing HFAs to triacylglycerol.
Abstract
The fatty acids content of castor (Ricinus communis L.) seedoil is 80-90% ricinoleic acid, which is a hydroxy fatty acid(HFA). The structures and functional groups of HFAs are different from those of common fatty acids and are useful forvarious industrial applications. However, castor seeds containthe toxin ricin and an allergenic protein, which limit theircultivation. Accordingly, many researchers are conducting studiesto enhance the production of HFAs in Arabidopsis thaliana, amodel plant for oil crops. Oleate 12-hydroxylase from castor(RcFAH12), which synthesizes HFA (18:1-OH), was transformedinto an Arabidopsis fae1 mutant, resulting in the CL37 lineproducing a maximum of 17% HFA content. In addition, castorphospholipid:diacylglycerol acyltransferase 1-2 (RcPDAT1-2),which catalyzes the production of triacylglycerol by transferringHFA from phosphatidylcholine to diacylglycerol, was transformed into the CL37 line to develop a P327 line that produces 25% HFA. In this study, we investigated changes in HFAcontent when endogenous Arabidopsis PDAT1 (AtPDAT1) ofthe P327 line was edited using the CRISPR/Cas9 technique. The successful mutation resulted in three independent lineswith different mutation patterns, which were transmitted untilthe T4 generation. Fatty acid analysis of the seeds showed thatHFA content decreased in all three mutant lines. These findings indicate that AtPDAT1 as well as RcPDAT1-2 in the P327line are involved in transferring and increasing HFAs to triacylglycerol.
참고문헌 (35)
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