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Abstract
The development of ω-3 fatty acid-rich vegetable oils is essential to enrich the production of functional foods. Sacha Inchi (Plukenetia volubilis L.) is a unique oilseed crop with much potential. Its seeds contain rich polyunsaturated fatty acids (PUFAs), especially linoleic acid (LA, C18:2) and α-linolenic acid (ALA, C18:3). Endoplasmic reticulum -located ω-6 and ω-3 fatty acid desaturases (FAD) are responsible for the biosynthesis of LA and ALA, respectively, in plant seeds. Here, we isolated two full-length FAD genes from Sacha Inchi, named PvFAD2 and PvFAD3, which encoded predicted amino acid residues of 384 and 379 in protein, respectively. Protein sequence and subcellular localization analysis revealed that they were located in the endoplasmic reticulum (ER). Heterologous expression in Saccharomyces cerevisiae confirmed that PvFAD2 and PvFAD3 could catalyze LA and ALA synthesis, respectively. The stability and catalytic efficiency of the PvFAD3 protein may be closely related to temperature. In transgenic tobacco, using seed-specific expression promoters, PvFAD2 and PvFAD3 significantly promotes the production of LA (from 68% to 70.5%) and ALA (from 0.7% to 3.1%) in seed oil. These results show that PvFAD2 and PvFAD3 do, indeed, function as crucial enzymes for PUFAs biosynthesis, and provide a key gene source for the sustainable production of lipids with tailored fatty acid compositions via genetic engineering in other oil crops.
Highlights
Sacha Inchi (Plukenetia volubilis L., Euphorbiaceae), which bears star-shaped fruit capsules, known as mountain peanut, Inca nut or Inca Inchi, is native to the Amazon region of South America, including parts of Peru and northwestern Brazil (Hamaker et al, 1992; Gillespie, 2007)
The full-length of the coding sequences of PvFAD2 and PvFAD3 were 1152 bp and 1137 bp, respectively
Studies have documented that the production of polyunsaturated fatty acids (PUFAs) in seed oil is usually controlled by several endoplasmic reticulum (ER)-located fatty acid desaturases (FAD) genes such as FAD2 and FAD3, and many FAD2 and FAD3 genes have been extensively identified in oilseed plants, relatively few FAD genes, for any given plant, have been functionally examined (Dar et al, 2017)
Summary
Sacha Inchi (Plukenetia volubilis L., Euphorbiaceae), which bears star-shaped fruit capsules, known as mountain peanut, Inca nut or Inca Inchi, is native to the Amazon region of South America, including parts of Peru and northwestern Brazil (Hamaker et al, 1992; Gillespie, 2007). The ratio of ω-6 to ω-3 fatty acids in Sacha Inchi seed oils ranges from 0.83 to1.09, which is considered to be an optimal proportion of functional oils for human health (Simopoulos, 2011; Simopoulos, 2008). The physiological and molecular mechanism that underlie the accumulation of the high content of ALA in Sacha Inchi seeds largely remain uncertain. Dissecting the molecular mechanism behind ALA biosynthesis in Sacha Inchi seeds and identifying key genes responsible for controlling or regulating the biosynthesis of ALA would facilitate the use of genetic engineering in crop breeding to produce lipids with tailored fatty acid compositions as well as the provision of healthy functional oils
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