Dataset Information

Increased expression of fatty acid and ABC transporters enhances seed oil production in camelina.

ABSTRACT:

Background

Lipid transporters play an essential role in lipid delivery and distribution, but their influence on seed oil production in oilseed crops is not well studied.

Results

Here, we examined the effect of two lipid transporters, FAX1 (fatty acid export1) and ABCA9 (ATP-binding cassette transporter subfamily A9) on oil production and lipid metabolism in the oilseed plant Camelina sativa. Overexpression (OE) of FAX1 and ABCA9 increased seed weight and size, with FAX1-OEs and ABCA9-OEs increasing seed length and width, respectively, whereas FAX1/ABCA9-OEs increasing both. FAX1-OE and ABCA9-OE displayed additive effects on seed oil content and seed yield. Also, OE of FAX1 and ABCA9 affected membrane lipid composition in developing pods, especially on phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. The expression of some genes involved in seed oil synthesis, such as DGAT2, PDAT1, and LEC1, was increased in developing seeds of FAX1- and/or ABCA9-OEs.

Conclusion

These results indicate that increased expression of FAX1 and ABCA9 can potentially be applied to improving camelina oil production.

SUBMITTER: Cai G

PROVIDER: S-EPMC7913393 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Publications

Increased expression of fatty acid and ABC transporters enhances seed oil production in camelina.

Cai Guangqin G Wang Geliang G Kim Sang-Chul SC Li Jianwu J Zhou Yongming Y Wang Xuemin X

Biotechnology for biofuels 20210227 1

<h4>Background</h4>Lipid transporters play an essential role in lipid delivery and distribution, but their influence on seed oil production in oilseed crops is not well studied.<h4>Results</h4>Here, we examined the effect of two lipid transporters, FAX1 (fatty acid export1) and ABCA9 (ATP-binding cassette transporter subfamily A9) on oil production and lipid metabolism in the oilseed plant Camelina sativa. Overexpression (OE) of FAX1 and ABCA9 increased seed weight and size, with FAX1-OEs and AB ...[more]

PMID: 33640013

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Project description:Metabolic engineering of the omega-3 (ω3) long chain polyunsaturated fatty acid biosynthesis pathway has generated fish oil-like levels of pharmaceutically and nutritionally important docosahexaenoic acid (DHA) in plant seeds. However, the majority of DHA has been accumulated at the sn-1 and sn-3 positions of triacylglycerol (TAG) in these engineered seeds, leaving only a minor amount (∼10%) at sn-2 position and indicating a strong discrimination (or, a very poor specificity) for DHA by seed lysophosphatidic acid acyltransferases (LPAATs), which mediate the acylation of sn-2 position of glycerol backbone. In order to increase the level of DHA at sn-2 position of TAG and to increase overall DHA level in seeds, we attempted to discover DHA-preferring LPAATs. Several LPAATs for acylation of the sn-2 position of the TAG glycerol backbone were investigated for substrate preference for DHA. In transiently expressing these LPAATs in Nicotiana benthamiana, a Mortierella alpina LPAAT had the highest substrate specificity for accumulating DHA onto oleoyl-lysophosphatidic acid (oleoyl-LPA), while the plant LPAATs tested showed lower preference for DHA. In a competition assay with a pool of four ω3 acyl-Coenzyme A (CoA) substrates involved in the DHA biosynthesis pathway, LPAATs from both M. alpina and Emiliania huxleyi showed a high preference for DHA-CoA acylation onto oleoyl-LPA. When docosahexaenoyl-LPA was used as the acyl receiver, M. alpina LPAAT also showed a high preference for DHA-CoA. Stable overexpression of M. alpina LPAAT in an Arabidopsis line that expressed the DHA biosynthesis pathway significantly increased both the total DHA levels and the distribution of DHA onto the sn-2 position of seed TAG. LC-MS analysis of the seed TAG species also confirmed that overexpression of M. alpina LPAAT increased di-DHA and tri-DHA TAGs, suggesting that the M. alpina LPAAT could enrich DHA at the TAG sn-2 position, leading to a metabolic engineering of oil seed for channeling DHA into the sn-2 position of TAG and to a higher DHA level.

Project description:BackgroundChloroplast and endoplasmic reticulum (ER)-localized fatty acid (FA) transporters have been reported to play important roles in oil (mainly triacylglycerols, TAG) biosynthesis. However, whether these FA transporters synergistically contribute to lipid accumulation, and their effect on lipid metabolism in microalgae are unknown.ResultsHere, we co-overexpressed two chloroplast-localized FA exporters (FAX1 and FAX2) and one ER-localized FA transporter (ABCA2) in Chlamydomonas. Under standard growth conditions, FAX1/FAX2/ABCA2 over-expression lines (OE) accumulated up to twofold more TAG than the parental strain UVM4, and the total amounts of major polyunsaturated FAs (PUFA) in TAG increased by 4.7-fold. In parallel, the total FA contents and major membrane lipids in FAX1/FAX2/ABCA2-OE also significantly increased compared with those in the control lines. Additionally, the total accumulation contribution ratio of PUFA, to total FA and TAG synthesis in FAX1/FAX2/ABCA2-OE, was 54% and 40% higher than that in UVM4, respectively. Consistently, the expression levels of genes directly involved in TAG synthesis, such as type-II diacylglycerol acyltransferases (DGTT1, DGTT3 and DGTT5), and phospholipid:diacylglycerol acyltransferase 1 (PDAT1), significantly increased, and the expression of PGD1 (MGDG-specific lipase) was upregulated in FAX1/FAX2/ABCA2-OE compared to UVM4.ConclusionThese results indicate that the increased expression of FAX1/FAX2/ABCA2 has an additive effect on enhancing TAG, total FA and membrane lipid accumulation and accelerates the PUFA remobilization from membrane lipids to TAG by fine-tuning the key genes involved in lipid metabolism under standard growth conditions. Overall, FAX1/FAX2/ABCA2-OE shows better traits for lipid accumulation than the parental line and previously reported individual FA transporter-OE. Our study provides a potential useful strategy to increase the production of FA-derived energy-rich and value-added compounds in microalgae.

Dataset Information

Increased expression of fatty acid and ABC transporters enhances seed oil production in camelina.

Background

Results

Conclusion

Publications

Increased expression of fatty acid and ABC transporters enhances seed oil production in camelina.

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