Abstract
Producing unusual fatty acids (FAs) in crop plants has been a long-standing goal of green chemistry. However, expression of the enzymes that catalyze the primary synthesis of these unusual FAs in transgenic plants typically results in low levels of the desired FA. For example, seed-specific expression of castor (Ricinus communis) fatty acid hydroxylase (RcFAH) in Arabidopsis (Arabidopsis thaliana) resulted in only 17% hydroxy fatty acids (HFAs) in the seed oil. In order to increase HFA levels, we investigated castor phospholipid:diacylglycerol acyltransferase (PDAT). We cloned cDNAs encoding three putative PDAT enzymes from a castor seed cDNA library and coexpressed them with RcFAH12. One isoform, RcPDAT1A, increased HFA levels to 27%. Analysis of HFA-triacylglycerol molecular species and regiochemistry, along with analysis of the HFA content of phosphatidylcholine, indicates that RcPDAT1A functions as a PDAT in vivo. Expression of RcFAH12 alone leads to a significant decrease in FA content of seeds. Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12 restored FA levels to nearly wild-type levels, and this was accompanied by a major increase in the mass of HFAs accumulating in the seeds. We show the usefulness of RcPDAT1A for engineering plants with high levels of HFAs and alleviating bottlenecks due to the production of unusual FAs in transgenic oilseeds.
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Claims 7
Claim · model-written summary
Castor hydroxylase expression produced only 17% hydroxy fatty acids in Arabidopsis seed oil.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianae0 - Organ / stage
- seede0
- Gene
- RcFAHe0
- Outcome
- hydroxy fatty acids (HFAs) in the seed oile0
- Direction
- onlye0
- Value
- 17%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“seed-specific expression of castor (Ricinus communis) fatty acid hydroxylase (RcFAH) in Arabidopsis (Arabidopsis thaliana) resulted in only 17% hydroxy fatty acids (HFAs) in the seed oil”
Claim · model-written summary
Three putative PDAT enzyme cDNAs were cloned from castor seed.
- Taxon
- castor
Ricinus communise0 - Gene
- PDATe0
- Outcome
- cDNAs encoding three putative PDAT enzymese0
- Value
- threee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We cloned cDNAs encoding three putative PDAT enzymes from a castor seed cDNA library”
Claim · model-written summary
RcPDAT1A raised hydroxy fatty acid levels to 27%.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- RcPDAT1Ae0
- Gene
- RcPDAT1Ae0
- Outcome
- HFA levelse0
- Direction
- increasede0
- Value
- 27%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“One isoform, RcPDAT1A, increased HFA levels to 27%”
t0“Arabidopsis”
Claim · model-written summary
Molecular analyses indicate RcPDAT1A acts as a PDAT in vivo.
- Taxon
- ArabidopsisArabidopsist0
- Gene
- RcPDAT1Ae0
- Outcome
- functions as a PDAT in vivoe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Analysis of HFA-triacylglycerol molecular species and regiochemistry, along with analysis of the HFA content of phosphatidylcholine, indicates that RcPDAT1A functions as a PDAT in vivo”
t0“Arabidopsis”
Claim · model-written summary
RcFAH12 expression alone significantly reduced seed fatty acid content.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- Expression of RcFAH12 alonee0
- Gene
- RcFAH12e0
- Outcome
- FA content of seedse0
- Direction
- decreasee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Expression of RcFAH12 alone leads to a significant decrease in FA content of seeds”
t0“Arabidopsis”
Claim · model-written summary
Coexpressing RcPDAT1A and RcDGAT2 with RcFAH12 restored fatty acid levels nearly to wild type.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12e0
- Outcome
- FA levelse0
- Direction
- restorede0
- Qualifier
- nearly wild-type levelse0
- Comparator
- wild-typee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12 restored FA levels to nearly wild-type levels”
t0“Arabidopsis”
Claim · model-written summary
The same coexpression substantially increased seed hydroxy fatty acid mass.
- Taxon
- ArabidopsisArabidopsist0
- Treatment
- Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12e1
- Outcome
- mass of HFAs accumulating in the seedse0
- Direction
- major increasee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“this was accompanied by a major increase in the mass of HFAs accumulating in the seeds”
t0“Arabidopsis”
t1“Arabidopsis”
e1“Coexpression of RcPDAT1A and RcDGAT2 (for diacylglycerol acyltransferase 2) with RcFAH12 restored FA levels to nearly wild-type levels”
The same questions in other papers
transgenic · molecular: metabolites · ArabidopsisPositive and negative results
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- no effect
- positive
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