ORCID Profile
0000-0003-4488-8147
Current Organisation
CSIRO Black Mountain Laboratories
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In Research Link Australia (RLA), "Research Topics" refer to ANZSRC FOR and SEO codes. These topics are either sourced from ANZSRC FOR and SEO codes listed in researchers' related grants or generated by a large language model (LLM) based on their publications.
Biochemistry and Cell Biology | Phycology | Plant Biology | Agricultural Molecular Engineering of Nucleic Acids and Proteins | Fermentation, Biotechnology And Industrial Microbiology | Aquaculture | Synthetic Biology
Other | Renewable energy not elsewhere classified (e.g. geothermal) | Expanding Knowledge in the Chemical Sciences | Environmentally Sustainable Animal Production not elsewhere classified | Aquaculture | Expanding Knowledge in the Biological Sciences |
Publisher: Springer Science and Business Media LLC
Date: 03-2006
DOI: 10.1007/BF02914050
Publisher: Elsevier BV
Date: 04-2011
Publisher: Elsevier BV
Date: 02-2007
Publisher: MDPI AG
Date: 26-05-2010
DOI: 10.3390/NU2060572
Publisher: Elsevier BV
Date: 03-2006
Publisher: Springer Science and Business Media LLC
Date: 20-05-2011
Abstract: Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T 1 progeny. An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium -infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T 1 seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T 1 progeny displayed Mendelian inheritance. This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications.
Publisher: Springer Science and Business Media LLC
Date: 31-10-2007
Publisher: Wiley
Date: 12-06-2013
DOI: 10.1016/J.FEBSLET.2013.06.003
Abstract: Demonstration of the function of the Arabidopsis thaliana acyl-CoA:diacylglycerol acyltransferase 2 (AtDGAT2) has remained elusive despite biochemical testing of genetic mutants and overexpression lines. We show that transiently expressed AtDGAT2 in the Nicotiana benthamiana leaf resulted in an increase in triacylglycerol twice as great as the increase observed following parallel expression of AtDGAT1. AtDGAT2 showed higher conversion from labeled diacylglycerol to triacylglycerol compared to AtDGAT1, and was acyl-CoA dependent. In addition, AtDGAT2 had different acyl-CoA substrate preference than AtDGAT1. These results allow us to conclude that AtDAGT2 is a functional acyl-CoA:diacylglycerol acyltransferase enzyme that can catalyse substantial increase in TAG synthesis.
Publisher: Springer Science and Business Media LLC
Date: 21-01-2015
DOI: 10.1007/S00299-015-1737-5
Abstract: Simultaneous gene silencing of both FAD2 genes in high linoleic acid flax leads to high level of oleic acid, which is stable across multiple generations. High oleic oil is one of the preferred traits in oil crop engineering due to its stability and multiple applications as an industrial feedstock. Flax possesses two isoforms of FAD2 enzymes that desaturate monounsaturated oleic acid to polyunsaturated linoleic acid. These two enzymes are encoded by two FAD2 genes. By simultaneous gene silencing both FAD2 genes in high linoleic acid flax, Linola, high level of oleic acid up to 80% was achieved in 69 silencing lines. The high oleic trait was stable across multiple generations with oleic acid reaching up to 77% in homozygote T3 progeny. The RNAi-mediated gene-silencing approach generated high oleic linseed oil, as well as a high oleic platform that can be exploited for further fatty acid engineering.
Publisher: Wiley
Date: 29-11-2012
DOI: 10.1111/PBI.12023
Abstract: Society has come to rely heavily on mineral oil for both energy and petrochemical needs. Plant lipids are uniquely suited to serve as a renewable source of high-value fatty acids for use as chemical feedstocks and as a substitute for current petrochemicals. Despite the broad variety of acyl structures encountered in nature and the cloning of many genes involved in their biosynthesis, attempts at engineering economic levels of specialty industrial fatty acids in major oilseed crops have so far met with only limited success. Much of the progress has been h ered by an incomplete knowledge of the fatty acid biosynthesis and accumulation pathways. This review covers new insights based on metabolic flux and reverse engineering studies that have changed our view of plant oil synthesis from a mostly linear process to instead an intricate network with acyl fluxes differing between plant species. These insights are leading to new strategies for high-level production of industrial fatty acids and waxes. Furthermore, progress in increasing the levels of oil and wax structures in storage and vegetative tissues has the potential to yield novel lipid production platforms. The challenge and opportunity for the next decade will be to marry these technologies when engineering current and new crops for the sustainable production of oil and wax feedstocks.
Publisher: Wiley
Date: 21-05-2015
DOI: 10.1111/PBI.12386
Abstract: High oleic oil is an important industrial feedstock that has been one of the main targets for oil improvement in a number of oil crops. Crambe (Crambe abyssinica) is a dedicated oilseed crop, suitable for industrial oil production. In this study, we down-regulated the crambe fatty acid desaturase (FAD) and fatty acid elongase (FAE) genes for creating high oleic seed oil. We first cloned the crambe CaFAD2, CaFAD3 and CaFAE1 genes. Multiple copies of each of these genes were isolated, and the highly homologous sequences were used to make RNAi constructs. These constructs were first tested in Arabidopsis, which led to the elevated oleic or linoleic levels depending on the genes targeted, indicating that the RNAi constructs were effective in regulating the expression of the target genes in nonidentical but closely related species. Furthermore, down-regulation of CaFAD2 and CaFAE1 in crambe with the FAD2-FAE1 RNAi vector resulted in even more significant increase in oleic acid level in the seed oil with up to 80% compared to 13% for wild type. The high oleic trait has been stable in subsequent five generations and the GM line grew normally in greenhouse. This work has demonstrated the great potential of producing high oleic oil in crambe, thus contributing to its development into an oil crop platform for industrial oil production.
Publisher: Springer Science and Business Media LLC
Date: 02-2013
Publisher: Frontiers Media SA
Date: 26-05-2014
Publisher: Wiley
Date: 08-01-2013
DOI: 10.1016/J.FEBSLET.2012.12.018
Abstract: Metabolic engineering approaches to increase plant oil levels can generally be ided into categories which increase fatty acid biosynthesis ('Push'), are involved in TAG assembly ('Pull') or increase TAG storage/decrease breakdown ('Accumulation'). In this study, we describe the surprising synergy when Push (WRI1) and Pull (DGAT1) approaches are combined. Co-expression of these genes in the Nicotiana benthamiana transient leaf expression system resulted in TAG levels exceeding those expected from an additive effect and biochemical tracer studies confirmed increased flux of carbon through fatty acid and TAG synthesis pathways. Leaf fatty acid profile also synergistically shifts from polyunsaturated to monounsaturated fatty acids.
Publisher: Springer Science and Business Media LLC
Date: 05-2011
DOI: 10.1007/S11248-011-9517-7
Abstract: We describe a transgenic microalgal Δ9-elongase pathway transformed in both Brassica napus and Arabidopsis thaliana seed resulting in the production of arachidonic acid (ARA). This pathway is noteworthy for both the production of ARA in seed tissue and the low levels of intermediate C20 fatty acids that accumulate. We also demonstrate that the arachidonic acid is naturally enriched at the sn2 position in triacylglycerol. This is the first report of ARA production by the Δ9-elongase pathway in an oilseed.
Publisher: CSIRO Publishing
Date: 2006
DOI: 10.1071/FP05297
Abstract: The transgenic production of unusual fatty acids in oil seed crops offers an alternative, renewable resource for industry. However, transgenic expression of genes catalysing the synthesis of unusual fatty acids has generally resulted in these fatty acids accumulating at levels significantly below the levels in the wild species from which the genes were sourced. This study reports expression of additional copies of any of three Δ12-desaturase genes (FAD2) from Crepis palaestina Bornm., cotton (Gossypium hirsutum L.) or Arabidopsis thaliana (L.) Heynh. with C. palaestina Δ12-epoxygenase gene (Cpal2), in an Arabidopsis mutant having a significantly higher level of linoleic acid substrate. This resulted in the highest levels of vernolic acid accumulation, 21% of total fatty acids, reported so far in any transgenic plant expressing the Δ12-epoxygenase. Similarly, the co-expression of C. palaestina Cpal2 and a transgenic copy of FAD2 in cotton seed that contains large amounts of linoleic acid substrate also resulted in greater accumulation of vernolic acid in seed than did expression of C. palaestina Cpal2 alone.
Publisher: Public Library of Science (PLoS)
Date: 16-04-2012
Publisher: Springer Science and Business Media LLC
Date: 21-06-2016
DOI: 10.1038/SREP28298
Abstract: As a resource allocation strategy, plant growth and defense responses are generally mutually antagonistic. Brassinosteroid (BR) regulates many aspects of plant development and stress responses, however, genetic evidence of its integrated effects on plant growth and stress tolerance is lacking. We overexpressed the Arabidopsis BR biosynthetic gene AtDWF4 in the oilseed plant Brassica napus and scored growth and stress response phenotypes. The transgenic B. napus plants, in comparison to wild type, displayed increased seed yield leading to increased overall oil content per plant, higher root biomass and root length, significantly better tolerance to dehydration and heat stress and enhanced resistance to necrotrophic fungal pathogens Leptosphaeria maculans and Sclerotinia sclerotiorum. Transcriptome analysis supported the integrated effects of BR on growth and stress responses in addition to BR responses associated with growth, a predominant plant defense signature, likely mediated by BES1/BZR1, was evident in the transgenic plants. These results establish that BR can interactively and simultaneously enhance abiotic and biotic stress tolerance and plant productivity. The ability to confer pleiotropic beneficial effects that are associated with different agronomic traits suggests that BR–related genes may be important targets for simultaneously increasing plant productivity and performance under stress conditions.
Publisher: Oxford University Press (OUP)
Date: 09-1984
DOI: 10.1104/PP.76.1.139
Publisher: Oxford University Press (OUP)
Date: 08-2002
DOI: 10.1104/PP.006353
Abstract: The endogenous Δ12-desaturase gene (FAD2) in Arabidopsis was targeted for silencing using seed-specific cosuppression (CS), hairpin (HP) RNA (hpRNA), and intron-spliced HP (iHP) constructs. The iHP construct, incorporating the 120-bp 3′-untranslated region of the FAD2 gene, gave the highest degree of silencing. In some iHP lines Δ12-desaturase activity was reduced to levels as low as those in the nullfad2-1 mutant, and every primary transformant showed a pronounced reduction in FAD2 activity. One highly silenced iHP line was propagated for five generations and showed no reversion or diminution in its degree of silencing. About 75% of plants transformed with the HP construct, targeting theFAD2 coding region, gave dramatically reduced Δ12-desaturase activity, whereas approximately 50% of plants transformed with the CS construct, containing the same coding region sequence, showed silencing at a much less profound level. In all three types of constructs, the degree of silencing was increased when the transgenes were homozygous, but this was much more pronounced for the CS constructs. All three types of construct could give a single locus that was capable of effective silencing, but in the one such CS line where this was the case, the locus had a complex insertion pattern. This is consistent with the concept that posttranscriptional gene silencing is induced by double-stranded, or self-complementary, RNA that is formed in cases of CS by complex insertion patterns at a single locus and that the most effective way of generating profoundly silenced plants is by the use of constructs that encode hpRNAs. Furthermore, these results demonstrate for the first time, to our knowledge, that iHP constructs targeted against an endogenous seed-expressed gene are clearly able to generate phenotypic changes that are inherited stably over several generations, making this approach a reliable technique for genetic modification of seed quality and possibly other traits in agricultural plants.
Publisher: Elsevier BV
Date: 12-2011
Publisher: Springer Science and Business Media LLC
Date: 10-10-2009
DOI: 10.1007/S10126-009-9230-1
Abstract: The production of long-chain polyunsaturated fatty acids from precursor molecules linoleic acid (LA 18:2omega6) and alpha-linolenic acid (ALA 18:3omega3) is catalysed by sequential desaturase and elongase reactions. We report the isolation of a front-end Delta6-desaturase gene from the microalgae Ostreococcus lucimarinus and two elongase genes, a Delta6-elongase and a Delta5-elongase, from the microalga Pyramimonas cordata. These enzymes efficiently convert their respective substrates when transformed in yeast (39-75% conversion for omega3 substrate fatty acids), and the Delta5-elongase in particular displays higher elongation efficiency (75% for conversion of eicosapentaenoic acid (20:5omega3) to docosapentaenoic acid (22:5omega3)) than previously reported genes. In addition, the Delta6-desaturase is homologous with acyl-CoA desaturases and shows a strong preference for the omega3 substrate ALA.
Publisher: Wiley
Date: 06-11-2009
DOI: 10.1111/J.1467-7652.2009.00453.X
Abstract: The assembly of multistep recombinant pathways in stably transformed plants is a cornerstone of crops producing new products yet can be a laborious and time-consuming process. Any heterologous expression platform capable of providing a rapid estimation of the functional assembly of an entire pathway would guide the design of such transgenic traits. In this study, we use a Nicotiana benthamiana transient leaf expression system to simultaneously express five genes, from five independent T(DNA) binary vectors, to assemble a complete recombinant pathway in five days. In this study, we demonstrate the production of long-chain polyunsaturated fatty acids (LC-PUFA) requiring five transgene-encoded reactions to convert endogenous fatty acids to LC-PUFA. The addition of a triacylglycerol assembly enzyme, Arabidopsis thaliana diacylglyceride-O-acyltransferase, and fractionation of the total lipid profile demonstrated that leaf oils contained 37% newly synthesised LC-PUFA, including 7% arachidonic acid (AA), 6% eicosopentaenoic acid and 3% docosahexaenoic acid. The calculation of enzymatic conversion efficiencies at each step of LC-PUFA synthesis suggests that this transient assembly of a complicated multistep pathway is highly efficient. Unlike experiments using stably transformed plants our assembly of an intricate pathway maintained full gene-for-gene interchangeability and required a fraction of the time and glasshouse space. Furthermore, an exogenous LC-PUFA fatty acid substrate, AA, was fed and metabolised by a transiently expressed Delta17-desaturase enzyme, and provided results similar to those obtained in yeast feeding experiments. Although the assay was ideal for LC-PUFA pathways, this assay format may become a powerful tool for the characterisation and step-wise improvement of other recombinant pathways and multigenic traits.
Publisher: MDPI AG
Date: 21-02-2014
DOI: 10.3390/NU6020776
Publisher: Wiley
Date: 24-10-2013
DOI: 10.1111/PBI.12131
Publisher: Elsevier BV
Date: 08-2013
DOI: 10.1016/J.PHYTOCHEM.2013.05.006
Abstract: We have characterised the FAD2 gene family from Hiptage benghalensis, a tropical plant that accumulates high levels of ricinoleic acid in its seeds. Functional characterisation of six FAD2 gene family members showed that two of them were capable of functioning as Δ12-hydroxylases while the other FAD2 members were confirmed to be Δ12-desaturases. The Δ12-hydroxylation function of these two genes was confirmed in yeast cells, using C16:1(Δ9) and C18:1(Δ9) monounsaturated fatty acids as substrates. These Δ12-hydroxylases, like the other Δ12-hydroxylases previously cloned from plants Ricinus communis (castor), Physaria fendleri and fungus Claviceps purpurea, also showed some Δ12-desaturase activity. The hydroxylation activity of the two Hiptage hydroxylases was further confirmed by their expression in the Arabidopsis fad2/fae1 double mutant where they were able to produce equivalent or higher levels hydroxylated fatty acids in the seed oil when compared with the other known hydroxylases.
Publisher: Frontiers Media SA
Date: 09-2014
Publisher: Frontiers Media SA
Date: 2013
Publisher: Wiley
Date: 26-07-2010
Publisher: Frontiers Media SA
Date: 22-12-2015
Publisher: CSIRO Publishing
Date: 2015
DOI: 10.1071/CP15009
Abstract: Sugarcane (Poaceae) has not undergone any commercial selection based upon seed characteristics. As the plant is grown from vegetative cuttings and the stalk harvested for its sucrose content, relatively little is known about its seed compared with other grass crops. The seeds of sugarcane were small, 1.8 × 0.8 mm, and the embryo comprised about one-third of the seed volume. Among the s les analysed, the seed contained on average 37%, 20% and 10% of the fresh weight as starch, protein and lipid, respectively. Histochemical staining showed that the starch was confined to the endosperm and the lipid to the embryo and aleurone layer. Protein was found in the embryo, endosperm and aleurone layer. There were small but significant differences between the sources of sugarcane seed. The wild relative S. spontaneum had significantly less starch than the commercial hybrid sugarcane seed. The lipid content was higher for sugarcane seed than for the seeds of many other grasses, possibly because of the high ratio of lipid-containing embryo to endosperm. Following artificial ageing, the observed decline in seed viability was not closely reflected by any significant changes in composition, although protein and sugars were reduced after 168 h. These results contribute to our understanding of the sexual reproductive biology of sugarcane, which is important for the science-based environmental risk evaluation of the release of genetically modified sugarcane.
Publisher: American Astronomical Society
Date: 05-06-2013
Publisher: Oxford University Press (OUP)
Date: 08-2002
DOI: 10.1104/PP.001933
Abstract: We have genetically modified the fatty acid composition of cottonseed oil using the recently developed technique of hairpin RNA-mediated gene silencing to down-regulate the seed expression of two key fatty acid desaturase genes, ghSAD-1-encoding stearoyl-acyl-carrier protein Δ9-desaturase andghFAD2-1-encoding oleoyl-phosphatidylcholine ω6-desaturase. Hairpin RNA-encoding gene constructs (HP) targeted against either ghSAD-1 or ghFAD2-1 were transformed into cotton (Gossypium hirsutum cv Coker 315). The resulting down-regulation of the ghSAD-1 gene substantially increased stearic acid from the normal levels of 2% to 3% up to as high as 40%, and silencing of the ghFAD2-1gene resulted in greatly elevated oleic acid content, up to 77% compared with about 15% in seeds of untransformed plants. In addition, palmitic acid was significantly lowered in both high-stearic and high-oleic lines. Similar fatty acid composition phenotypes were also achieved by transformation with conventional antisense constructs targeted against the same genes, but at much lower frequencies than were achieved with the HP constructs. By intercrossing the high-stearic and high-oleic genotypes, it was possible to simultaneously down-regulate both ghSAD-1 and ghFAD2-1to the same degree as observed in the in idually silenced parental lines, demonstrating for the first time, to our knowledge, that duplex RNA-induced posttranslational gene silencing in independent genes can be stacked without any diminution in the degree of silencing. The silencing of ghSAD-1 and/or ghFAD2-1 to various degrees enables the development of cottonseed oils having novel combinations of palmitic, stearic, oleic, and linoleic contents that can be used in margarines and deep frying without hydrogenation and also potentially in high-value confectionery applications.
Publisher: Elsevier BV
Date: 04-2005
DOI: 10.1016/J.PBI.2005.01.012
Abstract: Metabolic engineering of plants to express high levels of new fatty acids that are of nutritional and industrial importance has proven to be highly challenging. Significant advances have been made recently, however, particularly in the development of the first plant oils to contain long-chain polyunsaturated fatty acids, such as arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. Methods of increasing the accumulation of Delta12-modified fatty acids synthesized by transgenically expressed FAD2-like enzymes have also been investigated. Biochemical analyses of plants that express these introduced fatty-acid metabolic pathways have highlighted the central importance of ensuring the removal of novel fatty acids from their site of synthesis on phosphatidylcholine to enable their further modification, exclusion from membrane lipids and accumulation in seed triacylglycerols.
Publisher: Wiley
Date: 06-11-2008
DOI: 10.1111/J.1365-2583.2008.00841.X
Abstract: We report the first isolation and characterization of insect fatty acid Delta12-desaturase genes, AdD12Des from house cricket (Acheta domesticus) and TcD12Des from the red flour beetle (Tribolium castaneum), responsible for the production of linoleic acid from oleic acid. Sequence analysis shows the cricket and flour beetle Delta12-desaturase genes have evolved independently from all previously known Delta12-desaturases and are much more closely related to the archetypal stearoyl-Coenzyme A-acting desaturase from rat than to the phospholipid-acting Delta12-desaturases widely reported in plants. Phylogenetic and functional analysis indicates the cricket AdD12Des gene may have evolved from an ancestral Delta9-desaturase. By contrast, the beetle Delta12-desaturase is distantly related to the cricket genes and beetle Delta9-desaturases suggesting evolution by an independent route. Linoleic acid has key physiological roles in insects and this is the first report of genes capable of producing this essential fatty acid in higher animals.
Publisher: Elsevier BV
Date: 03-2007
DOI: 10.1016/J.PHYTOCHEM.2006.12.016
Abstract: The marine microalga Pavlova salina produces lipids containing approximately 50% omega-3 long chain polyunsaturated fatty acids (LC-PUFA) such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Three cDNA sequences, designated PsD4Des, PsD5Des, PsD8Des, were isolated from P. salina and shown to encode three front-end desaturases with Delta4, Delta5 and Delta8 specificity, respectively. Southern analysis indicated that the P. salina genome contained single copies of all three front-end fatty acid desaturase genes. When grown at three different temperatures, analysis of fatty acid profiles indicated P. salina desaturation conversions occurred with greater than 95% efficiency. Real-Time PCR revealed that expression of PsD8Des was higher than for the other two genes under normal growth conditions, while PsD5Des had the lowest expression level. The deduced amino acid sequences from all three genes contained three conserved histidine boxes and a cytochrome b(5) domain. Sequence alignment showed that the three genes were homologous to corresponding desaturases from other microalgae and fungi. The predicted activities of these three front-end desaturases leading to the synthesis of LC-PUFA were also confirmed in yeast and in higher plants.
Publisher: Springer Science and Business Media LLC
Date: 11-03-2010
Abstract: Metabolic engineering of seed biosynthetic pathways to ersify and improve crop product quality is a highly active research area. The validation of genes driven by seed-specific promoters is time-consuming since the transformed plants must be grown to maturity before the gene function can be analysed. In this study we demonstrate that genes driven by seed-specific promoters contained within complex constructs can be transiently-expressed in the Nicotiana benthamiana leaf-assay system by co-infiltrating the Arabidopsis thaliana LEAFY COTYLEDON2 (LEC2) gene. A real-world case study is described in which we first assembled an efficient transgenic DHA synthesis pathway using a traditional N. benthamiana Cauliflower Mosaic Virus (CaMV) 35S-driven leaf assay before using the LEC2-extended assay to rapidly validate a complex seed-specific construct containing the same genes before stable transformation in Arabidopsis . The LEC2-extended N. benthamiana assay allows the transient activation of seed-specific promoters in leaf tissue. In this study we have used the assay as a rapid preliminary screen of a complex seed-specific transgenic construct prior to stable transformation, a feature that will become increasingly useful as genetic engineering moves from the manipulation of single genes to the engineering of complex pathways. We propose that the assay will prove useful for other applications wherein rapid expression of transgenes driven by seed-specific constructs in leaf tissue are sought.
Publisher: Public Library of Science (PLoS)
Date: 21-01-2014
Publisher: Oxford University Press (OUP)
Date: 2011
Publisher: Wiley
Date: 02-12-2016
DOI: 10.1111/PBI.12506
Publisher: Elsevier BV
Date: 05-2010
DOI: 10.1016/J.YMBEN.2009.12.001
Abstract: Long-chain (> or = C20) polyunsaturated fatty acids (LC-PUFA) EPA and DHA (20:5(Delta5,8,11,14,17) and 22:6(Delta4,7,10,13,16,19)) have well-documented health benefits against coronary heart disease, rheumatoid arthritis and other disorders. Currently, the predominant sources of these fatty acids are marine fish and algal oils, but research is being conducted to ensure that a sustainable, land-based production system can be developed. We here describe the metabolic engineering of an artificial pathway that produces 26% EPA in leaf triacylglycerol using a newly-identified Delta6-desaturase from the marine microalga Micromonas pusilla. We also demonstrate that this enzyme appears to function as an acyl-CoA desaturase that has preference for omega3 substrates both in planta and in yeast. Phylogenetic analysis indicates that this desaturase shares highly conserved motifs with previously described acyl-CoA Delta6-desaturases.
Publisher: Springer Science and Business Media LLC
Date: 06-11-2009
DOI: 10.1007/S10126-008-9157-Y
Abstract: The marine microalga Pavlova salina (Haptophyta, Pavlovophyceae) produces lipids containing approximately 50% n-3 long-chain polyunsaturated fatty acids including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). A full-length cDNA sequence, designated PsElo5, was isolated from P. salina. Sequence alignment showed that the gene was homologous to corresponding ELO-type elongases from other microalgae. Heterologous expression of PsElo5 in yeast and in higher plants confirmed that it encodes a specific Delta5-elongase activity as predicted and, furthermore, within the n-3 pathway, the elongation activity was confined exclusively to EPA.
Publisher: CSIRO Publishing
Date: 2005
DOI: 10.1071/FP05084
Abstract: Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are nutritionally important long-chain (≥ C20) omega-3 polyunsaturated fatty acids (ω3 LC-PUFA) currently obtained mainly from marine sources. A set of genes encoding the fatty acid chain elongation and desaturation enzymes required for the synthesis of LC-PUFA from their C18 PUFA precursors was expressed seed-specifically in Arabidopsis thaliana. This resulted in the synthesis of DHA, the most nutritionally important ω3 LC-PUFA, for the first time in seed oils, along with its precursor EPA and the ω6 LC-PUFA arachidonic acid (ARA). The assembled pathway utilised Δ5 and Δ6 desaturases that operate on acyl-CoA substrates and led to higher levels of synthesis of LC-PUFA than previously reported with acyl-PC desaturases. This demonstrates the potential for development of land plants as alternative sources of DHA and other LC-PUFA to meet the growing demand for these nutrients.
Publisher: Springer Science and Business Media LLC
Date: 22-05-2013
DOI: 10.1007/S00122-013-2129-2
Abstract: There are two types of safflower oil, high oleic (HO) with 70-75 % oleic acid and high linoleic (HL) with about 70 % linoleic acid. The original HO trait in safflower, found in an introduction from India, is controlled by a partially recessive allele ol at a single locus (Knowles and Bill 1964). In the lipid biosynthesis pathway of developing safflower seeds, microsomal oleoyl phosphatidylcholine desaturase (FAD2) is largely responsible for the conversion of oleic acid to linoleic acid. In vitro microsomal assays indicated drastically reduced FAD2 enzyme activity in the HO genotype compared to conventional HL safflower. A previous study indicated that a single-nucleotide deletion was found in the coding region of CtFAD2-1 that causes premature termination of translation in the HO genotypes, and the expression of the mutant CtFAD2-1Δ was attenuated in the HO genotypes compared to conventional HL safflower (Guan et al. 2012). In this study, we hypothesise that down-regulation of CtFAD2-1 expression in the HO genotype may be explained by nonsense-mediated RNA decay (NMD). NMD phenomenon, indicated by gene-specific RNA degradation of defective CtFAD2-1Δ, was subsequently confirmed in Arabidopsis thaliana seed as well as in the transient expression system in Nicotiana benthamiana leaves. We have developed a perfect molecular marker corresponding to the olol mutation that can facilitate a rapid screening and early detection of genotypes carrying the olol mutation for use in marker-assisted selection for the management of the HO trait in safflower breeding programmes.
Publisher: MDPI AG
Date: 11-03-2014
DOI: 10.3390/NU6031063
Publisher: Public Library of Science (PLoS)
Date: 07-11-2012
Publisher: Frontiers Media SA
Date: 2012
Start Date: 03-2023
End Date: 03-2026
Amount: $423,000.00
Funder: Australian Research Council
View Funded ActivityStart Date: 01-2004
End Date: 12-2004
Amount: $10,000.00
Funder: Australian Research Council
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