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Li C.,National University of Singapore | Ng A.,National University of Singapore | Ng A.,Nanyang Technological University | Xie L.,National University of Singapore | And 7 more authors.
Plant Cell Reports | Year: 2016

Key message: Casbene is a precursor to phorbol esters and down-regulating casbene synthase effectively reduces phorbol ester biosynthesis.Seed-specific reduction of phorbol ester (PE) helps develop Jatropha seed cake for animal nutrition. Abstract: Phorbol esters (PEs) are diterpenoids present in some Euphorbiaceae family members like Jatropha curcas L. (Jatropha), a tropical shrub yielding high-quality oil suitable as feedstock for biodiesel and bio jet fuel. Jatropha seed contains up to 40 % of oil and can produce oil together with cake containing high-quality proteins. However, skin-irritating and cancer-promoting PEs make Jatropha cake meal unsuitable for animal nutrition and also raise some safety and environmental concerns on its planting and processing. Two casbene synthase gene (JcCASA163 and JcCASD168) homologues were cloned from Jatropha genome and both genes were highly expressed during seed development. In vitro functional analysis proved casbene synthase activity of JcCASA163 in converting geranylgeranyl diphosphate into casbene which has been speculated to be the precursor to PEs. A seed-specific promoter driving inverted repeats for RNAi interference targeting at either JcCASA163 or both genes could effectively down-regulate casbene synthase gene expression with concurrent marked reduction of PE level (by as much as 85 %) in seeds with no pleiotropic effects observed. Such engineered low PE in seed was heritable and co-segregated with the transgene. Our work implicated casbene synthase in Jatropha PE biosynthesis and provided evidence for casbene being the precursor for PEs. The success in reducing seed PE content through down-regulation of casbene synthase demonstrates the feasibility of intercepting PE biosynthesis in Jatropha seed to help address safety concerns on Jatropha plantation and seed processing and facilitate use of its seed protein for animal nutrition. © 2015, Springer-Verlag Berlin Heidelberg.


Yunping B.,National University of Singapore | Ngoc Ha B.T.,National University of Singapore | Eunice Y.,Nanyang Technological University | Loong Chueng L.,JOil S Pte Ltd | And 3 more authors.
Ecotoxicology and Environmental Safety | Year: 2012

Jatropha curcas (Jatropha) is a tropical shrub that is gaining popularity as a biofuel feedstock plant. Phorbol esters (PEs) are tetracyclic tiglian diterpenoids that are present in Jatropha seeds and other parts of plant. Epidermal cell irritating and cancer promoting PEs not only reduce commercial values of Jatropha seed cake but also cause some safety and environment concerns on PE leaching to soil. A simple bioassay of PE toxicity was conducted by incubating 48h old brine shrimp (Artemia salina) nauplii with Jatropha oil for 24h. 1-4% of Jatropha oil (corresponding to PE concentration of 25-100mgL-1) had mortality rate of 5-95%, with LC50 estimated to be 2.7% of oil or 67mgL-1 of PE. Jatropha oil was incubated with clay or black soil (autoclaved or non-autoclaved) in the darkness or under sunlight for different periods of time before oil was re-extracted and tested for PE content by HPLC and for remaining toxicity with the brine shrimp bioassay. Under sunlight, PE decreased to non-detectable level within six days. Toxicity reduced to less than 5% mortality rate that is comparable to rapeseed oil control within the same period. In contrast, PE level and toxicity remained little changed when Jatropha oil was incubated in the darkness. Such PE degradation/detoxification was also found independent of the presence of soil or soil microorganisms. We conclude that sunlight directly degrades and detoxifies PEs and this finding should alleviate the concern on long term environmental impact of PE leaching. © 2012 Elsevier Inc.


Yi C.,JOil S Pte Ltd | Zhang S.,JOil S Pte Ltd | Liu X.,National University of Singapore | Bui H.T.N.,National University of Singapore | And 2 more authors.
BMC Plant Biology | Year: 2010

Background: There is a growing interest in Jatropha curcas L. (jatropha) as a biodiesel feedstock plant. Variations in its morphology and seed productivity have been well documented. However, there is the lack of systematic comparative evaluation of distinct collections under same climate and agronomic practices. With the several reports on low genetic diversity in jatropha collections, there is uncertainty on genetic contribution to jatropha morphology.Result: In this study, five populations of jatropha plants collected from China (CN), Indonesia (MD), Suriname (SU), Tanzania (AF) and India (TN) were planted in one farm under the same agronomic practices. Their agronomic traits (branching pattern, height, diameter of canopy, time to first flowering, dormancy, accumulated seed yield and oil content) were observed and tracked for two years. Significant variations were found for all the agronomic traits studied. Genetic diversity and epigenetic diversity were evaluated using florescence Amplified Fragment Length Polymorphism (fAFLP) and methylation sensitive florescence AFLP (MfAFLP) methods. Very low level of genetic diversity was detected (polymorphic band <0.1%) within and among populations. In contrast, intermediate but significant epigenetic diversity was detected (25.3% of bands were polymorphic) within and among populations. More than half of CCGG sites surveyed by MfAFLP were methylated with significant difference in inner cytosine and double cytosine methylation among populations. Principal coordinates analysis (PCoA) based on Nei's epigenetic distance showed Tanzania/India group distinct from China/Indonesia/Suriname group. Inheritance of epigenetic markers was assessed in one F1 hybrid population between two morphologically distinct parent plants and one selfed population. 30 out of 39 polymorphic markers (77%) were found heritable and followed Mendelian segregation. One epiallele was further confirmed by bisulphite sequencing of its corresponding genomic region.Conclusion: Our study confirmed climate and practice independent differences in agronomic performance among jatropha collections. Such agronomic trait variations, however, were matched by very low genetic diversity and medium level but significant epigenetic diversity. Significant difference in inner cytosine and double cytosine methylation at CCGG sites was also found among populations. Most epigenetic differential markers can be inherited as epialleles following Mendelian segregation. These results suggest possible involvement of epigenetics in jatropha development. © 2010 Yi et al; licensee BioMed Central Ltd.


Trademark
JOil S Pte. Ltd. | Date: 2013-08-13

Industrial oils and greases; industrial lubricants; fuels; biodiesel fuel; illuminants, namely, illuminating grease, illuminating oil, illuminating wax and illuminating fuel; candles and wicks for candles for lighting. Agricultural, horticultural and forestry products in the nature of living plants and parts of living plants, namely, bulbs, seeds, roots, stems, leaves, flowers, fresh or raw fruits, and seedlings; live trees; live fruit trees; live plants; live flowering plants; natural flowers; flower seeds; seedlings; seeds for plant propagation; parts of living plants, namely, stem and root tubers for plant propagation; seedlings for plant propagation; live Jatropha Curcus plants; Jatropha Curcus seeds, seedlings and fresh fruits; live Jatropha Curcus plants and seedlings for propagation purposes. Marketing and promotion services; business management regarding marketing activities and launching of new products; advertising; business consultancy services; business consultancy services relating to commercial propagation of plants; retail store services featuring plants; wholesale stores featuring plants; the bringing together, for the benefit of others, of a variety of goods, enabling customers to conveniently view and purchase those goods from a retail or wholesale outlet or from a general merchandise catalogue by mail order, telephone, fax and/or from a computer database or the global communications network particularly specializing in the marketing of the sale of goods of others; business management; business administration; administration of the business affairs of laboratories and plantations; business assistance relating to the establishment and operation of laboratories and plantations; arranging and conducting trade shows and exhibitions relating to plants, horticulture and agriculture; information, advisory and consultancy services relating to all the aforesaid services.


Trademark
JOil S Pte. Ltd. | Date: 2011-01-18

Industrial oils and greases; industrial lubricants; fuels; biodiesel fuel; illuminants, namely, illuminating grease, illuminating oil, illuminating wax and illuminating fuel; candles and wicks for candles for lighting. Agricultural, horticultural and forestry products in the nature of living plants and parts of living plants, namely, bulbs, seeds, roots, stems, leaves, flowers, fresh or raw fruits, and seedlings; live trees; live fruit trees; live plants; live flowering plants; natural flowers; flower seeds; seedlings; seeds for plant propagation; parts of living plants, namely, stem and root tubers for plant propagation; seedlings for plant propagation; live Jatropha Curcus plants; Jatropha Curcus seeds, seedlings and fresh fruits; live Jatropha Curcus plants and seedlings for propagation purposes. Marketing and promotion services; business management regarding marketing activities and launching of new products; advertising; business consultancy services; business consultancy services relating to commercial propagation of plants; retail store services featuring plants; wholesale stores featuring plants; the bringing together, for the benefit of others, of a variety of goods, enabling customers to conveniently view and purchase those goods from a retail or wholesale outlet or from a general merchandise catalogue by mail order, telephone, fax and/or from a computer database or the global communications network particularly specializing in the marketing of the sale of goods of others; business management; business administration; administration of the business affairs of laboratories and plantations; business assistance relating to the establishment and operation of laboratories and plantations; arranging and conducting trade shows and exhibitions relating to plants, horticulture and agriculture; information, advisory and consultancy services relating to all the aforesaid services.


PubMed | JOil S Pte Ltd
Type: | Journal: BMC plant biology | Year: 2010

There is a growing interest in Jatropha curcas L. (jatropha) as a biodiesel feedstock plant. Variations in its morphology and seed productivity have been well documented. However, there is the lack of systematic comparative evaluation of distinct collections under same climate and agronomic practices. With the several reports on low genetic diversity in jatropha collections, there is uncertainty on genetic contribution to jatropha morphology.In this study, five populations of jatropha plants collected from China (CN), Indonesia (MD), Suriname (SU), Tanzania (AF) and India (TN) were planted in one farm under the same agronomic practices. Their agronomic traits (branching pattern, height, diameter of canopy, time to first flowering, dormancy, accumulated seed yield and oil content) were observed and tracked for two years. Significant variations were found for all the agronomic traits studied. Genetic diversity and epigenetic diversity were evaluated using florescence Amplified Fragment Length Polymorphism (fAFLP) and methylation sensitive florescence AFLP (MfAFLP) methods. Very low level of genetic diversity was detected (polymorphic band <0.1%) within and among populations. In contrast, intermediate but significant epigenetic diversity was detected (25.3% of bands were polymorphic) within and among populations. More than half of CCGG sites surveyed by MfAFLP were methylated with significant difference in inner cytosine and double cytosine methylation among populations. Principal coordinates analysis (PCoA) based on Neis epigenetic distance showed Tanzania/India group distinct from China/Indonesia/Suriname group. Inheritance of epigenetic markers was assessed in one F1 hybrid population between two morphologically distinct parent plants and one selfed population. 30 out of 39 polymorphic markers (77%) were found heritable and followed Mendelian segregation. One epiallele was further confirmed by bisulphite sequencing of its corresponding genomic region.Our study confirmed climate and practice independent differences in agronomic performance among jatropha collections. Such agronomic trait variations, however, were matched by very low genetic diversity and medium level but significant epigenetic diversity. Significant difference in inner cytosine and double cytosine methylation at CCGG sites was also found among populations. Most epigenetic differential markers can be inherited as epialleles following Mendelian segregation. These results suggest possible involvement of epigenetics in jatropha development.

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