Biotechnology for Biofuels | |
Development of marker-free transgenic Jatropha plants with increased levels of seed oleic acid | |
Jing Qu1  Hui-Zhu Mao1  Wen Chen1  Shi-Qiang Gao1  Ya-Nan Bai1  Yan-Wei Sun1  Yun-Feng Geng1  Jian Ye1  | |
[1] Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore | |
关键词: transgenic; marker free; Jatropha; high oleic acid; FAD2; Cre-lox recombination; Biodiesel; | |
Others : 798348 DOI : 10.1186/1754-6834-5-10 |
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received in 2011-10-27, accepted in 2012-02-29, 发布年份 2012 | |
【 摘 要 】
Background
Jatropha curcas is recognized as a new energy crop due to the presence of the high amount of oil in its seeds that can be converted into biodiesel. The quality and performance of the biodiesel depends on the chemical composition of the fatty acids present in the oil. The fatty acids profile of the oil has a direct impact on ignition quality, heat of combustion and oxidative stability. An ideal biodiesel composition should have more monounsaturated fatty acids and less polyunsaturated acids. Jatropha seed oil contains 30% to 50% polyunsaturated fatty acids (mainly linoleic acid) which negatively impacts the oxidative stability and causes high rate of nitrogen oxides emission.
Results
The enzyme 1-acyl-2-oleoyl-sn-glycero-3-phosphocholine delta 12-desaturase (FAD2) is the key enzyme responsible for the production of linoleic acid in plants. We identified three putative delta 12 fatty acid desaturase genes in Jatropha (JcFAD2s) through genome-wide analysis and downregulated the expression of one of these genes, JcFAD2-1, in a seed-specific manner by RNA interference technology. The resulting JcFAD2-1 RNA interference transgenic plants showed a dramatic increase of oleic acid (> 78%) and a corresponding reduction in polyunsaturated fatty acids (< 3%) in its seed oil. The control Jatropha had around 37% oleic acid and 41% polyunsaturated fatty acids. This indicates that FAD2-1 is the major enzyme responsible for converting oleic acid to linoleic acid in Jatropha. Due to the changes in the fatty acids profile, the oil of the JcFAD2-1 RNA interference seed was estimated to yield a cetane number as high as 60.2, which is similar to the required cetane number for conventional premium diesel fuels (60) in Europe. The presence of high seed oleic acid did not have a negative impact on other Jatropha agronomic traits based on our preliminary data of the original plants under greenhouse conditions. Further, we developed a marker-free system to generate the transgenic Jatropha that will help reduce public concerns for environmental issues surrounding genetically modified plants.
Conclusion
In this study we produced seed-specific JcFAD2-1 RNA interference transgenic Jatropha without a selectable marker. We successfully increased the proportion of oleic acid versus linoleic in Jatropha through genetic engineering, enhancing the quality of its oil.
【 授权许可】
2012 Qu et al; licensee BioMed Central Ltd.
【 预 览 】
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【 参考文献 】
- [1]Durrett TP, Benning C, Ohlrogge J: Plant triacylglycerols as feedstocks for the production of biofuels. Plant J 2008, 54:593-607.
- [2]Graef G, LaVallee BJ, Tenopir P, Tat M, Schweiger B, Kinney AJ, Van Gerpen JH, Clemente TE: A high-oleic-acid and low-palmitic-acid soybean: agronomic performance and evaluation as a feedstock for biodiesel. Plant Biotechnol J 2009, 7:411-421.
- [3]Frankel EN: Lipid Oxidation. Dundee, UK: Oily Press; 1998.
- [4]Tat ME, Wang PS, Gerpen JHV, Clemente TE: Exhaust emissions from an engine fueled with biodiesel from high-oleic soybeans. J Am Oil Chem Soc 2007, 84:865-869.
- [5]Carroll A, Somerville C: Cellulosic Biofuels. Annu Rev Plant Biol 2008, 60:165-182.
- [6]Joachim H (Ed): Physic nut. Jatropha curcas L Gatersleben, Rome: Institute of Plant Genetics and Crop Plant Research/International Plant Genetic Resources Institute; 1996.
- [7]Clemente TE, Cahoon EB: Soybean oil: genetic approaches for modification of functionality and total content. Plant Physiol 2009, 151:1030-1040.
- [8]Ye J, Qu J, Bui HT, Chua NH: Rapid analysis of Jatropha curcas gene functions by virus-induced gene silencing. Plant Biotechnol J 2009, 7:964-976.
- [9]Li MR, Li HQ, Jiang HW, Pan XP, Wu GJ: Establishment of an Agrobacteriuim-mediated cotyledon disc transformation method for Jatropha curcas. Plant Cell Tiss Organ Cult 2008, 92:173-181.
- [10]Lu C, Napier JA, Clemente TE, Cahoon EB: New frontiers in oilseed biotechnology: meeting the global demand for vegetable oils for food, feed, biofuel, and industrial applications. Curr Opin Biotechnol 2011, 22:252-259.
- [11]Sato S, Hirakawa H, Isobe S, Fukai E, Watanabe A, Kato M, Kawashima K, Minami C, Muraki A, Nakazaki N, et al.: Sequence analysis of the genome of an oil-bearing tree, Jatropha curcas L. DNA Res 2011, 18:65-76.
- [12]Dyer JM, Chapital DC, Kuan JC, Mullen RT, Turner C, McKeon TA, Pepperman AB: Molecular analysis of a bifunctional fatty acid conjugase/desaturase from tung. Implications for the evolution of plant fatty acid diversity. Plant Physiol 2002, 130:2027-2038.
- [13]Lu C, Fulda M, Wallis JG, Browse J: A high-throughput screen for genes from castor that boost hydroxy fatty acid accumulation in seed oils of transgenic Arabidopsis. Plant J 2006, 45:847-856.
- [14]Zuo J, Niu QW, Moller SG, Chua NH: Chemical-regulated, site-specific DNA excision in transgenic plants. Nat Biotechnol 2001, 19:157-161.
- [15]Guo HS, Fei JF, Xie Q, Chua NH: A chemical-regulated inducible RNAi system in plants. Plant J 2003, 34:383-392.
- [16]Schuler MA, Ladin BF, Pollaco JC, Freyer G, Beachy RN: Structural sequences are conserved in the genes coding for the alpha, alpha' and beta-subunits of the soybean 7S seed storage protein. Nucleic Acids Res 1982, 10:8245-8261.
- [17]Yi C, Zhang S, Liu X, Bui HT, Hong Y: Does epigenetic polymorphism contribute to phenotypic variances in Jatropha curcas L.? BMC Plant Biol 2010 2010., 10
- [18]Deng X, Fang Z, Liu Yh, Yu CL: Production of biodiesel from Jatropha oil catalyzed by nanosized solid basic catalyst. Energy 2011, 36:777-784.
- [19]Barua PK: Biodiesel from Seeds of Jatropha Found in Assam, India. International Journal of Energy, Information and Communications 2011, 2:53-65.
- [20]Bamgboye A, Hansen A: Prediction of cetane number of biodiesel fuel from the fatty acid methyl ester (FAME) composition. International Agrophysics 2008, 22:21-29.
- [21]Sarina A, Arorab R, Singhb NP, Sharmac M, Malhotra RK: Influence of metal contaminants on oxidation stability of Jatropha biodiesel. Energy 2009, 34:1271-1275.
- [22]Chapman EJ, Carrington JC: Specialization and evolution of endogenous small RNA pathways. Nat Rev Genet 2007, 8:884-896.
- [23]Scherder C, Fehr W: Agronomic and seed characteristics of soybean lines with increased oleate content. Crop Sci 2008, 48:1755-1758.
- [24]Bachlava E, Burton J, Brownie C, Wang S, Auclair J, Cardinal A: Heritability of oleic acid content in soybean seed oil and its genetic correlation with fatty acid and agronomics traits. Crop Sci 2008, 48:1764-1772.
- [25]Jones A, Davies HM, Voelker TA: Palmitoyl-acyl carrier protein (ACP) thioesterase and the evolutionary origin of plant acyl-ACP thioesterases. Plant Cell 1995, 7:359-371.
- [26]Wu PZ, Li J, Wei Q, Zeng L, Chen YP, Li MR, Jiang HW, Wu GJ: Cloning and functional characterization of an acyl-acyl carrier protein thioesterase (JcFATB1) from Jatropha curcas. Tree Physiol 2009, 29:1299-1305.
- [27]Pan B-Z, Xu Z-F: Benzyladenine Treatment Significantly Increases the Seed Yield of the Biofuel Plant Jatropha curcas. J Plant Growth Regul 2011, 30:166-174.
- [28]Mao HZ, Ye J, Chua NH: Genetic transformation of Jatropha curcas. 2009. International Application No.:PCT/SG2009/000479. U.S.A
- [29]Allen GC, Flores-Vergara MA, Krasynanski S, Kumar S, Thompson WF: A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide. Nat Protoc 2006, 1:2320-2325.
- [30]Sambrook J, Russell DW: Molecular cloning: a laboratory manual. 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 2001.