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Genetic Modification of Soybean with a Novel Grafting Technique: Downregulating the FAD2-1 Gene Increases Oleic Acid Content

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Abstract

A hairpin RNA-encoding construct targeting gmFAD2-1 was transformed into soybean, and an optimised Agrobacterium-mediated embryonic tip system was employed. A novel intergeneric grafting method using transgenic plantlets as scions was used instead of the conventional rooting method. Compared with traditional acclimatisation, the survival ratio of cleft grafts increased by 70%, and the culture period was shortened by about 40 days. The regeneration frequency of the grafted shoots in this embryonic tip system was approximately 74.6%. Soybean transformants were confirmed by Southern and Northern blot hybridisation analyses. The fatty acid composition of the T1 and T2 seeds from the transformed plants was determined by gas chromatography. The resulting downregulation of the gmFAD2-1 gene substantially increased the level of oleic acid from 16% to 55% as indicated by the oleic desaturation proportion (ODP). The ratio of plants with high ODP, moderate ODP and low ODP was about 1:2:1, which was consistent with a single-gene segregation pattern.

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References

  • Birch RG (1997) Plant transformation: problems and strategies for practical application. Annu Rev Plant Physiol Plant Mol Biol 48:297–326

    Article  PubMed  CAS  Google Scholar 

  • Buhr T, Sato S, Ebrahim F, Xing AQ, Zhou Y, Mathiesen M, Schweiger B, Kinney A, Staswick P, Clemente T (2002) Ribozyme termination of RNA transcripts down-regulate seed fatty acid genes in transgenic soybean. Plant J 30:155–163

    Article  PubMed  CAS  Google Scholar 

  • Chapman KD, Austin-Brown S, Sparace SA, Kinney AJ, Ripp KG, Pirtle IL, Pirtle RM (2001) Transgenic cotton plants with increased seed oleic acid content. J Am Oil Chem Soc 78:941–947

    Article  CAS  Google Scholar 

  • Chen Z, Wang ML, Barkley NA, Pittman RN (2010) A simple allele-specific PCR assay for detecting FAD2 alleles in both A and B genomes of the cultivated peanut for high-oleate trait selection. Plant Mol Biol Rep 28:542–548

    Article  CAS  Google Scholar 

  • Finer JJ, McMullen MD (1991) Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell Dev AN 27:175–182

    Article  Google Scholar 

  • Hamilton AJ, Brown S, Yuanhai H, Ishizuka M, Lowe A, Solis A-GA, Gierson D (2002) A transgene with repeated DNA causes high frequency, post-transcriptional suppression of ACC-oxidase gene expression in tomato. Plant J 15:737–746

    Article  Google Scholar 

  • Helfer A, Clément B, Michler P, Otten L (2003) The Agrobacterium oncogene AB-6b causes a graft-transmissible enation syndrome in tobacco. Plant Mol Biol 52:483–493

    Article  PubMed  CAS  Google Scholar 

  • Heppard EP, Kinney AJ, Stecca KL, Miao GH (1996) Developmental and growth temperature regulation of two different microsomal omega-6 desaturase genes in soybeans. Plant Physiol 110:311–319

    Article  PubMed  CAS  Google Scholar 

  • Hinchee MAW, Connor-Ward DV, Newell CA, McDonnell RE, Sato SJ, Gasser CS, Fischhoff DA, Re DB, Fraley RT, Horsch RB (1988) Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer. Nat Biotechnol 6:915–922

    Article  CAS  Google Scholar 

  • Hitz WD (1995) Plant lipid metabolism. Kluwer, Netherlands, pp 506–508

    Google Scholar 

  • Jiang BG (2004) Optimization of Agrobacterium mediated cotton transformation using shoot apices explants and quantitative trait loci analysis of yield and yield component traits in upland cotton (Gossypium hirsutum L.). Ph.D. Dissertation. Louisiana State University, Baton Rouge, LA 70803

  • Jin SX, Liang SG, Zhang XL, Nie Y, Guo XP (2006) An efficient grafting system for transgenic plant recovery in cotton (Gossypiumhirsutum L.). Plant Cell Tissue Organ 85:181–185

    Article  Google Scholar 

  • Kinny AJ (1998) Plants as industrial chemical factories—new oils from genetically engineered soybeans. Fett—Lipid 100:173–176

    Google Scholar 

  • Kinny AJ (2007) Development of genetically engineered soybean oils for food applications. J Food Lipids 3:273–292

    Article  Google Scholar 

  • Knowlton S, Elkton M (1999) Soybean oil having high oxidative stability. US Patent 5981781:1–14

    Google Scholar 

  • Ko TS, Lee S, Farrand SK, Korban SS (2004) A partially disarmed vir helper plasmid, pKYRT1, in conjunction with 2, 4-dichlorophenoxyactic acid promotes emergence of regenerable transgenic somatic embryos from immature cotyledons of soybean. Planta 218:536–541

    Article  PubMed  CAS  Google Scholar 

  • Lacombe S, Souyris I, Bervillé AJ (2009) An insertion of oleate desaturase homologous sequence silences via siRNA the functional gene leading to high oleic acid content in sunflower seed oil. Mol Genet Genomics 281:43–54

    Article  PubMed  CAS  Google Scholar 

  • Li MJ, Xia H, Zhao CZ, Li AQ, Li CS, Bi YP, Wan SB, Wang XJ (2010) Isolation and characterization of putative acetyl-CoA carboxylases in Arachis hypogaea L. Plant Mol Biol Rep 28:58–68

    Article  CAS  Google Scholar 

  • Liang YL, Zhao SJ, Zhang X (2009) Antisense suppression of cycloartenol synthase results in elevated ginsenoside levels in panax ginseng hairy roots. Plant Mol Biol Rep 27:298–304

    Article  CAS  Google Scholar 

  • Liu Q, Surinder S, Allen G (2002) High-steric and-oleic cottonseed oils produced by hairpin RNA-mediated post-transcriptional gene silencing. Plant Physiol 129:1–12

    Article  Google Scholar 

  • Liu HK, Yang C, Wei ZM (2004) Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system. Planta 219:1042–1049

    Article  PubMed  CAS  Google Scholar 

  • Loganathan M, Maruthasalam S, Shiu LY, Lien WC, Hsu WH, Lee PF, Yu CW, Lin CH (2010) Regeneration of soybean (Glycine max L. Merrill) through direct somatic embryogenesis from the immature embryonic shoot tip. In Vitro Cell Dev AN 46:265–273

    Article  Google Scholar 

  • Lou T, Deng WY, Zeng J, Zhang FL (2009) Cloning and characterization of a stearoyl-acyl carrier protein desaturase gene from Cinnamomum longepaniculatum. Plant Mol Biol Rep 27:13–19

    Article  Google Scholar 

  • Luo JH, Gould JH (1999) In vitro shoot-tip grafting improves recovery of cotton plants from culture. Plant Cell Tissue Organ 57:211–213

    Article  Google Scholar 

  • Mroczka A, Roberts PD, Fillatti JJ, Wiggins BE, Ulmasov T, Voelker T (2010) An intron sense suppression construct targeting soybean FAD2-1 requires a double-stranded RNA-producing inverted repeat T-DNA insert. Plant Physiol 153:882–891

    Article  PubMed  CAS  Google Scholar 

  • Nandeshwar SB, Moghe S, Chakrabarty PK, Deshattiwar MK, Kranthi K, Anandkumar P, Mayee CD, Khadi BM (2009) Agrobacterium-mediated transformation of cry1Ac gene into shoot-tip meristem of diploid cotton Gossypium arboreum cv. RG8 and regeneration of transgenic plants. Plant Mol Biol Rep 27:549–557

    Article  CAS  Google Scholar 

  • Olhoft PM, Lin K, Galbraith J, Nielsen NC, Somers DA (2001) The role of thiol compounds in increasing Agrobacterium-mediated transformation of soybean cotyledonary-node cells. Plant Cell Rep 20:731–737

    Article  CAS  Google Scholar 

  • Paz MM, Martinez JC, Kalvig AB, Fonger TM, Wang K (2006) Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Rep 25:206–213

    Article  PubMed  CAS  Google Scholar 

  • Schlueter JA, Vasylenko-Sanders IF, Deshpande S, Yi J, Siegfried M, Roe BA, Schlueter SD, Scheffler BE, Shoemaker RC (2007) The FAD2 gene family of soybean: insights into the structural and functional divergence of a paleopolyploid genome. Crop Sci 47:14–26

    Article  Google Scholar 

  • Singh SP, Green AG, Stoutjesdijk PA, Liu Q (2000) Inverted-repeat DNA: a new gene-silencing tool for seed lipid modification. Biochem Soc Trans 28:925–927

    Article  PubMed  CAS  Google Scholar 

  • Sleight P (1992) Cholesterol and coronary heart disease mortality. Intern Med J 22:576–579

    CAS  Google Scholar 

  • Somers DA, Samac DA, Olhoft PM (2003) Recent advances in legume transformation. Plant Physiol 131:892–899

    Article  PubMed  CAS  Google Scholar 

  • Stoutjesdijk PA, Hurlstone C, Singh SP, Green AG (2000) High-oleic acid Australian Brassica napus and B.juncea varieties produced by co-suppression of endogenous Δ12-desaturases. Biochem Soc Trans 28:938–940

    Article  PubMed  CAS  Google Scholar 

  • Stoutjesdijk PA, Singh SP, Liu Q, Hurlstone C, Waterhouse P, Green A (2002) HpRNA-mediated targeting of the arabidopsis FAD2 gene gives highly efficient and stable silencing. Plant Physiol 129:1723–1731

    Article  PubMed  CAS  Google Scholar 

  • Wang YX, Wang XF, Ma ZY, ZHang GY, Zhao JF (2007) A new high-efficient cotton graft technique and application. Sci Agricultura Sin 40:264–270

    Google Scholar 

  • Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, Green AG, Waterhouse PM (2001) Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J 27:581–590

    Article  PubMed  CAS  Google Scholar 

  • Xu YN, Wang ZN, Jiang GZ, Li LB, Kuang TY (2003) Effect of various temperature on phosphatidylglycerol biosynthesis in thylakoid membrane. Plant Physiol 118:57–63

    Article  CAS  Google Scholar 

  • Yin DM, Deng SZ, Zhan KH, Cui DQ (2007) High-oleic peanut oils produced by hpRNA-mediated gene silencing of oleate desaturase. Plant Mol Biol Rep 25:154–163

    Article  CAS  Google Scholar 

  • Zambre MA, Clercq JD, Vranová E, Montagu VM, Angenon G, Dillen W (1998) Plant regeneration from embryo-derived callus in Phaseolus vulgaris L. (common bean) and P. acutifolius A. Gray (tepary bean). Plant Cell Rep 17:626–630

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the project supporting programmes from the genetically modified organisms breeding major projects, China (no. 2008ZX08004-004). We thank Jilin Academy of Agricultural Sciences for kindly supplying seeds.

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Authors and Affiliations

  1. Key Laboratory of Vegetation Ecology, School of Life Sciences, Northeast Normal University, 130024, Changchun, China

    Wei Chen, Kai Song, Yirong Cai, Wangfeng Li & Lixia Liu

  2. Key Laboratory of Molecular Epigenetics of MOE, Northeast Normal University, 130024, Changchun, China

    Bao Liu

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  1. Wei Chen
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  2. Kai Song
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  3. Yirong Cai
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  4. Wangfeng Li
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  5. Bao Liu
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  6. Lixia Liu
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Correspondence to Lixia Liu.

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Chen, W., Song, K., Cai, Y. et al. Genetic Modification of Soybean with a Novel Grafting Technique: Downregulating the FAD2-1 Gene Increases Oleic Acid Content. Plant Mol Biol Rep 29, 866–874 (2011). https://doi.org/10.1007/s11105-011-0286-5

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