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MAM gene silencing leads to the induction of C3 and reduction of C4 and C5 side-chain aliphatic glucosinolates in Brassica napus

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Abstract

Methylthioalkylmalate (MAM) synthases and their associated genes that have been extensively investigated in Arabidopsis control the side-chain elongation of methionine during the synthesis of aliphatic glucosinolates. A Brassica homolog of the Arabidopsis MAM genes was used in this study to analyze the role of MAM genes in B. napus through RNA interference (RNAi). The silencing of the MAM gene family in B. napus canola and B. napus rapeseed resulted in the reduction of aliphatic glucosinolates and total glucosinolate content. The results indicated that RNAi has potential for reducing glucosinolate content and improving meal quality in B. napus canola and rapeseed cultivars. Interestingly, MAM gene silencing in B. napus significantly induced the production of 2-propenyl glucosinolate, a 3-carbon side-chain glucosinolate commonly found in B. juncea mustard. Most transgenic plants displayed induction of 2-propenyl glucosinolate; however, the absolute content of this glucosinolate in transgenic B. napus canola was relatively low (less than 1.00 μmol g−1 seed). In the high glucosinolate content progenies derived from the crosses of B. napus rapeseed and transgenic B. napus canola, MAM gene silencing strongly induced the production of 2-propenyl glucosinolate to high levels (up to 4.45 μmol g−1 seed).

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References

  • Bednarek P, Pislewska-Bednarek M, Svatos A, Schneider B, Doubsky J, Mansurova M, Humphry M, Consonni C, Panstruga R, Sanchez-Vallet A, Molina A, Schulze-Lefert P (2009) A glucosinolate metabolism pathway in living plant cells mediates broad-spectrum antifungal defense. Science 323:101–106

    Article  PubMed  CAS  Google Scholar 

  • Beekwilder J, van Leeuwen W, van Dam NM, Bertossi M, Grandi V, Mizzi L, Soloviev M, Szabados L, Molthoff JW, Schipper B, Verbocht H, de Vos RC, Morandini P, Aarts MG, Bovy A (2008) The impact of the absence of aliphatic glucosinolates on insect herbivory in Arabidopsis. PLoS ONE 3:e2068

    Article  PubMed  Google Scholar 

  • Benderoth M, Textor S, Windsor AJ, Mitchell-Olds T, Gershenzon J, Kroymann J (2006) Positive selection driving diversification in plant secondary metabolism. Proc Natl Acad Sci USA 103:9118–9123

    Article  PubMed  CAS  Google Scholar 

  • Celenza JL, Quiel JA, Smolen GA, Merrikh H, Silvestro AR et al (2005) The Arabidopsis ATR1 Myb transcription factor controls indolic glucosinolate homeostasis. Plant Physiol 137:253–262

    Article  PubMed  CAS  Google Scholar 

  • Chen SX, Glawischnig E, Jorgensen K, Naur P, Jorgensen B, Olsen CE, Hansen CH, Rasmussen H, Pickett JA, Halkier BA (2003) CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis. Plant J 33:923–937

    Article  PubMed  CAS  Google Scholar 

  • Clay NK, Adio AM, Denoux C, Jander G, Ausubel FM (2009) Glucosinolate metabolites required for an Arabidopsis innate immune response. Science 323:95–101

    Article  PubMed  CAS  Google Scholar 

  • de Kraker JW, Luck K, Textor S, Tokuhisa JG, Gershenzon J (2007) Two Arabidopsis genes (IPMS1 and IPMS2) encode isopropylmalate synthase, the branch point step in the biosynthesis of leucine. Plant Physiol 143:970–986

    Article  PubMed  Google Scholar 

  • Fahey JW, Zhang Y, Talalay P (1997) Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. Proc Natl Acad Sci USA 94:10367–10372

    Article  PubMed  CAS  Google Scholar 

  • Gigolashvili T, Berger B, Mock HP, Muller C, Weisshaar B et al (2007a) The transcription factor HIG1/MYB51 regulates indolic glucosinolate biosynthesis in Arabidopsis thaliana. Plant J 50:886–901

    Article  PubMed  CAS  Google Scholar 

  • Gigolashvili T, Yatusevich R, Berger B, Muller C, Flugge UI (2007b) The R2R3-MYB transcription factor HAG1/MYB28 is a regulator of methionine-derived glucosinolate biosynthesis in Arabidopsis thaliana. Plant J 51:247–261

    Article  PubMed  CAS  Google Scholar 

  • Grubb CD, Abel S (2006) Glucosinolate metabolism and its control. Trends Plant Sci 11:89–100

    Article  PubMed  CAS  Google Scholar 

  • Halkier BA, Gershenzon J (2006) Biology and biochemistry of glucosinolates. Annu Rev Plant Biol 57:303–333

    Article  PubMed  CAS  Google Scholar 

  • Hasan M, Friedt W, Pons-Kühnemann J, Freitag NM, Link K, Snowdon RJ (2008) Association of gene-linked SSR markers to seed glucosinolate content in oilseed rape (Brassica napus ssp. napus). Theor Appl Genet 116:1035–1049

    Article  PubMed  CAS  Google Scholar 

  • Hirai MY, Sugiyama K, Sawada Y, Tohge T, Obayashi T et al (2007) Omics based identification of Arabidopsis Myb transcription factors regulating aliphatic glucosinolate biosynthesis. Proc Natl Acad Sci USA 104:6478–6483

    Article  PubMed  CAS  Google Scholar 

  • Kliebenstein DJ, Kroymann J, Brown P, Figuth A, Pedersen D, Gershenzon J, Mitchell-Olds T (2001) Genetic control of natural variation in Arabidopsis thaliana glucosinolate accumulation. Plant Physiol 126:811–825

    Article  PubMed  CAS  Google Scholar 

  • Kraling K, Robbelen G, Thies W, Herrmann M, Ahmadi MR (1990) Variation in seed glucosinolates in lines of Brassica napus. Plant Breed 105:33–39

    Article  Google Scholar 

  • Kroymann J, Textor S, Tokuhisa JG, Falk KL, Bartram S, Gershenzon J, Mitchell-Olds T (2001) A gene controlling variation in Arabidopsis glucosinolate composition is part of the methionine chain elongation pathway. Plant Physiol 127:1077–1088

    Article  PubMed  CAS  Google Scholar 

  • Kroymann J, Donnerhacke S, Schnabelrauch D, Mitchell-Olds T (2003) Evolutionary dynamics of an Arabidopsis insect resistance quantitative trait locus. Proc Natl Acad Sci USA 100:14587–14592

    Article  PubMed  CAS  Google Scholar 

  • Li G, Quiros CF (2002) Genetic analysis, gene expression and molecular characterization of BoGSL-ELONG, a major gene involved in the aliphatic glucosinolate pathway of Brassica species. Genetics 162(4):1937–1943

    PubMed  CAS  Google Scholar 

  • Li G, Quiros CF (2003) The Brassica oleracea gene BoGSL-ALK regulates in planta alkenyl aliphatic glucosinolates in Arabidopsis thaliana. Theor Appl Genet 106:1116–1121

    PubMed  CAS  Google Scholar 

  • Li G, Gao M, Yang B, Quiros CF (2003) Gene to gene alignment between the Arabidopsis and Brassica oleracea genomes. Theor Appl Genet 107:168–180

    Article  PubMed  CAS  Google Scholar 

  • Magrath R, Herron C, Giamoustaris A, Mithen R (1993) The inheritance of aliphatic glucosinolates in Brassica napus. Plant Breed 111:55–72

    Article  CAS  Google Scholar 

  • Magrath R, Bano F, Morgner M, Parkin I, Sharpe A, Lister C, Dean C, Turner J, Lydiate D, Mithen R (1994) Genetics of aliphatic glucosinolates. 1. Side chain elongation in Brassica napus and Arabidopsis thaliana. Heredity 72:290–299

    Article  CAS  Google Scholar 

  • Matthiessen JN, Shackleton MA (2005) Biofumigation: environmental impacts on the biological activity of diverse pure and plant-derived isothiocyanates. Pest Manag Sci 61:1043–1051

    Article  PubMed  CAS  Google Scholar 

  • McVetty PBE, Fernando D, Li G, Tahir M, Zelmer C (2009) High-erucic acid, low-glucosinolate rapeseed (HEAR) cultivar development in Canada. In: Hou CT, Shaw JF (eds) Biocatalysis and agricultural biotechnology. CRC, Boca Raton, pp 43–61

    Google Scholar 

  • Moloney MM, Walker JM, Sharma KK (1989) High efficiency transformation of Brassica napus using Agrobacterium vectors. Plant Cell Rep 8:238–242

    Article  CAS  Google Scholar 

  • Nesi N, Delourme R, Brégeon M, Falentin C, Renard M (2008) Genetic and molecular approaches to improve nutritional value of Brassica napus L. seed. C R Biol 331:763–771

    Article  PubMed  CAS  Google Scholar 

  • Nour-Eldin HH, Hansen BG, Norholm MHH, Jensen JK, Halkier BA (2006) Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments. Nucleic Acids Res 34:e122

    Article  PubMed  Google Scholar 

  • Quiros CF, Grellet F, Sadowski J, Suzuki T, Li G, Wroblewski T (2001) Arabidopsis and Brassica comparative genomics: sequence, structure and gene content in the ABI1-Rps2-Ck1 chromosomal segment and related regions. Genetics 157:1321–1330

    PubMed  CAS  Google Scholar 

  • Reintanz B, Lehnen M, Reichelt M, Gershenzon J, Kowalczyk M, Sandberg G, Godde M, Uhl R, Palme K (2001) Bus, a bushy Arabidopsis CYP79F1 knockout mutant with abolished synthesis of short-chain aliphatic glucosinolates. Plant Cell 13:351–367

    Article  PubMed  CAS  Google Scholar 

  • Schuster J, Knill T, Reichelt M, Gershenzon J, Binder S (2006) BRANCHED-CHAIN AMINOTRANSFERASE4 is part of the chain elongation pathway in the isoynthesis of methionine derived glucosinolates in Arabidopsis. Plant Cell 18:2664–2679

    Article  PubMed  CAS  Google Scholar 

  • Sønderby IE, Hansen BG, Bjarnholt N, Ticconi C, Halkier BA et al (2007) A systems biology approach Identifies a R2R3 MYB gene subfamily with distinct and overlapping functions in regulation of aliphatic glucosinolates. PloS ONE 2:e1322

    Article  PubMed  Google Scholar 

  • Textor S, Bartram S, Kroymann J, Falk KL, Hick A, Pickett JA, Gershenzon J (2004) Biosynthesis of methionine-derived glucosinolates in Arabidopsis thaliana: recombinant expression and characterization of methylthioalkylmalate synthase, the condensing enzyme of the chain-elongation cycle. Planta 218:1026–1035

    Article  PubMed  CAS  Google Scholar 

  • Textor S, de Kraker JW, Hause B, Gershenzon J, Tokuhisa JG (2007) MAM3 catalyzes the formation of all aliphatic glucosinolate chain lengths in Arabidopsis. Plant Physiol 144:60–71

    Article  PubMed  CAS  Google Scholar 

  • Vinjamoori V, Byrum JR, Hayes T, Das PK (2004) Challenges and opportunities in the analysis of raffinose oligosaccharides, pentosans. J Anim Sci 82:319–328

    PubMed  CAS  Google Scholar 

  • Zang YX, Kim JH, Park YD, Kim DH, Hong SB (2008) Metabolic engineering of aliphatic glucosinolates in Chinese cabbage plants expressing Arabidopsis MAM1, CYP79F1, and CYP83A1. BMB Rep 41:472–478

    PubMed  CAS  Google Scholar 

  • Zang YX, Kim HU, Kim JA, Lim MH, Jin M, Lee SC, Kwon SJ, Lee SI, Hong JK, Park TH, Mun JH, Seol YJ, Hong SB, Park BS (2009) Genome-wide identification of glucosinolate synthesis genes in Brassica rapa. FEBS J 276:3559–3574

    Article  PubMed  CAS  Google Scholar 

  • Zhang GY, Talalay P, Cho C, Posner GH (1992) A major inducer of anticarcinogenic protective enzymes from broccoli: isolation and elucidation of structure. Proc Natl Acad Sci USA 89:2399–2403

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The research was supported by the Genome Canada/Genome Alberta and Genome Prairie project “Designing Oilseeds for Tomorrow’s Markets” and Manitoba Provincial Government. We thank Dr. Rakow from Agriculture Agri-Food Canada, Saskatoon who provided the material DH12075 for plant transformation.

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Authors and Affiliations

  1. The Department of Plant Science, University of Manitoba, Winnipeg, MB, R3T2N2, Canada

    Zheng Liu, Peter B. E. McVetty, Fouad Daayf & Genyi Li

  2. Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada

    Joe Hammerlindl

  3. Genome Prairie, 101-111 Research Drive, Saskatoon, SK, S7N 3R2, Canada

    Wilf Keller

  4. The Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Carlos F. Quiros

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  1. Zheng Liu
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Correspondence to Genyi Li.

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Liu, Z., Hammerlindl, J., Keller, W. et al. MAM gene silencing leads to the induction of C3 and reduction of C4 and C5 side-chain aliphatic glucosinolates in Brassica napus. Mol Breeding 27, 467–478 (2011). https://doi.org/10.1007/s11032-010-9444-y

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