Abstract
Molecular mapping and tagging of the erucic acid trait (C22:1) in Brassica juncea was done by a candidate gene approach. Two QTLs underlying the variation of seed erucic acid content were assigned to two linkage groups of a B. juncea map using a doubled haploid (DH) mapping population derived from high × low erucic acid F1 hybrid. Two consensus primers corresponding to the full-length Fatty Acid Elongase 1 (FAE1) gene, reported to be involved in the elongation of C18:1 to C22:1, were designed. PCR amplification and subsequent cloning and sequencing identified two FAE1 genes (FAE1.1 and FAE1.2) in both high and low erucic acid mustard lines. Sequence alignment of corresponding FAE1 genes between high and low erucic acid mustard lines identified four substitution type single nucleotide polymorphisms (SNPs) in FAE1.1 and three in FAE1.2. Using the SNuPE method of SNP genotyping, these two genes were mapped to two independent loci that co-segregated with the two QTLs governing the erucic acid trait. Association of wild (E1E2) and mutant (e1e2) haplotypes of two FAE1 genes with erucic acid variation in two segregating populations revealed that the e1e1e2e2 genotype identified low erucic acid individuals (<2%) and E1E1E2E2 identified individuals with highest erucic acid content (>40%). The E1e1E2e2 heterozygote was found to be intermediate in phenotype. The applicability of these SNPs in marker-assisted manipulation of the erucic acid trait was verified by genotyping a set of contrasting germplasm of B. juncea belonging to two distinct gene pools (Indian and east European) and other oil-yielding Brassica species.
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References
Barret P, Delourme R, Renard M, Domergue F, Lessire R, Delseny M, Roscoe TJ (1998) A rapeseed FAE1 gene is linked to the E1 locus associated with variation in the content of erucic acid. Theor Appl Genet 96:177–186
Batley J, Mogg R, Edwards D, O’Sullivan H, Edwards KJ (2003) A high-throughput SNuPE assay for genotyping SNPs in the flanking regions of Zea mays sequence tagged simple sequence repeats. Mol Breed 11:111–120
Beare JL, Campbell JA, Youngs CG, Craig BM (1963) Effects of saturated fat in rats fed rapeseed oil. Can J Biochem Physiol 41:605–612
Bhat MA, Gupta ML, Banga SK, Raheja RK, Banga SS (2002) Erucic acid heredity in Brassica juncea—some additional information. Plant Breed 121:456–458
Brar DS (2002) Molecular marker assisted breeding. In: Jain SM, Brar DS, Ahloowalia BS (eds) Molecular techniques in crop improvement. Kluwer Academic Press, Dordrecht, The Netherlands, pp 55–83
Chen X, Levine L, Kwok PY (1999) Fluorescence polarization in homogenous nucleic acid analysis. Genome Res 9:492–498
Fourmann M, Barret P, Renard M, Pelletier G, Delourme R, Brunel D (1998) The two genes homologous to Arabidopsis FAE1 co-segregate with the two loci governing erucic acid content in Brassica napus. Theor Appl Genet 96:852–858
James DW Jr, Dooner HK (1990) Isolation of EMS-induced mutants in Arabidopsis altered in seed fatty acid composition. Theor Appl Genet 80:241–245
James DW Jr, Lim E, Keller J, Plooy I, Ralston E, Dooner HK (1995) Directed tagging of the Arabidopsis Fatty Acid Elongation 1 (FAE1) gene with the maize transposon Activator. Plant Cell 7:309–319
Jourdren C, Barret P, Horvais R, Foisset N, Delourme R, Renard M (1996) Identification of RAPD markers linked to the loci controlling erucic acid level in rapeseed. Mol Breed 2:61–71
Kirk JTO, Hurlstone CG (1983) Variation and inheritance of erucic acid content in Brassica juncea. Z Pflanzenzuchtg 90:331–338
Kirk JTO, Oram RN (1981) Isolation of erucic acid-free lines of Brassica juncea: Indian mustard now a potential oilseed crop in Australia. J Aust Inst Agric Sci 47:51–52
Kunst L, Taylor DC, Underhill EW (1992) Fatty acid elongation in developing seeds of Arabidopsis thaliana. Plant Physiol Biochem 30:425–434
Lassner MW, Lardizabal K, Metz JG (1996) A jojoba β-ketoacyl-CoA synthase cDNA complements the canola fatty acid elongation mutation in transgenic plants. Plant Cell 8:281–292
Lemieux B, Miquel M, Somerville C, Browse J (1990) Mutants of Arabidopsis with alterations in seed lipid fatty acid composition. Theor Appl Genet 80:234-240
Meksem K, Ruben E, Hyten DL, Schmidt ME, Lightfoot DA (2001) High-throughput genotyping for a polymorphism linked to soybean cyst nematode resistance gene Rhg4 by using Taqman probes. Mol Breed 7:63–71
Potts D, Rakow G, Males DR (1999) Canola quality Brassica juncea, a new oilseed crop for the Canadian prairies.Proceedings of the 10th International Rapeseed Congress, Canberra, Australia 26–29 September, 1999
Pradhan AK, Gupta V, Mukhopadhyay M, Arumugam N, Sodhi YS, Pental D (2003) A high-density linkage map in Brassica juncea (Indian mustard) using AFLP and RFLP markers. Theor Appl Genet 106:607–614
Rogers SO, Bendich AJ (1994) Extraction of total cellular DNA from plants, algae and fungi. In: Gelvin SV, Shilperoot RA (eds) Plant molecular biology manual. Kluwer Academic Press, Dordrecht, The Netherlands, pp 1–8
See D, Kanazin V, Talbert H, Blake T (2000) Electrophoretic detection of single nucleotide polymorphisms. Biotechniques 28:710–716
Sodhi YS, Mukhopadhyay M, Arumugam N, Verma JK, Gupta V, Pental D, Pradhan AK (2002) Genetic analysis of total glucosinolate in crosses involving a high glucosinolate Indian variety and a low glucosinolate line of Brassica juncea. Plant Breed 121:508–511
Srivastava A, Gupta V, Pental D, Pradhan AK (2001) AFLP based genetic diversity assessment amongst agronomically important natural and some newly synthesized lines of Brassica juncea. Theor Appl Genet 102:193–199
Stam P (1993) Construction of integrated genetic linkage maps by means of a new computer package: JoinMap. Plant J 3:739–744
Stam P, Van Ooijen JW (1996) JoinMap version 2.0: software for the calculation of genetic linkage maps. CPRO-DLO, Wageningen, The Netherlands
Stefansson BR, Hougen FW (1964) Selection of rape plants (Brassica napus) with seed oil practically free from erucic acid. Can J Plant Sci 44:359–364
Syvänen AC (2001) Accessing genetic variation: Genotyping single nucleotide polymorphisms. Nat Rev Genet 2:930–942
Tanhuanpää P, Schulman A (2002) Mapping of genes affecting linolenic acid content in Brassica rapa ssp. oleifera. Mol Breed 10:51–62
Tanhuanpää P, Vikki J, Vihinen M (1998) Mapping and cloning of FAD2 gene to develop allele-specific PCR for oleic acid in spring turnip rape (Brassica rapa ssp. oleifera). Mol Breed 4:543–550
Thies W (1971) Schnelle and einfache analysen der fettsäurezusammensetzung in einzelnen rapskotyledonen. I. Gaschromatographische methoden. Z Pflanzenzuecht 65:181–202
U N (1935) Genomic analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Jpn J Bot 7:389 – 452
Van Ooijen JW, Malipaard C (1996) Map QTL (tm) version 3.0: Software for the calculation of QTL positions on linkage maps. CPRO-DLO, Wageningen,The Netherlands
Acknowledgements
This work was supported by the National Dairy Development Board (NDDB), India and its subsidiary, Dhara Vegetable Oil and Food Company Ltd (DOFCO). SNP analysis was done on sequencer provided by NMITLI Programme of Council of Scientific and Industrial Research, Government of India.
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Gupta, V., Mukhopadhyay, A., Arumugam, N. et al. Molecular tagging of erucic acid trait in oilseed mustard (Brassica juncea) by QTL mapping and single nucleotide polymorphisms in FAE1 gene. Theor Appl Genet 108, 743–749 (2004). https://doi.org/10.1007/s00122-003-1481-z
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DOI: https://doi.org/10.1007/s00122-003-1481-z