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The Low Linolenic Acid Soybean Line PI 361088B Contains a Novel GmFAD3A Mutation

USDA-ARS, Plant Genetics Research Unit, 110 Waters Hall, Univ. of Missouri, Columbia, MO 65211. Mention of a trademark, vendor, or proprietary product does not constitute a guarantee or warranty of the product by the USDA and does not imply its approval to the exclusion of other products or vendors that may also be suitable

View larger version (25K): [in this window] [in a new window]   Figure 1. Two rapid molecular marker assays to distinguish between PI 361088B and wild-type GmFAD3A alleles. (A) Sequence of nucleotides 301 to 330 of the GmFAD3A coding sequence from ‘Williams 82’ and PI 361088B DNA. Amino acid sequence from 101 to 110 is displayed below their corresponding codons. The XmnI recognition site is indicated by a black bar above the sequence. Note that the XmnI recognition site is disrupted by the two nucleotide insertion in the PI 361088B sequence. (B) A 200-bp sequence encompassing the region shown in Fig. 1A was amplified from Williams 82 or PI 361088B genomic DNA and treated with or without XmnI endonuclease. Products were then resolved on a 2% agarose gel (shown). When XmnI cleaves the Williams 82 polymerase chain reaction (PCR) product, bands of 140 and 60 bp are generated. (C) Melting curve analysis on Williams 82 and PI 361088B PCR products that have been digested with XmnI. The melting curve average was 77.4°C for the Williams 82 product and 79.4°C for the PI 361088B product.

View larger version (29K): [in this window] [in a new window]   Figure 2. An allele-specific assay to distinguish between PI 361088B and wild-type GmFAD3A alleles. (A) Sequence of nucleotides 301 to 330 of the GmFAD3A coding sequence from ‘Williams 82’ and PI 361088B DNA. Amino acid sequence is displayed below their corresponding codons. The position of the forward primers used in the assay is indicated by a black bar above the corresponding sequence. A dashed line is used where sequence is not shown. The "long GC" and "short GC" 5' tails are also indicated. (B) Melting curve analysis on Williams 82 and PI 361088B PCR products from the allele-specific polymerase chain reaction. The melting curve shows a melting temperature of 79.8°C for PI 361088B and 82.2°C for Williams 82.

View larger version (29K): [in this window] [in a new window]   Figure 3. An overview of GmFAD3A alleles and mutations found in low linolenic acid soybean lines. Schematic diagram of GmFAD3A alleles found in the low linolenic acid soybean lines A5, J18, M5, CX1512-44, C1640, and PI 361088B. Exons are indicated by black boxes and introns by horizontal lines. To the right of the gene schematics is a list of the amount (g linolenic acid kg–1 of oil) of linolenic acid levels in seeds of the various lines. Also indicated is whether a rapid molecular assay has been published to test for the presence of that particular allele. Relevant references for each line or mutant are also listed.