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Three Microsomal Omega-3 Fatty-acid Desaturase Genes Contribute to Soybean Linolenic Acid Levels

^a USDA-ARS, Plant Genetics Research Unit, University of Missouri, Columbia, MO 65211
^b Dep. of Agronomy, University of Missouri, Columbia, MO 65211

View larger version (13K): [in a new window]   Fig. 1. Phylogenetic tree of soybean and Arabidopsis omega-3 fatty-acid desaturases. The protein sequences for the soybean and Arabidopsis omega-3 fatty-acid desaturases were subjected to a ClustalW1.8 alignment and Phylip (no bootstrap) phylogenetic tree prediction (Brodsky et al., 1995). The bar represents 0.1 genetic distance units calculated as the average of the pairwise distances. Genes preceded by Gm are from soybean and those preceded by At are from Arabidopsis thaliana. The GenBank accession numbers for the sequences are GmFAD3A (AY204710), GmFAD3B (AY204711 and L22964), GmFAD3C (AY204712 and AB05125), GmL22964 (L22964), AtFAD3 (D26508), AtFAD7 (D14007), and AtFAD8 (L27158).

View larger version (99K): [in a new window]   Fig. 2. Alignment of soybean FAD3 homologs with Arabidopsis FAD3. The open reading frames for the Pana FAD3 homologs were translated and subjected to a ClustalW1.8 alignment with the Arabidopsis FAD3 gene (AtFAD3). Identical amino acid residues are shaded black and similar amino acids are shaded gray.

View larger version (18K): [in a new window]   Fig. 3. Relative expression of GmFAD3A, GmFAD3B, and GmFAD3C in soybean tissues. Steady state mRNA levels for each gene were quantitated by quantitative PCR following reverse transcription of total RNA from each tissue. The histograms represent the percent of each gene normalized to the housekeeping gene, EF1. Error bars are plus one standard deviation from the mean.

View larger version (100K): [in a new window]   Fig. 4. Amplification of GmFAD3A, GmFAD3B, and GmFAD3C sequences from Williams 82 and A5 genomic DNA. Genomic DNA (50 ng) was subjected to 40 cycles of PCR amplification with primers specific for GmFAD3A, GmFAD3B, or GmFAD3C and products were separated on an agarose gel. W82 lanes contained template DNA from Williams 82, A5 lines contained template DNA from A5, and NEG (negative) lanes contained water in place of template DNA as a control. The predicted product sizes were 149, 139, and 246 bp for GmFAD3A, GmFAD3B, and GmFAD3C, respectively. Molecular weight (MW) lanes contained 100 bp ladder MW markers.