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Two Genes from Soybean Encoding Soluble 9 Stearoyl-ACP Desaturases

^a Microbiology Dep., North Carolina State Univ., Raleigh, NC 27695
^b Crop Science Dep., North Carolina State Univ., Raleigh, NC 27695
^c USDA-ARS Soybean and Nitrogen Fixation Unit and Plant Pathology Dep., North Carolina State Univ., Raleigh, NC 27695

View larger version (100K): [in a new window]   Fig. 1. The soluble 9 stearoyl-ACP desaturase (SACPD) gene from soybean. A. Gene structural organization showing exon 1, encoding a transit peptide, exons 2 and 3, and introns 1 and 2. Exon 3 contains the region of amino acid sequence variability in G. max SACPDs. B. Alignment of Glycine max cultivar Dare SACPD-A and SACPD-B protein sequences with SACPD protein sequences from Arabidopsis Ab (AAB64035), Ricinus communis (castor bean) Cb (X56508), and a Glycine max EST, Gm (L34346).

View larger version (52K): [in a new window]   Fig. 2. SACPD exon 3 sequence alignments. A. Nucleotide (Blastn) and B. Protein (Blastx) sequences of a section of SACPD exon 3 from 9 Glycine lines showing nucleotide and amino acid sequence variation. Sequences of Bragg-A, N01–3544, and Dare A and B were determined in this study. AW 755581, an EST of cultivar Jack, BG 882246 and AI 941223, ESTs of cultivar Williams, BI 471396, an EST of cultivar Bragg-B, and BG 363272, an EST of cultivar Corolla are GenBank accessions.

View larger version (91K): [in a new window]   Fig. 3. Ethidium bromide stained gel showing amplicons produced by PCR using EcoR1 restricted Dare genomic DNA template with SACPD gene-specific primers (see Materials and Methods). Lane 1, 100bp DNA ladder; lane 2, 133-bp product produced with the SACPD-A primer pair; lane 3, 111-bp product produced with the SACPD-B primer pair.

View larger version (74K): [in a new window]   Fig. 4. Southern blots of Dare soybean cultivar genomic DNA hybridized with SACPD-A and SACPD-B exon 3 specific probes. Lane (1), genomic DNA digested with BamHI and lane (2), with EcoRI. Molecular size (kb) is indicated on the left.

View larger version (22K): [in a new window]   Fig. 5. Relative expression of cultivar Dare GmSACPD-A and GmSACPD-B at four stages of soybean seed development (between R5 and R6) at 18, 23, 28, and 35 DAF. Steady state mRNA levels were determined by real-time PCR after reverse transcription of total RNA. The values represent the percent of each gene normalized to the housekeeping gene actin. Bars indicate the standard error of the mean.