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Production of -Linolenic Acid and Stearidonic Acid in Seeds of Marker-Free Transgenic Soybean¹

^a Plant Science Initiative, Univ. of Nebraska, Lincoln, NE 68588
^b Stine-Haskell Research Center, DuPont Agricultural Products, Newark, DE 19714
^c Dep. of Agronomy and Horticulture, Univ. of Nebraska, Lincoln, NE 68583
^d DuPont Exp. Stn., Wilmington, DE 19880

View larger version (26K): [in a new window]   Fig. 1. Two T-DNA Binary Vector pPTN331. RB and LB refer to the right border and left border elements, respectively. The restriction sites shown reflect the enzymes used in the hybridization analyses.

View larger version (104K): [in a new window]   Fig. 2. Northern blot analysis on immature embryos derived from line 404-1. Top panel, northern blot. Lower panel, ethidium bromide gel of northern blot. Lane 1, wild-type soybean immature embryo. Lanes 2 to 4, T2 immature embryos derived from line 404-1.

View larger version (105K): [in a new window]   Fig. 3. Southern blot analysis on line 400-1. Upper panel, 6-desaturase probe. Lower panel, bar gene probe. Lane 1, parental (T0). Lanes 2 to 4, derived T1 individuals. Lane 5 (WT), wild-type soybean DNA. Lane 6 (C), 50 pg pPTN331 (see Fig. 1). DNAs digested with HindIII. Designation HT above the lane indicates the individual was herbicide tolerant.

View larger version (53K): [in a new window]   Fig. 4. (A) Southern blot analysis on herbicide-sensitive/modified fatty-acid profile (HS/GLA/STA) T1 individuals derived from lines 419-2 and 420-5. Top panel, 6 desaturase probe. Lower panel, bar gene probe. Lanes 1 to 4, T1 individuals from line 419-2. Lanes 5 to 8, T1 individual from line 420-5. Lane 9, wild-type (WT) control. Lane 10, 50 pg of pPTN331 positive control. DNAs digested with Sst I. Numbers above each column indicate percentage of GLA in the fatty-acid analysis. HT and HS refer to herbicide tolerant and herbicide sensitive, respectively. (B) Southern blot analysis on HS/GLA/STA T1 individuals derived from lines 411-5, 414-5, and 418-6. Figure description as in (A). Lanes 1 and 2, T1 individuals from line 411-5. Lanes 3 to 6, T1 individuals from line 414-5. Lanes 7 to 10, T1 individuals from line 418-6. Lane 11, wild-type (WT) control. Lane 12, 50 pg of pPTN331 positive control. (C) Southern blot analysis on GLA/STA T1 individuals derived from lines 414-2 and 414-3. Figure description as in (A). Lanes 1 to 4, T1 individuals from line 414-2. Lane 5, T1 individual from line 414-3. Lane 6, wild-type (WT) control. Lane 2, 50 pg of pPTN331 positive control.

View larger version (77K): [in a new window]   Fig. 5. Southern blot analysis on T2 individuals derived from marker-free T1 seed of line 420-5. Upper panel, 6 desaturase probe. Lower panel, bar gene probe. Lanes 1-6, T2 individuals from marker-free population of line 420-5. Lane 7, wild-type (WT) soybean. Lane 8, 50 pg pPTN331 positive control. DNAs digested with EcoRI. Numbers above the column refer to the percentage of GLA in the T2 seed sample. HS indicates all individuals were scored herbicide sensitive.