Specific for palmitoyl, stearoyl and oleoyl-alp thioesters nucleic acid fragments encoding acyl-acp thiosesterase enzymes and the use of these fragments in altering plant oil composition
First Claim
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1. An isolated nucleic acid fragment encoding an acyl-ACP thioesterase which catalyzes the hydrolysis of palmitoyl, stearoyl and oleoyl-ACP thioesters, wherein said isolated nucleic acid fragment hybridizes to nucleotides 271 to 1206 of SEQ ID NO:
- 1 or nucleotides 282 to 1217 of SEQ ID NO;
3 or nucleotides 210 to 1121 of SEQ ID NO;
23 under one of the following sets of conditions;
(a) hybridization in 6×
SSC, 1 mM EDTA, 1% SDS, and 100 μ
g/mL denatured salmon sperm DNA at 45°
C. for 10 h and wash 3 times at room temperature with 0.6×
SSC containing 0.1% SDS, then once at 50°
C. for 5 min in the same solution and two additional washes for 5 min each at 50°
C. in 0.2×
SSC, 0.1% SDS followed by a 1 min rinse under the same conditions;
or(b) hybridization in 6×
SSC, 1 mM EDTA, 1% SDS, and 100 μ
g/mL denatured salmon sperm DNA at 45°
C. for 10 h and wash twice at room temperature with 0.6×
SSC containing 0.1% SDS followed by two 5 minute washes and one 1 minute wash in the same solution and all at 52°
C.
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Abstract
Isolated nucleic acid fragments encoding an acyl-ACP thioesterase enzyme which catalyzes the hydrolysis of palmitoyl, stearoyl and oleoyl-ACP thioesters are described. Use of such fragments in altering plant oil composition is also described.
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Citations
18 Claims
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1. An isolated nucleic acid fragment encoding an acyl-ACP thioesterase which catalyzes the hydrolysis of palmitoyl, stearoyl and oleoyl-ACP thioesters, wherein said isolated nucleic acid fragment hybridizes to nucleotides 271 to 1206 of SEQ ID NO:
- 1 or nucleotides 282 to 1217 of SEQ ID NO;
3 or nucleotides 210 to 1121 of SEQ ID NO;
23 under one of the following sets of conditions;(a) hybridization in 6×
SSC, 1 mM EDTA, 1% SDS, and 100 μ
g/mL denatured salmon sperm DNA at 45°
C. for 10 h and wash 3 times at room temperature with 0.6×
SSC containing 0.1% SDS, then once at 50°
C. for 5 min in the same solution and two additional washes for 5 min each at 50°
C. in 0.2×
SSC, 0.1% SDS followed by a 1 min rinse under the same conditions;
or(b) hybridization in 6×
SSC, 1 mM EDTA, 1% SDS, and 100 μ
g/mL denatured salmon sperm DNA at 45°
C. for 10 h and wash twice at room temperature with 0.6×
SSC containing 0.1% SDS followed by two 5 minute washes and one 1 minute wash in the same solution and all at 52°
C. - View Dependent Claims (3, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18)
- 1 or nucleotides 282 to 1217 of SEQ ID NO;
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2. An isolated nucleic acid fragment of at least 41 base pairs, which is useful in antisense inhibition or cosuppression of endogenous thioesterase activity in a transformed plant, wherein said isolated nucleic acid fragment hybridizes to nucleotides 271 to 1206 of SEQ ID NO:
- 1 or nucleotides 282 to 1217 of SEQ ID NO;
3 under the following set of conditions;hybridization in 6×
SSC, 1 mM EDTA, 1% SDS, and 100 μ
g/mL denatured salmon sperm DNA at 45°
C. for 10 h and wash 3 times at room temperature with 0.6×
SSC containing 0.1% SDS, then once at 50°
C. for 5 min in the same solution and two additional washes for 5 min each at 50°
C. in 0.2×
SSC, 0.1% SDS followed by a 1 min rinse under the same conditions.
- 1 or nucleotides 282 to 1217 of SEQ ID NO;
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4. An isolated nucleic acid fragment encoding the Brassica species acyl-ACP thioesterase encoded by nucleotides 1 to 1412 of SEQ ID NO:
- 23.
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5. An isolated nucleic acid fragment encoding the Brassica species acyl-ACP thioesterase encoded by nucleotides 36 to 1121 of SEQ ID NO:
- 23.
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6. An isolated nucleic acid fragment encoding the Brassica species acyl-ACP thioesterase encoded by nucleotides 210 to 1121 of SEQ ID NO:
- 23.
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17. A method of producing plant seed oil comprising reduced levels of palmitic and stearic acids comprising:
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(a) transforming plant cells of an oil-producing species with a nucleotide sequence selected from the group consisting of SEQ ID NO;
1, SEQ ID NO;
3, and SEQ ID NO;
23, operably linked in the antisense orientation to transcription regulatory sequences, to produce transformed plant cells;(b) growing fertile plants from the transformed plant cells of step (a); (c) screening progeny seeds from the fertile plants of step (b) for the desired reduced levels of palmitic and stearic acids; and (d) processing the progeny seeds of step (c) to obtain oil comprising reduced levels of palmitic and stearic acids as compared to seeds of untransformed plants.
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Specification