Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent
<p>Abstract</p> <p>Background</p> <p>In soybean somatic embryo transformation, the standard selection agent currently used is hygromycin. It may be preferable to avoid use of antibiotic resistance genes in foods. The objective of these experiments was to develop a selec...
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BMC
2009-11-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/9/94 |
| _version_ | 1818643738696613888 |
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| author | Kwanyuen Prachuab Polisetty Raghuveer McConnell Matt Mamadou Lewamy Rao Suryadevara S Hildebrand David |
| author_facet | Kwanyuen Prachuab Polisetty Raghuveer McConnell Matt Mamadou Lewamy Rao Suryadevara S Hildebrand David |
| author_sort | Kwanyuen Prachuab |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>In soybean somatic embryo transformation, the standard selection agent currently used is hygromycin. It may be preferable to avoid use of antibiotic resistance genes in foods. The objective of these experiments was to develop a selection system for producing transgenic soybean somatic embryos without the use of antibiotics such as hygromycin.</p> <p>Results</p> <p>When tested against different alternate selection agents our studies show that 0.16 μg/mL glufosinate, 40 mg/L isopropylamine-glyphosate, 0.5 mg/mL (S-(2 aminoethyl)-L-cysteine) (AEC) and the acetolactate synthase (ALS) inhibitors Exceed<sup>® </sup>and Synchrony<sup>® </sup>both at 150 μg/mL inhibited soybean somatic embryo growth. Even at the concentration of 2 mg/mL, lysine+threonine (LT) were poor selection agents. The use of AEC may be preferable since it is a natural compound. Unlike the plant enzyme, dihydrodipicolinate synthase (DHPS) from <it>E. coli </it>is not feed-back inhibited by physiological concentrations of lysine. The <it>dapA </it>gene which codes for <it>E. coli </it>DHPS was expressed in soybean somatic embryos under the control of the CaMV 35S promoter. Following introduction of the construct into embryogenic tissue of soybean, transgenic events were recovered by incubating the tissue in liquid medium containing AEC at a concentration of 5 mM. Only transgenic soybeans were able to grow at this concentration of AEC; no escapes were observed.</p> <p>Conclusion</p> <p>Genetically engineered soybeans expressing a lysine insensitive DHPS gene can be selected with the non-antibiotic selection agent AEC. We also report here the inhibitory effects of glufosinate, (isopropylamine-glyphosate) (Roundup<sup>®</sup>), AEC and the ALS inhibitors Exceed<sup>® </sup>and Synchrony<sup>® </sup>against different tissues of soybean</p> |
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| issn | 1472-6750 |
| language | English |
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| spelling | doaj.art-0c39b3bfbfd949e3808315b19d4da39b2022-12-21T22:11:01ZengBMCBMC Biotechnology1472-67502009-11-01919410.1186/1472-6750-9-94Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agentKwanyuen PrachuabPolisetty RaghuveerMcConnell MattMamadou LewamyRao Suryadevara SHildebrand David<p>Abstract</p> <p>Background</p> <p>In soybean somatic embryo transformation, the standard selection agent currently used is hygromycin. It may be preferable to avoid use of antibiotic resistance genes in foods. The objective of these experiments was to develop a selection system for producing transgenic soybean somatic embryos without the use of antibiotics such as hygromycin.</p> <p>Results</p> <p>When tested against different alternate selection agents our studies show that 0.16 μg/mL glufosinate, 40 mg/L isopropylamine-glyphosate, 0.5 mg/mL (S-(2 aminoethyl)-L-cysteine) (AEC) and the acetolactate synthase (ALS) inhibitors Exceed<sup>® </sup>and Synchrony<sup>® </sup>both at 150 μg/mL inhibited soybean somatic embryo growth. Even at the concentration of 2 mg/mL, lysine+threonine (LT) were poor selection agents. The use of AEC may be preferable since it is a natural compound. Unlike the plant enzyme, dihydrodipicolinate synthase (DHPS) from <it>E. coli </it>is not feed-back inhibited by physiological concentrations of lysine. The <it>dapA </it>gene which codes for <it>E. coli </it>DHPS was expressed in soybean somatic embryos under the control of the CaMV 35S promoter. Following introduction of the construct into embryogenic tissue of soybean, transgenic events were recovered by incubating the tissue in liquid medium containing AEC at a concentration of 5 mM. Only transgenic soybeans were able to grow at this concentration of AEC; no escapes were observed.</p> <p>Conclusion</p> <p>Genetically engineered soybeans expressing a lysine insensitive DHPS gene can be selected with the non-antibiotic selection agent AEC. We also report here the inhibitory effects of glufosinate, (isopropylamine-glyphosate) (Roundup<sup>®</sup>), AEC and the ALS inhibitors Exceed<sup>® </sup>and Synchrony<sup>® </sup>against different tissues of soybean</p>http://www.biomedcentral.com/1472-6750/9/94 |
| spellingShingle | Kwanyuen Prachuab Polisetty Raghuveer McConnell Matt Mamadou Lewamy Rao Suryadevara S Hildebrand David Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent BMC Biotechnology |
| title | Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent |
| title_full | Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent |
| title_fullStr | Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent |
| title_full_unstemmed | Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent |
| title_short | Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent |
| title_sort | non antibiotic selection systems for soybean somatic embryos the lysine analog aminoethyl cysteine as a selection agent |
| url | http://www.biomedcentral.com/1472-6750/9/94 |
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