Non-antibiotic selection systems for soybean somatic embryos: the lysine analog aminoethyl-cysteine as a selection agent.

BMC Biotechnology
Suryadevara S RaoDavid Hildebrand

Abstract

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. When tested against different alternate selection agents our studies show that 0.16 microg/mL glufosinate, 40 mg/L isopropylamine-glyphosate, 0.5 mg/mL (S-(2 aminoethyl)-L-cysteine) (AEC) and the acetolactate synthase (ALS) inhibitors Exceed and Synchrony both at 150 microg/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 E. coli is not feed-back inhibited by physiological concentrations of lysine. The dapA gene which codes for E. coli 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 contain...Continue Reading

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Citations

Feb 16, 2013·Genetics and Molecular Biology·Milena Schenkel HomrichMaria Helena Bodanese-Zanettini
Apr 2, 2013·In Vitro Cellular & Developmental Biology. Plant : Journal of the Tissue Culture Association·Christell van der VyverJames Lloyd

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Datasets Mentioned

BETA
M12844
AF234293

Methods Mentioned

BETA
transgenic
PCR
transgenics
gene gun
chip

Software Mentioned

ChemStation
Exceed
Synchrony

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