Feedback-insensitive anthranilate synthase gene as a novel selectable marker for soybean transformation

Abstract

Because of the concern about escape of antibiotic- or herbicide-resistant transgenes from transgenic crops, selectable marker genes from plant origin would be an alternative choice for plant transformation. In this study, a feedback-insensitive anthranilate synthase gene ( ASA2 ) cloned from a tobacco cell line was tested for Agrobacterium-mediated transformation of axis tissue of soybean mature embryo, with a tryptophan analogue 5-methyltryptophan (5-MT) as the selective agent. Southern blot analysis of the To transgenic lines confirmed the integration of the ASA2 gene into the soybean genome. Northern blot analysis showed the ASA2 gene was also expressed in the leave tissue, and the free tryptophan content in the leaf tissue of transgenic soybean was about 59% to 123% more than that in the wild type. PCR analysis of the T1 progeny showed that the transgene was inherited in a Mendelian fashion. All these results indicate that this feedback-insensitive ASA2 gene can be used as a selectable marker gene for plant transformation. This work also demonstrated that the ASA2 gene coding for the a-subunits from one plant (tobacco) can interact with the n-subunits of a heterologous plant (soybean) to form an active anthranilate synthase enzyme. The use of this feedback-insensitive gene as a novel selectable marker for plant transformation is also discussed.