Abstract
Selectable markers help the transformed cell/tissue to survive in an otherwise lethal exposure to an antibiotic or herbicide. Unfortunately, almost all the traditional selectable markers are antibiotic and herbicide resistance genes, which are controversial on human health concerns and environmental impact. Novel plant-derived, non-antibiotic, and non-herbicide selectable markers are urgently needed in plant transformation. Our previous work showed that the seedlings of overexpression Arabidopsis lines of AtGASA6 survived on medium with a high concentration of sugar, which leads to the hypothesis that AtGASA6 could be a selectable marker on media with high or low sugar content. In this study, leaf explants of AtGASA6 overexpression tobacco lines regenerated shoots on sugar-free shooting medium while those of wild type could not. Moreover, the seeds of AtGASA6 overexpression tobacco lines germinated and grew into normal seedlings on sugar-free MS medium while those of WT could not. Attractively, no developmental defects were observed in AtGASA6 transgenic progenies. Using AtGASA6 as a selectable marker, overexpression tobacco lines of GAI, which restrains plant size, were created on sugar-free media. The GAI overexpression lines had a smaller plant size than that of control. Considering its plant-derived and non-antibiotic nature, GASA6 is promising to be used as a selectable marker in plant transformation.
Highlights
Genetic transformation and genome editing help us create crops with advanced characteristics, such as high yield, high abiotic stress tolerance, and high biotic stress resistance
It is unlikely that transgenic plants using an antibiotic selectable marker will be authorized in the European Union
Overexpression of AtGASA6 was found to help transgenic Arabidopsis seeds survive on a medium with a high concentration of sugar (Zhong et al 2015)
Summary
Genetic transformation and genome editing help us create crops with advanced characteristics, such as high yield, high abiotic stress tolerance, and high biotic stress resistance. The most widely used selectable markers are antibiotic genes that confer resistance to toxins, such as antibiotics and herbicides. These antibiotic genes include nptII which encodes the neomycin phosphotransferase II conferring resistance to kanamycin (Fraley et al 1983), hpt that encodes hygromycin phosphotransferase detoxifying hygromycin (Waldron et al 1985), and bar endowing resistance to phosphinothricin (Deblock et al 1989). Antibiotic genes assist the selection of transformants efficiently. Whether they harm human health and ecological balance is still unknown. Proteins coded by these antibiotic genes are always undesirable in the final products.
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