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Basal media including Murashige and Skoog (MS), Woody Plant Medium (WPM), and Driver and Kuniyuki (DKW) media were compared for use in guayule regeneration and propagation. Explants grown on MS basal medium showed large size calli with multiple shoots, but the calli produced on DKW medium were of higher quality, and longer true shoots were produced. Adventitious root formation was best in WPM basal medium. A comparison of macro and micro nutrient levels in MS, DKW, and WPM media revealed significant differences in the calcium levels. Under controlled conditions, guayule plants in vitro incorporated calcium into plant tissues in proportion to the amount available. Moreover, the addition of calcium nitrate tetrahydrate at moderate levels into MS medium significantly improved the emergence of single, elongated true shoots, suggesting a requirement for calcium in guayule tissue culture. We have further explored selection agent (antibiotic) concentration for the identification of transgenic plants using a spectinomycin resistance gene (aadA). Based on the results, we have improved tissue culture protocols for guayule including spectinomycin as the selection agent. [ABSTRACT FROM AUTHOR]

Antibiotic-resistance genes of bacterial origin are invaluable markers for plant genetic engineering. However, these genes are feared to pose possible risk to human health by horizontal gene transfer from transgenic plants to bacteria, potentially resulting in antibiotic-resistant pathogenic bacteria; this is a considerable regulatory concern in some countries. The Atwbc19 gene, encoding an Arabidopsis thaliana ATP-binding cassette transporter, has been reported to confer resistance to kanamycin specifically as an alternative to bacterial antibiotic-resistance genes. In this report, we transformed hybrid aspen ( Populus canescens × P. grandidentata) with the Atwbc19 gene. Unlike Atwbc19-transgenic tobacco that was only resistant to kanamycin, the transgenic Populus plants also showed resistance to three other aminoglycoside antibiotics (neomycin, geneticin, and paromomycin) at comparable levels to plants containing a CaMV35S- nptII cassette. Although it is unknown why the transgenic Populus with the Atwbc19 gene is resistant to all aminoglycoside antibiotics tested, the broad utility of the Atwbc19 gene as a reporter gene is confirmed here in a second dicot species. Because the Atwbc19 gene is plant-ubiquitous, it might serve as an alternative selectable marker to current bacterial antibiotic-resistance marker genes and alleviate the potential risk for horizontal transfer of bacterial-resistance genes in transgenic plants. [ABSTRACT FROM AUTHOR]