caa a22 4500 467901309 CHVBK 20180406152819.0 cr unu---uuuuu 170328e20060501xx s 000 0 eng 10.1007/s00299-005-0084-3 doi (NATIONALLICENCE)springer-10.1007/s00299-005-0084-3 MicroTom—a high-throughput model transformation system for functional genomics [Elektronische Daten] [Yinghui Dan, Hua Yan, Tichafa Munyikwa, Jimmy Dong, Yanling Zhang, Charles Armstrong] We have developed a high-throughput Agrobacterium-mediated transformation model system using both nptII and the 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens strain CP4 (cp4) based selections in MicroTom, a miniature rapid-cycling cherry tomato variety. With the NPTII selection system, transformation frequency calculated as independent transgenic events per inoculated explant ranged from 24 to 80% with an average of 56%, in industrial production scale transformation experiments. For CP4, with glyphosate selection, the average transformation frequency was 57%. Stable transformation frequency was positively correlated with transient expression (R=0.85), and variable with the genes of interest. DNA integration and germline transformation were confirmed by biological assay, Southern Blot analysis, and R1 phenotype segregation. Transgene expression was observed in leaf, root, stem, flower, and fruit tissues of the transgenic plants. Ninety-five percent of transgenic events coexpressed two introduced genes based on β-glucuronidase (GUS) and neonmycin phosphotransferase II (NPTII) expression. Seventy-five percent of transgenic events contained one to two copies of the introduced uidA (GUS) gene based on Southern analysis. Transgenic plants from the cotyledon explants to the transgenic plants transferred to soil were produced within about 2-3 months depending on the genes of interest. The utility of this MicroTom model transformation system for functional genomic studies, such as identification of genes related to important agricultural traits and gene function, is discussed. Springer-Verlag, 2005 Agrobacterium nationallicence High-throughput nationallicence Kanamycin nationallicence MicroTom nationallicence Transformation nationallicence Dan Yinghui Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Yan Hua Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Munyikwa Tichafa Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Dong Jimmy Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Zhang Yanling Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Armstrong Charles Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut Plant Cell Reports Springer-Verlag 25/5(2006-05-01), 432-441 0721-7714 25:5<432 2006 25 299 https://doi.org/10.1007/s00299-005-0084-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0084-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dan Yinghui Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 700 1- Yan Hua Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 700 1- Munyikwa Tichafa Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 700 1- Dong Jimmy Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 700 1- Zhang Yanling Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 700 1- Armstrong Charles Monsanto Company, 700 Chesterfield Parkway, 63017, Chesterfield, MO, USA aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/5(2006-05-01), 432-441 0721-7714 25:5<432 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer