caa a22 4500 467901058 CHVBK 20180406152818.0 cr unu---uuuuu 170328e20060801xx s 000 0 eng 10.1007/s00299-005-0081-6 doi (NATIONALLICENCE)springer-10.1007/s00299-005-0081-6 Rapid and reproducible Agrobacterium -mediated transformation of sorghum [Elektronische Daten] [Arlene Howe, Shirley Sato, Ismail Dweikat, Mike Fromm, Tom Clemente] A rapid and reproducible Agrobacterium-mediated transformation protocol for sorghum has been developed. The protocol uses the nptII selectable marker gene with either of the aminoglycosides geneticin or paromomycin. A screen of various A. tumefaciens strains revealed that a novel C58 nopaline chromosomal background carrying the chrysanthopine disarmed Ti plasmid pTiKPSF2, designated NTL4/Chry5, was most efficient for gene transfer to sorghum immature embryos. A NTL4/Chry5 transconjugant harboring the pPTN290 binary plasmid, which carries nptII and GUSPlus TM expression cassettes, was used in a series of stable transformation experiments with Tx430 and C2-97 sorghum genotypes and approximately 80% of these transformation experiments resulted in the recovery of at least one transgenic event. The transformation frequencies among the successful experiments ranged from 0.3 to 4.5%, with the average transformation frequency being approximately 1% for both genotypes. Over 97% of the transgenic events were successfully established in the greenhouse and were fully fertile. Co-expression of GUSPlus TM occurred in 89% of the transgenic T0 events. Seed set for the primary transgenic plants ranged from 145 to 1400 seed/plant. Analysis of T1 progeny demonstrated transmission of the transgenes in a simple Mendelian fashion in the majority of events. Springer-Verlag, 2005 Genetic engineering nationallicence Immature embryo nationallicence GUS Plus TM nationallicence nptII nationallicence Howe Arlene Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut Sato Shirley Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut Dweikat Ismail Department of Agronomy and Horticulture, University of Nebraska, Lincoln, NE, USA aut Fromm Mike Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut Clemente Tom Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 784-791 0721-7714 25:8<784 2006 25 299 https://doi.org/10.1007/s00299-005-0081-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0081-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Howe Arlene Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut NATIONALLICENCE 700 1- Sato Shirley Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut NATIONALLICENCE 700 1- Dweikat Ismail Department of Agronomy and Horticulture, University of Nebraska, Lincoln, NE, USA aut NATIONALLICENCE 700 1- Fromm Mike Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut NATIONALLICENCE 700 1- Clemente Tom Center for Biotechnology, University of Nebraska, N300 Beadle Center, Lincoln, NE, USA aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/8(2006-08-01), 784-791 0721-7714 25:8<784 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer