caa a22 4500 467900337 CHVBK 20180406152816.0 cr unu---uuuuu 170328e20060701xx s 000 0 eng 10.1007/s00299-005-0099-9 doi (NATIONALLICENCE)springer-10.1007/s00299-005-0099-9 A high throughput Agrobacterium tumefaciens -mediated transformation method for functional genomics of perennial ryegrass ( Lolium perenne L.) [Elektronische Daten] [Shivendra Bajaj, Yidong Ran, Jonathan Phillips, Gunaseelan Kularajathevan, Sunil Pal, Dan Cohen, Kieran Elborough, Sathish Puthigae] A robust and high throughput Agrobacterium genetic transformation procedure has been developed for perennial ryegrass (Lolium perenne L.). Embryogenic callus lines were selected and maintained as plants in vitro. Embryogenic calli derived from meristematic regions of the vegetative tillers were co-cultivated with Agrobacterium tumefaciens strain EHA101 carrying the plasmid pCAMBIA 1305.1 in the presence of acetosyringone for 3-4 days. The calli were grown under 94.8 and 151.6μM hygromycin selection, respectively for two cycles of 2-weeks each, followed by transfer to regeneration medium with 47.4μM hygromycin. Regenerated plants were rooted and successfully transferred to soil. The transgenic nature of the regenerated plants was confirmed by DNA gel blot analysis and gene expression demonstrated by GUS histochemical assay and/or reverse transcription PCR. After development of the transformation procedure, we used Agrobacterium strain EHA101 carrying a modified binary plasmid pMH bearing genes of interest. In the past 2 years, we have produced more than 1,000 plants with constructs encoding different genes of interest from perennial ryegrass. Springer-Verlag, 2005 Agrobacterium nationallicence Functional genomics nationallicence Genetic transformation nationallicence Lolium perenne nationallicence Perennial ryegrass nationallicence Transgenic plants nationallicence Bajaj Shivendra HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut Ran Yidong HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut Phillips Jonathan Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut Kularajathevan Gunaseelan HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut Pal Sunil HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut Cohen Dan HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut Elborough Kieran Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut Puthigae Sathish Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut Plant Cell Reports Springer-Verlag 25/7(2006-07-01), 651-659 0721-7714 25:7<651 2006 25 299 https://doi.org/10.1007/s00299-005-0099-9 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0099-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bajaj Shivendra HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut NATIONALLICENCE 700 1- Ran Yidong HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut NATIONALLICENCE 700 1- Phillips Jonathan Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut NATIONALLICENCE 700 1- Kularajathevan Gunaseelan HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut NATIONALLICENCE 700 1- Pal Sunil HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut NATIONALLICENCE 700 1- Cohen Dan HortResearch, Private Bag 92169, Mt Albert, Auckland, New Zealand aut NATIONALLICENCE 700 1- Elborough Kieran Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut NATIONALLICENCE 700 1- Puthigae Sathish Pastoral Genomics, P. O. Box 109185, Newmarket, Auckland, New Zealand aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/7(2006-07-01), 651-659 0721-7714 25:7<651 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer