Cultivar variability in the Agrobacterium‐rice cell interaction and plant regeneration

Abstract

Thirteen cultivars of rice (Oryza sativa) were tested for plant regeneration from calli initiated from the scutella of mature seeds by water stress treatment using a high concentration of agarose, and examined for their response to Agrobacterium tumefaciens LBA4404, carrying a plasmid pTOK233, harboring genes for kanamycin resistance (nptII), hygromycin resistance (hpt) and β‐glucuronidase (gus). Plant regeneration frequency was considerably increased in most of the cultivars when the calli were treated with water stress, as compared with untreated controls. In particular, the cultivars Dongjinbyeo, IR43, Nagdongbyeo and Sinseonchalbyeo showed an increased frequency of shoot regeneration. Expression of GUS was detected in all of the co‐cultivated cultivars. Based on GUS expression at 3 days after co‐cultivation with A. tumefaciens, three rice cultivars (Dongjinbyeo, Hwayoungbyeo and Nagdongbyeo) were judged highly susceptible to A. tumefaciens, while Milyang 23, Nonganbyeo and Samgangbyeo cultivars were weakly susceptible. Plantlets were readily regenerated when the hygromycin‐resistant calli were transferred to a regeneration medium containing hygromycin. Intense blue staining was observed in GUS assays of leaf segments, roots and flower organs from regenerated plants. Stable integration and expression of the introduced hpt and gus genes were confirmed by Southern blot analysis of the transformants. Therefore, Dongjinbyeo and Nagdongbyeo cultivars proved to be both highly susceptible to A. tumefaciens and highly responsive to plant regeneration.

Genome-wide analysis of RopGEF gene family to identify genes contributing to pollen tube growth in rice (Oryza sativa)

“…After, the plasmid was transformed into the Agrobacterium tumefaciens LBA4404. The transgenic rice plants were generated through the stable transformation via Agrobacterium-mediated co-cultivation [63]. All of the cloning primers we have used in our experiments are listed in Additional file 9.…”

Overexpression of AmRosea1 Gene Confers Drought and Salt Tolerance in Rice

“…The plasmid pCAMBIA1300- AmRosea1 was transformed into rice cultivar Nipponbare ( O ryza sativa L. ssp. japonica ) by the Agrobacterium -mediated co-cultivation method [ 59 , 60 ] and regenerated on MS medium containing 50 mg·L −1 hygromycin B. The seeds of different transgenic plants were harvested separately.…”

Over‐expressed rice ADP‐ribosylation factor 1 (RARF1) induces pathogenesis‐related genes and pathogen resistance in tobacco plants

“…Oryza sativa L. cv. Milyang 117 and NB15‐3 suspension cultures were raised and maintained in a modified MS medium as previously described (Lee et al 1999). Fungal spores were isolated from rice blast fungus Magnaporthe grisea strains KJ101 and KJ301 and inoculated into NB15‐3 cells (5 × 10 5 spores ml −1 ) as described by Kim et al (2000).…”