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Enhanced Agrobacterium-mediated Transformation of Embryogenic Calli of Upland Cotton via Efficient Selection and Timely Subculture of Somatic Embryos.
- Source :
-
Plant Molecular Biology Reporter . Sep2008, Vol. 26 Issue 3, p174-185. 12p. 2 Color Photographs, 2 Diagrams, 2 Charts, 1 Graph. - Publication Year :
- 2008
-
Abstract
- Agrobacterium- tumefaciens-mediated transformation of cotton embryogenic calli (EC) was enhanced by choosing appropriate EC and improving efficiency of coculture, selection cultivation, and plant regeneration. After 48-h cocultivation, the number of β-glucuronidase (GUS)-positive calli characterized by yellow, loose, and fine-grained EC was twofold greater than that of gray, brown, and coarse-granule EC. It indicated that efficiency of transient transformation was affected by EC morphology. And transient transformation efficiency was also improved by cocultivation on the medium adding 50 mg l−1 acetosyringone at 19°C for 48 h. Subculturing EC on the selection medium with low cell density was beneficial to production of more kanamycin-resistant (Km-R) calli lines. From an original 0.3-g EC, an average of 20 Km-R calli lines were obtained from a selection dish and the GUS-positive rate of Km-R clones was 81.97%. A large number of normal plants were rapidly regenerated on the differentiation medium with dehydration treatments and the GUS-positive rate of regeneration plants was about 72.60%. Polymerase chain reaction analysis of GUS-positive plantlets revealed a 100% positive detection rate for neomycin phosphotransferase II gene and uidA. Southern blot of transgenic plants regenerated from different Km-R calli lines demonstrated that the target gene, mostly with the low copy number, has been integrated into the cotton genome. [ABSTRACT FROM AUTHOR]
- Subjects :
- *AGROBACTERIUM
*COTTON
*SOMATIC embryogenesis
*PLANT cell culture
*RHIZOBIACEAE
Subjects
Details
- Language :
- English
- ISSN :
- 07359640
- Volume :
- 26
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- Plant Molecular Biology Reporter
- Publication Type :
- Academic Journal
- Accession number :
- 35077010
- Full Text :
- https://doi.org/10.1007/s11105-008-0032-9