Biolistic transformation of Arabidopsis root hairs: a novel technique to facilitate map-based cloning

Citation
A. Kemp et al., Biolistic transformation of Arabidopsis root hairs: a novel technique to facilitate map-based cloning, PLANT J, 27(4), 2001, pp. 367-371
Citations number
19
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT JOURNAL
ISSN journal
0960-7412 → ACNP
Volume
27
Issue
4
Year of publication
2001
Pages
367 - 371
Database
ISI
SICI code
0960-7412(200108)27:4<367:BTOARH>2.0.ZU;2-V
Abstract
The final stage of map-based gene isolation is complementation of the mutan t phenotype with wild-type DNA to determine the exact location of the gene of interest. This usually involves Agrobacterium tumefaciens-mediated trans formation, which is reliable and produces stable transformants. However, th e process of Agrobacterium transformation may take up to three months to co mplete. If the mutant phenotype can be seen in a single cell, and the wild- type copy of the gene can act cell autonomously, then complementation of th e whole plant is not strictly necessary. We have developed a technique for the biolistic transformation of Arabidopsis thaliana root hairs, and used t his to test large insert clones for complementation of two recessive mutant phenotypes, a procedure that takes less than a day. Our results show that biolistic transformation can be used with transient assays to conduct rapid tests for complementation by large insert clones.