A rapid assay for genetic complementation of nitrate reductase deficiency via bulk protoplast fusion

✍ Scribed by Biasini, Grazia ;Màrton, Làszlò

• Publisher: Springer
• Year: 1985
• Tongue: English
• Weight: 412 KB
• Volume: 198
• Category: Article
• ISSN: 0026-8925
• DOI: 10.1007/bf00383018

✦ Synopsis

Genetic complementation of different nitrate reductase-deficient (NR-) Nicotiana plumbaginifolia cell lines could be recognized in the described fusion disc technique as early as 5-10 days after protoplast fusion. Protoplasts of previously characterized mutant cell lines, belonging to four different complementation groups, were mixed in pairwise combinations and fused by the drop-wise fusion technique at very high protoplast densities (10 6 protoplasts in an 8-10 mm spot) which resulted in the formation of a 'fusion disc' on the bottom of the petri dish. After 5-10 days of incubation in K3 medium the restoration of NR activity could be detected directly in the original culture by the in vivo NR assay. Such a rapid test giving information about the genetic complementation of different auxotrophic cell lines has not previously been published for plants.