A method was developed for rapid plant regeneration from protoplasts of birdsfoot trefoil (Lotus corniculatus cv. Leo) using in vitro cotyledons and a new enzyme formula. Protoplasts of a transgenic Nicotiana tabacum cv. Xanthi line KCR were isolated from leaves, in vitro shoots, and calli by an enzyme formula consisting of 0.5 % Cellulase R-10 and 0.05 % Pectolyase Y23 with either 0.4 M sucrose or 0.5 M mannitol as the osmoticum. Nine asymmetric fusion experiments were conducted between iodoacetate inactivated birdsfoot trefoil and irradiated transgenic tobacco protoplasts. The fusion products underwent several divisions but no visible colonies were obtained. Resistance analyses revealed the inactivation of the herbicide resistant gene in the transgenic tobacco calli. A putative chlorsulfuron resistant line has been obtained by in vitro selection from birdsfoot trefoil cotyledonous protoplasts. Their regenerants had normal ploidy, and expressed improved tolerance when sprayed with chlorsulfuron at the rate of 30 g/ha.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.41010 |
| Date | January 1992 |
| Creators | Vessabutr, Suyanee |
| Contributors | Grant, W. F. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Plant Science.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001324229, proquestno: NN87847, Theses scanned by UMI/ProQuest. |
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