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Nsibande, Busie Esther , 2012. In vitro regeneration of four Hypoxis species and transformation of Camelina sativa and Crambe abyssinica. Second cycle, A2E. Alnarp: SLU, MOVIUM



The advance of science and biotechnology has made it possible to prevent the complete loss of plant species by providing propagation methods that are fast and can result in the mass production of exact copies of the targeted plant species. Genetic transformation has become a useful instrument in the introduction of new traits to plants such as increased yield and disease resistance.
In this study potential in vitro regeneration protocols for four Hypoxis species were successfully developed. Efficient indirect regeneration (100% with 8 shoots per explant) of Hypoxis filiformis was obtained when corm explants were directly cultured in MS supplemented with 1 mg/l NAA. However, the highest mean number of shoots per explant (17) was obtained in basal MS supplemented with 3 mg/l kinetin. Efficient direct regeneration of H. acuminata (100% with 2 shoots per explant) was achieved in corm explants cultured with a piece of shoot attached in MS supplemented with 3 mg/l kinetin. Up to 30% and 21% seed germination was obtained in H. argentea and H. filiformis respectively, when seed coats were crushed before culturing in half-strength MS without PGRs. In addition, the use of activated charcoal and 0.1% mercuric chloride, respectively, helped in controlling the build-up of phenolic exudates in the medium and infection of explants and shoots by endogenous micro-organisms.
Agrobacterium-mediated transformation of Camelina sativa and Crambe abyssinica flowers by vacuum infiltration with three gene constructs: AtHb2. BvHb2, and vhb in camelina; and the FWS 3-1 vector harbouring three genes: ScFAR, ScWS, and DsRed in crambe failed to produce transgenic seeds.

Main title:In vitro regeneration of four Hypoxis species and transformation of Camelina sativa and Crambe abyssinica
Authors:Nsibande, Busie Esther
Supervisor:Zhu, Li-Hua
Examiner:Hofvander, Per
Series:Självständigt arbete vid LTJ-fakulteten, SLU
Volume/Sequential designation:UNSPECIFIED
Year of Publication:2012
Level and depth descriptor:Second cycle, A2E
Student's programme affiliation:None
Supervising department:(LTJ, LTV) > MOVIUM
Keywords:Agrobacterium, biotechnology, Camelina sativa, Crambe abyssinica, Hypoxis, In vitro propagation, regeneration, transformation, vacuum infiltration
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Subject. Use of subject categories until 2023-04-30.:Plant genetics and breeding
Deposited On:15 Jan 2013 07:14
Metadata Last Modified:15 Jan 2013 07:14

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