iv
The objective of this study was to optimize tissue culture and regeneration
parameters of immature inflorescence culture of Triticum aestivum cv. Bezostaja-
01 and Triticum durum cv. Kiziltan-91. The effects of callus age and vernalisation
time of explants on regeneration success were evaluated. For determination of
optimum vernalisation time of immature inflorescence, plants subjected to 4 ° / C
for 1, 2, 3, 4, and 5 weeks, respectively. Tillers containing immature
inflorescences were collected at the same time. Percentage of inflorescence formed
tillers over total explants were reached the highest value, 79 %, at 4 weeks cold
treated Kiziltan cultivar and, 73 %, at 5 weeks cold treated Bezostaja cultivar.
Isolated immature inflorescences were put onto 2mg /L 2,4-dichlorophenoxyacetic
acid and picloram containing callus induction medium for Kiziltan and Bezostaja
cultures, respectively. Callus induction rate were found to be 100 % for Kiziltan
and Bezostaja. These explants were taken to regeneration after 6, 9, 12 and 15
weeks of dark incubation period. The regeneration capacities of calli were
determined as shooting percentage and data were collected after 4, 8, 12, and 15
week regeneration period. The highest shooting percentage of 69 %, were obtained
from 6 weeks old calli produced from 4 weeks vernalised explants in Kiziltan
cultures at the end of 15 weeks regeneration period. However, shooting percentage
was 57.2 % for 9 weeks old calli while it decreases to 37.6 % in 12 weeks old calli
and 44.2 % in 15 weeks old calli at the end of 15 weeks regeneration period. This
showed that prolonged dark incubation period decreased regeneration capacity of
the callus. However, there was no significant difference in regeneration capacities
of calli produced from Bezostaja immature inflorescence and the highest shooting
percentage was obtained from 9 weeks old calli produced from 5 weeks vernalised
explants, 27.4 %.
Besides regeneration studies, optimization of transformation parameters for winter
wheat cultivars Kiziltan and Bezostaja by using Agrobacterium tumefaciens AGLI
containing binary vector pALl56 was performed. Transformation efficiencies were
determined by monitoring the transient expression of uidA gene via histochemical
GUS assay. Three to four weeks old calli were found to be more responsive to
Agrobacterium-mediated transformation in Kiziltan cultures. However, four to five
weeks old calli were found to be more responsive to Agrobacterium-mediated
transformation in Bezostaja cultures. Different transformation protocols were used.
It was found that MGL based and MMA based protocols could be used for
Bezostaja and Kiziltan transformation, respectively. The highest GUS expression,
84%, was obtained from 28 weeks old calli produced from 5 weeks vernalised
explants in Bezostaja cultures.
| Identifer | oai:union.ndltd.org:METU/oai:etd.lib.metu.edu.tr:http://etd.lib.metu.edu.tr/upload/3/12606523/index.pdf |
| Date | 01 September 2005 |
| Creators | Kavas, Musa |
| Contributors | Yucel, Meral |
| Publisher | METU |
| Source Sets | Middle East Technical Univ. |
| Language | English |
| Detected Language | English |
| Type | M.S. Thesis |
| Format | text/pdf |
| Rights | To liberate the content for public access |
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