Citation
Wee C.Y., . and Marziah Mahmood, . and Kadzimin Saleh, . and Tan C.S., . (2005) Optimisation of biological parameters for transformation of Oncidium Taka via the biolistic method. [Proceedings Paper]
Abstract
The biolistic method was used in the genetic transformation of Oncidium Taka using protocorm-like bodies and calli as target tissues. In this study biological parameters were optimized by observed expression of green fluorescent protein as transient expression. Bombardment was carries out using 3 mg gold microcarriers coated with 5ug of DNA into both target tissues that were pre-cultures on the same day to prevent a decrease in the quality and quantity of transient expression. Strong green fluorescent signals were easily detected using a Leica Stereo-fluorescence System MZ FLIII. The parameters optimized were as follows: expression of five different types of modified green fluorescent protein gene maximum expression time after bombardment pre-culture period of target tissue prior to bombardment and age of target tissues. The effects of the green fluorescent protein expression on different target tissues were also examined. In this study synthetic GFP-TYP mutant P35S-sgfp-TYG-nos gave the best transient expression in both target tissues. The highest transient target green fluorescent protein expression was observed two days post-bombardment. In comparison to the single and multi-cellular system protocorm-like-bodies had significantly higher green fluorescent protein expression compared to calli. The physiological stages of the PLBs and calli may also influence the degree of expression. From this study seven- and fourteen-day cultures of both calli and protocorm-like-bodies respectively had the highest transient expression.
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Abstract
The biolistic method was used in the genetic transformation of Oncidium Taka using protocorm-like bodies and calli as target tissues. In this study biological parameters were optimized by observed expression of green fluorescent protein as transient expression. Bombardment was carries out using 3 mg gold microcarriers coated with 5ug of DNA into both target tissues that were pre-cultures on the same day to prevent a decrease in the quality and quantity of transient expression. Strong green fluorescent signals were easily detected using a Leica Stereo-fluorescence System MZ FLIII. The parameters optimized were as follows: expression of five different types of modified green fluorescent protein gene maximum expression time after bombardment pre-culture period of target tissue prior to bombardment and age of target tissues. The effects of the green fluorescent protein expression on different target tissues were also examined. In this study synthetic GFP-TYP mutant P35S-sgfp-TYG-nos gave the best transient expression in both target tissues. The highest transient target green fluorescent protein expression was observed two days post-bombardment. In comparison to the single and multi-cellular system protocorm-like-bodies had significantly higher green fluorescent protein expression compared to calli. The physiological stages of the PLBs and calli may also influence the degree of expression. From this study seven- and fourteen-day cultures of both calli and protocorm-like-bodies respectively had the highest transient expression.
Additional Metadata
| Item Type: | Proceedings Paper |
|---|---|
| Additional Information: | Summary En |
| AGROVOC Term: | ONCIDIUM |
| AGROVOC Term: | CULTURE MEDIA |
| AGROVOC Term: | AGE |
| AGROVOC Term: | GENES |
| Geographical Term: | MALAYSIA |
| Depositing User: | Ms. Norfaezah Khomsan |
| Last Modified: | 24 Apr 2025 05:28 |
| URI: | http://webagris.upm.edu.my/id/eprint/16713 |
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