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| Agrobacterium -mediated Transformation of Several Turfgrass Cultivars |
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Abstract It is very important to improve stress characteristics of turfgrass such as drought, cold, hot and sanity by genetic engineering approach. But, the transformation system of Agrobacterium mediated turfgrass has not been set up completely. More research on it is still needed for perfection. Embryonic calli derived from the mature embryos of four turfgrass cultivars including Accent, Quest, Tengyue and Goalkeeper were used as target materials to be infected by Agrobacterium harboring nptII selectable gene and GUS report gene. Some calli were tested by histochemical staining for GUS gene transient expression after three days co-cultivation. And the remaining calli was transferred onto G418 10.0-25.0mg/l containing medium for selection and regeneration. The resistant plantlets to the antibiotics were identified by ELISA, PCR and X-Gluc staining for the integration and expression of the foreign genes. The results indicated that the frequency of GUS gene transient expression in the four cultivars varied from 8.6% to 46.9%, the highest in Accent, the lowest in Quest, and the medium in other two genotypes. In 144 total regeneration plants, 45 plantlets were conformed to be positive, in which the expression of nptII gene was suggested by ELISA, and among which the expression of GUS gene in 43 plants and the silence in 2 plants were demonstrated by histochemical staining. Regeneration frequency of resistant plantlets to the antibiotics from the four cultivars was 0-43.5%, and transformation efficiency of the four genotypes was 0-21.5%. The results also suggested that the both frequencies transient expression and stable transformation of the alien gene were not coincident in turfgrass, and the former frequency cannot be used to evaluate the transformation ability of turfgrass for Agrobacterium.
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Received: 31 July 2006
Published: 10 May 2010
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