Expression of a carrot 36 kD antifreeze protein gene improves cold stress tolerance in transgenic tobacco |
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Authors: | Xu?Wen-li Liu?Mei-qin Shen?Xin Email author" target="_blank">Lu?Cun-fu?Email author |
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Institution: | College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, P. R. China |
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Abstract: | Antifreeze proteins (AFPs) enable organisms to survive under cold conditions, and have great potential in improving cold tolerance
of cold-sensitive plants. In order to determine whether expression of the carrot 36 kD antifreeze protein gene confers improved
cold-resistant properties to plant tissues, we tried to obtain transgenic tobacco plants which expressed the antifreeze protein.
Cold, salt, and drought induced promoter Prd29A was cloned using PCR from Arabidopsis. Two plant expression vectors based on pBI121 were constructed with CaMV35S:AFP and Prd29A:AFP. Tobacco plantlets were transformed
by Agrobacterium-medicated transformation. PCR and Southern blotting demonstrated that the carrot 36 kD afp gene was successfully integrated
into the genomes of transformed plantlets. The expression of the afp gene in transgenic plants led to improved tolerance to
cold stress. However, the use of the strong constitutive 35S cauliflower mosaic virus (CaMV) promoter to drive expression
of afp also resulted in growth retardation under normal growing conditions. In contrast, the expression of afp driven by the
stress-inducible Prd29A promoter from Arabidopsis gave rise to minimal effects on plant growth while providing an increased tolerance to cold stress condition (2°C). The results
demonstrated the prospect of using Prd29A-AFP transgenic plants in cold-stressed conditions that will in turn benefit agriculture.
Supported by the National Natural Science Foundation of China (Grant No. 30271067), Fok Ying Tung Education Foundation (Grant
No. 71030), Key Teachers Foundation of the Educational Ministry of China, and Graduate Training Grant of Beijing Forestry
University (Grant No. 03SW003)] |
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Keywords: | antifreeze protein gene stress inducible-promoter tobacco genetic transformation cold tolerance |
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