Construction and transformation for the antisense expression vector of the polyphenol oxidase gene in Yali pear |
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| Authors: | Guiqin Li Jing Qi Yuxing Zhang Zhihua Gao Dongqian Xu Huixuan Li Chenmin Huo |
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| Institution: | (1) Institute of Crop Science, National Wheat Improvement Centre/The National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences (CAAS), 12 Zhongguancun South Street, Beijing, 100081, China;(2) International Maize and Wheat Improvement Centre (CIMMYT) China Office, c/o CAAS, 12 Zhongguancun South Street, Beijing, 100081, China;(3) Department of Agriculture and Food Western Australia Centre for Comparative Genomics, Murdoch University, Perth, WA, 6150, Australia; |
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| Abstract: | To inhibit the browning process in fruits of Yali pear, in this paper, antisense gene techniques were used to reduce the expression
of PPO gene. A cDNA fragment of 450 bp, which is located at the 3′ terminal of the polyphenol oxidase (PPO) gene, was amplified from Yali pear using the RT-PCR method, then the antisense expression vector was constructed by inserting
the fragment of the Yali pear PPO gene between the CaMV promoter and NOS terminator of the expression vector pBI121 in a reverse orientation. After that, with
the agrobacterium-mediated method, the PPO antisense gene was transformed into Yali pear shoots. Northern blot analysis and enzyme activity assay showed that the PPO
activities in the transgenic Yali pear shoots were significantly decreased, compared with the non-transformed Yali pear shoots.
This lays a good foundation for breeding new varieties of pears with browning resistance in the future. |
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