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| Establishment of Plant Expression Vector Carrying AFⅠ Gene and Cultivation of Antifreeze Lily (Lilium brownii var. viridulum) |
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Abstract Abstract Lily (Lilium brownii var. viridulum) is one of the main fresh cut flowers in the world which possesses both high ornamental and economic value. However, the yield will be greatly reduced because the low temperature inhibits the growth of the lily. To improve the antifreeze function of Lily and breed the new characters, Antifreeze protein gene (AFPⅢ) originates from Anarhichas minor which was codon optimized according to the codon usage bias in plants and the antifreeze gene (AFⅠ) was synthesized by the company. Then AFⅠ was inserted into Pcambia1390 vector by enzyme digestion and ligation with the Peanut chlorotic streak caulimovirus (PCISV) promoter. Thus the plants expression vector carrying AFⅠ gene was constructed. And then the P1390-PCISV-AFⅠ plasmid was transformed into Agrobacterium EHA105 by electric shock. Subsequently, AFⅠ gene was introduced into lily callus mediated by Agroinfection, and the results of PCR confirmed that the AFⅠ gene had been integrated into Oriental Lily cultivar "Conca D or"．Restoring growth test was carried out to examine the antifreezing function of the positive transgenic plants, and the results of the test revealed that experimental group had a great antifreezing function in -2 ℃ compared with the control group, and the Semi-quantitative reverse transcription PCR demonstrated that the expression of AFⅠ gene leads to the antifreezing function of transgenic plants. This study provides an important reference for the cultivation of new varieties of antifreeze plants.
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Received: 17 August 2017
Published: 21 May 2018
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