Rd29A启动子驱动 AtCDPK1 基因转化马铃薯的研究

In order to obtain normal growth and resistant drought transgenic potato,using Arabidopsis thaliana(Columbia ecotype) as material,DNA sequence of AtRd29 A gene ATG upstream from + 83 bp to-1 441 bp is cloned with PCR and recombinant DNA technologies.The sequence of bases was entirely consistent with that of promoter of the known gene.The plant expression vector p CHFRd-CDPK1 of AtCDPK1 gene drived by Rd29 A promoter was constructed,and transformed p CHFRd-CDPK1 into virus-free miniature of potato cultivar‘Favorita'using Agrobacterium-mediated method.After plant selection and regeneration,regenerated plants with resistance were obtained successfully.The PCR and Southern blotting analysis proved that the AtCDPK1 gene has been integrated in the genome of the potato.After drought stress using 30% PEG,RT-PCR analysis proved that the transcript levels of AtCDPK1 gene drived by Rd29 A promoter in transgenic potato plants were significantly higher.AtCDPK1 gene drived by Rd29 A promoter in transgenic potato plants is barely detectable in the water treatment control.However,expression of AtCDPK1 gene drived by 35 S promoter did not make significant difference in transgenic potato plants between PEG stress treatment and water treatment.Morphological observation showed that transgenic plants can grow normally after 30% PEG stress,the plants growing was better than non-transgenic plants,and control plants occur slightly wilt.This research will lay the foundation for further improved stress tolerance of crops by expression of resistant gene drived by stress-inducible promoter in the crop.