Vector Construction to Overexpress AhPLDα Genes from Peanut and Genetic Transformation into Arabidopsis thaliana

doi:10.7668/hbnxb.2016.06.006

Abstract

Abstract: This study provides a good foundation for further elucidating the function and mechanism of AhPLDα1 and AhPLDα2 in responding to drought stress in peanut.The full-length CDS sequences of AhPLDα1 and AhPLDα2 were cloned from Jihua 4 peanut leaf with ABA treatment using RT-PCR.The AhPLDα1 and AhPLDα2 gene fragments were inserted into the expression vector pBar-F3 by forward ways.These constructs were transformed into Agrobacterium tumefaciens GV3101 by freeze-thawing method and then introduced into wild-type Arabidopsis thaliana using modified floral-dip method.Under the condition of drought,the authors observed phenotypic changes of wild type and transgenic plants.Colony PCR and enzyme digestion results showed that,the plant overexpressing recombinant plasmid pBar-AhPLDα driven by CaMV35S promoter was successfully constructed.Glyphosate resistance screening,PCR detection and gene expression analysis showed that the positive transgenic plants were obtained.Under water deprivation,overexpression of AhPLDα1 and AhPLDα2 in transgenic Arabidopsis plants resulted in significantly enhanced tolerance to drought stress.In conclusion, AhPLDα1 and AhPLDα2 have a certain relation with drought stress signal transduction in transgenic Arabidopsis plants,and are potential candidate genes on the way to modified crop drought resistance.

Key words: Peanut, AhPLDα gene, Plant expression vector, Transgene, Drought stress signaling

CLC Number:

S332.4
Q78

LIU Yijie, CHEN Silong, CHENG Zengshu, WANG Jin, SONG Yahui, HAO Junhui, ZHANG Pengjuan, LI Yurong. Vector Construction to Overexpress AhPLDα Genes from Peanut and Genetic Transformation into Arabidopsis thaliana[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(6): 31-38. doi: 10.7668/hbnxb.2016.06.006.

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