Transformation of AmDHN Gene Driven by Rd29A Promoter into Alfalfa (Medicago sativa)  

Lizhen Nie
Author    Correspondence author
Legume Genomics and Genetics, 2022, Vol. 13, No.   
Received: 01 Jan., 1970    Accepted: 01 Jan., 1970    Published: 13 Dec., 2022
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In order to obtain normal growth and resistant drought transgenic alfalfa (Medicago satiua) using Arabidopsis thaliana and Ammppiptanthus mongolicus as material, DNA sequence of AtRd29A gene ATG upstream 1 520 bp is cloned with PCR and recombinant DNA technologies. Open reading frame of 572 bp of AmDHN gene was cloned by PCR. We construct the plant expression vector of AmDHN gene driven by Rd29A promoter. The cotyledon and hypocotyl of alfalfa variety ‘Aohanmuxu’ was acted as the receptor material and the vector pCHF-RD-DHN was transformed into alfalfa by Agrobacterium-mediated method. After callusselection and regeneration, regenerated plants with resistance were obtained successfully. The PCR analysis proved that the AmDHN gene has been transformed into the genome of the alfalfa. After drought stress using 20% PEG, RT-PCR analysis proved that the alfalfa plants with 20% PEG stress, AmDHN gene expression in transgenic alfalfa strain quantity obviously increased, and under the condition of water treatment, AmDHN gene was microexpressed in transgenic plants. This study will provide a basis for the expression of resistant genes driven by stress-inducible promoters and genetic improvement for forage grass.

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Legume Genomics and Genetics
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