[Genetic transformation of buckwheat ( Fagopyrum esculentum Moench ) with AtNHX1 gene and regeneration of salt-tolerant transgenic plants].
The Arabidopsis thaliana tonoplast Na+ /H+ antiporter gene, AtNHX1, was transferred into buckwheat by Agrobacterium-mediated method. Transgenic buckwheat plants were regenerated and selected on MS basal medium supplemented with 2.0mg/L 6-BA, 1.0mg/L KT, 0.lmg/L IAA, 50mg/L kanamycin and 500mg/L carbenicillin. 426 seedlings from 36 resistant calli originated from 864 explants (transformed about at 4.17 percentage) exhibited resistance to kanamycin. The transformants were confirmed by PCR, Southern blotting, RT-PCR and Northern blotting analysis. After stress treatment for 6 weeks with 200mmol/L NaCl, transgenic plants survived, while wild-type plants did not. After 3 days of stress treatment through different concentrations of NaCl, transgenic plants accumulated higher concentration of Na+ and proline than the control plants. However, the K+ concentration of transgenic plants declined in comparison with the control plants. Moreover, the rutin content of the roots, stems and leaves of transgenic buckwheat increased than those of the control plants. These results showed that it could be possible to improve the salt-tolerance of crops with genetic technology.
Cheng
Li-Hong
LH
Shaanxi Provincial Key Laboratory of Biotechnology, Northwest University, Xi' an 710069, China.
Zhang
Bo
B
Xu
Zi-Qin
ZQ
D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000502:physiology*; D015152:Blotting, Northern; D015139:Blotting, Southern; D027682:Cation Transport Proteins / Q000235:genetics / Q000502:physiology*; D019612:Fagopyrum / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012431:Rutin / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000502:physiology*; D014170:Transformation, Genetic
D029681:Arabidopsis Proteins; D027682:Cation Transport Proteins; C117751:NHX1 protein, Arabidopsis; D017923:Sodium-Hydrogen Exchangers; D012965:Sodium Chloride; D012431:Rutin; D011392:Proline; D012964:Sodium; D011188:Potassium