ANALYSIS OF GENE TARGETING EVENTS IN BUCKWHEAT (Fagopyrum esculentum) TRANSFORMED b THE MUTANT STRAINS OF Agrobacterium tumefaciens THAT HAVE DEFICIENCY IN T-­??DNA INTEGRATION STEP INTO PLANT GENOME

  • Mineo Kojima
  • Putu Suparthana
  • Tsutomu Shimizu
  • Masahiro Nogawa

Abstract

Gene targeting events were analyzed in buckwheat (Fagopyrum esculentum, var. Shinano No. 1) that werein planta transformed (T1) by the three strains of Agrobacterium tumefaciens harboring a gene targetingvector for an endogenous gene (acc.no.AB327276); M-­??1 mutant (abvA::Tn5) and M-­??31 mutant (virA::Tn5)strains, both of which are capable of transferring its T-­??DNA into nucleus of host plant cell, but have adeficiency in T-­??DNA integration step into chromosome and LBA4404 strain, being a commonly usedstrain. The results of both phenotype examination and Southern blot analysis implied that a targetingconstruct(s) was integrated into a different locus (loci) each depending on the strain of A. tumefaciens usedfor transformation. Both 5’-­??end and 3’-­??end flanking DNA segments that were expected from preciseinsertion of targeting construct into the endogenous U-­??gene locus were obtained by PCR with 3 (23%)transformants out of the randomly selected 13 transformants by M-­??1 mutant strain, while only 3’-­??endflanking DNA segment was obtained with 3(21%) transformants out of 14 transformants by M-­??31 strain.Taken together, the results suggested the potential of usage of a mutant strain of A. tumefaciens for genetargeting which has a deficiency in T-­??DNA integration step into plant genome.

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How to Cite
KOJIMA, Mineo et al. ANALYSIS OF GENE TARGETING EVENTS IN BUCKWHEAT (Fagopyrum esculentum) TRANSFORMED b THE MUTANT STRAINS OF Agrobacterium tumefaciens THAT HAVE DEFICIENCY IN T-­??DNA INTEGRATION STEP INTO PLANT GENOME. International Journal of Biosciences and Biotechnology, [S.l.], v. 2, n. 1, dec. 2014. ISSN 2655-9994. Available at: <https://ojs.unud.ac.id/index.php/jbb/article/view/13307>. Date accessed: 24 apr. 2024.
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Articles

Keywords

Agrobacterium tumefaciens; gene targeting; buckwheat; Fagopyrum esculentum; mutant strains of A. tumefaciens