Abstract
Auxotrophic strains of Agrobacterium tumefaciens have been developed to address the Agrobacterium overgrowth issue in plant genetic transformation; however, their application remains limited. Here, we generated novel histidine and leucine auxotrophic strains of A. tumefaciens EHA105, namely EHA105hisD− and EHA105leuA−, as well as a dual auxotrophic strain EHA105hisD−leuA−, through gene deletion. The transient expression efficiency and survival rate of these three auxotrophic strains in Nicotiana benthamiana were significantly impaired but could be restored to wild-type EHA105 levels by supplementation with appropriate concentrations of the corresponding amino acids (CAAs). The use of these three auxotrophic strains for the genetic transformation of N. benthamiana resulted in a significant reduction in Agrobacterium overgrowth and achieved transformation efficiency comparable to wild-type EHA105, when appropriate exogenous concentrations of the CAAs were supplied during the co-cultivation stage. Furthermore, through incubation experiments on various plants and soil, it was confirmed that the incidence of surviving cells from these three auxotrophic strains was much lower than that observed for the wild-type EHA105. In summary, this study reports on the characteristics of the novel auxotrophic strains of A. tumefaciens along with the effective use of such auxotrophic A. tumefaciens strains in plant genetic transformation.
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Claims 8
Claim · model-written summary
Histidine-auxotrophic EHA105hisD− was generated by gene deletion.
- Taxon
- Agrobacterium tumefaciens
Agrobacterium tumefacienst0 - Outcome
- EHA105hisD−e0
- Comparator
- EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“generated novel histidine and leucine auxotrophic strains of A. tumefaciens EHA105, namely EHA105hisD− and EHA105leuA−, as well as a dual auxotrophic strain EHA105hisD−leuA−, through gene deletion”
t0“Agrobacterium tumefaciens Strains for Transformation”
Claim · model-written summary
Leucine-auxotrophic EHA105leuA− was generated by gene deletion.
- Taxon
- Agrobacterium tumefaciensAgrobacterium tumefacienst0
- Outcome
- EHA105leuA−e0
- Comparator
- EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“generated novel histidine and leucine auxotrophic strains of A. tumefaciens EHA105, namely EHA105hisD− and EHA105leuA−, as well as a dual auxotrophic strain EHA105hisD−leuA−, through gene deletion”
t0“Agrobacterium tumefaciens Strains for Transformation”
Claim · model-written summary
Dual auxotrophic EHA105hisD−leuA− was generated by gene deletion.
- Taxon
- Agrobacterium tumefaciensAgrobacterium tumefacienst0
- Outcome
- EHA105hisD−leuA−e0
- Comparator
- Agrobacterium tumefacienst0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a dual auxotrophic strain EHA105hisD−leuA−, through gene deletion”
t0“Agrobacterium tumefaciens Strains for Transformation”
Claim · model-written summary
Transient expression efficiency of all three strains in Nicotiana was significantly impaired but restored with amino acids.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Treatment
- supplementation with appropriate concentrations of the corresponding amino acids (CAAs)e0
- Outcome
- transient expression efficiencye0
- Direction
- restorede0
- Comparator
- wild-type EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The transient expression efficiency and survival rate of these three auxotrophic strains in Nicotiana benthamiana were significantly impaired but could be restored to wild-type EHA105 levels by supplementation with appropriate concentrations of the corresponding amino acids (CAAs).”
Claim · model-written summary
Survival rate of all three strains in Nicotiana was significantly impaired but restored with amino acids.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- supplementation with appropriate concentrations of the corresponding amino acids (CAAs)e0
- Interactor
- auxotrophic strainse0other
- Outcome
- survival rate of these three auxotrophic strainse0
- Direction
- restorede0
- Comparator
- wild-type EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The transient expression efficiency and survival rate of these three auxotrophic strains in Nicotiana benthamiana were significantly impaired but could be restored to wild-type EHA105 levels by supplementation with appropriate concentrations of the corresponding amino acids (CAAs).”
Claim · model-written summary
With amino acid supplementation, transformation using auxotrophic strains significantly reduced Agrobacterium overgrowth.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Treatment
- these three auxotrophic strainse0
- Interactor
- Agrobacterium
.jpg?width=60)
Agrobacteriume0other - Outcome
- Agrobacterium overgrowthe0
- Direction
- reductione0
- Comparator
- wild-type EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“The use of these three auxotrophic strains for the genetic transformation of N. benthamiana resulted in a significant reduction in Agrobacterium overgrowth and achieved transformation efficiency comparable to wild-type EHA105, when appropriate exogenous concentrations of the CAAs were supplied during the co-cultivation stage.”
Claim · model-written summary
With amino acid supplementation, transformation efficiency matched wild-type EHA105.
- Taxon
- N. benthamianaNicotiana benthamianae0
- Treatment
- these three auxotrophic strainse0
- Outcome
- transformation efficiencye0
- Direction
- comparablee0
- Comparator
- wild-type EHA105e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The use of these three auxotrophic strains for the genetic transformation of N. benthamiana resulted in a significant reduction in Agrobacterium overgrowth and achieved transformation efficiency comparable to wild-type EHA105, when appropriate exogenous concentrations of the CAAs were supplied during the co-cultivation stage.”
Claim · model-written summary
Auxotrophic strain cell survival on plants and soil was much lower than wild-type EHA105.
- Treatment
- these three auxotrophic strainse0
- Outcome
- incidence of surviving cellse0
- Direction
- much lowere0
- Comparator
- wild-type EHA105e0
- Setting
- various plants and soile0
Not reportedTaxon
Verbatim
e0“through incubation experiments on various plants and soil, it was confirmed that the incidence of surviving cells from these three auxotrophic strains was much lower than that observed for the wild-type EHA105.”