Abstract
wxocA,wxocB,wxocD,wxocE and wzt are genes for lipopolysaccharide(LPS)biosynthesis in Xanthomonas oryzae pv.oryzicola(Xooc),and avrXa3 is an avirulence(avr)gene in Xanthomonas oryzae pv.oryzae(Xoo).Plasmids pBCSK(-)and pKNG101 with replicon of ColE1 and R6K,respectively,were used to construct gene knock-out transformation units.Transformation units of pBCSK∷ΔwxocA,pBCSK∷ΔwxocD,pBCSK∷ΔwxocE and PBCSK∷Δwzt to produce Xooc strain PS105 corresponding gene mutants were constructed with two fragments:central portion of target gene and pBCSK(-) vector.Transformation units of pKNG101∷ΔwxocB and pKNG101∷Δavr to produce wxocB mutant of Xooc strain RS105 and avirulence gene mutant in Xoo strain PXO99 were constructed with four fragments :pKNG101 vector,two flanking sequences of corresponding gene,and cm gene.Transformation was conducted by electroporation to yield 5 lps gene mutants(MwxocA,MwxocB,MwxocD,MwxocE,Mwzt)and 1 avr gene mutant(PXO99Δavr),and homologous recombinations were confirmed by Southern blot.The analysis of LPS by SDS-PAGE showed that all the lps gene mutants had defects in O-antigen biosynthesis.Virulence tests indicated that pathogenicity of MwxocB and MwxocE were completely lost and Mwzt was reduced.And PXO99Δavr showed less virulence on IRBB50(Xa4/xa5)and Asominori(Xa17).Therefore,the vectors pBCSK(-)and pKNG101 were first confirmed to be successfully used to knock-out genes in Xooc and Xoo and be helpful to identify the functions of pathogenicity-related genes in Xooc and Xoo.
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Claims 8
Claim · model-written summary
Five LPS gene mutants and one avirulence mutant were produced and confirmed.
Verbatim
e0“electroporation to yield 5 lps gene mutants(MwxocA,MwxocB,MwxocD,MwxocE,Mwzt)and 1 avr gene mutant(PXO99Δavr),and homologous recombinations were confirmed by Southern blot”
Claim · model-written summary
All LPS gene mutants had defective O-antigen biosynthesis.
- Outcome
- defects in O-antigen biosynthesise0
Not reportedTaxon
Verbatim
e0“all the lps gene mutants had defects in O-antigen biosynthesis”
Claim · model-written summary
MwxocB lost pathogenicity completely.
Not reportedTaxon
Verbatim
e0“pathogenicity of MwxocB and MwxocE were completely lost”
Claim · model-written summary
MwxocE lost pathogenicity completely.
Not reportedTaxon
Verbatim
e0“pathogenicity of MwxocB and MwxocE were completely lost”
Claim · model-written summary
Mwzt had reduced pathogenicity.
Not reportedTaxon
Verbatim
e0“Mwzt was reduced”
Claim · model-written summary
PXO99Δavr had less virulence on IRBB50.
Not reportedTaxon
Verbatim
e0“PXO99Δavr showed less virulence on IRBB50(Xa4/xa5)and Asominori(Xa17)”
Claim · model-written summary
PXO99Δavr had less virulence on Asominori.
Not reportedTaxon
Verbatim
e0“PXO99Δavr showed less virulence on IRBB50(Xa4/xa5)and Asominori(Xa17)”
Claim · model-written summary
The vectors successfully knocked out genes in Xooc and Xoo.
Not reportedComparator
Verbatim
e0“vectors pBCSK(-)and pKNG101 were first confirmed to be successfully used to knock-out genes in Xooc and Xoo”