Abstract
【Objective】 The objective of this study is to analyze the biological phenotypes of the pathogenicity-defective mutant strain B-1015, to clone its T-DNA integration flanking sequences, and finally to reveal the functions of targeted genes in pathogenesis of Ustilaginoidea virens. The study will contribute to better understanding of the molecular pathogenesis of U. virens. 【Method】 Biological phenotypes of B-1015 were analyzed by testing growth rate, sporulation ability, spore germination rate and pathogenicity. The copy numbers of T-DNA inserted into B-1015 were identified by Southern blot. The flanking sequences of T-DNA were cloned by hiTail-PCR and the whole gene sequences of the T-DNA targeted genes were cloned by RACE PCR. The genes expression were detected by RT-PCR. 【Result】By comparing with wild type strain P1, the sporulation ability and growth rate of the B-1015 were significantly declined, but the spore germination rate had no significant difference. The inserted mutant lost its pathogenicity in rice. Molecular detection analysis confirmed that the B-1015 was a stable single T-DNA insertional mutant. The T-DNA flanking sequences were obtained by hiTail-PCR. Two genes, Uvt-1015R and Uvt-1015L, were cloned by RACE PCR. Gene sequence analysis showed that Uvt-1015R contained a 948 bp open reading frame, which could encode a 295 amino acid protein, a 341 bp 5′-UTR and a 271 bp 3′-UTR. Another gene, Uvt-1015L, comprised a 351 bp open reading frame coding a 116 amino acid protein, a 31 bp 5′-UTR and a 174 bp 3′-UTR. By searching in the known protein, the Uvt-1015R and Uvt-1015L genes showed no homology of any protein. Sequence alignment showed that the flanking sequences of T-DNA were non-adjacent in the wild type genome and the T-DNA disrupted the two genes sequence. RT-PCR analysis confirmed that compared to P1 strain, the transcripts of Uvt-1015R was significantly decreased, and the transcripts of Uvt-1015L was not detected any more. 【Conclusion】 In B-1015, both the T-DNA insertion and chromosomal rearrangement caused important biological phenotype mutant, thus leading to the loss of pathogenicity.
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Claims 13
Claim · model-written summary
B-1015 sporulation and growth rate declined significantly versus wild type P1.
- Taxon
- rice
Oryza sativae0 - Interactor
- B-1015e1pathogen
- Outcome
- sporulation ability and growth rate of the B-1015e1
- Direction
- declinede1
- Comparator
- wild type strain P1e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“By comparing with wild type strain P1, the sporulation ability and growth rate of the B-1015 were significantly declined”
Claim · model-written summary
B-1015 spore germination rate did not differ significantly from P1.
- Taxon
- riceOryza sativae0
- Interactor
- B-1015e2pathogen
- Outcome
- spore germination ratee1
- Direction
- no significant differencee1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“spore germination rate had no significant difference”
e2“growth rate of the B-1015 were significantly declined”
Claim · model-written summary
The insertional mutant lost pathogenicity in rice.
- Taxon
- riceOryza sativae0
- Interactor
- inserted mutante0pathogen
- Outcome
- pathogenicity in ricee0
- Direction
- loste0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The inserted mutant lost its pathogenicity in rice.”
Claim · model-written summary
B-1015 was confirmed as a stable single T-DNA insertional mutant.
- Taxon
- riceOryza sativae0
- Interactor
- B-1015e1pathogen
- Outcome
- stable single T-DNA insertional mutante1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“the B-1015 was a stable single T-DNA insertional mutant”
Claim · model-written summary
T-DNA flanking sequences were obtained by hiTail-PCR.
- Taxon
- riceOryza sativae0
- Outcome
- T-DNA flanking sequencese1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“The T-DNA flanking sequences were obtained by hiTail-PCR.”
Claim · model-written summary
Uvt-1015R and Uvt-1015L were cloned by RACE PCR.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015R and Uvt-1015Le1
- Outcome
- cloned by RACE PCRe1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“Two genes, Uvt-1015R and Uvt-1015L, were cloned by RACE PCR.”
Claim · model-written summary
Uvt-1015R had a 948 bp open reading frame encoding a 295 amino acid protein and 341 bp and 271 bp untranslated regions.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015Re1
- Outcome
- open reading framee1
- Value
- 948 bpe1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“Uvt-1015R contained a 948 bp open reading frame, which could encode a 295 amino acid protein, a 341 bp 5′-UTR and a 271 bp 3′-UTR.”
Claim · model-written summary
Uvt-1015L had a 351 bp open reading frame coding a 116 amino acid protein and 31 bp and 174 bp untranslated regions.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015Le1
- Outcome
- open reading framee1
- Value
- 351 bpe1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“Uvt-1015L, comprised a 351 bp open reading frame coding a 116 amino acid protein, a 31 bp 5′-UTR and a 174 bp 3′-UTR.”
Claim · model-written summary
Neither Uvt-1015R nor Uvt-1015L had homology to a known protein.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015R and Uvt-1015Le1
- Outcome
- no homology of any proteine1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“Uvt-1015R and Uvt-1015L genes showed no homology of any protein”
Claim · model-written summary
T-DNA flanking sequences were nonadjacent in the wild type genome, and T-DNA disrupted two gene sequences.
- Taxon
- riceOryza sativae0
- Outcome
- flanking sequences of T-DNAe1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“the flanking sequences of T-DNA were non-adjacent in the wild type genome and the T-DNA disrupted the two genes sequence”
Claim · model-written summary
Uvt-1015R transcripts decreased significantly compared with P1.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015Re1
- Outcome
- transcripts of Uvt-1015Re1
- Direction
- decreasede1
- Comparator
- P1 straine1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“compared to P1 strain, the transcripts of Uvt-1015R was significantly decreased”
Claim · model-written summary
Uvt-1015L transcripts were no longer detected.
- Taxon
- riceOryza sativae0
- Gene
- Uvt-1015Le1
- Outcome
- transcripts of Uvt-1015Le1
- Direction
- not detected any moree1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“the transcripts of Uvt-1015L was not detected any more”
Claim · model-written summary
The authors attributed B-1015 loss of pathogenicity to T-DNA insertion and chromosomal rearrangement.
- Taxon
- riceOryza sativae0
- Outcome
- loss of pathogenicitye1
- Qualifier
- T-DNA insertion and chromosomal rearrangemente1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“pathogenicity in rice”
e1“both the T-DNA insertion and chromosomal rearrangement caused important biological phenotype mutant, thus leading to the loss of pathogenicity”