Abstract
【Objective】 The paper aimed at investigating the stability of the antifungal protein produced by Bacillus subtilis CCTCC M207209 under different conditions and evaluating its antifungal activity on apples against Penicillium expansum,and thus providing information on the application of strain CCTCC M207209 in practice.【Method】 Based on the greatest antifungal activity,ammonium sulfate saturation degree 50% was used to precipitate the active protein from the cell-free liquid culture of strain CCTCC M207209.After dissolved in a 0.2 mol/L pH 7.0 sodium phosphate buffer,the precipitated protein was dialyzed by a membrane of molecular weight of 8-10 ku against the same sodium phosphate buffer.The protein buffer solution retained by the membrane was used in all the other tests on the antifungal stability under different conditions.A cup-plate method was used to test the antifungal activity.Treatments for testing the active stability included holding at 121 ℃ for 30 min,under different pH and in 0.05 mol/L different metal ion solutions for 8 h,and certain UV strength for different periods.After the treatments,the protein solution was adjusted to its original pH and temperature.The antifungal activity on apples was tested by injecting 15 μL P.expansum spores suspension in the same hole after 30 μL antifungal substances were inoculated in apples.The tested antifungal substances included the antifungal protein,the liquid culture of strain CCTCC M207209 with cells,the autoclaved liquid culture with cells,and the cell-free liquid culture.The above inoculated apples were cultivated at 28 ℃ for 7 d.At the end of the cultivation,the ratio of P.expansum contaminated holes in apples was calculated and the diameter of the rotten spots was measured.【Result】 Protein precipitated at 50% ammonium sulfate saturation showed the greatest antifungal activity.The antifungal activity of the crude protein extracts did not reduce after treatment at 121 ℃ for 30 min and pH 7 for 8 h,but decreased quickly at the first 3 h under UV irradiation followed by very slow drop with more than 50% antifungal activity left after 8 h.Na+ and Mg2+ stimulated,but Ca2+,Zn2+ and Fe3+ destroyed the antifungal activity of the active protein at concentration of 0.05 mol/L The liquid culture with 1×108 cells/mL B.subtilis completely inhibited the growth of P.expansum(1.5×105 spores /mL)in apples.The antifungal protein reduced the occurrence rate of P.expansum contamination by 55.6% and significantly inhibited the growth of P.expansum in apples.【Conclusion】 The antifungal protein produced by B.subtilis CCTCC M207209 showed good stability under different environmental conditions and kept activity after autoclave,and thus had good potential in biological control of P.expansum in practice.
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Claims 9
Claim · model-written summary
Protein precipitated at 50% ammonium sulfate had the greatest antifungal activity.
- Taxon
- apples
Malus domesticae1 - Treatment
- 50% ammonium sulfate saturatione0
- Outcome
- antifungal activitye0
- Direction
- greateste0
Not reportedInteractor
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Protein precipitated at 50% ammonium sulfate saturation showed the greatest antifungal activity.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
Crude protein antifungal activity remained after 121 ℃ for 30 min and pH 7 for 8 h.
- Taxon
- applesMalus domesticae1
- Treatment
- treatment at 121 ℃ for 30 min and pH 7 for 8 he0
- Outcome
- antifungal activitye0
- Direction
- did not reducee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The antifungal activity of the crude protein extracts did not reduce after treatment at 121 ℃ for 30 min and pH 7 for 8 h,but decreased quickly at the first 3 h under UV irradiation followed by very slow drop with more than 50% antifungal activity left after 8 h.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
UV irradiation rapidly decreased antifungal activity in the first 3 h.
- Taxon
- applesMalus domesticae1
- Treatment
- UV irradiatione0
- Outcome
- antifungal activitye0
- Direction
- decreased quicklye0
- Qualifier
- at the first 3 he0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The antifungal activity of the crude protein extracts did not reduce after treatment at 121 ℃ for 30 min and pH 7 for 8 h,but decreased quickly at the first 3 h under UV irradiation followed by very slow drop with more than 50% antifungal activity left after 8 h.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
More than 50% antifungal activity remained after 8 h UV irradiation.
- Taxon
- applesMalus domesticae1
- Treatment
- UV irradiatione0
- Outcome
- antifungal activitye0
- Value
- more than 50%e0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The antifungal activity of the crude protein extracts did not reduce after treatment at 121 ℃ for 30 min and pH 7 for 8 h,but decreased quickly at the first 3 h under UV irradiation followed by very slow drop with more than 50% antifungal activity left after 8 h.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
Na+ and Mg2+ stimulated active protein antifungal activity.
- Taxon
- applesMalus domesticae1
- Treatment
- Na+ and Mg2+e0
- Outcome
- antifungal activitye0
- Direction
- stimulatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Na+ and Mg2+ stimulated,but Ca2+,Zn2+ and Fe3+ destroyed the antifungal activity of the active protein at concentration of 0.05 mol/L”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
Ca2+, Zn2+ and Fe3+ destroyed active protein antifungal activity.
- Taxon
- applesMalus domesticae1
- Treatment
- Ca2+,Zn2+ and Fe3+e0
- Outcome
- antifungal activitye0
- Direction
- destroyede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Na+ and Mg2+ stimulated,but Ca2+,Zn2+ and Fe3+ destroyed the antifungal activity of the active protein at concentration of 0.05 mol/L”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
B. subtilis liquid culture completely inhibited P. expansum growth in apples.
- Taxon
- applesMalus domesticae0
- Treatment
- liquid culture with 1×108 cells/mL B.subtilise0
- Interactor
- P.expansum
Penicillium expansume0pathogen - Second interactor
- B.subtilis
Bacillus subtilise0biocontrol_agent - Outcome
- growth of P.expansume0
- Direction
- completely inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The liquid culture with 1×108 cells/mL B.subtilis completely inhibited the growth of P.expansum(1.5×105 spores /mL)in apples.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
Antifungal protein reduced P. expansum contamination in apples by 55.6%.
- Taxon
- applesMalus domesticae0
- Treatment
- antifungal proteine0
- Interactor
- P.expansumPenicillium expansume0pathogen
- Outcome
- occurrence rate of P.expansum contaminatione0
- Direction
- reducede0
- Value
- 55.6%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The antifungal protein reduced the occurrence rate of P.expansum contamination by 55.6% and significantly inhibited the growth of P.expansum in apples.”
e1“The antifungal activity on apples was tested”
Claim · model-written summary
Antifungal protein significantly inhibited P. expansum growth in apples.
- Taxon
- applesMalus domesticae0
- Treatment
- antifungal proteine0
- Interactor
- P.expansumPenicillium expansume0pathogen
- Outcome
- growth of P.expansume0
- Direction
- inhibitede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The antifungal protein reduced the occurrence rate of P.expansum contamination by 55.6% and significantly inhibited the growth of P.expansum in apples.”
e1“The antifungal activity on apples was tested”
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