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A rapid assay for patulin degradation by the basidiomycetous yeast Rhodotorula glutinis strain LS11
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- Wright, Sandra, A. I., 1962 (författare)
- Gothenburg University,Göteborgs universitet,Institutionen för växt- och miljövetenskaper,Department of Plant and Environmental Sciences
- Ianiri, Giuseppe (författare)
- De Felice, Dario (författare)
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- Castoria, Raffaello (författare)
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- (creator_code:org_t)
- 2008
- 2008
- Engelska.
- Ingår i: COST Action 924. Novel approaches for the control of postharvest diseases and disorders.. ; , s. 19-29
- Konferensbidrag (övrigt vetenskapligt/konstnärligt)
Abstract
Ämnesord
Stäng
- The mycotoxin patulin is produced by the green mould pathogen Penicillium expansum in rotting apples during postharvest storage. Strains of epiphytic yeasts and yeast-like organisms have the capacity to protect apples from infection by green mould. One of these, Rhodotorula glutinis strain LS11 can degrade patulin to two non-toxic products, of which one is desoxypatulinic acid. The aim of the work was to develop a rapid, high-throughput assay for patulin degradation in order to enable efficient screening of mutants and library clones to identify genes of LS11 that are involved in patulin degradation. Escherichia coli is highly sensitive to patulin (6-10 μg/ml) but is insensitive to desoxypatulinic acid. This information was utilized to test if a method for patulin detection that was based on inhibition zones in a lawn seeded with E. coli could replace the existing TLC plate method, which is cumbersome and time-consuming. Indeed, an assay for patulin based on the sensitivity of E. coli was developed and validated. In addition, a degradation assay using 96-well microtiter plates was made more efficient by cutting the assay time from ten to three days, by shaking the culture, reducing the assay volume and reducing the amount of patulin utilized. R. glutinis LS11 was mutagenized by random insertion of a hygromycin B resistance cassette, through Agrobacterium-mediated transformation. The putative transformants were tested for stability and assayed for patulin degradation using the method developed. Some of the mutants did not degrade patulin and all of these also had lost the ability to grow in the presence of this mycotoxin. The results show that the screening method developed was suited to detecting mutants, library clones or strains with altered patulin degradation profiles in liquid culture.
Ämnesord
- LANTBRUKSVETENSKAPER -- Lantbruksvetenskap, skogsbruk och fiske -- Livsmedelsvetenskap (hsv//swe)
- AGRICULTURAL SCIENCES -- Agriculture, Forestry and Fisheries -- Food Science (hsv//eng)
Nyckelord
- apple
- mycotoxins
- desoxypatulinic acid
- mutagenesis
- TLC
Publikations- och innehållstyp
- vet (ämneskategori)
- kon (ämneskategori)
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