Conference Portal, 2018 International Conference on Pure and Applied Science

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Cultivation and Detection of Viable But Nonculturable (VBNC) Fungi in Soil by Using Yoghurt Whey Infusion Agar (YWIA)
Wijdan A. Ali Al Mohamdi, Bahrouz M.A. Al-jaff, Sajid S.S. AL–Saeedi

##manager.scheduler.building##: Presidency Building Conference Auditorium
Date: 2018-04-23 03:05 PM – 03:25 PM
Last modified: 2018-08-15


Nonculturable and nonsporulating fungi represent a great challenge in studying biotrophic,endotrophic and mycorrhizal fungal groups. In this study a novel culture dependent technique complementedwith PCR was used to cultivate and identify nonculturable fungi in soil. To develop techniques for cultivationof nonculturable fungi for the purpose of expanding studies on fungal biodiversity in soil, yoghurt wheyinfusion agar (YWIA) was developed. By this technique, from higher dilutions of soil two tiny colonies (lessthan 1 mm in diameter) had grown on YWIA (1.0% and 2.0% filtration sterilized yoghurt whey) afterincubation for 1-3 months at 28˚C, but had not shown growth when replicated on traditional mycologicalmedia, were selected, purified, studied culturally and microscopically, and identified by molecular methods.Growth of the two isolates on PDA, but not Sabouraud Dextrose Agar, Malt extract agar, or Czapek DoxAgar, happened after three weeks of incubation when a loopful of a colony grown after several successivesubculturing on YWIA was transferred to PDA. The growth on PDA has been empirically proven due to theinoculum size effect and the cultural and microscopical features were more resemble Aspergillus spp. Twofungal specific primer sets (EF4F / ITS4r and EF60F / ITS4r) were used to amplify partial sequences of fungalrRNA gene included ITS sequences. The two partial sequences of the two isolates were aligned through theBLASTN phylogenetic analysis against NCBI database which revealed higher identities with Aspergillusterreus. The blast tree showed that both isolates are neighbor joined to Aspergillus spp. These results led us toconsider the isolates as viable but nonculturable fungi (VBNC fungi) like the common phenomenon found insome bacterial species.


Published 01 August 2018



Non-culturable fungi; polymerase chain reaction; soil, viable but non-culturable fungi; yoghurt whey infusion agar

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