Use of pyrosequencing to quantify incidence of a specific Aspergillus flavus strain within complex fungal communities associated with commercial cotton crops.
作者： Modan K. Das , Kenneth C. Ehrlich , Peter J. Cotty
关键词: Biotechnology 、 Pyrosequencing 、 Veterinary medicine 、 Aspergillus flavus 、 Cottonseed 、 Complementation 、 Fiber crop 、 Mycotoxin 、 Fungi imperfecti 、 Biology 、 Aflatoxin
摘要: ABSTRACT Atoxigenic strains of Aspergillus flavus have been used as aflatoxin management tools on over 50,000 hectares commercial crops since 2000. To assess treatment efficacy, atoxigenic strain incidence is routinely monitored by vegetative compatibility analyses (VCA) that require culturing, generation auxotrophs, and complementation with tester mutants. Two pyrosequencing assays (PA) no culturing were developed for monitoring incidences ginned cottonseed. The assays, which quantify frequencies characteristic single nucleotide polymorphisms (SNPs) in the aflR pksA genes, validated against standard VCA cottonseed collected from gins South Texas, Arizona, Southern California where AF36 to manage contamination. Cottonseed washings subjected both PA. PA was performed directly DNA isolated particulates pelleted wash water centrifugation. Addition CaCl2 a...
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