Quantification of trichothecene-producing Fusarium species in harvested grain by competitive PCR to determine efficacies of fungicides against fusarium head blight of winter wheat
Quantification of trichothecene-producing Fusarium species in harvested grain by competitive PCR to determine efficacies of fungicides against fusarium head blight of winter wheat
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DOI：
10.1128/aem.67.4.1575-1580.2001
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发表时间：
2001-04-01
影响因子：
4.4
通讯作者：
Jenkinson, P
中科院分区：
文献类型：
--
作者：
Edwards, SG;Pirgozliev, SR;Jenkinson, P
我们开发了一种基于PCR的检测方法，以定量的基础上，从β-二烯合酶基因（Tri 5）的引物产生β-二烯镰刀菌。对一系列镰刀菌头枯病（FHB）（也称为赤霉病）病原体的引物进行了测试，发现特异性扩增260 bp的产物，从25个菌株属于六个产二孢霉素镰刀菌物种。定量测定了田间试验收获的谷物中产生腐殖酸的镰刀菌和腐殖酸真菌毒素脱氧雪腐镰刀菌烯醇（DON）的量，该田间试验旨在测试杀真菌剂叶菌唑、嘧菌酯和戊唑醇控制FHB的效力。在收获的谷物中，FHB严重程度与DON之间没有发现相关性，但是在谷物中存在的产生二孢霉素的镰刀菌的量与DON之间存在良好的相关性。与未处理的对照组相比，嘧菌酯没有影响产孢镰刀菌的水平。叶菌唑和戊唑醇显着降低收获的粮食中的镰刀菌的数量。我们假设，杀真菌剂影响FHB严重程度和收获的谷物中的DON量之间的关系，通过改变FHB疾病复合体内的产二孢霉素镰刀菌的比例，而不是通过改变DON生产率。Tri 5定量PCR检测将有助于减少食品和动物饲料中的青霉烯真菌毒素的研究。
We developed a PCR-based assay to quantify trichothecene-producing Fusarium based on primers derived from the trichodiene synthase gene (Tri5). The primers were tested against a range of fusarium head blight (FHB) (also known as scab) pathogens and found to amplify specifically a 260-bp product from 25 isolates belonging to six trichothecene-producing Fusarium species. Amounts of the trichothecene-producing Fusarium and the trichothecene mycotoxin deoxynivalenol (DON) in harvested grain from a field trial designed to test the efficacies of the fungicides metconazole, azoxystrobin, and tebuconazole to control FHB were quantified. No correlation was found between FHB severity and DON in harvested grain, but a good correlation existed between the amount of trichothecene-producing Fusarium and DON present within grain. Azoxystrobin did not affect levels of trichothecene-producing Fusarium compared with those of untreated controls. Metconazole and tebuconazole significantly reduced the amount of trichothecene-producing Fusarium in harvested grain. We hypothesize that the fungicides affected the relationship between FHB severity and the amount of DON in harvested grain by altering the proportion of trichothecene-producing Fusarium within the FHB disease complex and not by altering the rate of DON production. The Tri5 quantitative PCR assay will aid research directed towards reducing amounts of trichothecene mycotoxins in food and animal feed.