应用定量聚合酶链反应(qPCR)测定小麦穗镰刀菌感染。

V Rossi, V Terzi, F Moggi, C Morcia, P Faccioli, M Haidukowski, M Pascale
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引用次数: 22

摘要

定量聚合酶链式反应法定量测定小麦收获粒和头部组织中产生毛霉烯的镰刀菌和禾谷镰刀菌DNA的准确性,并与籽粒感染的发生率和脱氧雪腐镰刀菌醇水平进行了比较。在第一个试验中,6个硬粒小麦和面包小麦品种随机种植2年,在抽穗至面团成熟之间的6个生育期接种大分生镰刀菌,获得了广泛的赤霉病发病率。真菌DNA与脱氧雪腐镰刀菌醇含量之间存在密切的关系,这种关系在镰刀菌属、小麦种、品种以及广泛的赤霉病感染中都是一致的。在第二个试验中,在环境控制条件下种植盆栽小麦植株,并在开花时接种两种镰刀菌;接种前和接种后6 h ~ 12 d采集头样,采用定量聚合酶链反应法处理。该试验可以在感染发生后检测真菌入侵植物的动态,并在疾病症状出现之前筛选出感染的存在:镰孢镰刀菌在18-24小时内检测到感染,而谷草镰刀菌在2-9天内检测到感染。
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Assessment of Fusarium infection in wheat heads using a quantitative polymerase chain reaction (qPCR) assay.

The accuracy of a quantitative polymerase chain reaction assay in quantifying the DNA of trichothecene-producing F. culmorum and F. graminearum within harvested wheat grains and head tissue was evaluated in comparison with incidences of infected kernels and deoxynivalenol levels. In a first experiment, six durum and bread wheat varieties were grown in randomized plots for a 2-year period, and inoculated with Fusarium macroconidia at six growth stages between heading and dough ripening, to obtain a wide range of Fusarium head blight incidences. There was a close relationship between fungal DNA and the amount of deoxynivalenol, and this relationship was consistent over Fusarium species, wheat species and varieties, and over a wide range of Fusarium head blight infection. In a second experiment potted wheat plants were grown under environmentally controlled conditions and inoculated with the two Fusarium species at full flowering; head samples were collected before inoculation and after 6 h to 12 days, and processed by the quantitative polymerase chain reaction assay. This assay made it possible to detect the dynamic of fungal invasion in planta after infection had occurred, and to single out the presence of infection before the onset of the disease symptoms: A robust detection of the infection occurred within 18-24 h for F. culmorum, and within 2-9 days for F. graminearum.

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