Gerin Donato, Pollastro Stefania, Raguseo Celeste, De Miccolis Angelini Rita M, Faretra Francesco
Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, Bari, Italy.
Public Laboratory of Research SELGE Network No. 14, Bari, Italy.
Front Microbiol. 2018 Aug 31;9:2073. doi: 10.3389/fmicb.2018.02073. eCollection 2018.
strain icc012 and strain icc080, the microbial active ingredients of Remedier (ISAGRO, Novara, Italy), are biocontrol agents (BCAs) employable for crop protection against a wide range of fungal pathogens, including soil-borne pathogens and fungi involved in grapevine trunk disease. In this study, single and duplex real-time quantitative PCR (qPCR) methods to detect and quantify and were developed. Primers/probe sets were designed on the and genes and tested for specificity on a panel of microorganisms commonly associated with grape wood and soil. No differences were observed comparing single- and duplex-qPCR assays on different BCAs, 1 pg of target DNA was detected approximately at = 34. -values and the efficiency were always equal to 0.99 and >80%, respectively. The detection limit of the duplex-qPCR assay on artificially inoculated samples was 2 × 10 and 4 × 10 conidia g of grape wood tissue and soil, respectively. The methods will be useful to better schedule BCA application in the field and in grapevine nurseries, as well as for investigating the dynamic of BCA populations.
菌株ICC012和菌株ICC080是Remedier（意大利诺瓦拉的ISAGRO公司）的微生物活性成分，它们是生物防治剂（BCAs），可用于保护作物免受多种真菌病原体侵害，包括土传病原体和与葡萄树干病害有关的真菌。在本研究中，开发了用于检测和定量[具体物质未明确写出]的单重和双重实时定量PCR（qPCR）方法。基于[具体基因未明确写出]基因设计了引物/探针组，并在一组通常与葡萄木和土壤相关的微生物上测试了其特异性。在不同生物防治剂上比较单重和双重qPCR检测时未观察到差异，在约 = 34时检测到1 pg的目标DNA，[具体指标未明确写出]值和效率分别始终等于0.99和>80%。在人工接种样品上双重qPCR检测的检测限分别为葡萄木组织和土壤中2×10和4×10个分生孢子/克。这些方法将有助于更好地安排生物防治剂在田间和葡萄苗圃中的应用，以及用于研究生物防治剂种群动态。
