A real time loop-mediated isothermal amplification (RealAmp) assay for rapid detection of Pleurostoma richardsiae in declining olive plants

IF 1.9 3区农林科学 Q2 AGRONOMY

Phytopathologia Mediterranea Pub Date : 2022-07-19 DOI:10.36253/phyto-12748

Abdraouf Sadallah, S. Minutillo, F. Valentini, M. Raimondo, F. Lops, A. Carlucci, A. Ippolito, A. D’ONGHIA

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引用次数: 0

Abstract

Pleurostoma richardsiae is associated with host trunk diseases, known to cause dieback, cankers and wilting of woody trees, and human infections. This fungus was isolated from wood tissues of declining olive trees and grapevines showing esca disease symptoms, in the Apulia region of Italy. Fungus detection has been based on morphological and molecular features, which are time-consuming to identify and require well-trained personnel. Improvement of Pl. richardsiae detection in olive was achieved through development of real time loop-mediated isothermal amplification targeting the intergenic spacer (IGS) region of the fungus. Specificity of the assay was confirmed using ten Pl. richardsiae strains and 36 other fungus strains of species usually isolated from declining olive trees. The achieved limit of detection was 7.5 × 10-2 ng μL-1 of Pl. richardsiae genomic DNA. A preliminary validation of RealAmp was also performed using material from infected olive plants artificially inoculated in a greenhouse.

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实时环介导等温扩增（RealAmp）法快速检测衰退橄榄植物中的厚胸胸膜瘤

richardepleurostoma与宿主树干疾病有关，已知会引起枯死、溃烂和木本树木枯萎，以及人类感染。这种真菌是从意大利阿普利亚地区表现出埃斯卡病症状的橄榄树和葡萄藤的木材组织中分离出来的。真菌检测是基于形态学和分子特征，这是费时的识别和需要训练有素的人员。通过开发针对菌种间间隔区(IGS)的实时环介导等温扩增技术，改进了橄榄中Pl. richardsiae的检测方法。用10株plicrichardsiae菌种和36株通常从凋落橄榄树中分离的菌种证实了该方法的特异性。该方法的检出限为7.5 × 10-2 ng μL-1。RealAmp的初步验证还使用了在温室中人工接种的受感染橄榄植株的材料。

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来源期刊

Phytopathologia Mediterranea 生物-植物科学

CiteScore

4.40

自引率

8.30%

发文量

审稿时长

6-12 weeks

期刊介绍： Phytopathologia Mediterranea is an international journal edited by the Mediterranean Phytopathological Union. The journal’s mission is the promotion of plant health for Mediterranean crops, climate and regions, safe food production, and the transfer of new knowledge on plant diseases and their sustainable management. The journal deals with all areas of plant pathology, including etiology, epidemiology, disease control, biochemical and physiological aspects, and utilization of molecular technologies. All types of plant pathogens are covered, including fungi, oomycetes, nematodes, protozoa, bacteria, phytoplasmas, viruses, and viroids. The journal also gives a special attention to research on mycotoxins, biological and integrated management of plant diseases, and the use of natural substances in disease and weed control. The journal focuses on pathology of Mediterranean crops grown throughout the world. The Editorial Board of Phytopathologia Mediterranea has recently been reorganised, under two Editors-in-Chief and with an increased number of editors.