The fungus was later named as Pilidiella granati and then C. granati was first described by Saccardo in 1876 as Phoma granati, isolated from pomegranate plants from Italy. This pathogen may induce many symptoms leading to twig blight and crown rots and death of the plant (Mirabolfathy et al., 2012). granati (Sacc.) is one of the most deleterious diseases involved in a serious threat to pomegranate cultivation all around the world including China (Chen et al., 2014 Mirabolfathy et al., 2012 Thomidis, 2015 Kc and Vallad, 2016). Twig blight and crown rot of pomegranate caused by C. punicae, producing various disease symptoms (Palou et al., 2013 Munhuweyi et al., 2016). Pomegranate, covering five continents, is unfortunately damaged by a range of pathogenic fungi, including Botrytis cinerea, Alternaria alternata, Penicillium implicatum, Coniella granati, Aspergillus niger, and bacterial pathogens such as Xanthomonas axonopodis pv. The pomegranate fruit and its peel are especially known for their high nutritional contents, essential vitamins, minerals, and antioxidant properties, their ability to reduce cholesterol level, heart diseases, and cancer risk (Ismail et al., 2012 Neyrinck et al., 2013). The various parts of the plant including leaves, twigs, flowers, and fruits are important due to their high medicinal values and health benefits (Bekir et al., 2013 Ismail et al., 2012). China is one of the world's major pomegranate producing countries with about 1.2 million tons annual production and 120,000 hm 2 plantation area (Niu et al., 2014). Pomegranate ( Punica granatum L.) is an important shrub plant of tropical and subtropical climatic regions of the world. Thus, this LAMP assay can be used for the simple, fast, and precise diagnosis of twig blight and crown rot disease in suspected and infected parts of pomegranate plants. The resultant amplicons were easily detected by a visual examination in vials, using SYBR Green I, where they produced a yellowish green color for positive reactions, and were also identifiable by electrophoresis on the agarose gel. The assay was successfully evaluated for the minimum detection limit of 1 pg of crude DNA, which showed better sensitivity results using underoptimized LAMP reaction condition, at 65 ☌ for 60 min, which was superior to using conventional PCR. specifically among 23 fungal species isolated from different fruits including pomegranate in the Anhui Province of China. The assay is based on the use of four primers recognizing six distinct areas of translation elongation factor 1 alpha ( Tef-1α ) region (204 bp) of genomic DNA (gDNA) of Coniella spp. In this study, a sensitive loop-mediated isothermal amplification (LAMP) assay was developed for the rapid diagnosis of twig blight and crown rot disease of pomegranate caused by Coniella spp. Twig blight and crown rot of pomegranate ( Punica granatum L.) is a destructive disease, caused by Coniella granati, a plant pathogenic fungus, leading to severe losses by damaging plant health, lowering fruit production, and deteriorating fruit quality.
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