Development of a nested-PCR assay for the rapid detection of Pilidiella granati in pomegranate fruit

Yang, Xue; Hameed, Uzma; Zhang, Aifang; Zang, Huadong; Gu, Chen; Chen, Yu; Xu, Yongsheng

doi:10.1038/srep40954

Cited by 21 publications

(11 citation statements)

References 45 publications

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“…Finally, the assay proposed herein could provide a disease monitoring in a couple of hours, through the analysis of flowers, branches, shoots, and mummies, and thus the enrichment of epidemiological knowhow, favoring prevention practices. As such, it might represent an improvement of the detection methods already described in the literature [33].…”

Section: Discussionmentioning

confidence: 98%

The Effect of Polyphenols on Pomegranate Fruit Susceptibility to Pilidiella granati Provides Insights into Disease Tolerance Mechanisms

et al. 2020

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Pilidiella granati, also known as Coniella granati, is the etiological agent of pomegranate fruit dry rot. This fungal pathogen is also well-known as responsible for both plant collar rot and leaf spot. Because of its aggressiveness and the worldwide diffusion of pomegranate crops, the selection of cultivars less susceptible to this pathogen might represent an interesting preventive control measure. In the present investigation, the role of polyphenols in the susceptibility to P. granati of the two royalties-free pomegranate cultivars Wonderful and Mollar de Elche was compared. Pomegranate fruit were artificially inoculated and lesion diameters were monitored. Furthermore, pathogen DNA was quantified at 12–72 h post-inoculation within fruit rind by a real time PCR assay setup herein, and host total RNA was used in expression assays of genes involved in host-pathogen interaction. Similarly, protein extracts were employed to assess the specific activity of enzymes implicated in defense mechanisms. Pomegranate phenolic compounds were evaluated by HPLC-ESI-MS and MS2. All these data highlighted ‘Wonderful’ as less susceptible to P. granati than ‘Mollar de Elche’. In the first cultivar, the fungal growth seemed controlled by the activation of the phenylpropanoid pathway, the production of ROS, and the alteration of fungal cell wall. Furthermore, antifungal compounds seemed to accumulate in ‘Wonderful’ fruit following inoculation. These data suggest that pomegranate polyphenols have a protective effect against P. granati infection and their content might represent a relevant parameter in the selection of the most suitable cultivars to reduce the economic losses caused by this pathogen.

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Section: Discussionmentioning

confidence: 98%

The Effect of Polyphenols on Pomegranate Fruit Susceptibility to Pilidiella granati Provides Insights into Disease Tolerance Mechanisms

et al. 2020

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“…C. granati is an important fungal agent that causes preand post-harvest fruit decay and poses a new threat to the rapidly growing pomegranate industry in many parts of the World [11]. Post-harvest disease management for pomegranate fruit remains an important problem [37].…”

Section: Discussionmentioning

confidence: 99%

“…Crown rot disease causes tree decline, blight on branches and post-harvest fruit decay. Presence of this disease has been identified in Brazil, Cyprus, Italy, Spain, Korea, Florida, Israel, Iran, China, North Carolina, Pakistan, Netherlands, Mexico, Greece, Turkey and the United States [11].…”

Section: Introductionmentioning

confidence: 99%

Biological Control of Coniella granati Saccardo in Pomegranate

Tekiner¹,

Kotan²,

Tozlu³

et al. 2020

ujar

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Coniella granati Saccardo (Synonym Pilidiella granati) is a fungal pathogen that causes fruit brown rot, cankers on shoots and crown rot of pomegranate trees. Although cultural and chemical control is recommended against C. granati; cultural control is not enough and limited number of advisable fungicides used in chemical control against this pathogen. Therefore, alternative strategies are needed for this pathogen control. In this context, it was aimed to investigate the effect of some bacterial biocontrol agents against C. granati under in vitro conditions. Dual culture of eleven bacterial biocontrol agents [1 Bacillus megaterium (TV 3D), 3 Bacillus substilis (TV 6F, TV 17C, CP1), 1 Bacillus cereus (TV 85D), 1 Paenibacillus polymxa (TV 12E), 2 Pseudomonas fluorescens (MF 3, AR 9), 1 Burkholderia cepacia (BA 7) 1 Pantoea agglomerans (MF 1) and 1 Bacillus thuringiensis (BAB 420)] were tested for antagonistic properties against C. granati. Percent inhibition rate values changed from 11.90% to 66.67% in dual culture. B. cereus (TV 85D, 66.67%) was the most effective strains against C. granati respectively by B. substilis (TV 17C, 64.29%; TV 6F, 60.71%) in in vitro. As a result, promising results were obtained from these isolates in in vitro conditions. These isolates should be tested in vivo conditions for controlling the post-harvest decay of pomegranate fruits caused by C. granati.

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“…Twig blight and crown rot of pomegranate are emerging diseases in pomegranate cultivation that are caused by Pilidiella granati . A nested PCR assay improved both sensitivity and detection of P. granati and made it possible to diagnose the causative agent when the sample contained DNA as low as 10 pg of P. granati ( Yang X. et al., 2017 ). Great yam disease caused by Colletotrichum gloeosporioides ( Raj et al., 2013 ) and eucalyptus dieback disease caused by Cylindrocladium scoparium ( Qiao et al., 2016 ) were also detected by this technique.…”

Section: Molecular Tools For Detection Of Fungimentioning

confidence: 99%

Recent Advances in Molecular Diagnostics of Fungal Plant Pathogens: A Mini Review

Hariharan¹,

Prasannath²

2021

Front. Cell. Infect. Microbiol.

118

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Phytopathogenic fungal species can cause enormous losses in quantity and quality of crop yields and this is a major economic issue in the global agricultural sector. Precise and rapid detection and identification of plant infecting fungi are essential to facilitate effective management of disease. DNA-based methods have become popular methods for accurate plant disease diagnostics. Recent developments in standard and variant polymerase chain reaction (PCR) assays including nested, multiplex, quantitative, bio and magnetic-capture hybridization PCR techniques, post and isothermal amplification methods, DNA and RNA based probe development, and next-generation sequencing provide novel tools in molecular diagnostics in fungal detection and differentiation fields. These molecular based detection techniques are effective in detecting symptomatic and asymptomatic diseases of both culturable and unculturable fungal pathogens in sole and co-infections. Even though the molecular diagnostic approaches have expanded substantially in the recent past, there is a long way to go in the development and application of molecular diagnostics in plant diseases. Molecular techniques used in plant disease diagnostics need to be more reliable, faster, and easier than conventional methods. Now the challenges are with scientists to develop practical techniques to be used for molecular diagnostics of plant diseases. Recent advancement in the improvement and application of molecular methods for diagnosing the widespread and emerging plant pathogenic fungi are discussed in this review.

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Development of a nested-PCR assay for the rapid detection of Pilidiella granati in pomegranate fruit

Cited by 21 publications

References 45 publications

The Effect of Polyphenols on Pomegranate Fruit Susceptibility to Pilidiella granati Provides Insights into Disease Tolerance Mechanisms

The Effect of Polyphenols on Pomegranate Fruit Susceptibility to Pilidiella granati Provides Insights into Disease Tolerance Mechanisms

Biological Control of Coniella granati Saccardo in Pomegranate

Recent Advances in Molecular Diagnostics of Fungal Plant Pathogens: A Mini Review

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