Loop-mediated isothermal amplification (LAMP) detection of Phytophthora hibernalis, P. syringae and P. cambivora

Li, Guanrong; Huang, Genghua; Zhu, Lin-Hui; Lv, Dajin; Cao, Baohong; Liao, Fang; Luo, Jiafeng

doi:10.1007/s42161-018-0136-5

Cited by 12 publications

(11 citation statements)

References 18 publications

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“…Sun et al were reported that the LAMP diagnostic assay contributes to the rapid and accurate detection of soft-rot disease in Amorphophallus konjac at an early stage [46]. LAMP-based detection showed to be more sensitive than PCR in detecting Phytophthora hibernalis, P. syringae, and P. cambivora [47]. The comparison of LAMP assays with direct bacterial cultivation showed high sensitivity and high specificity.…”

Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification: a rapid molecular technique for early diagnosis of Pseudomonas syringae pv. syringae of stone fruits

Goudarzi

Mortazavi

2020

Journal of Genetic Engineering and Biotechnology

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Background Pathogenic bacteria cause significant economic damages in agriculture. The detection of such bacteria is considered as a continual interest for plant pathologists to prevent disease dissemination. Pseudomonas syringae pv. syringae is one of the most important bacterial pathogens infecting yield and quality of stone fruits throughout the world. Biochemical assays such as a LOPAT and GATTa are common methods to detect this pathogen. Serological tests and culturing on King’s B selective medium also used to isolate this bacterium. Selective media is composed of specific and effective ingredients to inhibit the growth of certain species of microbes in a mixed culture while allowing others to grow. These are used for the growth of only selected microorganisms. King’s B medium can be used as a general medium for the non-selective isolation cultivation and pigment production of Pseudomonas species from foods, cosmetic samples, plants, etc. Nevertheless, the mentioned methods are not enough accurate to differentiate the strains. On the other hand, PCR-based techniques are sensitive and efficient in detecting plant diseases. However, these techniques are not practicable for those researchers who do not have access to a thermal cycler. We have used loop-mediated isothermal amplification to couple with a target. The amplification of syrD gene using loop and bumper primers can be used to prevent disease dissemination. Results The outcome of this investigation indicated more sensitivity of LAMP in comparison to PCR. The direct addition of SYBR Gold in microtube is more sensitive than gel in both LAMP and PCR byproducts so we can eliminate gel electrophoresis, while the LAMP showed high sensitivity and high specificity in comparison to results obtained by cultivation. The described molecular test could detect Pseudomonas syringae pv. syringae type in nearly 1 h, and this is the first time that Lamp molecular detection of Pseudomonas syringae pv. syringae particularly on stone fruits is described and introduced. Conclusions The obtained data confirmed that LAMP is a fast, cheap, and high specific method for the rapid detection of Pseudomonas syringae pv. syringae to the comparison of PCR and culture.

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Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification: a rapid molecular technique for early diagnosis of Pseudomonas syringae pv. syringae of stone fruits

Goudarzi

Mortazavi

2020

Journal of Genetic Engineering and Biotechnology

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“…It also occasionally causes diseases of other crops such as tomato and apple (Erwin & Ribeiro, 1996). Due to its potential impact on the horticultural industry, this exotic pathogen has been listed as a quarantine pest in China since 2007 (Li et al, 2019). In 2015, P. hibernalis was detected in citrus fruit shipments from Tulare County, CA, USA to Shanghai, China, leading to a suspension of importing CA citrus products to China from February to November 2015 (Hao et al, 2018; Jiao et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

“…With the advent of molecular technology, other faster and more cost-effective techniques including isothermal diagnostic assays have become the mainstream in rapid detection of many Phytophthora species (Miles, Martin & Coffey, 2015) such as P. infestans (Hansen et al, 2016; Khan et al, 2017; Si Ammour et al, 2017), P. sojae (Dai et al, 2012; Dai, Zheng & Wu, 2015; Rojas et al, 2017), P. nicotianae (Li et al, 2015), P. cinnamomi (Dai et al, in press). A loop-mediated isothermal amplification (LAMP) assay for detecting P. hibernalis was developed by Li et al (2019).…”

Section: Introductionmentioning

confidence: 99%

A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis

Dai

Yang

et al. 2019

PeerJ

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Phytophthora hibernalis, the causal agent of brown rot of citrus fruit, is an important worldwide pathogen and a quarantine pest in China. Current diagnosis of the disease relies on disease symptoms, pathogen isolation and identification by DNA sequencing. However, symptoms caused by P. hibernalis can be confused with those by other Phytophthora and fungal species. Moreover, pathogen isolation, PCR amplification and sequencing are time-consuming. In this study, a rapid assay including 20-min recombinase polymerase amplification targeting the Ypt1 gene and 5-min visualization using lateral flow dipsticks was developed for detecting P. hibernalis. This assay was able to detect 0.2 ng of P. hibernalis genomic DNA in a 50-µL reaction system. It was specific to P. hibernalis without detection of other tested species including P. citrophthora, P. nicotianae, P. palmivora and P. syringae, four other important citrus pathogens. Using this assay, P. hibernalis was also detected from artificially inoculated orange fruits. Results in this study indicated that this assay has the potential application to detect P. hibernalis at diagnostic laboratories and plant quarantine departments of customs, especially under time- and resource-limited conditions.

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“…It also occasionally causes diseases of other crops such as tomato and apple (Erwin & Ribeiro 1996). Due to its potential impact on the horticultural industry, this exotic pathogen has been listed as a quarantine pest in China since 2007 (Li et al 2019). In 2015, P. hibernalis was detected in citrus fruit shipments from Tulare County, California, USA to Shanghai, China, leading to a suspension of importing Californian citrus PeerJ reviewing PDF | (2019:07:39552:1:1:NEW 7 Oct 2019)…”

Section: Introductionmentioning

confidence: 99%

“…With the advent of molecular technology, other faster and more cost-effective techniques including isothermal diagnostic assays have become the mainstream in rapid detection of many Phytophthora species (Miles et al 2015) such as P. infestans (Hansen et al 2016;Khan et al 2017;Si Ammour et al 2017), P. sojae (Dai et al 2012; Dai et al 2015;Rojas et al 2017), P. nicotianae (Li et al 2015), P. cinnamomi (Dai et al 2019). A loop-mediated isothermal amplification (LAMP) assay for detecting P. hibernalis was developed by Li et al (2019).…”

Section: Introductionmentioning

confidence: 99%

Peer Review #1 of "A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis (v0.1)"

Ioos¹

2019

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Phytophthora hibernalis, the causal agent of brown rot of citrus fruit, is an important worldwide pathogen and a quarantine pest in China. Current diagnosis of the disease relies on disease symptoms, pathogen isolation, and identification by DNA sequencing. However, symptoms caused by P. hibernalis can be confused with those by other Phytophthora and fungal species. Moreover, pathogen isolation, PCR amplification, and sequencing are timeconsuming. In this study, a rapid assay including 20-min recombinase polymerase amplification targeting the Ypt1 gene and 5-min visualization using lateral flow dipsticks was developed for detecting P. hibernalis. This assay was able to detect 0.2 ng of P. hibernalis genomic DNA in a 50-µL reaction system. It was specific to P. hibernalis without detection of other tested species including P. citrophthora, P. nicotianae, P. palmivora, and P. syringae, four other important citrus pathogens. Using this assay, P. hibernalis was also detected from artificially inoculated orange fruits. Results in this study indicated that this assay has the potential application to detect P. hibernalis at diagnostic laboratories, and plant quarantine departments of customs, especially under time-and resource-limited conditions.

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Loop-mediated isothermal amplification (LAMP) detection of Phytophthora hibernalis, P. syringae and P. cambivora

Cited by 12 publications

References 18 publications

Loop-mediated isothermal amplification: a rapid molecular technique for early diagnosis of Pseudomonas syringae pv. syringae of stone fruits

Loop-mediated isothermal amplification: a rapid molecular technique for early diagnosis of Pseudomonas syringae pv. syringae of stone fruits

A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis

Peer Review #1 of "A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis (v0.1)"

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