scholarly journals Detection of Potato Virus Y (Pvy) by Reverse-Transcription Loop-Mediated Nucleic Acid Amplification (Rt-Lamp)

2017 ◽  
Vol 75 (1) ◽  
pp. 77-85
Author(s):  
Krzysztof Treder ◽  
Joanna Chołuj ◽  
Bogumiła Zacharzewska ◽  
Mateusz Mielczarek
Keyword(s):  
Potato Virus ◽  
Reverse Transcription ◽  
Plant Pathogens ◽  
Potato Virus Y ◽  
Potato Crop ◽  
Lamp Assay ◽  
Good Alternative ◽  
Plant Viruses ◽  
Nucleic Acid Amplification ◽  
Simple Protocol

Abstract Potato virus Y (PVY), a type member of the genus Potyvirus (family Potyviridae), is currently the most important virus infecting the potato crop. PVY is also a dangerous pathogen of the tomato, pepper, and tobacco. The reverse transcription loop-mediated amplification (RT-LAMP) is gaining recognition as a good alternative to RT-PCR in diagnosing plant viruses. Here, we provide a detailed description of a simple protocol for fast and sensitive detection of PVY by the RT-LAMP assay, which can be easily adapted to detect other plant pathogens, harboring both RNA and DNA genomes.

scholarly journals Survival and Transmission of Potato Virus Y, Pepino Mosaic Virus, and Potato Spindle Tuber Viroid in Water

2013 ◽  
Vol 80 (4) ◽  
pp. 1455-1462 ◽  
Author(s):  
N. Mehle ◽  
I. Gutiérrez-Aguirre ◽  
N. Prezelj ◽  
D. Delić ◽  
U. Vidic ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Potato Virus ◽  
Plant Pathogens ◽  
Potato Virus Y ◽  
Potato Spindle Tuber Viroid ◽  
Plant Viruses ◽  
Detection Methods ◽  
Mechanical Transmission ◽  
Pepino Mosaic Virus ◽  
Viroid Infection

ABSTRACTHydroponic systems and intensive irrigation are used widely in horticulture and thus have the potential for rapid spread of water-transmissible plant pathogens. Numerous plant viruses have been reported to occur in aqueous environments, although information on their survival and transmission is minimal, due mainly to the lack of effective detection methods and to the complexity of the required transmission experiments. We have assessed the role of water as a source of plant infection using three mechanically transmissible plant pathogens that constitute a serious threat to tomato and potato production: pepino mosaic virus (PepMV), potato virus Y (PVY), and potato spindle tuber viroid (PSTVd). PepMV remains infectious in water at 20 ± 4°C for up to 3 weeks, PVY (NTN strain) for up to 1 week, and PSTVd for up to 7 weeks. Experiments using a hydroponic system show that PepMV (Ch2 genotype) and PVY (NTN strain) can be released from plant roots into the nutrient solution and can infect healthy plants through their roots, ultimately spreading to the green parts, where they can be detected after a few months. In addition, tubers developed on plants grown in substrate watered with PSTVd-infested water were confirmed to be the source of viroid infection. Our data indicate that although well-known pathways of virus spread are more rapid than water-mediated infection, like insect or mechanical transmission through leaves, water is a route that provides a significant bridge for rapid virus/viroid spread. Consequently, water should be taken into account in future epidemiology and risk assessment studies.

scholarly journals Rapid Detection of SARS-CoV-2 Using Reverse transcription RT-LAMP method

2020 ◽  
Author(s):  
Weihua Yang ◽  
Xiaofei Dang ◽  
Qingxi Wang ◽  
Mingjie Xu ◽  
Qianqian Zhao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcription ◽  
Virus Disease ◽  
Lamp Assay ◽  
Nucleic Acid Amplification ◽  
N Gene ◽  
Nucleic Acid Detection ◽  
Lamp Method ◽  
Rt Pcr ◽  
Rapid Amplification

AbstractCorona Virus Disease 2019 (COVID-19) is a recently emerged life-threatening disease caused by SARS-CoV-2. Real-time fluorescent PCR (RT-PCR) is the clinical standard for SARS-CoV-2 nucleic acid detection. To detect SARS-CoV-2 early and control the disease spreading on time, a faster and more convenient method for SARS-CoV-2 nucleic acid detecting, RT-LAMP method (reverse transcription loop-mediated isothermal amplification) was developed. RNA reverse transcription and nucleic acid amplification were performed in one step at 63 °C isothermal conditions, and the results can be obtained within 30 minutes. ORF1ab gene, E gene and N gene were detected at the same time. ORF1ab gene was very specific and N gene was very sensitivity, so they can guarantee both sensitivity and specificity for SARS-CoV-2. The sensitivity of RT-LAMP assay is similar to RT-PCR, and specificity was 99% as detecting 208 clinical specimens. The RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection, which is useful for the rapid and reliable clinical diagnosis of SARS-CoV-2.

scholarly journals Identification of Potato Virus Y (Pvy) and Its Economic Importance on Potato Crop

2014 ◽  
Vol 2 (2) ◽  
pp. 304-309
Author(s):  
Nabeel Kassim ◽  
Zulaykha Nerway ◽  
Kurdistan Yousif
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Potato Crop ◽  
Economic Importance

scholarly journals Reverse Transcription Loop-Mediated Isothermal Amplification of DNA for Detection of Potato virus Y

Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 605-610 ◽  
Author(s):  
Xianzhou Nie
Keyword(s):  
Potato Virus ◽  
Reverse Transcription ◽  
Potato Virus Y ◽  
Dna Amplification ◽  
Isothermal Amplification ◽  
Rt Pcr ◽  
Potato Leaf ◽  
Time Saving ◽  
Loop Mediated Isothermal Amplification ◽  
One Step

A reverse transcription loop-mediated isothermal amplification of DNA (RT-LAMP) for detection of Potato virus Y (PVY) was developed. In this procedure, a set of four primers matching a total of six sequences of the coat protein (CP) gene of PVY were designed in such a way that a loop could be formed and elongated during DNA amplification. Using PVY CP complementary DNA clones as templates, the LAMP reaction was optimized by adjusting the concentrations of MgSO4, dNTPs, and Bst DNA polymerase. The effects of fragment length of target DNA on LAMP also were investigated. Two-step and one-step RT-LAMPs were performed using RNA extracts of various PVY cultures, and the results were correlated with two-step reverse transcription polymerase chain reaction (RT-PCR) for detection of PVY. Further, the turbidity caused by precipitation of magnesium pyrophosphate formed in positive RT-LAMP reactions was used to measure the amplification by utilizing a time-saving spectrophotometric method. The one-step RT-LAMP-turbidity method gave results comparable with the two-step RT-PCR method for detection of PVY from potato leaf and tuber samples. Of the total 240 samples, 234 were diagnosed similarly by both methods.

scholarly journals Detection of Potato virus Y associated with African nightshade leafy vegetable (Solanum scabrum miller) in Kenya

2020 ◽  
Vol 12 (3) ◽  
pp. 255-259
Author(s):  
S.L. Kimaru ◽  
D.C. Kilalo ◽  
J.W. Kimenju ◽  
W.M. Muiru
Keyword(s):  
Potato Virus ◽  
Food Systems ◽  
Potato Virus Y ◽  
Plant Viruses ◽  
Molecular Techniques ◽  
Sub Saharan Africa ◽  
Small Scale ◽  
Molecular Testing ◽  
Ecological Zones ◽  
Field Samples

Abstract. The African nightshades (ANS) have been part of the food systems in sub-Saharan Africa for generations. They are particularly attractive to small scale farmers because the risks of crop losses are much lower compared to the exotic vegetables. Plant viruses are economically important pathogens affecting African nightshade production. More than 200 plant viruses are reported to infest solanaceous family crops. This study was carried out to detect Potato virus Y associated with African nightshade. Symptomatic leaf samples were obtained from four agro-ecological zones in Kenya. The viral pathogens were detected using serology and molecular techniques. Laboratory experiments were carried out at the University of Nairobi and Kenya Plant Health Inspectorate Service (KEPHIS) from February 2017 to August, 2018. Field samples had the highest mean viral percentage frequency of 44.8% compared to greenhouse samples having 29.8% using serology methods. Molecular testing revealed that greenhouse and farm field sample were positive for the three Potato virus Y strains PVYN:O (necrotic recombinant), PVYO (ordinary) and PVYNTN (necrotic). Detection of Potato virus Y in ANS suggests that it is prevalent in the sampled regions and could cause problems to other solanaceous crops.

scholarly journals First Report of the Necrotic Strain of Potato virus Y (PVYN) on Potatoes in the Northwestern United States

Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1177-1177 ◽  
Author(s):  
J. M. Crosslin ◽  
P. B. Hamm ◽  
K. C. Eastwell ◽  
R. E. Thornton ◽  
C. R. Brown ◽  
...  
Keyword(s):  
United States ◽  
Monoclonal Antibodies ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Plant Viruses ◽  
Enzyme Analysis ◽  
Rt Pcr ◽  
Veinal Necrosis ◽  
Pcr Products ◽  
Plant Pathol

More than 50 isolates of Potato virus Y (PVY) with characteristics of strains that cause tobacco veinal necrosis (PVYN) were obtained from potatoes (Solanum tuberosum L.) grown in the northwestern United States. These isolates are being characterized at the biological and molecular levels. Isolate RR1 was obtained from leaves of potato cv. Ranger Russet showing distinct mottling and leaf deformity, which is in contrast to the leaf-drop and necrosis usually observed with ordinary strains of PVY (PVYO) in this variety. Isolate AL1 was obtained from tubers of potato cv. Alturas showing distinct internal light brown rings and blotches. When RR1 and AL1 were transmitted to tobacco (Nicotiana tabacum L. cvs. Samsun NN and 423), they caused systemic veinal necrosis, including stem and petiole lesions typical of PVYN strains (2). Symptoms induced by RR1 and AL1 on tobacco appeared 9 to 11 days after inoculation, whereas some other isolates caused delayed veinal necrosis. All isolates that produced veinal necrosis on tobacco were detectable with PVY polyclonal antisera. Potato virus X was not detected by enzyme-linked immunosorbent assay in tobacco plants showing veinal necrosis. Some isolates, including AL1, failed to react in serological tests using PVYN-specific monoclonal antibodies obtained from three commercial sources. Other isolates, including RR1, were detectable with these monoclonal antibodies. Reverse transcription-polymerase chain reaction (RT-PCR) products obtained with primers specific for the coat protein (CP) open reading frame (ORF) were cloned and sequenced. AL1 possesses a CP more closely related to PVYO type isolates, which would account for its failure to react with PVYN monoclonal antibodies. In this regard, AL1 is similar to the PVYN-Wilga isolate (1). Other isolates that are detectable with the PVYN monoclonal antibodies possess a CP more consistent with N strains of the virus. Results of RT-PCR tests using primers derived from the P1 ORF sequence (3), and the restriction enzyme analysis and sequencing of the RT-PCR products, all suggest that AL1 and RR1 are related to European-type members of PVY tuber necrotic (NTN) or N strains. However, other isolates under investigation appear to be more closely related to previously reported North American NTN types (3). The symptomatology of these viruses on tobacco and potato, and the serological and molecular data clearly show that at least two distinct variants of PVYN have been found for the first time in a major potato production area of the United States, and pose a potential threat to the potato industry. References: (1) B. Blanco-Urgoiti et al. Eur. J. Plant Pathol. 104:811, 1998. (2) J. A. de Bokx and H. Huttinga. Potato virus Y. Descriptions of Plant Viruses. No. 242, CMI/AAB, Surrey, England, 1981. (3) R. P. Singh et al. Can J. Plant Pathol. 20:227, 1998.

scholarly journals Potato virus Y and Potato leafroll virus management under climate change in sub-Saharan Africa

2020 ◽  
Vol 116 (11/12) ◽  
Author(s):  
Kerstin Krüger ◽  
Jacquie E. van der Waals
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Production Systems ◽  
Plant Viruses ◽  
Potato Leafroll Virus ◽  
Climatic Conditions ◽  
Cultural Control ◽  
Sub Saharan Africa ◽  
Sub Saharan ◽  
Leafroll Virus

Potato has increased in importance as a staple food in sub-Saharan Africa, where its production is faced with a multitude of challenges, including plant disease development and spread under changing climatic conditions. The economically most important plant viruses affecting potatoes globally are Potato virus Y (PVY) and Potato leafroll virus (PLRV). Disease management relies mostly on the use of insecticides, cultural control and seed certification schemes. A major obstacle in many sub-Saharan Africa countries is the availability of disease-free quality seed potatoes. Establishment and implementation of quality control through specialised seed production systems and certification schemes is critical to improve seed potato quality and reduce PVY and PLRV sources. Seed could be further improved by breeding virus-resistant varieties adapted to different environmental conditions combined with management measures tailored for smallholder or commercial farmers to specific agricultural requirements. Innovative technologies – including more sensitive testing, remote sensing, machine learning and predictive models – provide new tools for the management of PVY and PLRV, but require support for adoption and implementation in sub-Saharan Africa.

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