Evaluation of PCR, IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

2001 ◽  
Vol 57 (11) ◽  
pp. 1068-1074 ◽  
Author(s):  
Said K Ibrahim ◽  
Stephen T Minnis ◽  
Anthony D P Barker ◽  
Mike D Russell ◽  
Patrick P J Haydock ◽  
...  
2019 ◽  
Vol 15 (2) ◽  
pp. 77-84
Author(s):  
Auliya Selamet ◽  
Supramana Supramana ◽  
Meity Suradji Sinaga ◽  
Ali Nurmansyah ◽  
Kikin Hamzah Mutaqin

The Morphology and Morphometry of Potato Cyst Nematodes (Globodera spp.) from Dataran Tinggi Dieng, Central JavaIn Indonesia, the potato cyst nematode (PCN) was first reported in 2003, at potato plantation in Batu, East Java. Golden cyst (Globodera rostochiensis) was detected in Bandung, West Java; Banjarnegara, Central Java; Batu, East Java; and Gowa, South Sulawesi, whereas the pale cyst (G. pallida) has been reported at limited distribution in Dataran Tinggi Dieng (Central Java). The aim of this research was to identify the Globodera species of Dataran Tinggi Dieng based on morphological and morphometrical methods. Soil samples were collected from Dieng plateau at 30 selected potato plantations ranging from 1 100 – 2 100 m above sea level. The identification of PCN was done by observing the morphological and morphometrical key characters of cysts and second juveniles (J2). Two species of Globodera, that are G. rostochiensis and G. pallida, were identified from soil samples. The key characters of G. pallida including distinct forward projection of J2 stylet knob, granek’s ratio value less than 3, and number of cuticular ridges between vulval basin and anus are less than 12 were identified. There were 25 specimens that having granek’s ratio less than 3, and 10 specimens having cuticular ridges number less than 12. G. rostochiensis which has the shape of a stylet knob protrudes posteriorly and the value of the granek’s ratio greater than 3.5 was identified in 30 specimens. 


2002 ◽  
Vol 140 (2) ◽  
pp. 187-195 ◽  
Author(s):  
S T MINNIS ◽  
P P J HAYDOCK ◽  
S K IBRAHIM ◽  
I G GROVE ◽  
K EVANS ◽  
...  

Author(s):  
Alex Reid ◽  
Fiona Evans ◽  
Vincent Mulholland ◽  
Yvonne Cole ◽  
Jon Pickup

Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 883-883
Author(s):  
I. Ostojić ◽  
D. Grubišić ◽  
M. Zovko ◽  
T. Miličević ◽  
T. Gotlin Čuljak

The golden potato cyst nematode, Globodera rostochiensis (Wollenweber, 1923) Behrens, 1975, is the most problematic pest of potato (Solanum tuberosum L. subsp. tuberosum) worldwide and it is recognized as an A1 plant health quarantine species. In Bosnia and Herzegovina, there have been no surveys for the presence of potato cyst nematodes (PCN) in ware potato lands, although they have been documented in the nearby countries of Serbia and Croatia (3). To determine the presence and distribution of PCN and to prevent further spread or increase of PCN populations, a survey in ware potato land was undertaken in 2008, 2009, and 2010. Soil samples consisting of 100 cores of 4 to 5 ml of soil were taken in a grid pattern throughout the plots. Samples of approximately 500 g were processed in the laboratory. The samples were air dried and cysts were extracted using a Spears's flotation method (4) from a 100-ml subsample. Species identification was based on morphometrical characters (2) and real-time PCR where the ribosomal internal transcribed spacer (ITS5, PITSr3) region was amplified and detected by the Sequence Dection Software v1.3 (Applied Biosystems, Carlsbad, CA) (1). In 2008, soil sampling was conducted in five locations and PCN were not detected in any of 10 soil samples taken. In June 2009, 17 soil samples were collected from seven locations. Two viable cysts of G. rostochiensis were detected in one soil sample from Tihaljina (Zaside; E 6447659.560, N 4797392.101, altitude 130 m). A total of 110 soil samples were collected from 90 locations in May and June 2010. Five viable cysts of G. rostochiensis were detected in one soil sample taken from Čapljina (Opličići – Turajlovina; E 6485169, N 4775964, altitude 222 m). On the basis of the survey results, G. rostochiensis is not widely spread but the cysts are detectable, indicating that the infestation is relatively recent and that there is a threat of further spread of this quarantine species. Consequently, producers need to grow resistant potato cultivars and implement additional control measures to keep the nematode populations below damaging levels. References: (1) J. Bacic et al. Russ. J. Nematol. 16:1, 61, 2008. (2) C. C. Fleming and T. O. Powers. Potato Cyst Nematodes: Biology, Distribution and Control. R. J. Marks and B. B. Brodie, eds.CAB International, Wallingford, 1998. (3) D. Grubišic et al. J. Pest Sci. 80:21, 2007. (4) J. F. Spears. Agr. Handb. No. 353. U.S. Dep. Agric., 1968.


Nematology ◽  
2007 ◽  
Vol 9 (3) ◽  
pp. 431-435 ◽  
Author(s):  
Thomas Been ◽  
Thea van Beers ◽  
Petra van Bekkum ◽  
Annelies Beniers

AbstractIn order to process large soil samples containing potato cyst nematodes, the Seinhorst (1964) cyst elutriator was scaled up to process both sandy and marine-clay soils in batches of up to 2.5 kg. Several modifications were implemented. To maintain the required upward current of 3.01 min−1, an inflow of 8.01 min−1 was necessary in the enlarged, 7.5 cm diam. sedimentation tube. Also water inflow is now regulated using a flow meter with pressuriser. Several experiments were undertaken, using artificial sandy soil and marine-clay soil, both naturally infested with potato cyst nematodes. In the final experiment, using the 8.01 min−1 inflow for 4 min, there was a loss of 0.65% and 0.74% of cysts, and eggs and juveniles, respectively. This indicates that the yield (99%) of the scaled-up elutriator matches that of the original Seinhorst elutriator and, therefore, is suitable for statutory and scientific research.


2020 ◽  
Vol 21 (1) ◽  
pp. 54-64
Author(s):  
Sleiman Kachacha ◽  
◽  
Said Ibrahim ◽  
Georges Kachacha ◽  
Loudmilla Ibrahim ◽  
...  

Our study was conducted in Bekaa area (Lebanon) to determine the presence and distribution of the potato cyst nematode, Globodera spp. A total of 80 soil samples were collected from 30 different areas. Theoverallegg counts revealed that 39 (79.6%) out of 49, cyst affected soil samples,collected contained eggs. In Zahlé, the count of nematodes in the soil samples revealed that 9 out of 11 samples tested (81.8%) contained cysts nematodes, but only 7 (77.7%) of the cyst contained eggs. The level of infestation of soil sample ranged between 0 and 5.3 eggs/g soil. No nematodes infection was detected in samples collected from Aana, Kamed El Laouz, Kefraya and Khiara. The identity of a newly discovered population of potato cyst nematodeGlobodera pallidaassociated with potato in West Bekaa was established by molecular methods.A set ofthreepolymerase chain reaction (PCR)primers readily identifiedthe presence of G. rostochiensis and G. pallida fromfield samples. The origin of this infestationis unknown. The results of the occurrence and distribution of Globodera spp in each area are discussed.


Nematology ◽  
2001 ◽  
Vol 3 (1) ◽  
pp. 75-83 ◽  
Author(s):  
Peter Jones ◽  
John Byrne ◽  
Ken Devine

AbstractThe fractionation of root leachates from tomato and potato on a Sephadex G-10 column revealed many similarities between the elution profiles of hatching activities towards potato cyst nematodes (PCN). When aliquots of either tomato or potato root leachate were mixed with different soil samples, hatching factors exhibiting G. pallida-selective hatching activity showed less affinity for the soil matrix than did those selective for G. rostochiensis . This was confirmed by the earlier elution of G. pallida-selective hatching factors from columns in which soil was used as the solid phase in low pressure liquid chromatography. The selectivity of the earliest-eluting hatching factors towards G. pallida, relative to G. rostochiensis, was found to increase significantly as the percentage soil organic matter content increased.


Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1125-1125 ◽  
Author(s):  
A. Przetakiewicz

Potato (Solanum tuberosum L. subsp. tuberosum) is one of the most important crops in Poland. During vegetation season, pests and pathogens including a quarantine-significant nematode Globodera rostochiensis (Woll.) Behrens can infect growing plants. Regional surveys have revealed potato cyst nematodes (PCN) to be widely distributed in Poland, with the highest infestation throughout the northwest of the country. Since 1946, only pathotype Ro1 of G. rostochiensis has been recorded. Previous data revealed the occurrence of pathotype Ro5 in three separate provinces. In recent surveys, 22 soil samples from infested fields in northern and western parts of Poland were collected. Each sample comprised 10 kg of soil collected in a grid pattern. Samples of approximately 2 kg were dried for extracting nematodes. The cysts were obtained with an automatic cyst extractor (Mekku Gmbh). A molecular test was performed as a first step to discriminate G. rostochiensis from G. pallida. DNA from 30 cysts per sample was extracted with DNeasy Blood and Tissue KIT (Qiagen). Multiplex PCR analysis with two pairs of primers, GroR-GroF and PaR-PaF (1), was employed. The PCR resulted in a single 315-bp DNA fragment specific for G. rostochiensis. No amplification was shown with primers specific for G. pallida. Bioassays on differential genotypes of potato were carried out to identify the pathotype of G. rostochiensis. Five genotypes of potato: S. vernei hybr. 58.1642/4, S. vernei hybr. 62.33.3, S. vernei hybr. 65.346/19, S. multidissectum hybr. P 55/7, S. kurtzianum hybr. 60.21.19, and two varieties, Desiree and Maris Piper, were inoculated with 22 populations of G. rostochiensis cysts (2). Tests were performed in three replications. Eighteen of the cyst populations were pathotype Ro1 and four were pathotype Ro5. The abundance of cysts in locations where the new pathotype was detected suggests that there is a threat of further spread of G. rostochiensis. Consequently, producers and farmers need to grow resistant potato cultivars to keep nematode population below damaging level. To our knowledge, this is the first report of Globodera rostochiensis Ro5 in Poland. References: (1) A. Fullaondo et al. Nematology 1:157, 1999. (2) J. Kort et. al. Nematologica 23:333, 1977.


Sign in / Sign up

Export Citation Format

Share Document