scholarly journals Development of a Species-Specific SCAR-PCR Assay for Direct Detection of Sugar Beet Cyst Nematode (Heterodera schachtii) from Infected Roots and Soil Samples

Life ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1358
Author(s):  
Chen Jiang ◽  
Yingdong Zhang ◽  
Ke Yao ◽  
Sulaiman Abdulsalam ◽  
Guangkuo Li ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Cyst Nematode ◽  
Soil Samples ◽  
Heterodera Schachtii ◽  
Cyst Nematodes ◽  
Pcr Assay ◽  
Beet Cyst Nematode ◽  
Field Samples ◽  
Species Specific ◽  
Sugar Beet Cyst Nematode

Sugar beet cyst nematode (SBCN, Heterodera schachtii) is an important nematode that causes significant yield losses of 25–50% or more in most areas of sugar beet production worldwide. Rapid and accurate identification of this species is essential to support decisions on pest management. However, the difference between H. schachtii and other Heterodera spp. based on morphology is a challenging task. In the present study, a SCAR-PCR assay was developed to identify and differentiate H. schachtii in infected root and soil samples. H. schachtii-species-specific SCAR-PCR primers OPA06-HsF and OPA06-HsR were designed from the randomly amplified polymorphic DNA (RAPD) marker amplified with random primer OPA06. The developed primers specifically amplify a 922-bp fragment from the target populations but did not amplify DNA from non-target cyst nematodes including Heterodera, Globodera, Cactodera, and other related species tested in this study. The sensitivity detection indicated that 5 × 10−4 of a single cyst, 1/320 of a single second-stage juvenile (J2), or 10 pg of genomic DNA could be detected. The assay accurately identifies the different stages of H. schachtii in sugar beet and oilseed rape roots as well as a single J2 in 10 g of soil. Finally, the SCAR-PCR assay detected H. schachtii in seven samples out of the fifteen field samples. The assay will not only be useful for differentiating H. schachtii from mixed populations of Heterodera spp. but also for effective detection of the species directly from infested samples. The assay also requires no expertise in the taxonomy and morphology of the species but serves to improve the diagnosis of H. schachtii in infested fields.

scholarly journals First Confirmed Report of Sugar Beet Cyst Nematode, Heterodera schachtii, in North Dakota

Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 772-772 ◽  
Author(s):  
B. D. Nelson ◽  
M. D. Bolton ◽  
H. D. Lopez-Nicora ◽  
T. L. Niblack ◽  
L. del Rio Mendoza
Keyword(s):  
Sugar Beet ◽  
North Dakota ◽  
Body Length ◽  
Cyst Nematode ◽  
Soil Samples ◽  
The State ◽  
28S Rdna ◽  
Heterodera Schachtii ◽  
Beet Cyst Nematode ◽  
Sugar Beet Cyst Nematode

Sugar beet (Beta vulgaris L.) and canola (Brassica napus L.) are major crops in North Dakota, with sugar beet production primarily in the eastern part of the state in the Red River Valley and canola production across the northern half of the state. Both crops are hosts of sugar beet cyst nematode (SBCN), Heterodera schachtii Schmidt. In April 2011, soil samples were collected from four sugar beet fields belonging to three growers who believed the fields were infested with SBCN. The fields were located in a 65-km2 area in the Yellowstone Valley of western North Dakota. Cysts were extracted by sieving and Heterodera-like cysts with eggs were observed in all four soil samples. Population densities in the four fields ranged from 100 to 1,750 eggs/100 cm3 soil. Sugar beet seedlings (cv. M832224) were grown in a potting mix for 6 weeks in the greenhouse and then transferred to conetainers (type D40; volume 656 ml) containing autoclaved river sand. Conetainers were placed in sand in plastic pots immersed in a water bath at 27°C. Three plants were each infested with 800 eggs from field No. 2. After 55 days of incubation, the average number of females was 115 per plant. A similar experiment was conducted with canola cvs. Hyclass 940, Caliber 30, and Westar, which were inoculated with 500 eggs each from field No. 2. After 53 days of incubation, there was an average of 39, 20, and 30 females for each respective cultivar. Flask-shaped cysts (n = 26) from canola roots were light to dark brown; the vulval cone was ambifinestrate with dark brown, molar-shaped bullae positioned underneath the vulval bridge. Body length (excluding neck) ranged from 600 to 850 μm (mean 701.2 μm); body width, 350 to 580 μm (mean 469.2 μm); and length/width ratio, 1.2 to 1.8 (mean 1.5). Second-stage juvenile (J2) (n = 21) body length ranged from 400 to 485 μm (mean 437.1 μm); stylet length was 25 μm (no variation) with forwardly directed knobs; conical tail with rounded tip ranged from 37.5 to 55.0 μm long (mean 46.6 μm) with hyaline region from 20.0 to 32.5 μm (mean 27.3 μm); and lateral field presented four incisures. These morphometrics were used to identify H. schachtii according to Subbotin et al. (4). Confirmation of identification was by amplification and sequencing of a 28S rDNA gene fragment (1) from individual females (GenBank Accession No. JQ040526), which was 100% identical to H. schachtii 28S rDNA sequence (GenBank Accession No. GU475088). To our knowledge, this is the first confirmed report of H. schachtii in North Dakota. A 1958 report of SBCN in North Dakota (2) was not subsequently confirmed (3). Because there is extensive canola production across the northern part of the state bordering western and eastern sugar beet- production areas, canola may serve as a bridge for movement of SBCN from west to east. SBCN is a potential threat to these two important crops. References: (1) A. Amiri et al. Eur. J. Plant Pathol. 108:497, 2002. (2) F. Caveness. J. Sugar Beet Res. 10:544, 1958. (3) P. Donald and R. Hosford. Plant Dis. 64:45, 1980. (4) S. A. Subbotin et al. Systematics of Cyst Nematodes (Nematoda: Heteroderinae). Nematology Monographs and Perspectives. Vol. 8B. Brill, The Netherlands. 2010.

Sugar beet nematode Heterodera schachtii distribution and harmfulness in Ukraine

2016 ◽  
Vol 3 (3) ◽  
pp. 3-11 ◽  
Author(s):  
L. Pylypenko ◽  
K. Kalatur ◽  
J. Hallmann
Keyword(s):  
Sugar Beet ◽  
Wide Distribution ◽  
Cyst Nematode ◽  
Control Measures ◽  
Heterodera Schachtii ◽  
Prevention Measures ◽  
Economic Threshold ◽  
Beet Cyst Nematode ◽  
Effi Ciency ◽  
Sugar Beet Cyst Nematode

Aim. To determine the distribution of sugar beet cyst nematode Heterodera schachtii in Ukraine, to defi ne its population density in soil, the degree of damage and the effi ciency of control measures. Methods. Field survey for soil samples, cyst extraction by fl otation and sieving method, light microscopy. Results. The examination of 12,130 ha of agricultural fi elds in six regions of Ukraine, conducted in 2010–2015, recorded the sugar beet cyst nematode to occur in fi ve of them (Kyiv, Chernihiv, Cherkasy, Khmelnytsky, and Vinnytsia), at a total area of 2,572 ha. Taking into consideration the results of previous nematological surveys, the presence of sugar beet cyst nematode has now been documented for 18 regions of Ukraine. In most regions H. schachtii was present in 10–11 % of the examined sugar and seed producing farms with nematode population densities reaching up to 142,000 eggs and juveniles in 100 cc of soil (710-fold above the economic threshold). Conclusions. The main reasons for this wide distribution of H. schachtii in Ukraine are seen in the negligence of prevention measures, unavailability of documented data on its occurrence (missing surveys), crop rotations with over 20 % of host plants, and unavailability of effi cient nematicides and domestic nematode-resistant sugar beet cultivars.

First Report of Sugar Beet Cyst Nematode, Heterodera schachtii on Beetroot and Broccoli in Mexico

Plant Disease ◽  
2019 ◽  
Vol 103 (6) ◽  
pp. 1434-1434
Author(s):  
I. M. Escobar-Avila ◽  
Y. Cruz-Alvarado ◽  
A. Tovar-Soto ◽  
S. A. Subbotin
Keyword(s):  
Sugar Beet ◽  
Cyst Nematode ◽  
Heterodera Schachtii ◽  
First Report ◽  
Beet Cyst Nematode ◽  
Sugar Beet Cyst Nematode

Partial isolation of a water soluble pheromone from the sugar beet cyst nematode, Heterodera schachtii, using a novel bioassay

Nematology ◽  
2001 ◽  
Vol 3 (1) ◽  
pp. 55-64 ◽  
Author(s):  
Ekaterini Riga ◽  
John Potter ◽  
Michael Jonz ◽  
A. Joffre Mercier
Keyword(s):  
Sugar Beet ◽  
High Performance ◽  
Cyst Nematode ◽  
Water Soluble ◽  
Heterodera Schachtii ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Beet Cyst Nematode ◽  
Partial Isolation ◽  
Sugar Beet Cyst Nematode

AbstractThe sex pheromone of female Heterodera schachtii, the sugar beet cyst nematode, was partially isolated using high performance liquid chromatography (HPLC). Female-conditioned medium (FCM), produced by incubating 20 H. schachtii females in 1 ml of ultrapure water, attracted males in a concentration dependent manner. The attractive material was not contained within the female gelatinous matrix and did not attract second stage juveniles (J2). Characterisation of FCM revealed that the attractive material was water soluble and heat stable. Male H. schachtii exhibited stylet thrusting behaviour upon exposure to FCM, and this behaviour was used as a novel indicator of biological activity.

scholarly journals Population Dynamics of Sugar-Beet Cyst Nematode (Heterodera Schachtii) On Spring and Winter Oilseed Rape Crops

2008 ◽  
Vol 48 (2) ◽  
pp. 237-245 ◽  
Author(s):  
Renata Dobosz ◽  
Stefan Kornobis
Keyword(s):  
Population Dynamics ◽  
Sugar Beet ◽  
Oilseed Rape ◽  
Second Generation ◽  
Cyst Nematode ◽  
Heterodera Schachtii ◽  
Winter Oilseed Rape ◽  
Beet Cyst Nematode ◽  
Adult Females ◽  
Sugar Beet Cyst Nematode

Population Dynamics of Sugar-Beet Cyst Nematode (Heterodera Schachtii) On Spring and Winter Oilseed Rape Crops The influence of spring as well as winter oilseed rape on the change in population density of H. schachtii was investigated in microplot experiment in the years 2002-2005. The spring oilseed rape caused the increase of nematode population (Pf/Pi were 1.31-1.79), in contrast to winter oilseed rape where the population distinctly decreased (Pf/Pi were 0.49-0.59). No statistically important differences between winter oilseed rape crops and fallow were observed. Comparative observations of the life cycle were carried out in pot experiments outdoors. On spring oilseed rape only one generation was completed. The development of the second generation was interrupted because of crop harvesting. In winter oilseed rape J3 and J4 occurred in roots in the autumn where they probably did not over-winter, except of the years 2006-2007. In the spring there was a second emergence of larvae and adult females were observed in June and July. The development of the second generation was interrupted because of harvesting. In winter oilseed rape sown in the spring one generation was completed.

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