Cereal Cyst Nematode Heterodera Research Articles

Occurrence, identification and phylogenetic analyses of cereal cyst nematodes (Heterodera spp.) in Turkey

AbstractPlant-parasitic nematodes are very common on cereal crops and cause economic losses via reduction in grain quality and quantity. During 2014, 83 soil samples were collected from wheat and barley fields in 21 districts of 13 provinces across five regions (Central Anatolia, Marmara, Aegean, Southeast Anatolia, and Black Sea Region) of Turkey. Cyst-forming nematodes were found in 66 samples (80%), and the internal transcribed spacer (ITS) sequencing and species-specific PCR identified the species in 64 samples as Heterodera filipjevi, Heterodera latipons, and Heterodera avenae. The predominant pathogenic cereal cyst nematode was H. filipjevi, which was found in all five regions surveyed. H. avenae was only detected in Southeast Anatolia whereas H. latipons was detected in Southeast Anatolia and Central Anatolia. ITS-rDNA phylogenetic analyses showed that H. avenae isolates from China clustered with H. australis, and Turkish isolates were closely related to European and USA isolates of this species. H. filipjevi from Turkey and China were clustered closely with those from the UK, Germany, Russia, and the USA. The density of many of these populations exceeded or approached the maximum threshold level for economic loss. To our knowledge, this is the first report of H. filipjevi in Diyarbakir, Edirne, and Kutahya provinces, and the first report of H. avenae in Diyarbakir Province. These results exhibit the most rigorous analysis to date on the occurrence and distribution of Heterodera spp. in Turkey's major wheat-producing areas, thus providing a basis for more specific resistance breeding, as well as other management practices.

Ijuhya vitellina sp. nov., a novel source for chaetoglobosin A, is a destructive parasite of the cereal cyst nematode Heterodera filipjevi.

Cyst nematodes are globally important pathogens in agriculture. Their sedentary lifestyle and long-term association with the roots of host plants render cyst nematodes especially good targets for attack by parasitic fungi. In this context fungi were specifically isolated from nematode eggs of the cereal cyst nematode Heterodera filipjevi. Here, Ijuhya vitellina (Ascomycota, Hypocreales, Bionectriaceae), encountered in wheat fields in Turkey, is newly described on the basis of phylogenetic analyses, morphological characters and life-style related inferences. The species destructively parasitises eggs inside cysts of H. filipjevi. The parasitism was reproduced in in vitro studies. Infected eggs were found to harbour microsclerotia produced by I. vitellina that resemble long-term survival structures also known from other ascomycetes. Microsclerotia were also formed by this species in pure cultures obtained from both, solitarily isolated infected eggs obtained from fields and artificially infected eggs. Hyphae penetrating the eggshell colonised the interior of eggs and became transformed into multicellular, chlamydospore-like structures that developed into microsclerotia. When isolated on artificial media, microsclerotia germinated to produce multiple emerging hyphae. The specific nature of morphological structures produced by I. vitellina inside nematode eggs is interpreted as a unique mode of interaction allowing long-term survival of the fungus inside nematode cysts that are known to survive periods of drought or other harsh environmental conditions. Generic classification of the new species is based on molecular phylogenetic inferences using five different gene regions. I. vitellina is the only species of the genus known to parasitise nematodes and produce microsclerotia. Metabolomic analyses revealed that within the Ijuhya species studied here, only I. vitellina produces chaetoglobosin A and its derivate 19-O-acetylchaetoglobosin A. Nematicidal and nematode-inhibiting activities of these compounds have been demonstrated suggesting that the production of these compounds may represent an adaptation to nematode parasitism.

Development and characterization of EST-derived SSR markers in the cereal cyst nematode Heterodera avenae

The cereal cyst nematode Heterodera avenae is a pathogen of cereal crops and causes high yield losses worldwide. In this study, a collection of 37,348 expressed sequence tags (ESTs) of H. avenae was mined for EST-based simple sequence repeat (SSR) markers, which resulted in the identification of 5604 SSRs. A total of 210 pairs of SSR primers were further developed and used for validation of the amplification rate and assessment of the polymorphism. Eight SSR markers were finally identified and analyzed using 96 individual cysts of H. avenae sampled from three provinces in China. These loci were found to be moderately polymorphic with 3–8 alleles per locus. The observed and expected heterozygosity across the three populations ranged from 0.000 to 0.594 and from 0.000 to 0.731, respectively. The polymorphism information content (PIC) was medium and ranged from 0.080 to 0.562, with a mean of 0.409. The F ST ranged from 0.1034 to 0.1550, indicating moderate genetic differentiation among the three H. avenae populations. These EST-SSR markers can be used to study population genetic diversity and the dispersal route of H. avenae in China.

Cereal cyst nematodes: importance, distribution, identification, quantification, and control

Small grain cereals, such as wheat, barley and oats are considered among the most important food sources. Plant-parasitic nematodes play a considerable role in decreasing cereal yields. The three-major species of cereal cyst nematodes (CCN) Heterodera avenae, H. latipons, and H. filipjevi are distributed worldwide and cause considerable damage. This review provides information regarding the global distribution of these nematode species, yield loss due to CCN, their biology and pathogenic relation to plants, identification and control through agricultural practices, biological agents and resistance breeding. As morphological identification of CCN is difficult and time-consuming, several molecular techniques for the identification of these CCN species have been developed in recent years. The restrictions on the use of nematicides demand for resistance to CCN. Resistance genes in several lines are known and are used in numerous breeding programmes against CCN; pyramiding these resistance genes into high yielding cultivars that could become commercially available for farmers is progressing.

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English

Morphological and molecular characterizations of cereal cyst nematode Heterodera avenae Wollenweber, 1924 from the Czech Republic

The cereal cyst nematode, Heterodera avenae Wollenweber, 1924, is a major pest of cereal crops throughout the world and causes serious yield losses, especially of wheat. Previous studies have shown that this species is widely distributed in the Czech Republic. In this study, seven populations of H. avenae were molecularly studied, and one population was morphologically described. Three regions (18S, 28S, and internal transcribed spacer 1) of ribosomal DNA were sequenced and the analysis of the 18S gene of six populations did not reveal any variation, whereas the internal transcribed spacer 1 and 28S sequences of six populations differed by only two nucleotides from a population in Žilina. Precise and quick identification of cereal cyst nematodes is important for effective control measures and ribosomal sequence analyses of seven populations in this study will be useful in future phylogenetic studies of Heterodera spp. occurring in the Czech Republic.

Efficacy Evaluation of Seed-Coating Compounds Against Cereal Cyst Nematodes and Root Lesion Nematodes on Wheat

Cereal cyst nematodes (Heterodera avenae and H. filipjevi) and root lesion nematodes (Pratylenchus spp.) have been found to infect cereals in 16 provinces of China. To develop a nematicide that effectively controls nematodes, two novel chemical products, methylene bis thiocyanate (MBT) and MBT + thiamethoxam (MTT); four common pesticides, fipronil + chlorpyrifos (FIC), emamectin benzoate, imidacloprid, and Bacillus thuringiensis; and one fungicide, iprodione, were tested as seed coatings for the control of cereal cysts and root lesion nematodes from 2013 to 2015. Wheat seeds were treated with these seven seed coatings before sowing, and changes in the numbers of Heterodera spp. and Pratylenchus spp. were recorded during three different growth stages. Wheat yields were also compared after harvest. All treatments reduced the numbers of Pratylenchus in wheat and of cysts and eggs of Heterodera in the soil compared with the untreated control. Among the treatments, application of MTT or FIC was more effective than that of the other treatments for nematode control, and the other treatments had similar effects. The results of this study have demonstrated that MTT and FIC applied as seed treatments effectively reduce the number of cysts, inhibit the reproduction of Heterodera and Pratylenchus, and enhance wheat yields. MTT and FIC are thus suitable for controlling nematodes on wheat under natural field conditions.

Biochemical changes of the cereal cyst nematode, Heterodera avenae, at low temperatures

Cereal cyst nematode (Heterodera avenae) diapause is induced by high temperatures and is broken by low temperatures. In this study, metabolic responses were monitored in diapause and non-diapause H. avenae during exposure to 4°C for 10 weeks. The results showed that there was no difference in total carbohydrate content. The content of glycogen and glycerol at 0 week was relatively high but decreased with increased storage time at 4°C. The content of trehalose of the nematode at 10 weeks was significant lower than that at 5 weeks at 4°C. Protein content increased significantly after incubation for 5 and 10 weeks. Esterase and trehalase activity increased with the increasing period at 4°C and showed a significant difference between treatments for esterase activity but there was no significant difference between 5 and 10 weeks for trehalase activity. The SDS-PAGE pattern indicated that a 15.5 kDa protein was absent at 10 weeks and present at 0 and 5 weeks. Esterase isoenzyme patterns of H. avenae showed that at 10 weeks there were four bands: EST 0.21, EST 0.24, EST 0.30 and EST 0.34 (Rf values). EST 0.24 was the common band in the three treatments. Biochemical tests were conducted to correlate with hatching experiments using the same treatments. 2-DE patterns of H. avenae showed that diapause and non-diapause nematodes had 409 and 412 protein spots, respectively, and 19 protein spots were unique: 11 distinct proteins in non-diapause and eight distinct proteins in diapause. This information could be helpful in understanding the diapause mechanism of the cereal cyst nematode.

GC-MS Analysis of Nematicidal Essential Oil of Mentha canadensis Aerial Parts against Heterodera avenae and Meloidogyne incognita

The essential oil of Mentha canadensis aerial parts was obtained by hydrodistillation and analyzed by gas chromatography (GC) and gas chromaotography-mas spectrometry (GC-MS). A total of 36 components of the essential oil were identified. The major constituents in the essential oil of M. canadensis were menthol (28.8 %), α-pinene (16.4 %), menthone (12.7 %), α-terpineol (6.3 %) and limonene (5.5 %). Four active constituents (limonene, menthol, α-pinene and α-terpinol) were isolated from the essential oil of M. canadensis based on bioactivity-directed fractionation. The essential oil of M. canadensis aerial parts exhibited nematicidal activity against cereal cyst nematodes (Heterodera avenae) and root-knot nematodes (Meloidogyne incognita) with LC values of 385.7 μg/ml and 139.0 μg/ml, respectively. The isolated constituents, menthol and α-terpinol possessed nematicidal activity against H. avenae and M. incognita with LC50 values of 242.5 μg/ml, 190.3 μg/ml and 147.4 μg/ml, 115.2 μg/ml, respectively. The study indicated that the essential oil of M. canadensis aerial parts and its two major constituents, α-terpineol and menthol have a potential to be developed to a natural nematicides for the control of nematodes.

Influence of temperature and storage conditions on the hatching behavior of cereal cyst nematodes (Heterodera avenae Wollenweber) from Egypt

The cereal cyst nematode Heterodera avenae has been reported in wheat fields in Egypt; however no information is available on the hatch of second-stage juveniles (J2) from such populations of H. avenae. The aim of this study was to subject five Egyptian populations of H. avenae that were obtained from different regions in the Ismailia Province to different temperatures and storage durations to determine the effects on J2 emergence. The Egyptian populations of H. avenae were compared to a population originating from a different climatic zone in Germany (Grafenreuth, Bavaria). Results showed that the J2 hatching pattern of the Egyptian H. avenae populations did not differ from each other. Although the Egyptian and the German populations of H. avenae hatched primarily between 5 and 20 °C, significant differences in J2 hatching have been observed among them. Highest emergence of J2 of the Egyptian populations was observed between 10 and 15 °C, whereas the highest level of J2 emergence of the German population was between 5 and 10 °C. Storing cysts to 5 °C before incubation at 10 °C stimulated the hatch of the German population of H. avenae significantly compared to the control. Whereas, storing cysts at 20 °C, before incubation at 10 °C, significantly stimulated J2 hatching of the Egyptian populations compared to the control. The J2 hatching behavior of the Egyptian populations of H. avenae was in agreement with reports for Mediterranean ecotypes which have winter activity, while the hatching pattern of the German population was similar to those of the Northern ecotypes with spring activity.

Isolation and characterization of a fatty acid- and retinoid-binding protein from the cereal cyst nematode Heterodera avenae

A gene encoding fatty acid- and retinoid-binding protein was isolated from the cereal cyst nematode Heterodera avenae and the biochemical function of the protein that it encodes was analysed. The full-length cDNA of the Ha-far-1 gene is 827 bp long and includes a 22- nucleotide trans-spliced leader sequence (SL1) at its 5-end. The genomic clone of Ha-far-1 consists of eight exons separated by seven introns, which range in size from 48 to 186 bp. The Ha-far-1 cDNA contains an open reading frame encoding a 191 amino acid protein, with a predicted secretory signal peptide. Sequence analysis showed that Ha-FAR-1 has highest similarity to the Gp-FAR-1 protein from the potato cyst nematode, Globodera pallida and that the protein was grouped with all homologues from other plant-parasitic nematodes in a phylogenetic analysis. Fluorescence-based ligand binding analysis confirmed that the recombinant Ha-FAR-1 protein was able to bind fatty acids and retinol. Spatial and temporal expression assays showed that the transcripts of Ha-far-1 accumulated mainly in the hypodermis and that the gene is most highly expressed in third-stage juveniles of H. avenae. Fluorescence immunolocalization showed that the Ha-FAR-1 protein was present on the surface of the infective second-stage juveniles of H. avenae. Nematodes treated with dsRNA corresponding to Ha-far-1 showed significantly reduced reproduction compared to nematodes exposed to dsRNA from a non-endogenous gene, suggesting that Ha-far-1 may be an effective target gene for control of H. avenae using an RNAi strategy.

Detection of Dual Heterodera avenae Resistance plus Tolerance Traits in Spring Wheat.

The cereal cyst nematode Heterodera avenae reduces wheat yield in the Pacific Northwest. Resistance and tolerance traits among spring wheat cultivars were poorly defined. Screening trials were conducted with 39 cultivars over a 2-year period in irrigated commercial fields that were infested by H. avenae. Comparisons were made between drill strips treated or untreated with aldicarb at the time of planting. Root sampling at the time of plant anthesis indicated that cultivars differed greatly in susceptibility to H. avenae, with numbers of newly produced white H. avenae females ranging from <5 to 70 per plant. Aldicarb reduced mean numbers of white females as much as 99% on the most susceptible cultivar ('Glee') and increased mean grain yield as much as 77% for the least tolerant cultivar ('Cataldo'). Density of H. avenae eggs in untreated soil following harvest was significantly higher than the density in aldicarb-treated plots. Agronomically acceptable traits of resistance plus tolerance were identified in one cultivar of hard red spring wheat ('WB-Rockland') and two cultivars of hard white spring wheat ('Klasic' and 'LCS Star') but in none of the soft white spring wheat cultivars. This is the first report of spring wheat cultivars expressing the dual traits of resistance plus tolerance to H. avenae.

Genetic characterization of resistance to stripe rust, leaf rust, Karnal bunt and cereal cyst nematode in a multiple disease resistant wheat stock W8627

Genetic analysis of multiple disease resistance was carried out in segregating populations of bread wheat line W8627 and PBW343 against stripe rust (Puccinia striiformis), leaf rust (Puccinia triticina), Karnal bunt (Tilletia indica) isolates and cereal cyst nematode (Heterodera avenae). Seedling response of W 8627, PBW 343 and F1 against 78S84 race of Puccinia striiformis, 77-5 race of P. triticina reflected that the wheat line W8627 possessed seedling resistance genes against both the races. Based on the segregation pattern of F2 generation and F3 families, two complementary recessive genes for resistance to 78S84 race of Puccinia striiformis and one recessive gene each for resistance to 77-5 race P. triticina, mixture of Tilletia indica isolates and Ha 41 biotype of H. avenae were identified. Co-segregation studies revealed no linkage between concerned resistance genes.

Identification and characterisation of resistance to the cereal cyst nematode Heterodera filipjevi in winter wheat

The aim of this study was to search for new sources of resistance against the cereal cyst nematode,Heterodera filipjevi, in a collection of 290 wheat accessions. The plants were inoculated with juveniles and assessed for the number of females and cysts. One percent of the wheat accessions were ranked as resistant, 16% as moderately resistant, 41% as moderately susceptible, 26% as susceptible and 15% as highly susceptible. The infection rate and the number of females and cysts per plant were significantly lower in the resistant accession Nudakota and three moderately resistant accessions Ekonomka, Katea and Lantian 12 compared with susceptible cv. Bezostaya 1. Nematode development was reduced in resistant and moderately resistant accessions. The size of females and the total number of eggs and second-stage juveniles were reduced only in Ekonomka. No significant difference in plant height, plant weight, root length, root weight and root volume were recorded for inoculated plants compared to non-inoculated plants. This study has identified four resistant wheat accessions offering new material for breeding the resistance toH. filipjevi.

Response of wheat to a Jordanian isolate of Mediterranean cereal cyst nematode (Heterodera latipons)

A total of 52 wheat genotypes comprising landraces of hard (durum), common bread wheat cultivars, and synthetic hexaploid wheat were evaluated for their reaction against a Jordanian isolate of the Mediterranean Cereal Cyst Nematode Heterodera latipons (MCCN). Three genotypes obtained from International Center for Agricultural Research in the Dry Areas (ICARDA), the Australian bread wheat cultivars, Drysdale, Gladius, GS50A and Silverstar, and the synthetic hexaploids, Langdon*Aus18913, Langdon*Aus18964, Langdon*Aus181-1472, Langdon*CPI-110649 and Langdon*CPI-110756 showed resistance to MCCN. Further, 19 out of the 23 Australian synthetic hexaploid genotypes were also resistant. The characterisation of Cre genes showed that the synthetic haploids, Langdon*Aus18913, Langdon*Aus18964, Aus 34262, Aus 34423 and Aus 34448 genotypes, carrying Cre3 resistant gene, expressed high levels of resistance to MCCN while the genotypes CPI133859, CPI133872, Aus 10894 and Silverstar, carrying Cre1 resistant gene, varied in their levels of resistance to MCCN and ranged from moderately susceptible to resistant. The genotype, Frame carrying Cre8 was moderately resistant. We were not able to identify those wheat genotypes carrying Cre5 or Cre8 because of the lack of reproducibility of the PCR and the poor linkage of the markers associated with resistance to the two genes. On the other hand, some commercial cultivars and improved genotypes were resistant to MCCN and do not have Cre3 or Cre1. They may possess other uncharacterised Cre genes against MCCN and thus represent potential new sources of resistance genes that could be used for wheat improvement against H. latipons.

RNA-Seq Based Identification of Candidate Parasitism Genes of Cereal Cyst Nematode (Heterodera avenae) during Incompatible Infection to Aegilops variabilis.

One of the reasons for the progressive yield decline observed in cereals production is the rapid build-up of populations of the cereal cyst nematode (CCN, Heterodera avenae). These nematodes secrete so-call effectors into their host plant to suppress the plant defense responses, alter plant signaling pathways and then induce the formation of syncytium after infection. However, little is known about its molecular mechanism and parasitism during incompatible infection. To gain insight into its repertoire of parasitism genes, we investigated the transcriptome of the early parasitic second-stage (30 hours, 3 days and 9 days post infection) juveniles of the CCN as well as the CCN infected tissue of the host Aegilops variabilis by Illumina sequencing. Among all assembled unigenes, 681 putative genes of parasitic nematode were found, in which 56 putative effectors were identified, including novel pioneer genes and genes corresponding to previously reported effectors. All the 681 CCN unigenes were mapped to 229 GO terms and 200 KEGG pathways, including growth, development and several stimulus-related signaling pathways. Sixteen clusters were involved in the CCN unigene expression atlas at the early stages during infection process, and three of which were significantly gene-enriched. Besides, the protein-protein interaction network analysis revealed 35 node unigenes which may play an important role in the plant-CCN interaction. Moreover, in a comparison of differentially expressed genes between the pre-parasitic juveniles and the early parasitic juveniles, we found that hydrolase activity was up-regulated in pre J2s whereas binding activity was upregulated in infective J2s. RT-qPCR analysis on some selected genes showed detectable expression, indicating possible secretion of the proteins and putative role in infection. This study provided better insights into the incompatible interaction between H. avenae and the host plant Ae. varabilis. Moreover, RNAi targets with potential lethality were screened out and primarily validated, which provide candidates for engineering-based control of cereal cyst nematode in crops breeding.

Large-scale identification of wheat genes resistant to cereal cyst nematode Heterodera avenae using comparative transcriptomic analysis.

BackgroundCereal cyst nematode Heterodera avenae, an important soil-borne pathogen in wheat, causes numerous annual yield losses worldwide, and use of resistant cultivars is the best strategy for control. However, target genes are not readily available for breeding resistant cultivars. Therefore, comparative transcriptomic analyses were performed to identify more applicable resistance genes for cultivar breeding.MethodsThe developing nematodes within roots were stained with acid fuchsin solution. Transcriptome assemblies and redundancy filteration were obtained by Trinity, TGI Clustering Tool and BLASTN, respectively. Gene Ontology annotation was yielded by Blast2GO program, and metabolic pathways of transcripts were analyzed by Path_finder. The ROS levels were determined by luminol-chemiluminescence assay. The transcriptional gene expression profiles were obtained by quantitative RT-PCR.ResultsThe RNA-sequencing was performed using an incompatible wheat cultivar VP1620 and a compatible control cultivar WEN19 infected with H. avenae at 24 h, 3 d and 8 d. Infection assays showed that VP1620 failed to block penetration of H. avenae but disturbed the transition of developmental stages, leading to a significant reduction in cyst formation. Two types of expression profiles were established to predict candidate resistance genes after developing a novel strategy to generate clean RNA-seq data by removing the transcripts of H. avenae within the raw data before assembly. Using the uncoordinated expression profiles with transcript abundance as a standard, 424 candidate resistance genes were identified, including 302 overlapping genes and 122 VP1620-specific genes. Genes with similar expression patterns were further classified according to the scales of changed transcript abundances, and 182 genes were rescued as supplementary candidate resistance genes. Functional characterizations revealed that diverse defense-related pathways were responsible for wheat resistance against H. avenae. Moreover, phospholipase was involved in many defense-related pathways and localized in the connection position. Furthermore, strong bursts of reactive oxygen species (ROS) within VP1620 roots infected with H. avenae were induced at 24 h and 3 d, and eight ROS-producing genes were significantly upregulated, including three class III peroxidase and five lipoxygenase genes.ConclusionsLarge-scale identification of wheat resistance genes were processed by comparative transcriptomic analysis. Functional characterization showed that phospholipases associated with ROS production played vital roles in early defense responses to H. avenae via involvement in diverse defense-related pathways as a hub switch. This study is the first to investigate the early defense responses of wheat against H. avenae, not only provides applicable candidate resistance genes for breeding novel wheat cultivars, but also enables a better understanding of the defense mechanisms of wheat against H. avenae.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-2037-8) contains supplementary material, which is available to authorized users.

Gas Chromatography-Mass Spectrometric Analysis of Nematicidal Essential Oil ofValeriana amurensis P Smirn ex Kom (Valerianaceae) Roots and its Activity against Heterodera avenae

Purpose: To investigate the chemical composition and nematicidal activity of the essential oil of Valeriana amurensis roots against cereal cyst nematodes (Heterodera avenae).Methods: The essential oil of V. amurensis roots was obtained by hydrodistillation and analyzed by gas chromatography (GC) and gas chromaotography-mass spectrometry (GC-MS). The nematicidal activity of the essential oil and its major constituents was determined against second stage juveniles of H. avenae.Results: A total of 33 components of the essential oil were identified. The major constituents were bornyl acetate (12.5 %), patchoulol (11.6 %), caryophyllene (8.2 %), 3-methylvaleric acid (7.3 %) and isovaleric acid (6.5 %). The essential oil exhibited nematicidal activity against H. avenae with a medium lethal concentration (LC50) value of 311.6 μg/mL. The major constituents, isovaleric acid and 3- methylvaleric acid, exhibited nematicidal activity against H. avenae with LC50 of 218.2 and 683.8 μg/mL, respectively.Conclusion: The study indicates that the essential oil of V. amurensis roots and its two major constituents, isovaleric acid and 3-methylvaleric acid, have a potential to be developed to natural nematicides for the control of cereal cyst nematodes.Keywords: Valeriana amurensis, Heterodera avenae, Nematicidal activity, Isovaleric acid, 3- Methylvaleric acid, Essential oil, Cereal cyst nematodes

Distribution of the Cereal Cyst Nematodes (Heterodera spp.) in Wheat and Barley Fields in North-Eastern Regions of Syria

Cereal cyst nematodes (CCN) are important plant-parasitic nematodes of wheat and barley, and exist in most of the cereal growing regions of the world. As there is limited information on the nature and distribution of CCN species in Syria, a survey was conducted in north-eastern Syria to assess the distribution of CCN in the main wheat and barley growing areas. In the summer of 2009, a total of 167 composite soil samples were collected from 167 wheat and barley fields. Cysts were extracted from soil using the Fenwick can technique, then quantified and identified up to the species level by using both morphological and molecular methods. The study revealed that 62% of the fields were infested with the three Heterodera species: H. avenae, H. filipjevi and H. latipons. However, the most prevalent species was H. latipons, which was present in 76% of the infested samples, while it occurred alone in 67% of those samples. Heterodera avenae was detected singly in 20% of the samples, while 11% consisted of mixed populations with other species. Heterodera filipjevi was never detected alone; it was found mixed with H. avenae and/or H. latipons in 9% of the samples. The high number of extracted cysts (up to 116 cysts × 200 g−1 of soil) is most likely related to the monoculture practiced in that region and the fact that growers are unaware of the existence of resistant varieties.

Biological control of the cereal cyst nematode (Heterodera filipjevi) by Achromobacter xylosoxidans isolate 09X01 and Bacillus cereus isolate 09B18

Two bacterial strains, Achromobacter xylosoxidans (09X01) and Bacillus cereus (09B18), were isolated from cereal cyst nematodes (Heterodera filipjevi) in Henan, China. The two bacterial strains were subsequently evaluated for their biological control potential. The results showed that the culture filtrate of the two strains caused high mortality of the second stage juvenile (J2) nematodes and reduced in vitro egg hatch. Results of greenhouse and field trials demonstrated that treatments with bacterial suspensions of strains 09B18 and 09X01 significantly reduced the numbers of white females in wheat roots. The rate of reduction was 75.9% by 09B18 and 70.2% by 09X01 in greenhouse trials, and 43.5% by 09B18 and 51.1% by 09X01 in field trials. These treatments also increased wheat yields by 15.9% (09B18) and 13.2% (09X01) compared to untreated control. It is concluded that the two bacterial strains can be regarded as potential biocontrol agents.