Heterodera Filipjevi Research Articles

Dactylfungins and Tetralones: Bioactive Metabolites from a Nematode-Associated Laburnicola nematophila.

A chemical investigation of Laburnicola nematophila, isolated from cysts of the plant parasitic nematode Heterodera filipjevi, affored three dactylfungin derivatives (1-3) and three tetralone congeners (4-6). Dactylfungin C (1), laburnicolin (4), and laburnicolenone (5) are previously undescribed natural products. Chemical structures of the isolated compounds were determined based on 1D and 2D NMR spectroscopic analyses together with HR-ESI-MS spectrometry and comparison with data reported in the literature. The relative configurations of compounds 1, 2, and 4-6 were determined based on their ROESY data and analysis of their coupling constants (J values). The absolute configurations of 4-6 were determined through the comparison of their measured and calculated TDDFT-ECD spectra. Compounds 1-3 were active against azole-resistant Aspergillus fumigatus.

Chromosome-level genome assembly of the cereal cyst nematode Heterodera flipjevi

As an economically important plant parasitic nematode (PPN), Heterodera filipjevi causes great damage on wheat, and now it was widely recorded in many countries. While multiple genomes of PPNs have been published, high-quality genome assembly and annotation on H. filipjevi have yet to be performed. This study presents a chromosome-scale genome assembly and annotation for H. filipjevi, utilizing a combination of Illumina short-read, PacBio long-read, and Hi-C sequencing technologies. The genome consists of 9 pseudo-chromosomes that contain 134.19 Mb of sequence, with a scaffold N50 length of 11.88 Mb. In total, 10,036 genes were annotated, representing 75.20% of the total predicted protein-coding genes. Our study provides the first chromosome-scale genome for H. filipjevi, which is also the inaugural high-quality genome of cereal cyst nematodes (CCNs). It provides a valuable genomic resource for further biological research and pest management of cereal cyst nematodes disease.

Laburnicotides A-F: Acyclic N-Acetyl Oligopeptides from the Nematode-Cyst-Associated Fungus Laburnicola nematophila.

Nematode-associated fungi revealed the potential to produce a broad spectrum of chemical scaffolds. In this study, a mycelial extract of Laburnicola nematophila, a fungal strain derived from the cereal cyst nematode Heterodera filipjevi, was chemically explored and afforded six unprecedentedly reported acylic N-acetyl oligopeptides, laburnicotides A-F (1-6). Structure elucidation of the isolated compounds was established based on comprehensive 1D and 2D NMR spectroscopic analyses together with the acquired HR-ESI-MS spectrometric data. The absolute configuration of amino acid residues in 1-6 was established by performing advanced Marfey's derivatization method. All isolated compounds were assessed for their cytotoxic, antimicrobial, antiviral, and nematicidal activities with no potential activity observed.

Omnipolyphilins A and B: Chlorinated Cyclotetrapeptides and Naphtho-α-pyranones from the Plant Nematode-Derived Fungus Polyphilus sieberi.

Chemical exploration for two isolates of the recently described ascomycete species Polyphilus sieberi, derived from the eggs of the plant parasitic nematode Heterodera filipjevi, afforded the identification of many compounds that belong to various metabolite families: two previously undescribed chlorinated cyclotetrapeptides, omnipolyphilins A (1) and B (2), one new pyranonaphthoquinone, ventiloquinone P (3), a 6,6'-binaphto-α-pyranone dimer, talaroderxine D (4) in addition to nine known metabolites (5-13) were isolated from this biocontrol candidate. All isolated compounds were characterized by comprehensive 1D, 2D NMR, and HR-ESI-MS analyses. The absolute configurations of the cyclotetrapeptides were determined by a combination of advanced Marfey's method, ROE correlation aided by conformational analysis, and TDDFT-ECD calculations, while ECD calculations, Mosher's method, and experimental ECD spectra were used for ventiloquinone P (3) and talaroderxine D (4). Among the isolated compounds, talaroderxine D (4) showed potent antimicrobial activities against Bacillus subtilis and Staphylococcus aureus with MIC values of 2.1 and 8.3 μg mL-1, respectively. Additionally, promising inhibitory effects on talaroderxine D (4) against the formation of S. aureus biofilms were observed up to a concentration of 0.25 μg mL-1. Moreover, ophiocordylongiiside A (10) showed activity against the free-living nematode Caenorhabditis elegans.

First record of cyst nematode (Heterodera filipjevi) on winter wheat from Shanxi Province in North China.

Heterodera avenae, H. filipjevi, and H. laptipons are considered to be the major cyst nematode pathogens affecting most cereals and causing severe crop losses (Smiley and Yan 2015). In China, H. filipjevi was first recorded in Xuchang, Henan Province (Peng et al. 2010). Recently, H. filipjevi has been found in Anhui, Hebei, Shandong and Xinjiang provinces of China (Cui et al. 2021). To further understand the latest occurrence and distribution of H. filipjevi in China, a survey of cyst nematodes was conducted in the wheat planting area of Shanxi Province of North China from June 2018 to November 2020. White female cysts (5.8 ± 2.99 cysts per plant) were found on wheat roots in the sandy soil, and wheat was displaying symptoms of dwarfing, yellowing, and had few tillers in Licheng of Changzhi (N36°32´010´´, E113°27´039´´; N36°29´050´´, E113°23´023´´; N36°29´035´´, E113°22´020´´) and Zezhou of Jincheng (N35°33´057´´, E112°56´020´´) in Shanxi Province, and second-stage juveniles (J2s) were obtained from 13 soil samples using the sieving-decanting method. Four of the 13 samples were identified as H. filipjevi on the basis of morphological and molecular studies of female cysts and J2s. Morphologically, the cysts were lemon shaped and featured a pronounced vulval cone. The color ranged from light to dark brown. The white female shell was covered with a white crystalline layer. The vulval cone was bifenestrate with horseshoe-shaped bullae numerous and distinct, and a strongly developed underbridge. The main measurements (mean ± SD, range) of cysts (n = 13) were as follows: body length including neck 780.5 ± 53.9 μm (692 to 843 μm); body width 527.3 ± 55.5 μm (435 to 620 μm); length/width ratio 1.50 ± 0.21 (1.20 to 1.93); fenestra length 55.5 ± 4.1 μm (49 to 61 μm); fenestra width 24.8 ± 2.2 μm (21.1 to 28.8 μm); vulval slit length 9.0 ± 0.7 μm (7.8 to 9.6 μm); and underbridge length 66.8 ± 5.0 μm (61 to 77 μm). The measurements of J2s (n = 13) were as follows: body length 554.4 ± 23.4 μm (520to 587 μm); stylet length 22.7 ± 0.7 μm (21.5 to 23.8 μm); tail length 61.0 ± 5.5 μm (51.2 to 68.9 μm); and hyaline tail terminus length 37.3 ± 2.7 μm (33.4 to 42.3 μm). These morphological measurements are within the range characteristic of H. filipjevi (Peng et al. 2010). Genomic DNA was extracted from individual cyst (n = 6) and the rDNA internal transcribed spacer (ITS) sequence was amplified using the universal primers TW81 and AB28 (Joyce et al. 1994). The PCR test for each sample was repeated five times. The obtained ITS sequences (GenBank accession No. OQ421499 to OQ421502, 1054 bp) showed more than 99.5% similarity to those of H. filipjevi from the United States (GU079654 and KP878490), Turkey (KR704304 and KR704292), and China (MW789611, KY448473 and KT314234). The results were confirmed again by the species-specific primers HfF1 and HfR1of H. filipjevi and the target PCR fragments of 646 bp were obtained (Peng et al. 2013). The pathogenicity of H. filipjevi was verified by infesting winter wheat (Triticum aestivum 'Wenmai 19') and studying nematode developmentand reproduction with growth chamber (Cui et al. 2015). Eggs were hatched at 14-16°C, and freshly hatched J2s were used to inoculate wheat plants when the roots were approximately 1-centimeter long. Fifteen wheat plants were inoculated with 200 J2s, and three wheat plants without J2s were set as controls (Cui et al. 2021). Parasitic J2s and third- and fourth-stage juveniles were found in roots stained with acid fuchsin at 5, 15, and 25 days after inoculation (DAI), adult females were detected at 50 DAI, and a mean of 23.7 cysts per pot were extracted at 70 DAI (Cui et al. 2015). The morphological and molecular characteristics of the new cysts were identical to those of the H. filipjevi cysts from the original field samples, and no cysts formed in the control groups. Wheat is the main food and economic crop in Shanxi, and H. filipjevi, a potential threat to cereal crop production in Shanxi, should arouse sufficient attention. H. filipjevi is major cyst nematode pathogens of wheat and shows high prevalence in China. The loss of wheat production due to H. filipjevi is as high as 32.3% when the initial density ≥ 64 eggs/mL in soil (Li 2018). To the best of our knowledge, this is the first report of H. filipjevi in Shanxi Province of North China.

Recombinase Polymerase Amplification Coupled with CRISPR-Cas12a Technology for Rapid and Highly Sensitive Detection of Heterodera avenae and Heterodera filipjevi.

The cereal cyst nematodes Heterodera avenae and Heterodera filipjevi are recognized as cyst nematodes that infect cereal crops and cause severe economic losses worldwide. Rapid, visual detection of cyst nematodes is essential for more effective control of this pest. In this study, recombinase polymerase amplification (RPA) combined with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a (formerly known as cpf1) was developed for the rapid detection of H. avenae and H. filipjevi from infested field samples. The RPA reaction was performed at a wide range of temperatures from 35 to 42°C within 15 min. There was no cross-reactivity between H. avenae, H. filipjevi, and the common closely related plant-parasitic nematodes, indicating the high specificity of this assay. The detection limit of RPA-Cas12a was as low as 10-4 single second-stage juvenile (J2), 10-5 single cyst, and 0.001 ng of genomic DNA, which is 10 times greater than that of RPA-lateral flow dipstick (LFD) detection. The RPA-Cas12a assay was able to detect 10-1 single J2 of H. avenae and H. filipjevi in 10 g of soil. In addition, the RPA-LFD assay and RPA-Cas12a assays could both quickly detect H. avenae and H. filipjevi from naturally infested soil, and the entire detection process could be completed within 1 h. These results indicated that the RPA-Cas12a assay developed herein is a simple, rapid, specific, sensitive, and visual method that can be easily adapted for the quick detection of H. avenae and H. filipjevi in infested fields.

Transcriptome-proteomic analysis associated with resistance to wheat cyst nematode Heterodera filipjevi

Cereal-cyst-nematode-(CCN), Heterodera filipjevi, is a destructive nematode on cereal-crops globally. Limited information on CCN infection, epidemiology, and lack of adequate resistant cultivars hamper H. filipjevi management significantly. Thus, in this-study, the-reactions of resistant-and-susceptible-wheat cultivars, Triticum aestivum L,- to H. filipjevi,-Isfahan-pathotype, were investigated to determine changes in expression of gene-markers, Cre and PRX-groups, using Real-time-polymerase-chain-reaction-(qPCR). We also measured the-activity of POX and CAT, the-two-important-antioxidant enzymes in defense-mechanisms providing a better insight into defense responses of antioxidant enzymes in wheat against H. filipjevi. RNA was extracted from the-roots, 4 and 7-days-post-inoculation (dpi); and from the-leaves 6 and 7-weeks-post-inoculation-(wpi). Expression of Cre3, peroxidases-(PRX)-TaPrx111 and TaPrx112-genes increased at 4 dpi by 6.32, 20 and 8.71-fold in the roots, respectively. However, at 7-dpi, the expression of Cre3, TaPrx111 and TaPrx112-genes increased by-15.98, 5.93, and 16.59, respectively. Similarly, an increasing-trend was recorded in the-leaves. The-expression of Cre3-gene in the-wheat-leaves increased-by 8.85 and 9.77-fold in 6 and 7-wpi compared to the-controls, respectively, as well as by-3.74 and 5.93-fold for TaPrx111-gene; and 4.16 and 6.11-fold for TaPrx112-gene in 6 and 7-wpi, respectively. Our findings demonstrated that the activity of -POX and CAT increased by-1.45 to 2.54 times with CCN infected in resistance-genotypes. Surprisingly, PRX-gene-marker, basically resistant to abiotic-stresses such as high-salinity and drought, and also regulating the ROS-balance in plants, similarly modulates-immunity against-H. filipjevi. Conclusively, the accessions used in this-study that exhibited resistance-and-moderate-resistance offer additional-genetic-resources for wheat-breeding-programs.

Polydomus karssenii gen. nov. sp. nov. is a dark septate endophyte with a bifunctional lifestyle parasitising eggs of plant parasitic cyst nematodes (Heterodera spp.)

In this study fungal strains were investigated, which had been isolated from eggs of the cereal cyst nematode Heterodera filipjevi, and roots of Microthlaspi perfoliatum (Brassicaceae). The morphology, the interaction with nematodes and plants and the phylogenetic relationships of these strains originating from a broad geographic range covering Western Europe to Asia Minor were studied. Phylogenetic analyses using five genomic loci including ITSrDNA, LSUrDNA, SSUrDNA, rpb2 and tef1-α were carried out. The strains were found to represent a distinct phylogenetic lineage most closely related to Equiseticola and Ophiosphaerella, and Polydomus karssenii (Phaeosphaeriaceae, Pleosporales) is introduced here as a new species representing a monotypic genus. The pathogenicity tests against nematode eggs fulfilled Koch’s postulates using in vitro nematode bioassays and showed that the fungus could parasitise its original nematode host H. filipjevi as well as the sugar beet cyst nematode H. schachtii, and colonise cysts and eggs of its hosts by forming highly melanised moniliform hyphae. Light microscopic observations on fungus-root interactions in an axenic system revealed the capacity of the same fungal strain to colonise the roots of wheat and produce melanised hyphae and microsclerotia-like structure typical for dark septate endophytes. Confocal laser scanning microscopy further demonstrated that the fungus colonised the root cells by predominant intercellular growth of hyphae, and frequent formation of appressorium-like as well as penetration peg-like structures through internal cell walls surrounded by callosic papilla-like structures. Different strains of the new fungus produced a nearly identical set of secondary metabolites with various biological activities including nematicidal effects irrespective of their origin from plants or nematodes.

Identification of Genetic Resistance to the Crown and Root Rot Caused by Fusarium culmorum and Cereal Cyst Nematode (Heterodera filipjevi) in the Turkish Cereal Varieties

Soilborne pathogens associated with cereals cause significant yield losses throughout the world. Among soilborne pathogens Heterodera filipjevi and Fusarium culmorum are two main destructive causal agents that attack the roots and crowns of cereals and have been reported as the most prevalent species in Türkiye. A total of 245 Turkish cereal varieties consisting of wheat, oat, triticale and rye which were registered between 1931 to 2013; were tested for their resistance to Fusarium culmorum and Heterodera filipjevi. The varieties were screened under controlled and/or field conditions for two successive growing seasons. The bread wheat ‘Murat-1’ was the most resistant variety for both pathogens. The durum wheats ‘Yelken 2000’ and ‘Yılmaz 98’ were found the most promising varieties for resistance to F. culmorum and H. filipjevi, respectively. The triticale ‘Umranhanim’ ranked moderately resistant at adult plant stage while ‘Presto’ and ‘Melez 2001’ were ranked moderately resistant at seedling stage for Fusarium culmorum. The findings of this study are important for the global wheat breeding programs as those genotypes are still being used in crosses for new genotype development.

Transcriptome-based analysis of resistance mechanisms to Bipolaris sorokiniana, a common wheat root-rot disease.

In common root and crown rot (CRR), Bipolaris sorokiniana (Sace.) is one of the important in wheat, causing considerable yield losses globally. Sources of resistance can provide a feasible and effective method of control for plant disease management. However, knowledge on mechanisms of resistance is scarce. We screened 33 wheat genotypes against B. sorokiniana under greenhouse and field conditions. In addition, real-time quantitative PCR (qPCR) analysis using ten novel candidate gene markers, Cre3, EDS1, LTP5, PGIP, PR-1, PIEP1, TLP, UGT, Stb6 and PFT, was conducted on leaves and roots, along with changes in activity of antioxidant enzymes, peroxidase, catalase, β-1,3-glucanase, and phenolic content for their involvement in disease impact mechanisms. Lowest disease severity was in 'Alvand', followed by 'Baharan' and 'Bam' as resistant genotypes. Quantitative gene expression showed that, although the candidate defence genes were upregulated 1.24- to 3.5-fold in wheat roots and leaves inoculated with B. sorokiniana, they were highly regulated in resistant varieties 'Alvand', 'Mehregan' and 'Bam'. Cre3, a resistance gene to cereal cyst nematode Heterodera filipjevi, was regulated in cultivars resistant to B. sorokiniana. Similar results were obtained for Stb6, a gene resistant to Septoria tritici blotch, EDS1 resistant to powdery mildew, Blumeria graminis, and the genes PR-1 and UGT resistant to leaf rust, Puccinia triticina. Antioxidant enzyme activity also showed the highest increases in resistant genotypes. In conclusion, the T. aestivum-B. sorokiniana interaction in resistant wheat cultivars uses defence-related genes and enzymes that protect wheat towards sustainable development. Further such studies will shed light on simultaneous resistance to other diseases in wheat cultivars.

Molecular diagnosis and direct quantification of cereal cyst nematode (Heterodera filipjevi) from field soil using TaqMan real-time PCR

Heterodera filipjevi continues to be a major threat to wheat production worldwide. Rapid detection and quantification of cyst nematodes are essential for more effective control against this nematode disease. In the present study, a TaqMan-minor groove binder (TaqMan-MGB) probe-based fluorescence quantitative real-time PCR (qPCR) was successfully developed and used for quantifying H. filipjevi from DNA extracts of soil. The primers and probe designed from the obtained RAPD-SCAR marker fragments of H. filipjevi showed high specificity to H. filipjevi using DNA from isolates-confirmed species of 23 Heterodera spp., 1 Globodera spp. and 3 Pratylenchus spp. The qPCR assay is highly sensitive and provides improved H. filipjevi detection sensitivity of as low as 4–3 single second-stage juvenile (J2) DNAs, 10–3 female DNAs, and 0.01 μg μL–1 genomic DNAs. A standard curve relating to the threshold cycle and log values of nematode numbers was generated and validated from artificially infested soils and was used to quantify H. filipjevi in naturally infested field soils. There was a high correlation between the H. filipjevi numbers estimated from 32 naturally infested field soils by both conventional methods and the numbers quantified using the qPCR assay. qPCR potentially provides a useful platform for the efficient detection and quantification of H. filipjevi directly from field soils and to quantify this species directly from DNA extracts of field soils.

Diversity and Identification of Plant-Parasitic Nematodes in Wheat-Growing Ecosystems.

Several nematode species can be found in different densities in almost any soil ecosystem, and their diversity in those ecosystems depends on numerous reasons, such as climatic conditions and host presence. Cereals are one of the main hosts of plant-parasitic nematodes (PPN), chiefly root-lesion nematodes (RLN, Pratylenchus spp.) and cereal cyst nematodes (CCN, Heterodera spp.). These nematodes are known as major parasites of the cereal crops; however, agricultural areas accommodate various nematodes showing biological variation. The diversity of parasitic nematodes on cereals in the Sakarya provinces of Türkiye, where cereals are intensively grown and located in the middle of two climatic zones, has not been well studied. Therefore, in this study, we aimed to determine the diversity, identification, and molecular phylogeny of PPNs in wheat-growing ecosystems in the Hendek, Pamukova, Geyve, Akyazı, and Central districts of Sakarya. The diversity of PPNs was calculated using the Shannon diversity index. Thirteen PPN genera were detected in 92% of soil samples. Heterodera filipjevi was identified in 24% of the soil samples using morphological, morphometrical, and molecular tools. In the morphological and molecular analyses, intraspecific polymorphism was observed in H. filipjevi populations. The result indicated that the high infestation rate of H. filipjevi was recorded from Geyve and Pamukova, followed by Hendek and Akyazı; however, a low infestation rate was detected in the Central district. The moderate value of the Shannon index of migratory nematode species was obtained in wheat fields as 2.31, whereas the value of evenness was 0.93, implying moderate diversity and high evenness of nematodes. This study is the first comprehensive report on H. filipjevi from wheat cropping areas in the Sakarya province. Intensified cereal cropping systems with/without non-cereal rotations increased the risk of plant-parasitic nematodes, especially RLNs and H. filipjevi infection of wheat production areas in the province.

The Effect of Drought Stresses, Fusarium Culmorum and Heterodera Filipjevi and their Interactions on the Expression Pattern of Transcription Factor Gene NAC69-3 in Bread Wheat

The Effect of Drought Stresses, Fusarium Culmorum and Heterodera Filipjevi and their Interactions on the Expression Pattern of Transcription Factor Gene NAC69-3 in Bread Wheat

Interactions among cereal cyst nematode Heterodera filipjevi, dryland crown rot Fusarium culmorum, and drought on grain yield components and disease severity in bread wheat

The cereal cyst nematode (CCN) Heterodera and crown rot caused by Fusarium species limit cereal production and yield potential. Losses increase when CCN and crown rot occur together, especially under water stress conditions. The objective of this study was to investigate the interactions among Heterodera filipjevi, Fusarium culmorum, and drought on a set of wheat germplasm with differing levels of resistance/tolerance to CCN, crown rot and drought. Plant emergence was significantly reduced (56% seedling death) when seeds were planted in a water stress environment where F. culmorum and H. filipjevi were present. Crown rot was more severe under water stress compared with Fusarium inoculation alone. The number of cysts fell significantly when H. filipjevi was co-inoculated with F. culmorum, revealing an antagonistic interaction between the nematode and fungus. The highest number of cysts was found on the susceptible accession ‘Seri’ under water stress conditions. Water stress caused a significant reduction in plant height, while F. culmorum inoculation did not, unless combined with water stress. Yield components were significantly reduced by each of the single stresses and losses were greater when plants were exposed to double or triple stresses. Water stress and F. culmorum inoculation caused a significant reduction in spike weight and seeds per spike. The chlorophyll content of wheat leaves was negatively affected by water stress and inoculation by both nematode and fungus. In conclusion, water stress exacerbates the damage caused by CCN and crown rot, and planting drought-resistant varieties would be an ideal solution to reduce losses.

Morphology and DNA phylogeny differentiate several anamorphic ascomycetes associated with a cereal cyst nematode, Heterodera filipjevi, and new records for the mycobiota of Iran

Morphology and DNA phylogeny differentiate several anamorphic ascomycetes associated with a cereal cyst nematode, Heterodera filipjevi, and new records for the mycobiota of Iran

Resistance Response of Drought and Heat Tolerant Spring Wheat Lines against the Cereal Cyst Nematode Heterodera filipjevi

Wheat (Triticum aestivum L.) is extremely affected by several abiotic and biotic stresses. Drought and/or heat alongside the parasitism of cereal cyst nematodes of the Heterodera genera can have a combined destructive impact on wheat. Solely, the cereal cyst nematode species Heterodera filipjevi can cause wheat yield losses of up to 50%. Several control measures have been implemented, yet, the most economical and convenient control strategy is the use of resistant hosts. Therefore, the main aim of this study was to evaluate the resistant response of 257 spring wheat lines obtained from CIMMYT to Heterodera filipjevi that might contain novel sources of resistance and be added as genetic resources for future breeding programs. Also, provide a base for future research to understand the relationship between nematode resistances and drought and heat tolerance. The results indicated that 11 lines (4%) and 36 lines (14%) were resistant and moderately resistant, respectively. High frequency of susceptible and highly susceptible lines and low frequency of resistant lines within this set were also recorded. The linear regression analysis between the number of cysts formed and the resistance response grouping showed a strong, positive, linear correlation. Log-linear regression analysis showed that there is a weak positive correlation between the yield of heat tolerant wheat lines and their resistance to the cyst nematodes as these lines showed tolerance, while there was a weak negative correlation of formed cyst nematodes on the yield of drought tolerant lines. This study was able to add new genetic sources of resistance to Heterodera filipjevi for upcoming breeding programs.

First report of Heterodera filipjevi on winter wheat from Hebei Province in North China.

First report of Heterodera filipjevi on winter wheat from Hebei Province in North China.

Diversity of Mycobiota Associated with the Cereal Cyst Nematode Heterodera filipjevi Originating from Some Localities of the Pannonian Plain in Serbia.

Simple SummaryHeterodera filipjevi, the cereal cyst nematode, is one of the most globally recognized and economically important nematodes on wheat. As some other cyst nematodes that are plant root parasites, the cysts of H. filipjevi survive in soil for years and shelter a large number of microbes. The aims of this study were to investigate the diversity of mycobiota associated with the cereal cyst nematode H. filipjevi, to infer phylogenetic relationships of the found mycobiota, and to explore the ecological connection between fungi and the field history, including the potential of fungi in bioremediation and the production of novel bioactive compounds. The study showed that the fungal species associated with the H. filipjevi cysts belong to diverse phyla, including Ascomycota, Basidiomycota, and Mucoromycota. The members of Ascomycota (Fusarium avenaceum, Sarocladium kiliense, Setophoma terrestris) are plant parasites, indicating that crops were host plants for fungal infection of recent origin. The members of Basidiomycota (Bjerkandera adusta, Cerrena unicolor, Trametes hirsuta, etc.) are wood-decay fungi, the presence of which in agricultural soil indicates that forests were the preceding plants.Cereals, particularly wheat, are staple food of the people from the Balkans, dating back to the Neolithic age. In Serbia, cereals are predominantly grown in its northern part between 44° and 45.5° N of the Pannonian Plain. One of the most economically important nematodes on wheat is the cereal cyst nematode, Heterodera filipjevi. Cysts of H. filipjevi survive in soil for years and shelter a large number of microorganisms. The aims of this study were to investigate the diversity of mycobiota associated with the cereal cyst nematode H. filipjevi, to infer phylogenetic relationships of the found mycobiota, and to explore the ecological connection between fungi and the field history, including the potential of fungi in bioremediation and the production of novel bioactive compounds. Cysts were isolated from soil samples with a Spears apparatus and collected on a 150-µm sieve. The cysts were placed on potato dextrose agar, and maintained for two weeks at 27°C. Following fungal isolation and colony growing, the fungal DNA was extracted, the ITS region was amplified, and PCR products were sequenced. The study showed that the isolated fungal species belong to diverse phyla, including Ascomycota, Basidiomycota, and Mucoromycota. Ascomycota is represented by the families Clavicipitaceae, Sarocladiaceae, Nectriaceae, and Phaeosphaeriaceae. Basidiomycota is represented by the families Cerrenaceae, Polyporaceae, Phanerochaetaceae, and Meruliaceae, and the order Cantharellales. The family Mortierellaceae represents Mucoromycota. The members of Ascomycota and Basidiomycota both depict the field history. Ascomycota indicate the fungal infection is of recent origin, while Basidiomycota point toward the preceding host plants, enabling the plant field colonization history to be traced chronologically.

Plant-Parasitic Nematodes Associated With Wheat in Central, Eastern, and Southeastern Kazakhstan.

Kazakhstan is one of the biggest wheat producers, however, its wheat production is far below the average international wheat production standard due to biotic and abiotic stressors. Plant-parasitic nematodes are devastating for cereal production systems worldwide. A comprehensive survey was conducted in 2019 to identify plant-parasitic nematodes associated with wheat in different locations of central, eastern, and southeastern Kazakhstan. The results revealed 33 root-lesion and 27 cyst nematode populations from the 77 localities sampled. These two genera occurred in separate or in mixed populations. The root-lesion populations were identified as Pratylenchus neglectus and P. thornei while all cyst nematodes were identified as Heterodera filipjevi. The identification of nematodes was firstly performed based on morphological and morphometric features and confirmed by BLAST and phylogenetic analyses based on the internal transcribed spacer and the D2-D3 expansion located in the 28S gene of ribosomal DNA for CCN and RLN populations, respectively. Pratylenchus neglectus and P. thornei populations from Kazakhstan showed a high similarity with the American, European, and Asian populations. Heterodera filipjevi populations formed a well-supported cluster with the corresponding populations from different countries and showed a slightly intraspecific polymorphism. Kazakhstan populations of H. filipjevi may have multiple introductions in Kazakhstan due to the divergence among them. The results of this study are of great importance for breeding programs and will enable awareness for extension advisors to develop measures to control these nematodes in cereal cropping areas in Kazakhstan.

Phytophagous nematodes in cereal fields in Niğde Province, Turkey

This study evaluated the occurrence and incidence of phytophagous nematodes and identified the cereal cyst nematode species by morphological and molecular tools in the main cereal-growing areas in Nigde in 2018-2019. Phytophagous nematodes within twelve genera were detected in 95% of soil samples. The most common phytophagous nematodes in cereal soil were in the genera Heterodera, Ditylenchus , Merlinius , Pratylenchus , Aphelenchus , Aphelenchoides , Tylenchus , Helicotylenchus , Trophurus , Pratylenchoides , Filenchus and Xiphinema (in decreasing order of incidence). In particular, 75% of the soil samples from surveyed fields were infested with the cereal cyst nematodes (Heterodera spp.). Morphological characteristics of cysts and second-stage juveniles were calculated within the expected ranges for Heterodera filipjevi (Madzhidov, 1981) Stelter, 1984, however, two populations from Camardi was determined as Heterodera latipons Franklin, 1969 (Tylenchida: Heteroderidae). Intraspecific variation was not observed within the populations of H. filipjevi which could be in the same genotypic group. In addition to the high incidence of these Heterodera spp., intensive cereal cropping systems with/without non-cereal rotations in wheat production areas of Nigde also resulted in high incidence of root lesion nematode, Pratylenchus species.