Cloning of Parasitism Genes from the Mediterranean Cereal Cyst Nematode, Heterodera Latipons * (original) (raw)
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Development of a species-specific PCR to detect the cereal cyst nematode, Heterodera latipons
Nematology, 2013
Several Heterodera species can reduce the yield of wheat and barley, among which H. avenae, H. filipjevi and H. latipons are economically the most important. Their identification, based on morphological characteristics, is not straightforward but can be made easier using molecular techniques. In this study, we developed species-specific primers for the detection of H. latipons. The actin gene of eight Heterodera species was partially sequenced and, after purifying and sequencing the PCR products, all sequences were aligned to find unique sites. The alignment showed moderate to very high similarities between the species. However, a small fragment of the actin gene was suitable for the construction of a potentially useful species-specific primer for H. latipons. The optimised PCR was subsequently tested with several populations of 14 Heterodera species and a single population of Punctodera punctata. Heterodera latipons was represented by 16 populations originating from six different countries. The primer set (Hlat-act), designed using AlleleID 7.73, was shown to be very specific. To test its sensitivity further, the PCR was conducted on DNA extracted from five second-stage juveniles (J2) of H. latipons mixed with five or 100 J2 belonging to H. avenae. The PCR was able to detect up to 1:10 dilution of the DNA obtained from five J2. The results showed that a specific and sensitive H. latipons species-specific PCR was constructed.
De Novo Transcriptome Sequencing and Analysis of the Cereal Cyst Nematode, Heterodera avenae
PLoS ONE, 2014
The cereal cyst nematode (CCN, Heterodera avenae) is a major pest of wheat (Triticum spp) that reduces crop yields in many countries. Cyst nematodes are obligate sedentary endoparasites that reproduce by amphimixis. Here, we report the first transcriptome analysis of two stages of H. avenae. After sequencing extracted RNA from pre parasitic infective juvenile and adult stages of the life cycle, 131 million Illumina high quality paired end reads were obtained which generated 27,765 contigs with N50 of 1,028 base pairs, of which 10,452 were annotated. Comparative analyses were undertaken to evaluate H. avenae sequences with those of other plant, animal and free living nematodes to identify differences in expressed genes. There were 4,431 transcripts common to H. avenae and the free living nematode Caenorhabditis elegans, and 9,462 in common with more closely related potato cyst nematode, Globodera pallida. Annotation of H. avenae carbohydrate active enzymes (CAZy) revealed fewer glycoside hydrolases (GHs) but more glycosyl transferases (GTs) and carbohydrate esterases (CEs) when compared to M. incognita. 1,280 transcripts were found to have secretory signature, presence of signal peptide and absence of transmembrane. In a comparison of genes expressed in the pre-parasitic juvenile and feeding female stages, expression levels of 30 genes with high RPKM (reads per base per kilo million) value, were analysed by qRT-PCR which confirmed the observed differences in their levels of expression levels. In addition, we have also developed a user-friendly resource, Heterodera transcriptome database (HATdb) for public access of the data generated in this study. The new data provided on the transcriptome of H. avenae adds to the genetic resources available to study plant parasitic nematodes and provides an opportunity to seek new effectors that are specifically involved in the H. avenae-cereal host interaction.
The predicted secretome in the transcriptome of cereal cyst nematode, Heterodera avenae
Objective: The cereal cyst nematode (CCN, Heterodera avenae) is a major pest of wheat (Triticum spp) crops in many countries. Cyst nematodes are obligate endoparasites that become sedentary within plant roots and induce the transformation of normal plant cells into elaborate feeding cites with the help of secreted proteins. These effectors are secreted as molecular tools to manipulate host plant cell cycle machinery to facilitate its infestation. Here we report the transcriptome analysis of H. avenae to identify putative effectors of parasitism. Methodology: Herein, we have sequenced the transcriptome by Illumina GAIIx platform of pre-parasitic juvenile and parasitic female stage. Raw reads derived from H. avenae transcriptome were assembled using Abyss, Trinity and Velvet. Sequences were scanned for prediction of signal peptides and Subcellular localization with multiple algorithms SignalP 4.0, TargetP 1.1, Wolf PSort and SecretomeP2.0. Further functional annotation was performed w...
TURKISH JOURNAL OF ZOOLOGY, 2018
The Mediterranean cereal cyst nematode, Heterodera latipons, is an important plant parasitic nematode that negatively impacts cereal production worldwide, particularly in the Mediterranean Basin. Successful control of cereal cyst nematodes requires knowledge of the nematode species/pathotypes and their biology. However, little information is currently available concerning the phenotypic and genotypic variability of H. latipons populations in Turkey. This study analyzed the morphological/morphometric and genetic characteristics of Turkish H. latipons populations to define intraspecific distinctions between them. Morphological and morphometric studies focused on the pattern of the perineal region of cysts and second stage juveniles (J2), while the molecular analyses used sequenced ITS-rDNA regions targeting the internal transcribed spacer (ITS) region of rDNA. The data from these investigations demonstrate that nematode populations from Adana and Hatay provinces differ from the populations collected in Kilis, Gaziantep, and Mardin provinces. These phenotypic and genotypic differences between H. latipons populations may indicate that they are heterogenic, with at least two pathotypes. To date, this study is the most comprehensive analysis identifying H. latipons populations in major wheatproducing areas of Turkey.
Turkish Journal of Entomology, 2020
The Mediterranean cereal cyst nematode, Heterodera latipons Franklin, 1969 (Tylenchida: Heteroderidae) is a destructive plant-parasitic nematode on cereal crops in particularly wheat and barley. It has a global distribution with a severe negative impact on yield quantity. In this study, a survey was conducted to identify plant-parasitic nematodes in cereal-growing areas in Cyprus. Forty-five samples including roots and soil from the root zone of plants were collected from cereal fields located in Gazimağusa, Girne, Güzelyurt and Lefkoşa Provinces before crop harvesting from late-May and early-June in 2017. Cyst-forming nematodes were determined by Fenwick's flotation and decanting techniques from 37 soil samples (82%). The internal transcribed spacer (ITS) regions of the ribosomal DNA of isolates were amplified and sequenced and subjected to a BLASTn search of the NCBI database for species identification, and the analyses showed that all samples were identified as H. latipons. Phylogenetic analyses based on ITS sequences revealed that H. latipons isolates from Northern Cyprus were closely related to isolates obtained from Morocco, Russia, Syria and Turkey. Data of this study demonstrated for the first time the presence of H. latipons in the cereal fields of Gazimağusa, Girne, Güzelyurt and Lefkoşa Provinces, where the nematode most likely causes serious economic problems in the cereal production. These results were the most up-to-dated analyses on the occurrence of H. latipons in cereal fields of Northern Cyprus and provided basic data for breeding programs to improve the resistant levels in the local cultivars.
Heterodera latipons on Barley in Jordan
Phytopathologia Mediterranea
A survey of major barley-growing areas in Jordan was conducted to determine the prevalence and degree of infestation of the Mediterranean cereal cyst nematode (MCCN) Heterodera latipons. The nematode was found in several locations in the northern and southern Mediterranean and the eastern desert areas of Jordan, but it was not detected in the northern Jordan valley and the southern desert areas. In those areas where the cyst nematode occurred, the incidence varied between areas from 30 to 100%. The degree of infestation varied from moderate to very severe. Infestation was most severe in the fields of the Northern Mediterranean area. The MCCN completed one life cycle per growing season. Studies on three isolates of the nematode from three areas, Ar-Ramtha, Madaba, and Al-Karak, revealed that the morphometrics varied little among these isolates. These three isolates as a whole varied in their virulence to two commonly used barley cultivars, Rum and Acsade 106, being more virulent on c...
International Journal of Agriculture and Biology
Cereal cyst nematodes are one of the most important soil-borne pathogens of cereals throughout the world. This group of nematodes is considered the most economically damaging pathogens of wheat and barley in Iran. In the present study, a series experiments were conducted during 2007-2010 to determine the distribution and population density of cereal cyst nematodes and to examine the biology of Heterodera latipons in the winter wheat cv. Sardari in a microplot under rain-fed conditions over two successive years in Markazi province in central Iran. Results of field survey showed that 40% of the fields were infested with at least one species of either Heterodera filipjevi or H. latipons. H. filipjevi was most prevalent in Farmahin, Tafresh and Khomein, with H. latipons being found in Khomein and Zarandieh regions. Female nematodes were also observed in Bromus tectarum, Hordeum disticum and Secale cereale, which are new host records for H. filipjevi. Also, H. filipjevi and H. latipons w...
Turkish journal of entomology, 2012
Summary The Heterodera avenae group includes 12 species feeding on roots of cereals. Three species, Heterodera avenae Wollenweber, 1924, Heterodera filipjevi (Madzhidov, 1981) Stelter, 1984 and Heterodera latipons Franklin, 1969 are among the most economically important cyst nematode pests of cereals cultivated in different parts of Turkey. In this study, forty seven cereal cyst nematode isolates collected from cereal growing areas of the South
Scientific Reports, 2023
Cereal cyst nematode (CCN) is a major threat to cereal crop production globally including wheat (Triticum aestivum L.). In the present study, single-locus and multi-locus models of Genome-Wide Association Study (GWAS) were used to find marker trait associations (MTAs) against CCN (Heterodera avenae) in wheat. In total, 180 wheat accessions (100 spring and 80 winter types) were screened against H. avenae in two independent years (2018/2019 "Environment 1" and 2019/2020 "Environment 2") under controlled conditions. A set of 12,908 SNP markers were used to perform the GWAS. Altogether, 11 significant MTAs, with threshold value of −log10 (p-values) ≥ 3.0, were detected using 180 wheat accessions under combined environment (CE). A novel MTA (wsnp_Ex_ c53387_56641291) was detected under all environments (E1, E2 and CE) and considered to be stable MTA. Among the identified 11 MTAs, eight were novel and three were co-localized with previously known genes/QTLs/MTAs. In total, 13 putative candidate genes showing differential expression in roots, and known to be involved in plant defense mechanisms were reported. These MTAs could help us to identify resistance alleles from new sources, which could be used to identify wheat varieties with enhanced CCN resistance. Wheat (Triticum aestivum L.) is one of the most widely grown crop, cultivated on ~ 220 million ha, with a worldwide annual production of over 700 million tons, and global annual export value of around US$50 billion 1. China was the top wheat producing country in 2022 with production volume of over 137 million metric tons followed by India, Russia, Australia, Canada, Pakistan, and Ukraine 2. Plant-parasitic nematodes (PPNs) are a serious global problem to wheat production 3-5. Root-knot nematodes (RKNs; Meloidogyne spp.), cereal cyst nematodes (CCNs; Heterodera spp.), and root lesion nematodes (RLNs; Pratylenchus spp.) are the most common PPNs responsible for reduction in crop yield worldwide 6. Heterodera avenae is an economically significant PPN in temperate wheat-producing regions like North and South Africa, East and West Asia, Australia, Europe, the Indian Subcontinent, the Middle East, and North America 7. Cultivation of susceptible cultivars is the main cause of severe yield losses due to H. avenae infection in majority of wheat growing regions 5,8. In order to reduce the threat posed by rising H. avenae population densities and to keep the losses incurred below economic threshold, it is crucial to develop efficient disease management strategies. Crop rotation, chemical, and biological management strategies are some of the common approaches that could help to reduce the damage caused by these nematodes to some extent. Few nematicides such as aldicarb, oxamyl, abamectin are known to be effective in decreasing the nematode population and increasing yield in wheat and barley 8-10. Despite their benefits several nematicides are banned due to their high risk for human health and environment. Therefore, finding the host plant resistance is one of the most effective, economically and environmentally sustainable method to prevent or reduce nematode multiplication 11 .