Two new root endophyte and nematode cyst parasite species of the widely distributed genus Laburnicola

Fungal root endophytes, including the common form group of dark septate endophytes (DSEs), represent different taxonomic groups with potentially diverse life strategies. During surveys of DSE communities and of nematode cysts colonizing fungi, isolates representing Laburnicola ( Didymosphaeriaceae , Pleosporales ) lineages were discovered. Here we carried out a comprehensive study of the phylogenetic relationships and taxonomy of fungi collected from plant roots in Hungary, Mongolia, and Kazakhstan and from eggs of the cereal cyst nematode Heterodera filipjevi in Turkey. In addition to the study of the morphology and culture characteristics of the strains, four loci (internal transcribed spacer, partial large and small subunit regions of nuclear ribosomal DNA and partial translation elongation factor 1-alpha) were used to infer the molecular phylogenetic relationships of the strains within Laburnicola . The isolates were found to represent two distinct lineages, which are described here as novel species, Laburnicola nematophila and L. radiciphila . The interaction of the strains with plants and nematodes was examined using in vitro bioassays, which revealed endophytic interactions with the plant roots and parasitic interactions with the nematode eggs. Analyses of similar ITS sequences found in public databases revealed that members of the genus Laburnicola are widely distributed characteristic members of the plant microbiome, and they are reported as parasites of plant parasitic cyst nematodes here for the first time

Les antagonistes naturels d’Heterodera avenae dans diverses conditions de cultures de céréales en Algérie

A study was carried out to determine the diversity of the microflora associated with the cereal cyst nematode, Heterodera avenae, in four cereal production regions of Algeria. The isolation was mainly targeted toward fungi associated to cyst nematodes. The isolation resulted in the identification of six fungi: Fusarium sp., Penicillium spp., Verticillium spp., Rhizomucor spp., Rhizopus spp.and Ulocladium spp. In all the prospected regions, Fusarium sp. is most frequently associated with cysts. The highest diversity is observed at Dahmouni with the genera Aspergillus spp., Ulocladium spp., Rhizopus spp. and Rhizomucor spp. The in vitro test showed that the metabolites of Fusarium spp., Penicillium spp.and Aspergillus spp. parasitize the eggs at a high rate and thus constitute potential agents for the control of H. avenae. Keywords: Antagonists, fungi, bacteria, metabolites, cyst nematode, biological controlUn travail de recherche a été conduit pour mettre en évidence la diversité de la microflore associée au nématode à kyste des céréales, Heterodera avenae, dans quatre régions céréalières d’Algérie. L’isolement des micro-organismes associés a visé principalement les champignons inféodés aux kystes. L’isolement a abouti à l’identification de six champignons: Fusarium spp., Penicillium spp., Verticillium spp., Rhizomucor spp., Rhizopus spp. et Ulocladium spp. Dans toutes les régions prospectées, Fusarium sp. est le plus fréquemment associé aux kystes. La diversité la plus élevée est observée à Dahmouni avec les genres Aspergillus spp., Ulocladium spp., Rhizopus spp. et Rhizomucor spp. L’essai in vitro a montré que les métabolites de Fusarium spp., Penicillium spp.et Aspergillus spp. parasitent à un taux élevé les œufs et constituent ainsi des agents potentiels de lutte contre H. avenae. Mots-clés: Antagonistes, champignons, bactéries, métabolites, nématode à kyste, lutte biologiqu

Resistance response of drought and heat tolerant spring wheat lines against the cereal cyst nematode, heterodera filipjevi

Spot blotch caused by Bipolaris sorokiniana is a destructive disease of wheat worldwide. This study investigated the aggressiveness of B. sorokiniana isolates from different wheat-growing areas of Bolu province in Turkey on the cultivar Seri-82. Host susceptibility of 55 wheat cultivars was evaluated against the most aggressive isolate. Our results indicated that the cultivars Anafarta and Koç-2015 were the most resistant. A specific and sensitive qPCR assay was developed for detecting the pathogen in plant tissues and evaluating wheat plants with different resistance levels. Three primer sets, BsGAPDHF/BsGAPDHR, BsITSF/BsITSR, and BsSSUF/BsSSUR, were designed based on glyceraldehyde-3-phosphate dehydrogenase, internal transcribed spacers, and 18S rRNA loci of B. sorokiniana with detection limits of 1, 0.1, and 0.1 pg of pathogen DNA, respectively. The qPCR assay was highly sensitive and did not amplify DNA from the other closely related fungal species and host plants. The protocol differentiated wheat plants with varying degrees of resistance. The assay developed a useful tool for the quantification of the pathogen in the early stages of infection and may provide a significant contribution to a more efficient selection of wheat genotypes in breeding studies. In the present study, expression levels of PR proteins, phenylalanine ammonia-lyase, catalase, ascorbate peroxidase, and superoxide dismutase enzymes were upregulated in Anafarta (resistant) and Nenehatun (susceptible) cultivars at different post-infection time points, but more induced in the susceptible cultivar. The results showed considerable variation in the expression levels and timing of defense genes in both cultivars

Genome-Wide Association Study for Multiple Biotic Stress Resistance in Synthetic Hexaploid Wheat

Genetic resistance against biotic stress is a major goal in many wheat breeding programs. However, modern wheat cultivars have a limited genetic variation for disease and pest resistance and there is always a possibility of the evolution of new diseases and pests to overcome previously identified resistance genes. A total of 125 synthetic hexaploid wheats (SHWs; 2n = 6x = 42, AABBDD, Triticum aestivum L.) were characterized for resistance to fungal pathogens that cause wheat rusts (leaf; Puccinia triticina, stem; P. graminis f.sp. tritici, and stripe; P. striiformis f.sp. tritici) and crown rot (Fusarium spp.); cereal cyst nematode (Heterodera spp.); and Hessian fly (Mayetiola destructor). A wide range of genetic variation was observed among SHWs for multiple (two to five) biotic stresses and 17 SHWs that were resistant to more than two stresses. The genomic regions and potential candidate genes conferring resistance to these biotic stresses were identified from a genome-wide association study (GWAS). This GWAS study identified 124 significant marker-trait associations (MTAs) for multiple biotic stresses and 33 of these were found within genes. Furthermore, 16 of the 33 MTAs present within genes had annotations suggesting their potential role in disease resistance. These results will be valuable for pyramiding novel genes/genomic regions conferring resistance to multiple biotic stresses from SHWs into elite bread wheat cultivars and providing further insights on a wide range of stress resistance in wheat

Monitoring of host suitability and defense-related genes in wheat to Bipolaris sorokiniana

Spot blotch caused by Bipolaris sorokiniana is a destructive disease of wheat worldwide. This study investigated the aggressiveness of B. sorokiniana isolates from different wheat-growing areas of Bolu province in Turkey on the cultivar Seri-82. Host susceptibility of 55 wheat cultivars was evaluated against the most aggressive isolate. Our results indicated that the cultivars Anafarta and Koç-2015 were the most resistant. A specific and sensitive qPCR assay was developed for detecting the pathogen in plant tissues and evaluating wheat plants with different resistance levels. Three primer sets, BsGAPDHF/BsGAPDHR, BsITSF/BsITSR, and BsSSUF/BsSSUR, were designed based on glyceraldehyde-3-phosphate dehydrogenase, internal transcribed spacers, and 18S rRNA loci of B. sorokiniana with detection limits of 1, 0.1, and 0.1 pg of pathogen DNA, respectively. The qPCR assay was highly sensitive and did not amplify DNA from the other closely related fungal species and host plants. The protocol differentiated wheat plants with varying degrees of resistance. The assay developed a useful tool for the quantification of the pathogen in the early stages of infection and may provide a significant contribution to a more efficient selection of wheat genotypes in breeding studies. In the present study, expression levels of PR proteins, phenylalanine ammonia-lyase, catalase, ascorbate peroxidase, and superoxide dismutase enzymes were upregulated in Anafarta (resistant) and Nenehatun (susceptible) cultivars at different post-infection time points, but more induced in the susceptible cultivar. The results showed considerable variation in the expression levels and timing of defense genes in both cultivars

Genome-wide association study of root-lesion nematodes Pratylenchus species and crown rot Fusarium culmorum in bread wheat

Triticum aestivum L., also known as common wheat, is affected by many biotic stresses. Root diseases are the most difficult to tackle due to the complexity of phenotypic evaluation and the lack of resistant sources compared to other biotic stress factors. Soil-borne pathogens such as the root-lesion nematodes caused by the Pratylenchus species and crown rot caused by various Fusarium species are major wheat root diseases, causing substantial yield losses globally. A set of 189 advanced spring bread wheat lines obtained from the International Maize and Wheat Improvement Center (CIMMYT) were genotyped with 4056 single nucleotide polymorphisms (SNP) markers and screened for root-lesion nematodes and crown rot resistance. Population structure revealed that the genotypes could be divided into five subpopulations. Genome-Wide Association Studies were carried out for both resistances to Pratylenchus and Fusarium species. Based on our results, 11 different SNPs on chromosomes 1A, 1B, 2A, 3A, 4A, 5B, and 5D were significantly associated with root-lesion nematode resistance. Seven markers demonstrated association with P. neglectus, while the remaining four were linked to P. thornei resistance. In the case of crown rot, eight different markers on chromosomes 1A, 2B, 3A, 4B, 5B, and 7D were associated with Fusarium crown rot resistance. Identification and screening of root diseases is a challenging task; therefore, the newly identified resistant sources/genotypes could be exploited by breeders to be incorporated in breeding programs. The use of the identified markers in marker-assisted selection could enhance the selection process and cultivar development with root-lesion nematode and crown rot resistance

Biology and biotechnology of Trichoderma

Fungi of the genus Trichoderma are soilborne, green-spored ascomycetes that can be found all over the world. They have been studied with respect to various characteristics and applications and are known as successful colonizers of their habitats, efficiently fighting their competitors. Once established, they launch their potent degradative machinery for decomposition of the often heterogeneous substrate at hand. Therefore, distribution and phylogeny, defense mechanisms, beneficial as well as deleterious interaction with hosts, enzyme production and secretion, sexual development, and response to environmental conditions such as nutrients and light have been studied in great detail with many species of this genus, thus rendering Trichoderma one of the best studied fungi with the genome of three species currently available. Efficient biocontrol strains of the genus are being developed as promising biological fungicides, and their weaponry for this function also includes secondary metabolites with potential applications as novel antibiotics. The cellulases produced by Trichoderma reesei, the biotechnological workhorse of the genus, are important industrial products, especially with respect to production of second generation biofuels from cellulosic waste. Genetic engineering not only led to significant improvements in industrial processes but also to intriguing insights into the biology of these fungi and is now complemented by the availability of a sexual cycle in T. reesei/Hypocrea jecorina, which significantly facilitates both industrial and basic research. This review aims to give a broad overview on the qualities and versatility of the best studied Trichoderma species and to highlight intriguing findings as well as promising applications

Effect of temperatures on incubation duration of Heterodera avenae Wollenweber (Tylenchida: Heteroderidae) cysts and optimization of some materials and methods [Heterodera avenae Wollenweber (Tylenchida: Heteroderidae) yumurtalarínín inkübasyonu üzerine sícaklí gín etkisi ile en uygun deneme yöntem ve materyallerinin araştírílmasí]

In this study, the effect of temperatures on the incubation duration of the cereal cyst nematode cysts, Heterodera avenae Wollenweber and determining of the optimum methods and materials was investigated under invitro conditions. The effect of tempratures on incubation duration of Heterodera avenae cysts was studied on two different incubation stages. First incubation exposured to all cysts was set up 66 days at 4°C after each samples transfered different temperatures (5, 10, 15, 20 and 25°C). The cumulative hatching percent was significantly effected by different temperatures at the end of 252 days. Hatching was greater at lower temperatures (5, 10 15 °C) compared to at higher temperatures of (20, 25°C) ranging between 46.7, 82.3 and 45.9 % vs and 30.7 and 19.0%, respectively. Although the highest cumulative hatching of 82.3% was obtained at a constant 10°C at 252 days, the lowest cumulative hatching of 19.0% was obtained to 25°C at 252 days. The most suitable soil type for the in-vitro conditions was found the sandy soil (70 % sand: 29% field soil: 1% organic matter), best inoculation rate was 2,5 second stage juveniles per gram soil and inoculation time at planting day

Morphological and molecular identification of cyst nematode species (Heterodera spp.) in Algerian cereal fields

Cereal cyst nematodes (Heterodera spp.) are distributed globally and cause severe production losses of small grain cereals. To investigate the occurrence of cereal cyst nematodes in wheat-growing areas of Algeria, a survey was conducted and 27 cereal cyst nematode populations were collected. The populations were initially identified based on their morphological and morphometric characters, followed by molecular methods using speciesspecific primers, complemented by ITS-rDNA sequences. The morphological and morphometric features of second-stage juveniles (J2s) and cysts supported the presence of three Heterodera species: H. avenae, H. filipjevi and H. hordecalis. All morphological values of these distinct populations were very similar to those previously described for these species. Using species-specific primers for H. avenae and H. filipjevi, the specific bands of 109 bp and 646 bp confirmed the morphological identification of both species, respectively. In addition, the internal transcribed spacer (ITS) regions were sequenced to study the diversity of the 27 populations. These sequences were compared with those of Heterodera species available in the GenBank database (www.ncbi.nlm.nih.gov) and re-confirmed the identity of the species. Nineteen sequences of ITS-rDNA were similar (99–100%) to the sequences of H. avenae published in the GenBank, six sequences were similar (99–100%) to H. hordecalis, and two were similar (98–99%) to H. filipjevi. The results of this study are of great value to breeding programs and extension services, where they will contribute to the design of control measures to keep damaging nematodes in check