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Many Pythium spp. are causal agents of diseases of turfgrasses. Pythium spp. disseminate through irrigation systems in agricultural settings, and this study provides evidence that Pythium spp. also disseminate through golf course irrigation. Water samples were collected from irrigation heads and water sources at 10 golf courses in Missouri and Kansas, U.S.A. Samples were collected from 2018 to 2019 in April, July, and October. Phosphorus, nitrogen, and chloride concentrations were measured from irrigation head samples to determine if these parameters influence frequency of Pythium spp. detected. Pythium spp. were detected in samples through baiting and membrane filtration. Cultures were isolated on PARP media, and DNA was extracted from putative Pythium isolates. The internal transcribed spacer region was PCR-amplified and sequenced. Phylogenetic trees were constructed using representative sample sequences, sequences from seven morphologically identified reference isolates of Pythium, and similar GenBank accessions. Detected oomycete species include Lagenidium giganteum, Pythium biforme, Pythium insidiosum, Pythium marsipium, Pythium plurisporium, and Saprolegnia hypogyna. Twenty-one clades lacked species-level resolution, and 14 of these clades were associated with Pythium species. Clades A, C, D, E, I, and M contain Pythium species that cause root and crown rot on creeping bentgrass. Detected Pythium communities were dependent on the detection method used and sampling source. Pythium frequency and diversity were highest in April 2019. Sample temperature, sampling site, and chloride and nutrient concentrations did not influence Pythium frequency in samples. Irrigation systems using surface water sources contained at least three Pythium spp. over the course of 2 years.

The purpose of the current study is to report and provide the first record of the macroalgal diversity observed along the underrated northern coast of the Gulf of Kachchh (GoK), India, backed by its molecular study. The biodiversity data existing for the northern belt of the GoK, encompassing the Kachchh coast, is quite flimsy in contrast to the substantial amount of seaweed diversity data obtainable for its southern counterpart. Four Chlorophytes were collected from the selected sites and subjected to molecular characterization using internal transcribed spacer (ITS) regions, (ITS1 & ITS2). The ITS1 and ITS2 regions were amplified using ITS1 and ITS4 primers. The purified PCR products were sequenced. The G + C ratio of the ITS region ranged from 40.76 to 57.34%. The purity ratio ranged from 1.68 to 2.10, while the DNA concentration varied from 30 to 168 ng/µL for 4 different species tested. The average yield of the DNA recorded for these Chlorophytes was 5.45 µg. The phylogenetic tree was created by aligning the ITS sequence using MEGA 7 software following the maximum likelihood method. The taxonomic identity of all these 4 macroalgal species was successfully determined based on PCR results. The GenBank accession numbers [(MT452260), (MT452261), (MT484071), and (MT452262)] were obtained for all the four sequences. Overall, the northern region is species-poor in comparison with the southern belt of the Gulf of Kachchh, yet it bears some economically viable macroalgal species making it an important ecological area to be studied and explored.

Introduction: Toxigenic aspergilli are storage contaminants of poultry feeds and feed ingredients which produce secondary metabolites known as mycotoxins in these products. This research is aimed at detecting and characterizing mycotoxin producing aspergillus species present in feeds given to poultry birds and in the ingredients used in the production of these feeds. Method: A total of 102 samples of feed (n=30) and feed ingredients (n=72) were collected across 5 agro-ecological zones of Nigeria and analyzed for toxigenic Aspergilli using morphometric and molecular genomic characterization techniques. Different mycological media was used for isolation, by deploying pour plate method. The Aspergilli isolates morphometric identity were characterized using macroscopic and microscopic observed features. The isolated Aspergillus species were further characterized molecularly using genomic characterization protocols. Results: The results of the morphometric characterization indicated that nine (9) Aspergillus species were detected in the feed and feed ingredients, with Aspergillus flavus having the highest rate of occurrence among the isolates. Furthermore, the genomic typing using internal transcribe spacer (ITS) as a barcode, as well as comparison of the fungal isolates with other Aspergillus species in the genebank showed 99-100% similarity. Also, toxigenic gene typing indicated that some of the A. flavus were toxigenic with the detection of the following genes Nor-1, ver-1, OmtB and AflR. Conclusion: The findings of this study showed that toxigenic Aspergilli isolates are present in both poultry feed and feed ingredients from 5 agro-ecological zones in Nigeria and poses great public health issues.

In Brazil, Spigelia anthelmia is an herbaceous weedy plant normally used for the treatment of helminths in humans and animals. Leaves and stems of S. anthelmia showing black spots were observed in cities in the Northeast region of Brazil. Two fungal species were isolated from symptomatic tissues. The preliminary identification of the fungal isolates was based on morphological characteristics. To confirm the identification of the isolates, the ITS region and partial sequences of the ACT and GAPDH genes were amplified and sequenced. By using morphological and phylogenetic analyses of multilocus genes, the fungi were identified as Colletotrichum karstii and Colletotrichum siamense. All isolates were pathogenic to S. anthelmia seedlings. To the best of our knowledge, this is the first report of C. karstii and C. siamense causing anthracnose on S. anthelmia in the world.

The present study was aimed to characterize the Trichoderma species prevailing in the major pulse growing areas of India. The Trichoderma isolates (54) were collected from the rhizospheric soils of chickpea, pigeonpea and rice-pulse cropping system in Madhya Pradesh, Uttar Pradesh and Maharashtra states of India. The isolates were characterized using their culture-morphological and molecular characteristics. Compact mycelial growth of Trichoderma spp. was seen in PDA medium, less dense growth in malt extract agar and powdery growth in oatmeal agar. Colony colour in media was generally green to dark green and occasionally yellowish green to dark green with white tinge; and the pigmentation in the reverse side varied from white, yellowish or faint yellow. Phialides were ampulliform to sub-globose or lageniform divergent or crowded whorls of 2-5, the middle of phialides were markedly swollen, conidia were subglobose to ovoid, the conidiophores highly branched having intercalary or terminal chlamydospores of sub-globose to ellipsoid or pyriform shape. Based on PCR amplification of ITS region sequences 44 isolates were characterized at molecular level. The partial ITS sequences were analyzed using MegaBLAST with NCBI GenBank database to confirm their identity. The study revealed the presence of 5 species of Trichoderma viz., T. viride, T. asperelloides, T. koningiopsis, T. asperellum and T. harzianum.

The paper presents the results of phytopathogenic fungi determination in bare-root forest nurseries of Middle Siberia. Genetic analysis of pathogenic microflora of Pinus sylvestris L., Pinus sibirica Du Tour and Picea obovata Ledeb. seedlings allowed identification of 17 genera of micromycetes: Phoma Sacc., Lophodermium Chevall., Sclerophoma Hohn. (teleomorph Sydowia Bres.), Cladosporium Link, Alternaria Nees, Typhula (Pers.) Fr. etc. Most frequently detected fungi represented genera Phoma (23.7 %) and Lophodermium (23.6 %). Pinus sylvestris L. seedlings harboured the highest diversity of fungal taxa. Seven genera of microscopic fungi (Phoma sp., Didymella sp., Alternaria sp., Cladosporium sp., Lophodermium sp., Gremmenia sp., Sclerophoma sp.) were detected in all studied forest zones: taiga, forest-steppe and Southern-Siberian mountain. The obtained results demonstrate the usefulness of DNA analysis for the identification of phytopathogenic fungi in forest nurseries of Middle Siberia with several implications for increasing the efficacy of forest management.

Fungal diseases are one of the major factors responsible for severe agricultural losses with a deep impact to the economy. An early detection of these diseases, preferably at the agriculture site itself could be useful so that appropriate counter measures can be taken against the pathogens. In our paper, we have focused on analyzing and characterizing phytopathogens. Foldscope—an origami-based paper microscope—was used for visualization of diseased leaves, fruits, vegetable parts, pathogenic fungal pure cultures, etc. We characterized fungal isolates acquired from various vegetable, fruit, and ornamental plants. Our main objective was to evaluate whether the paper microscope could be used as a preliminary tool for onsite observation of fungal diseases in crops, so that immediate measures can be taken by farmers against fungal infestation prevailing in their agricultural fields. This will thus ensure reduction in crop and revenue losses for the farmers. Ours is the first report wherein phytopathogenic fungi from broccoli, gerbera, guava, okra, and orange as host plants have been successfully identified with the Foldscope as a tool and studied to the extent where cell wall–degrading enzymes, pathogenicity, and ITS region sequencing have been established for these fungi.

Western Ghats is one of the hottest hot spot in the world. Cleisostoma tenuifolium is an epiphytic orchid from the heart of this region. We have isolated and identified four endophytic fungi from this plant. Characterization of these fungi were done by analyzing the sequences. Genome were quantified and sequenced. Sequence similarity matrix were prepared by comparing the ITS region sequence of obtained fungi with its closely related fungal species from NCBI database. Phylogenetic tree were also constructed for further comparing the homological features of these fungi. From the study, the obtained fungi were Aspergillus flavus MT464452, Cladosporium xanthochromaticum MT464453, Colletotrichum cymbidicola MT464454, Aspergillus aculeatus MT464455. The wide variety of endophytic fungi isolated from the plant C. tenuifolium that belongs to Orchidaceae family further highlights the importance of orchids as reservoirs of fungal biodiversity and its conservation.

En la Sierra Nevada de Santa Marta los cultivos de cacao están conformados mayoritariamente por cultivares híbridos comerciales y, aunque se encuentran cacaos nativos, estos son poco cultivados. Dada la necesidad de verificar si estos cultivares de cacao encontrados en la Sierra pertenecen al grupo genético tipo Criollo, se realizó una caracterización fenotípica y genotípica de cacaos del municipio Dibulla, La Guajira. Para esto, se muestrearon 11 cultivares en Mingueo. Los rasgos fenotípicos se evaluaron empleando descriptores UPOV para cacao. Los parámetros cualitativos y cuantitativos se cotejaron por análisis de conglomerado y análisis de componentes principales (ACP), y las variables cuantitativas se compararon a través de la prueba no paramétrica test de Mann-Whitney. Para evaluar las relaciones genéticas, se estandarizaron protocolos de biología molecular y se secuenció la región ITS. A partir de las secuencias, se realizaron agrupamientos por métodos de distancia y filogenéticos. Finalmente, se encontraron diferencias significativas entre las semillas (p = 0,01), y resalta la coloración blanca del cotiledón de los criollos en contraste con la coloración púrpura oscura de los híbridos. Asimismo, los análisis de conglomerados, ACP y los análisis de secuencias demostraron diferencias entre el grupo de los cacaos nativos y los híbridos comerciales cultivados; además, los cacaos nativos se emparentan con el grupo de cacao tipo Criollo.

Chaetomium species are saprophytic fungi, havethe ability to degrade cellulose with hydrolytic enzymes. Hydrolytic enzymes are a cluster of enzymes, which have the flexibility of breaking down complicated molecules into smaller molecules. The objective of this study was to clarify the genetic diversity among the isolated Chaetomium population by using PCR-based methods, along with the study of their cellulolytic activities. Chaetomium spp. were isolated from twenty-eight samples of Cuminum cyminum and Pimpinella anisum, and identified morphologically and molecularly by ITS1 and ITS4 primers. The sequencing indicated that all the isolates of Chaetomium had a 99% sequence identity with Chaetomium globosum sequences from GenBank. Molecular techniques with Internal Transcribed Spacer (ITS) region sequencing and specific genes random primers polymerase chain reaction (SGRP-PCR) showed the existence of high DNA polymorphism of Chaetomium globosum. All isolates were tested for their ability of cellulases production. Nine of the thirteen isolates of Chaetomium globosumcould produce cellulases. There are variations between C. globosum isolates and the source of the isolates and their habitat do not correlate to their ability to secrete cellulases.

This study evaluates the morphological concept of Crepidotus variabilis and similar taxa based on basidiospore and cheilocystidia characters. A phylogenetic analysis of the ITS region clearly distinguished C. variabilis var. variabilis from the two other distinct taxa of the same lineage, C. variabilis var. trichocystis and C. neotrichocystis. We also recognised one additional morphologically similar and undescribed species that belongs to the C. variabilis lineage. All four taxa can also be recognised morphologically using basidiospore and cheilocystidia characters. Our detailed statistical comparison of these characters improves the knowledge of their variability and specifies more precisely the morphological circumscription of the taxa within this group.

Objective: In the present investigation, Trichoderma spp., isolated from rhizospheric soil, has been identified by Internal Transcribed Spacer (ITS) region sequencing technique and its antagonistic activity was evaluated against A. niger. Methods: The sequencing analysis was done with its ITS1 region of the rRNA gene. Using the ITS1 amplified products for all isolated fungi, a bi-directional DNA sequencing was done with high quality bases (>98% - 100%). Antagonistic activity was done using dual culture technique. Results: All of the ITS1 nucleotide sequences obtained in this study matched 97% - 100% with the published sequence of Trichoderma spp. The results confirmed the strains as T. asperellum and T. viride with gene bank accession no. (ZTa); MK937669 and (ZTv); MK503705, respectively. When phylogenetic analysis was done for the isolates, the optimal tree with the sum of branch length = 0.69585023 and 0.10077756 for T. asperellum and T. viride, respectively, was observed. There were a total of 678 and 767 for T. asperellum and T. viride positions in the final dataset, respectively. Antagonistic activity was done for the isolated strains of Trichoderma spp. against A. niger, and it was found that T. asperellum showed maximum antagonistic activity (79.33±7.09%). Conclusion: The findings prolong the genome availability for relative investigations pointing out phenotypic variances to compare with Trichoderma genetic diversity. The present investigation delivered the Bases of future studies for better knowledge in understanding the complicated connections of Trichoderma spp. to be used as an effective biocontrol agent.

Crocus asymmetricus (Iridaceae) is described as a new species endemic to the southern part of the Anatolian diagonal in Turkey. It is phylogenetically related to C. vitellinus and morphologically to C. antalyensis, but differs from these species in showing an asymmetric corm and a single point of root emergence. Both C. antalyensis and C. asymmetricus are illustrated and compared in this paper. A Bayesian phylogenetic tree of the nuclear rDNA ITS region confirms the affiliation of C. asymmetricus to C. ser. Flavi, and its close relationship to C. vitellinus. A new identification key to the species of C. ser. Flavi occurring in Turkey is also presented.

Saksenaea vasiformis is a species of the order Mucorales rarely reported as a cause of human mucormycosis. We report an unusual case of S. vasiformis otitis occurring in a diabetic woman after penetration of an insect in the right ear. Direct microscopic examination of the clinical sample showed hyaline and non septate hyphae belonging to the order Mucorales. Fungal identification was performed by sequencing the ITS region of the rDNA. To our knowledge, this is the first report of S. vasiformis infection in Tunisia. Keywords: Mucormycosis, Malignant otitis externa, Saksenea vasiformis

A total of 21 yeast isolates were recovered as part of a research project on biodiversity of yeasts in traditional dairy products in Alborz province, Iran. Standard protocols were used to carry out phenotypic, biochemical, physiological characterization and the phylogenetic analysis of combined the D1/D2 domain of the large ribosomal subunit (26S or LSU) and ITS region sequences. Five strains represented a potential new ascomycetous yeast species. Ascospore formation was not observed in these strains, and they did not ferment the examined carbon sources. Phylogenetic analysis placed these isolates in a well-supported sub-clade in the genus Saccharomycopsis. Here, we describe this novel yeast as Saccharomycopsis oxydans sp. nov.

Although many primers have been proposed for arbuscular mycorrhizal fungi (AMF)–specific sequencing, only a few studies have evaluated suitable primer pairs for MiSeq analysis. Here, we analyzed AMF community compositions using several primer pairs and evaluated which pair can detect a more diverse AMF community. Three AMF-specific primer pairs targeting the 18S and 28S rRNA genes and a fungal universal primer pair targeting the ITS region were examined using soil DNA as the template. The effect of nested PCR was also evaluated. Only a few AMF sequences were obtained from a fungal universal primer targeting the ITS region. On the other hand, use of AMF-specific primers improved AMF detection rates (35–98%). Particularly, almost all of the sequences obtained with the nested PCR were AMF-related sequences. However, the AMF community compositions were differentiated among the primer pairs. In addition, the number of AMF taxa and diversity decreased with the nested PCR, suggesting that it affected AMF community analysis.

The aim of this study is to identify fungal samples from Ankara (Turkey). Identification of the samples was made according to conventional and molecular methods (ITS region of the rDNA). Based on the high sequence similarity of the new record (hereafter will be referred to as ‘Ank Akata & Altuntaş 166’) with Calocybe persicolor (Fr.) Singer, the relevant agaric is considered to be C. persicolor. This result is also supported by the morphological data and proves the presence and distribution of this species in Turkey.

Ballast water is a vector for global translocation of microorganisms, and should be monitored to protect human and environmental health. This study utilizes high throughput sequencing (HTS) and machine learning to examine the bacterial and fungal microbiomes of ballast water to identify associations between 16S and 18S rRNA genes and the fungal ITS region. These sequencing regions were examined using the SILVA v132 and UNITE reference databases. The highest correlation was found between the communities in Silva_16S and UNITE_ITS (0.74). There was a higher proportion of positive inter-kingdom correlations than positive intra-kingdom interactions (p = 0.032). Understanding the reasons for this difference requires additional research under more controlled conditions. Finally, a machine learning model was used to examine the classification accuracy when using each sequencing region and reference database to identify ballast residence time and ballast sample location. There was significantly higher accuracy using SILVA (0.843) compared to UNITE (0.614) (p

Background Spodoptera litura Fabricius (Lepidoptera: Noctuidae) is an important pest causing severe damage to many cultivating plants such as corn worldwide, including Indonesia. This study was performed to obtain and identify entomopathogenic fungi (EPF) of S. litura collected from corn fields in 4 corn producing regions of Lampung, Indonesia, as well as to investigate the damage caused by this pest on corn in Lampung Province. Results Three corn fields in each region were selected for collecting soil samples. Soil samples were collected from 5 corn plant rhizospheres, at each field in six months of survey. Ten larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae) were laid on each soil sample as a bait, covered with a filter paper and incubated at room temperature. The emerging fungi from T. molitor cadaver were transferred onto Potato Dextrose Agar (PDA) medium and incubated for 7 days at room temperature. Pathogenicity test was determined against 3rd instar of S. litura larvae. Identification was performed based on the sequence of Internal Transcribed Spacer (ITS) Region. Observations on the corn damage caused by S. litura were conducted at all corn producing areas in Lampung. Twelve fungal isolates were obtained causing 0–75% of mortality of S. litura. Four fungal isolates (NKPT, SKHJ, SDHJ and RAHJ), which caused mortality more than 20%, were further identified. One isolate (NKPT) was confirmed as Beauveria bassiana and the other 3 isolates (SKHJ, SDHJ and RAHJ) were Aspergillus oryzae. S. litura generally caused slight damages to the corn which was found in every observation year performed during 2010–2019. Medium plant damage was observed in 2010–2012 and 2018–2019, severe damage was found in 2011 and crop failure was recorded in 2018. Conclusions Aspergillus oryzae and B. bassiana were the EPF recorded infecting S. litura in corn in Lampung Province. This was the first report on the isolates of A. oryzae as EPF of S. litura in Indonesia. Slight damages with S. litura were always recorded in every observation year but not for those of medium and severe damages and crop failure.

Schistidium foraminis-martini is described as a new species from the Austrian and Swiss Alps. The taxon is characterized by a warm brown coloration, ovate, concave and muticous leaves with rounded apices, ellipsoid capsules, constricted at mouth, the presence of stomata and a reduced, truncate peristome. Based on sequences of the ITS region the new species is phylogenetically rather isolated, exhibiting the closest relationship to S. agassizii which shares the muticous leaves but differs in having narrowly lingulate-lanceolate, almost flat leaves, cyathiform capsules and a perfect peristome. Schistidium foraminis-martini grows on moist calcareous rocks in the alpine and nival belt.

Hevea brasiliensis es la especie de mayor importancia económica del género Hevea, de la cual se obtiene 99 % de la producción mundial de hule natural. En México, durante 2018 se registró una producción total de 75 922 toneladas que se obtuvieron de 28 172 ha plantadas en cinco estados: Chiapas, Oaxaca, Tabasco, Veracruz y Puebla. De ellos, Veracruz ocupa el primer lugar, con 66 % de la producción. Las enfermedades en el árbol del hule afectan la óptima producción de látex en todo el mundo. En 2018, se detectó la incidencia en vivero de una pudrición en tocones de hule injertados con el clon IAN-873 en el municipio Martínez de la Torre; por lo anterior, el objetivo de esta investigación fue identificar el agente asociado a la pudrición de tocones en dicho lugar. Se cultivaron muestras de tejido leñoso con pudrición y micelio en medio Extracto de Malta Agar (MEA) y se incubaron a 28 ± 1 °C por 72 h; se desarrollaron colonias algodonosas, con micelio color blanco-crema. Con base en la morfología del cultivo en el medio MEA y el análisis filogenético cono la secuencia de la región ITS, obtenida mediante la amplificación con los primers ITS5/ITS4 del aislamiento, se identificó a Schizophyllum commune como el hongo asociado a la pudrición de los tocones de H. brasiliensis, lo que corresponde al primer registro para México.

Camels were formerly considered resistant to most of the diseases commonly affecting livestock. Recently, camels were found to be susceptible to a large number of pathogenic agents. The aim of this work is to identify the causative agent of meat lesion encountered in one humped male camels carcasses at abattoir in Matrouh governorate, Egypt. Out of 50 carcass of slaughtered male camel, 12 meat lesions were macroscopically found. Blackening and necrotic area lesions were exposed to routine mycological examination. This revealed the presence of Cryptococcus in one lesion. Cryptococcus infection is an opportunistic infection that occurs primarily among patients who are immunosuppressed or getting access through accidental penetration of skin barriers. The isolated fungi was tested by PCR amplification of the ITS region and sequencing of 28S ribosomal RNA gene. The alignment of the fungal isolates 28S region sequences revealed 94% identity with Cryptococcus cerealis in the gene bank library. For the best of our knowledge, this is the first isolation of C. cerealis from camel meat lesion in Egypt.

Background Enterocytozoon bieneusi has been increasingly reported to infect domestic animals and humans, with human infections primarily reported as zoonotic in origin. The aim of the present study was to determine the presence and genotype of E. bieneusi in humans and domestic animals in central Thailand by testing stool samples of 200 apparently healthy humans, 73 goats, 60 cattle and 65 pigs using nested-PCR/ sequence analysis based on the ITS region of SSU rRNA genes. Results E. bieneusi tested positive in 2 (1%) of the 200 stool samples collected from humans and 56 (28.3%) of the 198 stool samples collected from domestic animals. The highest prevalence of E. bieneusi was observed in pigs (39/65, 60%), followed by goats (14/73, 19.2%) and cattle (3/60, 5%). Seven novel E. bieneusi genotypes were identified, which were named GoatAYE1–4 and PigAYE1–3 and clustered in either zoonotic Group 1 or Group 2. Moreover, eleven previously described E. bieneusi genotypes were also identified (O, D, H, SX1, CHC8, CHG3, CS-10, SHZC1, LW1, WildBoar5, and EbpC). All novel genotypes exhibited zoonotic potential from a phylogenetic analysis of ITS region. Conclusion Our data showed that the prevalence of E. bieneusi is low in apparently healthy individuals and higher in pigs than cattle and goats. This study provides baseline data useful for controlling and preventing E. bieneusi infection in farm communities, where pigs and goats appear to be the major reservoir of E. bieneusi. The results of our study support the view that E. bieneusi is a zoonotic pathogen that should be considered a potential public health threat.

The results of morphological and molecular genetic analysis confirming the hybrid nature of Salix × zhataica Efimova, Shurduk et Ahti are presented. In particular, according to the complex of phenotypic features (on the basis of discriminant analysis), the hybrid, which resulted, apparently, from interspecific hybridization of S. brachypoda (Trautv. et C.A. Mey.) Kom. and S. pyrolifolia Ledeb., is closer to S. brachypoda, indirectly indicating that this “parent” can be maternal. The polymorphism of molecular-genetic markers such as nuclear microsatellite loci and nucleotide sequences of the ITS region was analyzed in order to verify the hybrid and its parent species, S. brachypoda (Trautv. et C.A. Mey.) Kom. and S. pyrolifolia Ledeb. For instance, in the ITS region, two single nucleotide transitions were found, which indicate that the hybrid borrowed the corresponding nucleotides from both parent species. The given data indicate that S. × zhataica is indeed a hybrid between S. brachypoda and S. pyrolifolia.

Rice sheath blight disease caused by Rhizoctonia solani is considered a distractive soil-borne disease of rice production worldwide. The study aimed to determine the causal organism of sheath bligh...

The contamination of soil with petrogenic hydrocarbons represents a significant human health concern as a result of the carcinogenic and mutagenic properties of a number of compounds commonly found in petroleum products such as polycyclic aromatic hydrocarbons (PAHs). Although it is known that soil fungal communities play an important role in the degradation of highly branched petroleum hydrocarbons and high molecular weight PAHs, little information is available regarding the impact of petrogenic contamination on the structure and diversity of the fungal community and the impact of soil type on the fungal community response. Here, the aim of this study was to evaluate the impact of petrogenic contamination on fungal communities in two soil types. In sandy soils, the most contaminated test pit had the highest fungal diversity. Fungal profiles were dominated by species from the class Eurotiomycetes and included the well-known hydrocarbon-degrading species of Aspergillus and Eurotiales. The dominance of these species did not change with contamination concentration,suggesting a level of adaptability to multiple carbon sources. There appeared to be no correlation between fungal species diversity and contaminant concentration in the clayey soil. Similar dominant fungal species were identified in the clay and sandy soils, all of which were part of the phylum Ascomycota. The clayey soils had a higher species diversity and range-weighted richness compared to sandy soils, which may be a result of the pore connectivity theory i.e. as a result of low water connectivity in soils the formation of diverse communities is promoted through creation of microhabitats. Refereed/Peer-reviewed

Jewels of Opar (T. paniculatum) belongs to Talinum genus, Portulacaceae family which contains secondary metabolites such as phytosterols, saponins, flavonoids, tannins, steroids. These organic compounds have anti-viral effects and are very effective on Herpes’ disease and skin infections. Besides, Jewels of Opar can also be used as a supporting medicine for Parkinson’s disease, heart disease and for lowering blood cholesterols. Currently, the identification of T. paniculatum has been mainly based on morphological analysis. However, this method often encounters obstacles when T. paniculatum has been completely or partially processed. In this work, we present the results of morphological characteristics, taxonomy and sequences characterisation of ITS region and rpoC1, rpoB genes of T. paniculatum in Northern provinces of Vietnam. Tuberous roots of T. paniculatum are cylindrical with many small roots. The stems are upright and divided into several branches. The stems are upright and divided into several branches. The leaves are staggered, generally oval, ovate-oblong, or egg back shaped; thick, glossy with wavy veins, without hairs. The flowers of the plants have five reddish purple wings, two sepals, more than ten stamens, and a spherical ovary. The fruits are small, and the ripe fruit is ash gray in color. The seeds are very small, slightly flat, and black. Internal transcribed spacer (ITS) region and two partial sequences of rpoC1 and rpoB genes isolated from T. paniculatum plants are 643 bp, 595 bp and 518 bp in length, respectively. Based on the combination of the characteristics of morphology and nucleotide sequences of ITS region, rpoC1 and rpoB genes, the Jewels of Opar samples collected in some northern provinces of Vietnam were determined to belong to T. paniculatum species, Talinum genus, Portulacaceae family. Characteristics of sequences of ITS region and rpoC1, rpoB genes are valuable for exploiting DNA barcodes to identify T. paniculatum in Vietnam.

In this study, three chloroplast genes (matK, rbcL and rpoC1) and one nuclear gene (ITS) were used to assess the nucleotide diversity for nine individuals of Canarium nigrum species in Bac Giang, Hoa Binh and Phu Tho provinces (three individuals from each province). The nucleotide sequence of the four regions (ITS, matK, rbcL and rpoC1) of C. nigrum were determined with the results showing their size to be 696 bp, 798 bp, 702 bp and 522 bp, respectively. Results of nucleotide sequence comparison for the studied samples exhibited 100% similarity for all the four gene regions. Sequence comparison with other species in the Canarium genus available in the GenBank revealed that the nucleotide diversity level (π) was the highest for the ITS gene (0.02), followed by matK (0.007), and the lowest for rbcL (0.003). The phylogenetic tree of C. nigrum with the species in Canarium genus indicated that the separation of species was the clearest for the rpoC1 gene, followed by rbcL, matK and ITS gene, with the bootrap values obtained from the branching nodes of each species ranging from 98 to 99%, 35 to 67%, 65 to 98% and 98 to 99%, respectively. The species C. nigrum had the closest nucleotide similarity to C. tramdenum (KP093200) and C. album (KP093198) for the ITS gene and to species C. subulatum (KR530509), C. acutifolium (KR530512) for matK gene region. These results suggests the rpoC1 gene region could be used as barcode for the species in genus Canarium.

El mildiu velloso causado por Peronospora sparsa Berkeley es una de las enfermedades más importantes del rosal en México, causando pérdidas de 100%. Su manejo se basa con fungicidas en continuas aplicaciones que tienden a generar poblaciones resistentes. La búsqueda de alternativas es indispensable. En el presente trabajo se confirmó la identidad morfométrica y molecular del agente asociado con el mildiu del rosal, se evaluó la eficacia del fosfito de potasio (K3PO3), quitosano, silicio y mefenoxam para el manejo de la enfermedad y se determinó su efecto en la longitud y diámetro de tallo floral. El estudio se realizó en verano y otoño de 2013 en Tenancingo, Estado de México. La caracterización morfométrica se realizó bajo microscopio compuesto y electrónico de barrido. Para la caracterización molecular, se amplificó el ADN ribosomal de la región ITS con los primers PS3 y PS1. Los tratamientos fueron: fosfito de potasio (K3PO3), quitosano, silicio, mefenoxam y testigo y se aplicaron a intervalos semanales. El diseño experimental fue de bloques al azar y la comparación de medias fue por Tukey (α= 0.05). Datos morfométricos y moleculares correspondieron a Peronospora sparsa. El K3PO3 y silicio redujeron la incidencia y severidad con respecto al testigo. El tratamiento con K3PO3 mostró incrementos de 24.8 y 97.5% en la longitud de tallos con diámetros de 7.5 y 6.2 mm en verano y otoño respectivamente, comparando con el testigo. Así, el K3PO3 y silicio pueden ser alternativas en el manejo del mildiu velloso del rosal en condiciones de invernadero.

Soil management practices have the potential to modify the diversity and function of microbes in agricultural fields. The aim of this study was to investigate bacterial and fungal diversity in a 15 yr wheat–pea rotation tillage experiment. The treatments included conventional tillage with stubble removed, no-till with stubble removed, no-till with stubble retained (NTS), and conventional tillage with stubble incorporated. Illumina high-throughput sequencing platform was employed to sequence bacteria 16S rRNA (V3V4) and fungi internal transcribed spacer (ITS2) region genes in 0–10 cm and 10–30 cm of soil sampled. The dominant bacterial and fungal phyla identified at 97% similarity cutoff across both depths of treatments were Proteobacteria (26.3%), Actinobacteria (25.1%), Acidobacteria (15.0%), Gemmatimonadetes (8.8%), Ascomycota (85.8%), and Basidiomycota (8.0%). NTS had significantly (p 3-N, and NH4-N at 0–10 cm depth. Tillage and stubble effects had a significant correlation with some phyla such as Proteobacteria, Actinobacteria, Gemmatimonadetes, JL-ETNP-Z39, Nitrospirae, Chloroflexi, Firmicutes, and other identified and unidentified minor microbial phyla. No-till and residue retention practices influenced fungal and bacterial species diversity through improved soil chemical properties, which have potential to affect the habitat and activity of soil microbes. Therefore, no-till and stubble retention could improve soil quality and promote sustainable agriculture in the rainfed Loess Plateau.

A comprehensive study of critical South Asian species Seseli diffusum (Umbelliferae—Apioideae—Apieae) was carried out to determine the most suitable taxonomic position of the species, based on morphology (carpoanatomy, micromorphology of the fruit indumentum and some vegetative characters) and molecular data from nrDNA ITS region sequencing. Comparison with certain other species of Seseli and several provisionally related Apieae genera showed a close affinity of S. diffusum to the genus Psammogeton and more distant relationships with some species of Trachyspermum, clearly polyphyletic genus. No species of Seseli, even in broad sense, was found to be closely related to S. diffusum. It was proposed to transfer this species into Psammogeton, while separating a new section, Pseudoseseli, within the latter.

Noedoost F, Vaezi J, Nikzat Siahkolaee S. 2021. Taxonomic investigation of the Xanthium strumarium L. complex (Asteraceae) distributed in Iran inferred from morphological, palynological and molecular data. Biodiversitas 22: 1961-1974. Unclear species boundaries due to high morphological similarities lead to different opinions on Xanthium classification. To solve this problem, we conducted investigation on morphological, palynological and molecular (ITS sequencing) of three species of Xanthium (i.e., X. strumarium, X. sibiricum, and X. brasilicum) forming Xanthium strumarium complex in Iran. We randomly collected 110 plant individuals of the Xanthium strumarium complex as well as X. spinosum from 30 geographical populations in eight provinces of Iran. Pollen grains of 19 populations were studied by scanning electron microscope (SEM). Results obtained from the analysis of morphological characters showed that the three species, X. strumarium, X. brasilicum and X. sibiricum were placed nearly together specially X. strumarium and X. brasilicum are obviously overlapped. Palynological results indicated the pollen grains of section Acanthoxanthium (i.e., X. spinosum) and Xanthium (i.e., X. strumarium, X. sibiricum, and X. brasilicum) are clearly differing by their LM and SEM characters. However, no significant difference found between the pollen of X. strumarium and X. brasilicum. The Bayesian phylogenetic tree of the ITS region of rDNA sequences revealed that X. strumarium, X. sibiricum and X. brasilicum could not be discriminated as independent species. Overall, the morphological, palynological and molecular data provided evidence to synonymize X. brasilicum and X. strumarium with X. orientale and X. sibiricum with X. strumarium.

Khotimah S, Suharjono, Ardyati T, Nuraini Y. 2021. The potential of cellulose-degrading fungi at various peat maturities in Teluk Bakung Peat Area, Kubu Raya District, Indonesia. Biodiversitas 22: 1981-1990. Fungi play important roles in degrading cellulose in the maturation process of peat. This study aimed to evaluate the potential of cellulose-degrading fungi at various peat maturities in the forest. Peat soil samples were collected from Teluk Bakung Village, Kubu Raya District, West Kalimantan using a random sampling method. Peat soil samples were obtained at six sampling sites in forests and palm oil plantations. The fungi were cultured by the pour plate method on 1% CMC media. The cellulose-degrading fungi were determined based on a clear zone formation around the colony in CMC agar media. The cellulolytic fungal population in the fibric, hemic, and sapric peats of the secondary forest were 3.2 × 103, 4.6 × 103, and 5.6 × 103 CFU/g, respectively, whereas at fibric, hemic, and sapric of the shrubs were 5.6 x 104, 3.9 × 105 and 4.7 × 104 CFU/g, respectively. Among the 25 cellulolytic fungal isolates, 5 isolates (HS1.5, HS3.4.1, HB2.2, SS2.4.1, and SB2.5.1) showed high average clear zone diameter (21-28 mm). The HS3.4.1 and HS1.5 isolates had cellulase enzyme activity of 132.16 and 127.3 U/mL, respectively. These isolates can grow and degrade cellulose at 30°C to 35°C and pH of 2, 3, 4, and 5. The HS3.4.1 and HS1.5 isolate were closely related to Penicillium citrinum based on the phylogenetic tree analysis.

Background Banana fruit scarring beetle (BFSB), Basilepta subcostata (Jac.) (Chrysomelidae:Coleoptera), is an important insect pest feeds on leaf and fingers, which affects the cosmetic value of the fruit. The pest is distributed in Assam, Bihar, West Bengal, Chhattisgarh, and North-eastern Hill regions of India. Results The pest is currently managed by foliar spray with insecticides. In order to identify eco-friendly control of the pest, attempts were made to isolate microbial agent and evaluate their potential to control the pest. A total of 27 entomopathogenic fugal isolates were obtained from Odoiporus longicollis (Oliver), Cosmopolites sordidus (Germar), Basilepta subcostata (Jac), and other insect Galleria mellonella (Fabr). Based on colony morphology, the collected fungal isolates were identified as Metarhizium spp. (17) and Beauveria spp. (10). Through ITS sequencing, the fungal isolates were further characterized at species level as B. bassiana (8), B. brongniartii (2), M. anisopliae (8), M. robertsii (6), M. guizhouense (2), and M. pinghaense (1). Their sequences were submitted in GenBank and obtained accession numbers. Among 27 isolates tested against B. subcostata under laboratory conditions, 3 isolates (M. anisopliae NRCBEPF-36, M. pinghaense NRCBEPF-7 and B. brongniartii NRCBEPF-27) recorded 100% beetle mortality, followed by 11 isolates with 95–99% and 13 isolates with 88–93% within 8 days of treatment. Conclusion This study highlights the two native North East India isolates B. brongniartii NRCBEPF-27 (MT151781) and M. anisopliae NRCBEPF-36 (MT140308) showed the significance to use as potential biocontrol agents against banana fruit scarring beetle B. subcostata. Further experiments under field conditions are required to evaluate their biological control efficacy against the pest.

The advent of high-throughput sequencing methods allowed researchers to fully characterize microbial community in environmental samples, which is crucial to better understand their health effects upon exposures. In our study, we investigated bacterial and fungal community in indoor and outdoor air of nine classrooms in three elementary schools in Seoul, Korea. The extracted bacterial 16S rRNA gene and fungal ITS regions were sequenced, and their taxa were identified. Quantitative polymerase chain reaction for total bacteria DNA was also performed. The bacterial community was richer in outdoor air than classroom air, whereas fungal diversity was similar indoors and outdoors. Bacteria such as Enhydrobacter, Micrococcus, and Staphylococcus that are generally found in human skin, mucous membrane, and intestine were found in great abundance. For fungi, Cladosporium, Clitocybe, and Daedaleopsis were the most abundant genera in classroom air and mostly related to outdoor plants. Bacterial community composition in classroom air was similar among all classrooms but differed from that in outdoor air. However, indoor and outdoor fungal community compositions were similar for the same school but different among schools. Our study indicated the main source of airborne bacteria in classrooms was likely human occupants; however, classroom airborne fungi most likely originated from outdoors.

In this study, faecal samples of four American Staffordshire terrier dogs (used for illegal fighting) were analysed by DNA extraction, molecular-phylogenetic and parasitological methods, in order to examine the occurrence of protozoan, apicomplexan parasites. In one sample, the DNA of Sarcocystis morae was shown to be present. This species was identified based on 100% identity with already reported sequences of S. morae from cervids in Lithuania and Spain. The result was also confirmed by phylogenetic analysis. The sporocysts of the canine S. morae isolate measured 14.95 × 9.75 μm on average. This is the first molecular evidence in support of the final host role of domestic dogs in the life cycle of S. morae. The most likely source of the infection was raw meat given to the examined dog to increase its physical achievement. In conclusion, under similar circumstances dogs may participate in the life cycle of S. morae in a ‘natural way’, shedding sporocysts/oocysts when used for hunting or taken to walks in forested areas.

The continuously increasing concentrations of atmospheric CO2 is predicted to affect biological processes at many levels of organisms. Yet, no study exists in the literature attempting to describe that the elevated atmospheric CO2 (eCO2) concentration may cause an evolutionary response on nucleotide sequences of ribosomal DNA of Myzus persicae [(Sulzer) (Hemiptera: Aphididae)]. Here, we provide a preliminary study to understand how the insect ribosomal DNA sequences are influenced under the elevated CO2 levels after several generations. Four M. persicae populations were established for 35 days under ambient CO2 (a CO2) (400 ppm), e CO2 (600 ppm), e CO2 (800 ppm) and e CO2 (1000 ppm) at 29ºC in moisture-controlled greenhouse chambers. Intraspecific variation of M. periscae was assessed by the sequencing and analyzing the internal transcribed spacer (ITS) region of Myzus persicae (Sulzer) under elevated atmospheric CO2 pressure. Based on our results, the phylogenetic analysis of ITS sequences differentiated the individuals grown at 800 ppm CO2 level. The alignment of ITS sequences of all specimens revealed several single-nucleotide substitutions on the nucleotide sequence of M. persicae samples grown at 800 ppm CO2 level. Overall results show that the elevated atmospheric CO2 levels could be a powerful evolutionary force than expected on M. persicae reared on eggplants

The rarest species of relict ironwoodC. caucasicaWilld. is naturally occurred on the Ile-Alatau Mountains but very little is known about genetic diversity and distribution and size of its populations in Kazakhstan. In during the sampling expedition were found two additional plant populations in Dzungarian and Kyrgyz Alatau Ranges. The objective of this work was targeted towards sequencing ITS region ofC. caucasicaand compare the obtained nucleotide sequence with available data on NCBI GeneBank for confident species identification. The identity ofC. caucasicasequence and availableC. australisandC. bungeanasequences was 93.87% and less. It could be associated with absence sequences producing significant alignments with the studied ironwood sequence and important deposited sequences of GenBank lacking Latin binominals is from environmental samples. Clarifying taxonomical status species and subspecies is difficult by morphological data and molecular markers should be used to correct identifying an endangered species ofC. caucasicagrowing in the east-southern and the south regions of Kazakhstan and providing direction to the conservation management of the plant.

Crocus adamioides Kernd. et Pasche, as it is currently known, was originally treated as C. biflorus Mill. subsp. adamii B. Mathew in the flora of Bulgaria by Mathew (1982) and verified by Rukšāns (2017). The taxon was afterwards described as a separate species (Kerndorff et al. 2012), the holotype being collected in Kırklareli Province, European Turkey. The species was for the first time mapped in two floristic regions of Bulgaria. The diagnostic characters based on the general morphology and leaf anatomy were defined from the natural populations of the species and compared to the type specimen and relevant data from the literature. The phylogenetic position of the species was clarified by sequencing of the internal transcribed spacer region (ITS: ITS1 + 5.SsrDNA + ITS2) and comparison of the obtained sequence with those annotated in NCBI. A phylogenetic tree was built using Bayesian phylogeny. Results have shown the highest degree of phylogenetic similarity with C. adamioides from Turkey. The closest relative C. ranjeloviciorum Kernd., Pasche, Harpke et Raca remains in the proximity. Our morphological, anatomical and molecular analyses have revealed that the Bulgarian population shows a peculiar combination of characters specific to C. adamioides.

A better understanding of factors shaping the rhizosphere microbiota is important for sustainable crop production. We hypothesized that the effect of agricultural management on the soil microbiota is reflected in the assemblage of the rhizosphere microbiota with implications for plant performance. We designed a growth chamber experiment growing the model plant lettuce under controlled conditions in soils of a long-term field experiment with contrasting histories of tillage (mouldboard plough vs cultivator tillage), fertilization intensity (intensive standard nitrogen (N) + pesticides/growth regulators vs extensive reduced N without fungicides/growth regulators), and last standing field crop (rapeseed vs winter wheat). High-throughput sequencing of bacterial and archaeal 16S rRNA genes and fungal ITS2 regions amplified from total community DNA showed that these factors shaped the soil and rhizosphere microbiota of lettuce, however, to different extents among the microbial domains. Pseudomonas and Olpidium were identified as major indicators for agricultural management in the rhizosphere of lettuce. Long-term extensive fertilization history of soils resulted in higher lettuce growth and increased expression of genes involved in plant stress responses compared to intensive fertilization. Our work adds to the increasing knowledge on how soil microbiota can be manipulated by agricultural management practices which could be harnessed for sustainable crop production.

Powerful DNA barcodes have been much more difficult to define in plants than in animals. In 2009, the international Consortium for the Barcoding Of Life (CBOL) chose the combination of the chloroplast genes (rbcL + matK) as the proposed official barcode for plants. However, this system has got important limits. First, any barcode system will only be useful if there is a clear barcode gap and if species are monophyletic. Second, chloroplast and mitochondrial (COI gene used for animals) barcodes will not be usable for discriminating hybrid species. Moreover, it was also shown that, using chloroplast regions, maximum species discrimination would be around 70% and very variable among plant groups. This is why many authors have more recently advocated for the addition of the nuclear ITS region to this barcode because it reveals more variations and allows the resolution of hybrid or closely related species. We tested different chloroplast genes (rbcL, matK, psaB, psbC) and the nuclear ITS region in the genus Vanilla, a taxonomically complex group and therefore a good model to test for the efficiency of different barcode systems. We found that the CBOL official barcode system performed relatively poorly in Vanilla (76% species discrimination), and we demonstrate that adding ITS to this barcode system allows to increase resolution (for closely related species and to the subspecies level) and to identify hybrid species. The best species discrimination attained was 96.2% because of one paraphyletic species that could not be resolved.

Simple Summary Yeasts are microorganisms presented naturally in meat products microbiota, which carry out an essential role in the maturation process of fermented sausages. The maturation of this type of product could be better controlled by the addition of a starter culture or a mix of strains that could standardize the process in diverse factories. Moreover, it would be favorable to find cultures that, not only conduct the curing process, but also present an added value such as the meat product protection (antioxidant capability and biocontrol activity). Yeasts also have an important role in the meat product aroma development. Therefore, it would be interesting to select a pleasant-aroma forming strain in order to elaborate a future starter culture for fermented sausage maturation. Abstract The wild yeast community was studied in fermented sausages from pork and game meat (deer and wild boar) during the maturation process from different curing rooms. Although the biotechnological importance of yeasts in the maturation process of pork sausages is known, there is a lack of information for sausage maturation involving game meat. A total of 123 yeasts were isolated and, by amplifying and sequencing of the ITS region, were classified in 14 species. Debaryomyces hansenii, Kazachstania servazzii, and Wickerhamomyces anomalus were isolated in both pork and game samples. The PCR-RAPD technique differentiated between 26 and 18 strains from pork and game meat sausages, respectively. The physicochemical parameters and their relationship with the yeast community were also studied. The antioxidant and anti-lipid peroxidation capability were analyzed and the 70% and 50% of the tested strains showed these abilities, respectively. Moreover, the biocontrol capability against mycotoxigenic molds was found in 19 strains, but better results were observed in game meat yeasts. On the other hand, almost 30% of strains produce a pleasant olfactory aroma, and volatile compounds associated with the yeast pathway metabolic during the maturation process have been characterized such as esters, aldehydes, fusel alcohols, etc. This study has allowed a better understanding of the biodiversity of this type of food, as well as selecting potential yeast strains for their future use as starters.

Aswan is an ancient city in Egypt; it has many potential issues to be a sustainable and advanced city in its region. On the other hand, it has many environmental problems according to the sustainability criteria such as air and water pollution, noisy, and unsatisfied transportation systems. So, this research aims to solve these problems and tries to keep the environment clean and increase the public satisfaction by promoting new and advanced transportation systems and enhancing the infrastructures in Aswan city without compromising the city heritage. The paper case study is renovating the main city Cornish and the ancient touristic market through studying the existing case and presenting a proposed renovation processes which included new public transportation systems according to public questionnaire results. The proposed renovation process is compared with Curitiba and Masdar city throughout their sustainable urban programs especially public transport and mobility systems. These systems will serve the Aswan people and tourists which reduces the people suffering and preserves the city heritage and touristic places.

Ancylostoma ceylanicum is a zoonotic helminth that is commonly found in domestic dogs and cats throughout Asia but is largely neglected in many countries. This study aimed to confirm the species of hookworm in dogs and soil environments and investigate the evolutionary analyses of A. ceylanicum among Thai and Asian populations. In a total of 299 dog fecal samples and 212 soil samples from 53 temples, the prevalence rates of hookworm infection by microscopic examination were 26.4% (79/299) and 10.4% (22/212) in dog and soil samples, respectively. A PCR-RFLP targeting the ITS region was then utilized to identify the hookworm species. In dogs, A. ceylanicum was the main hookworm species, and the rates of A. ceylanicum and A. caninum infections were 96.6% and 3.5%, respectively. The genetic characterization and diversity indices of the A. ceylanicumcox1 gene among Thai and Asian populations were evaluated. Nine haplotypes were identified from Thai A. ceylanicum, in which the haplotype diversity and the nucleotide diversity were 0.4436 and 0.0036, respectively. The highest nucleotide diversity of Chinese A. ceylanicum populations suggested that it could be the ancestor of the populations. Pairwise fixation indices indicated that Thai A. ceylanicum was closely related to the Malaysian population, suggesting a gene flow between these populations. The temples with hookworm-positive dogs were associated with the presence of hookworm-contaminated soil, as these levels showed an approximately four-fold increase compared with those in temples with hookworm-negative dogs (OR = 4.38, 95% CI: 1.55&ndash, 12.37). Interestingly, the genotypes of A. ceylanicum in the contaminating soil and infecting dogs were identical. Therefore, increased awareness and concern from the wider public communities with regard to the responsibility of temples and municipal offices to provide proper deworming programs to community dogs should be strongly endorsed to reduce the risk of the transmission of this zoonotic disease. In addition, parasitic examination and treatment should be strongly implemented before dogs are imported and exported worldwide.