Molecular Phylogenetic Characterization Research Articles

Characteristics of bast fibers derived from the bark of three paper-mulberry cultivars in Korea

Bast fiber from paper-mulberry (Broussonetia papyrifera) has long been utilized for handmade paper in Asia, serving as a prominent non-wood cellulose material. When restoring paper cultural assets, selecting a similar paper to the original ensures longevity and prevents deterioration issues like warping or tearing. The production of handmade paper involves several processes, including refining, steaming, and bleaching of raw materials. Once formed into sheets, the physical properties of the paper are dictated by these materials and remain unchanged unless the paper is newly made. Handmade paper production is labor-intensive and time-consuming, emphasizing the need for careful raw material selection to match desired characteristics. This study evaluated the dendrological, morphological, anatomical, and chemical compositions of three paper-mulberry wood types cultivated in Korea and inferred their suitability as pulp for papermaking or repair of archival paper specimens. Paper-mulberry wood showed differences in fiber length according to molecular phylogenetic characteristics, but there were no differences in anatomical characteristics owing to the immaturity of the wood. This study investigated the characteristics of the bast fibers from the bark of paper-mulberry trees with different genetic traits to identify favorable factors likely to affect the pulp and papermaking process and paper quality.

Discovery of Gibellula floridensis from Infected Spiders and Analysis of the Surrounding Fungal Entomopathogen Community.

Characterization of fungal spider pathogens lags far behind their insect counterparts. In addition, little to nothing is known concerning the ecological reservoir and/or fungal entomopathogen community surrounding infection sites. Five infected spider cadavers were identified in the neo-tropical climate of north-central Florida, USA, from three of which viable cultures were obtained. Multi-locus molecular phylogenetic and morphological characterization identified one isolate as a new Gibellula species, here named, Gibellula floridensis, and the other isolates highly similar to Parengyodontium album. The fungal entomopathogen community surrounding infected spiders was sampled at different habitats/trophic levels, including soil, leaf litter, leaf, and twig, and analyzed using ITS amplicon sequencing. These data revealed broad but differential distribution of insect-pathogenic fungi between habitats and variation between sites, with members of genera belonging to Metarhizium and Metacordyceps from Clavicipitaceae, Purpureocillium and Polycephalomyces from Ophiocordyceps, and Akanthomyces and Simplicillium from Cordycipitaceae predominating. However, no sequences corresponding to Gibellula or Parengyodontium, even at the genera levels, could be detected. Potential explanations for these findings are discussed. These data highlight novel discovery of fungal spider pathogens and open the broader question regarding the environmental distribution and ecological niches of such host-specific pathogens.

Molecular phylogenetic characterization of L-asparaginase-producing endophytic fungi inhabiting Prunus africana and Periploca linearifolia: Effect of incubation time and pH on enzyme production

The therapeutic use of L-asparaginase derived from bacterial sources has been constrained by various challenges, including toxicity and repression. This has prompted the exploration of alternative sources, particularly eukaryotic microorganisms like fungi, to enhance the safety and effectiveness of the L-asparaginase enzyme. This study aims to investigate the fungal endophytes inhabiting Periploca linearifolia and Prunus africana as a potential source of novel L-asparaginase. Additionally, the study seeks to examine the impact of incubation time and pH on L-asparaginase production. Standard surface sterilization techniques were used for isolation of endophytic fungi. Based on the research findings, 24 % of the fungal endophytes demonstrated a positive reaction for L-ASNase activity and were identified as Colletotrichum sydowii, Fusarium sporotrichioides, Fusarium solani, Cercospora canescens, Penicillium crustosum, Penicillium pancosmium, Phoma sp, Penicillium ubiquetum, septoria sp and Penicillium commune. A significant variation in the production of L-asparaginase based on the time of incubation and pH was observed. Most endophytic fungal isolates exhibit optimal enzyme activity on the 6th day of incubation. However, both Septoria sp and Colletotrichum sydowii recorded the highest L-asparaginase activity on the 9th day of incubation. Fusarium solani demonstrated peak activity of 12.4 ± 1.12 UI/mL on the 12th day of incubation. The optimal pH for L-asparaginase production by fungal endophytes was found to be between 5 and 6. The fungal endophytes isolated from medicinal plants have the potential to serve as sources of novel L-asparaginase. Furthermore, it is evident that fungal endophytes display considerable variation in L-asparaginase production, influenced by the duration of incubation and pH conditions.

Molecular detection and phylogenetic characterization of Rickettsia in ticks collected from leopard tortoise (Geochelone pardalis) in rural Zambia

In sub-Saharan Africa, limited studies have investigated zoonotic pathogens that may be harboured by ticks infesting reptiles such as tortoises. Here, we report the presence of pathogenic Rickettsia in ticks (Amblyomma marmoreum) collected from the leopard tortoise (Geochelone pardalis) in rural Zambia. Using polymerase chain reaction, 56% (49/87) of ticks were positive for the Rickettsia outer membrane protein (ompB) gene. Multi-locus sequence and phylogenetic analysis based on the ompB, ompA, and citrate synthase (gltA) genes showed that the ticks carried R. africae, and other Rickettsia spp. closely related to R.raoultii, R.massiliae, R.tamurae and R.monacensis. Given the proximity between humans, livestock, and wildlife in these habitats, there exists a considerable risk of transmission of zoonotic Rickettsia to human populations in this rural setting. These results call for heightened awareness and further research into the dynamics of tick-borne diseases in regions where humans and animals coexist, particularly in the context of tortoise-associated ticks as vectors. Understanding and addressing these potential disease vectors is crucial for effective public health measures and the prevention of Rickettsia zoonoses.

Morphological and molecular phylogenetic characterization of Sarcocystis kani sp. nov. and other novel, closely related Sarcocystis spp. infecting small mammals and colubrid snakes in Asia

We investigated the morphology and phylogenetic relationships of novel and previously recognized Sarcocystis spp. infecting small mammals and colubrid snakes in Asia. The nuclear 18S rRNA and mitochondrial cox1 of Sarcocystis sp.1 from mangrove snakes (Boiga dendrophila) in Thailand and Sarcocystis sp.2 from a ricefield rat (Rattus argentiventer) in Sumatra were partially sequenced. Sporocysts of Sarcocystis sp.1 induced development of sarcocysts in experimentally infected rats, which showed a unique ultrastructure that was observed previously by S.P. Kan in rats from Malaysia; therefore, we describe this species as Sarcocystis kani sp. nov. Its integration into the 18S rRNA phylogeny of Sarcocystis spp. cycling between small mammals and colubrid snakes helped clarify relationships among the so-called S. zuoi-complex of molecularly cryptic species: Sarcocystis kani sp. nov., S. sp.2, S. attenuati, S. scandentiborneensis, and S. zuoi were all included in this clade. Tree topology was resolved into dichotomies congruent with the morphological disparities between the taxa. However, cox1 gene sequencing (including newly sequenced S. singaporensis and S. zamani) revealed that Sarcocystis kani, S. attenuati, and S. scandentiborneensis were identical suggesting a recent, common ancestry. To identify other distinctive features, lineage-specific molecular patterns within both genes were examined revealing that all 18S rRNA sequences of the S. zuoi – complex possess a unique, 7-nt long motif in helix 38 of domain V7 that was different in S. clethrionomyelaphis which branched off basally from the complex. Three-dimensional homology modelling of COX1 protein structure identified amino acid substitutions within the barcode area specific for the S. zuoi-complex and substantial divergence in structurally important amino acids between Sarcocystis species of snakes as definitive hosts and other lineages of the Sarcocystidae. We discuss the utility of selected genes for species delimitation of the Sarcocystis spp. under investigation, which probably evolved during recent radiations of their intermediate and definitive hosts.

Molecular prevalence, phylogenetic characterization, and epidemiological risk factors of pulmonary nematodes in domestic cats in Türkiye

Aelurostrongylus abstrusus, Oslerus rostratus, and Troglostrongylus brevior are nematodes found in the respiratory system of domestic cats and cause a variety of symptoms. All three parasites use the same hosts and niches, and the morphological measurements of their L1s excreted in faeces overlap with each other. In this study, 300 cats brought to Ondokuz Mayıs University Veterinary Teaching Animal Hospital were screened for lungworms by morphological measurements and molecular analyses. The prevalence of the lungworms was found as 1.33% (4/300) for A. abstrusus, 0.66% (2/300) for O. rostratus, and 0.33% (1/300) for T. brevior. Molecular identification of A. abstrusus, T. brevior, and O. rostratus in domestic cats was carried out for the first time in Türkiye within the present study. This study also reveals the risk factors of lungworm positivity in domestic cats in Türkiye.

Integrative taxonomy revealing a troglobitic new genus of Diplommatinidae from Jiangxi, China (Gastropoda: Caenogastropoda: Cyclophoroidea)

A new subterranean diplommatinid snail is described and illustrated from Jiangxi, China. The species is diagnosed by its cylindrical-fusiform shell and absence of internal columellar lamellae and parietal folds. The morphological and molecular phylogenetic characterization of the new species supports the erection of a new genus, Sohtsuia Z.-Y. Chen, gen. nov. A molecular phylogeny of representative East Asian continental species of Diplommatina Benson, 1949 is provided and the relationships of the new genus are discussed. Sohtsuia diting sp. nov. is described as new species.

Phylogenetic Characterization of HIV-1 Sub-Subtype A1 in Karachi, Pakistan.

(1) Background: HIV-1 sub-subtype A1 is common in parts of Africa, Russia, former Soviet Union countries, and Eastern Europe. In Pakistan, sub-subtype A1 is the predominant HIV-1 subtype. Preliminary evidence suggests that distinct strains of HIV-1 sub-subtype A1 are circulating in Pakistan; however, an in-depth molecular phylogenetic characterization of HIV-1 sub-subtype A1 strains in Pakistan have not been presented. We performed a detailed characterization of the HIV-1 sub-subtype A1 epidemic in Pakistan using state-of-the-art molecular epidemiology and phylodynamics. (2) Methods: A total of 143 HIV-1 sub-subtype A1 gag sequences, including 61 sequences generated specifically for this study from PLHIVs part of our cohort, representing all sub-subtype A1 gag sequences from Pakistan, were analyzed. Maximum-likelihood phylogenetic cluster analysis was used to determine the relationship between Pakistani sub-subtype A1 strains and pandemic sub-subtype A1 strains. Furthermore, we used signature variation, charge distribution, selection pressures, and epitope prediction analyses to characterize variations unique to Pakistani HIV-1 strains and establish the association between signature variations and Gag epitope profile. (3) Results: The HIV-1 sub-subtype A1 sequences from Pakistan formed three main clusters: two that clustered with Kenyan sequences (7 and 10 sequences, respectively) and one that formed a Pakistan-specific cluster of 123 sequences that were much less related to other sub-subtype A1 sequences available in the database. The sequences in the Pakistan-specific cluster and the Kenyan reference strains exhibited several signature variations, especially at amino acid positions 312, 319, 331, 372, 373, 383, and 402. Structural protein modeling suggested that amino acid changes in these positions result in alterations of the Gag protein structure as well as in Gag-specific T-cell epitopes. (4) Conclusions: Our results suggest that the majority of the Pakistan HIV-1 sub-subtype A1 strains were unique to Pakistan and with a specific mutation pattern in Gag.

Molecular identification and phylogenetic characterization of A-strain isolates of maize streak virus from western Ethiopia.

The A-strain of maize streak virus (MSV) causes maize streak disease (MSD), which is a major biotic threat to maize production in sub-Saharan Africa. Previous studies have described different MSV strains of economic importance from southern and eastern African countries and how eastern African regions are hubs for MSV diversification. Despite these efforts, due to a lack of extensive sampling, there is limited knowledge about the MSV-A diversity in Ethiopia. Here, field sampling of maize plants and wild grasses with visible MSD symptoms was carried out in the western Ethiopian regions of Gambela, Oromia, and Benishangul-Gumuz during the maize-growing season of 2019. The complete genomes of MSV isolates (n = 60) were cloned and sequenced by the Sanger method. We used a model-based phylogenetic approach to analyse 725 full MSV genome sequences available in the GenBank database together with newly determined genome sequences from Ethiopia to determine their subtypes and identify recombinant lineages. Of the 127 fields accessed, MSD prevalence was highest, at 96%, in the Gambela region and lowest in Oromia, at 66%. The highest mean symptom severity of 4/5 (where 5 is the highest and 1 the lowest) was observed in Gambela and Benishangul-Gumuz. Our results show that these newly determined MSV isolates belong to recombinant lineage V of the A1 subtype, with the widest dissemination and greatest economic significance in sub-Saharan Africa and the adjacent Indian Ocean islands.

Recoding Amino Acids to a Reduced Alphabet may Increase or Decrease Phylogenetic Accuracy.

Common molecular phylogenetic characteristics such as long branches and compositional heterogeneity can be problematic for phylogenetic reconstruction when using amino acid data. Recoding alignments to reduced alphabets before phylogenetic analysis has often been used both to explore and potentially decrease the effect of such problems. We tested the effectiveness of this strategy on topological accuracy using simulated data on four-taxon trees. We simulated alignments in phylogenetically challenging ways to test the phylogenetic accuracy of analyses using various recoding strategies together with commonly used homogeneous models. We tested three recoding methods based on amino acid exchangeability, and another recoding method based on lowering the compositional heterogeneity among alignment sequences as measured by the Chi-squared statistic. Our simulation results show that on trees with long branches where sequences approach saturation, accuracy was not greatly affected by exchangeability-based recodings, but Chi-squared-based recoding decreased accuracy. We then simulated sequences with different kinds of compositional heterogeneity over the tree. Recoding often increased accuracy on such alignments. Exchangeability-based recoding was rarely worse than not recoding, and often considerably better. Recoding based on lowering the Chi-squared value improved accuracy in some cases but not in others, suggesting that low compositional heterogeneity by itself is not sufficient to increase accuracy in the analysis of these alignments. We also simulated alignments using site-specific amino acid profiles, making sequences that had compositional heterogeneity over alignment sites. Exchangeability-based recoding coupled with site-homogeneous models had poor accuracy for these data sets but Chi-squared-based recoding on these alignments increased accuracy. We then simulated data sets that were compositionally both site- and tree-heterogeneous, like many real data sets. The effect on the accuracy of recoding such doubly problematic data sets varied widely, depending on the type of compositional tree heterogeneity and on the recoding scheme. Interestingly, analysis of unrecoded compositionally heterogeneous alignments with the NDCH or CAT models was generally more accurate than homogeneous analysis, whether recoded or not. Overall, our results suggest that making trees for recoded amino acid data sets can be useful, but they need to be interpreted cautiously as part of a more comprehensive analysis. The use of better-fitting models like NDCH and CAT, which directly account for the patterns in the data, may offer a more promising long-term solution for analyzing empirical data. [Compositional heterogeneity; models of evolution; phylogenetic methods; recoding amino acid data sets.].

Molecular Prevalence and Phylogenetic Characterization of Blastocystis in Cattle in Kayseri Province, Turkey

Blastocystis is one of the most common emerging zoonotic parasites in humans and animals. This study aimed to determine the molecular prevalence and subtype of Blastocystis in cattle (Bos taurus). A total of 150 fresh fecal samples were collected from slaughtered cattle from various slaughterhouses in Kayseri Province, Central Anatolia. Genomic DNA was extracted from all samples and used in PCR analyses of the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis. Blastocystis positive samples were sequenced for identify subtypes. Obtained sequences were assembled with suitable genetic software, then phylogenetic relationships were revealed. According to PCR analyses, overall prevalence of Blastocystis was determined as 58.7%. The sequence analyses of the PCR product gene revealed the presence of one known livestock-specific subtype, ST10. Phylogenetic analysis revealed that ST10 isolates the characterized in the study were clustered with isolates identified previously from cattle. Molecular characterization and subtype of Blastocystis sp. in slaughtered cattle in slaughterhouses were obtained data with this study.

2-Methylisoborneol (2-MIB) Excretion by Pseudanabaena yagii under Low Temperature.

Outbreaks of 2-methylisoborneol (2-MIB) contamination in drinking water sources cause inconvenient odor issues in the water distribution system. In this study, microscopy-based isolation with physiological and molecular phylogenetic characterization were performed to investigate and characterize the 2-MIB odor producers that caused an odor problem in the freshwater system of the North Han River in the autumn of 2018. A benthic cyanobacterium was isolated from 2-MIB odor-issue freshwater samples and was found to be phylogenetically affiliated with Pseudanabaena yagii (99.66% sequence similarity), which was recorded in South Korea for the first time. The 2-MIB synthesis gene sequences from the odor-issue freshwater samples showed 100% similarity with those in the P. yagii strains. Protein sequences of 2-MIB synthase observed in the genome of the isolated strain showed structural and functional characteristics similar to those observed in other Pseudanabaena species. The 2-MIB production rate increased slowly during mat formation on the vessel wall; however, it rapidly increased after the temperature dropped. The 2-MIB gene was continuously expressed regardless of the temperature changes. These results suggest that the 2-MIB odor issue in the North Han River might be caused by the release of 2-MIB from the mat-forming P. yagii species in a low-temperature freshwater environment.

Molecular detection and phylogenetic characterization of Theileria equi in horses (Equus caballus) from a peri-urban area of Argentina

To investigate the presence of Theileria equi in an endemic area of equine piroplasmosis 42 horses (Equus caballus) from Corrientes City, Argentina were sampled. Eighty-one percent (34 blood samples) of the analyzed horses were tested positive to the presence of piroplasmid 18S rDNA. All these samples could be identified as T. equi by amplifying the specific EMA-1 (merozoite antigen 1) gene of this species. Phylogenetic analysis of an obtained 18S rDNA complete sequence from one strain resulted in the identification of this sample as T. equi sensu stricto (genotype A). This study presents the first molecular detection and characterization of T. equi by the complete 18S rDNA sequence in Argentina. Based on these results further studies should be carried out to investigate the distribution and heterogeneity of presented genotypes of T. equi in Argentina, which is essential for the diagnosis, prevention and treatment of equine piroplasmosis.

First report of molecular and phylogenetic analysis of Physaloptera praeputialis in naturally infected stray cats from India.

Nematodes of the genus Physaloptera are globally distributed and infect a multitude of hosts. Their life cycle involves orthopterans and coleopterans as intermediate hosts. The morphological characters alone are inadequate to detect and differentiate Physaloptera spp. from its congeners. Moreover, molecular studies are limited to compare them precisely. The present communication reports the first molecular phylogenetic characterization of feline Physaloptera spp. from India based on mitochondrial cytochrome c oxidase subunit 1 (COX1) and small subunit ribosomal DNA (18S rDNA). The nematodes were first isolated from the stomach of adult stray cats during necropsy examination. Based on the gross and microscopic characters, the worms were identified as P. praeputialis. Morphological identification was further confirmed through PCR targeting the barcode region of the mitochondrial cytochrome c oxidase subunit I (MT-COI) gene, using nematode-specific primers cocktail followed by species specific primers targeting partial COX1 and 18S rRNA genes. Generated sequences were submitted in NCBI GenBank (MW517846, MW410927, MW411349), and phylogenetic trees were constructed using the maximum likelihood method. When compared with other sequences of Physaloptera species across the globe, the present isolates showed 85.6-97.7% and 97.3-99% nucleotide homology based on COX1 and 18S rRNA gene, respectively. BLASTn analysis revealed a strong identity to other Physaloptera spp., and the phylogenetic tree placed all Physaloptera spp. in the same cluster. This study again indicates the usefulness of molecular techniques to substantiate the identity of species that may lack adequate descriptions and impart new insight for the potentially overlooked significance of P. praeputialis infections in felines.

Babesia gibsoni emerging with high prevalence and co-infections in “fighting dogs” in Hungary

Babesia gibsoni is considered as an emerging protozoan parasite of dogs in North America and Europe. However, no data have been published on its prevalence, molecular-phylogenetic characteristics and associated co-infections in dogs used for illegal fighting (i.e. predisposed to acquiring this piroplasm via biting) in Europe. In this study, blood samples from 79 American Staffordshire Terrier dogs, confiscated for illegal dog fights, were molecularly analyzed for tick-borne pathogens. Babesiagibsoni was detected in 32 dogs, i.e. with a prevalence of 40.5%. In addition, Babesia vulpes was found in 8 samples (prevalence of 10.1%), for the first time in dogs in Hungary. Canine hemoplasmas were also identified in 49 samples (62%): only Mycoplasma haemocanis in 32 (40.5%) dogs, only “Candidatus Mycoplasma haematoparvum” in 9 (11.4%) dogs, and both hemoplasmas in 8 (10.1%) dogs. Thus, hemoplasma infections also showed a particularly high prevalence in this dog population. Based on a partial fragment of the 18S rRNA gene, B. gibsoni from Hungary exhibited complete sequence identity with conspecific strains reported from Europe and Asia. The cytochrome c oxidase subunit 1 (cox1) gene sequence of this isolate showed the closest identity with B. gibsoni reported from Japan but had a nonsynonymous mutation (M33I). Furthermore, the 11 B. gibsoni-positive samples analyzed for sequence variants of the cytochrome b (cytb) gene showed the presence of a common mutation (P310S). Most importantly, B. gibsoni had two further nonsynonymous mutations, M121I and F258L, in a dog with severe and relapsing anemia following atovaquone treatment. Phylogenetically, both cytb sequence variants clustered together, with a clear geographical pattern showing the closest relationship of both haplotypes identified in Hungary with those from China and Japan. To the best of our knowledge, this is the first cox1 and cytb characterization of B. gibsoni in Europe, as well as the first report on the emergence of this piroplasm and hemoplasmas with high prevalence among “fighting dogs” north of the Mediterranean Basin.

Molecular identification and phylogenetic characterization of influenza A virus at a wildlife–livestock interface in Mexico

Influenza A virus (IAV) outbreaks constitute a constant threat to public health and pose a remarkable impact on socio-economic systems worldwide. Interactions between wild and domestic birds, humans and swine can lead to spillover events. Backyard livestock systems in proximity to wetlands represent high-risk areas for viral spread. However, some gaps remain in our knowledge of IAV transmission at the wildlife-livestock interface in Mexico. Hence, the study aimed at molecular identification and phylogenetic characterization of IAV in the wild duck-backyard livestock interface at a wetland of Mexico. A total of 875 animals were tested by real-time RT-PCR (qRT-PCR). We detected IAV in 3.68% of the wild ducks sampled during the winter season 2016-2017. Nonetheless, the samples obtained from backyard poultry and swine tested negative. The highest IAV frequency (11.10%) was found in the Mexican duck (Anas diazi). Subtypes H1N1, H3N2 and H5N2 were detected. Phylogenetic analyses revealed that IAV detected in wild birds from the Lerma wetlands was mostly related to swine and poultry IAV strains previously isolated in the United States and Mexico. Except, the UIFMVZ377/H5N2 related to North American waterbirds. In conclusion, the co-circulation of three IAV subtypes in wild ducks close to backyard farms in Mexico, as well as the local identification of influenza viruses genetically related to Mexican and North American IAV strains, highlights the importance of the Lerma marshes for influenza surveillance given the close interaction among wild birds, poultry, pigs and humans.

An analogy of Canavalia lectins by molecular phylogenetic characterization

Concanavalin is a highly revered plant lectin with wide biological significance. Canavalia lectins of the Diocleinae subtribe possess increased structural similarity, but vary in their biological roles and applications. The lectin gene sequences of C. cathartica (ConC), C. ensiformis (ConA), C. gladiata (ConG) and C. rosea (ConM) were sequenced, PCR amplified, collated and compared with members of Phaseoleae tribe. Comparison of nucleotide sequences reveals changes in few bases. All Canavalia spp. diverged from C. bonariensis in the phylogenetic trees constructed according to the nucleotide sequences and the corresponding protein sequence of the legume lectins. The fluctuations in root mean square deviation derived by superimposition of monomers and tetramers substantiate variability in sugar specificity and biological activities.

Vertical Transmission of a Nematode from Female Lizards to the Brains of Their Offspring.

Parasites have evolved a diversity of lifestyles that exploit the biology of their hosts. Some nematodes that parasitize mammals pass via the placenta or milk from one host to another. Similar cases of vertical transmission have never been reported in avian and nonavian reptiles, suggesting that egg laying may constrain the means of parasite transmission. However, here we report the first incidence of transovarial transmission of a previously undescribed nematode in an egg-laying amniote, the common wall lizard (Podarcis muralis). Nematodes enter the developing brain from the female ovary early in embryonic development. Infected lizard embryos develop normally and hatch with nematodes residing in their braincase. We present a morphological and molecular phylogenetic characterization of the nematode and suggest that particular features of lizard biology that are absent from birds and turtles facilitated the evolutionary origin of this novel life history.

An assessment of biological control of the banana pseudostem weevil Odoiporus longicollis (Olivier) by entomopathogenic fungi Beauveria bassiana

Abstract Banana (Musa sp.) is the most imperative staple food crop for all types of people worldwide, which is commonly grown in Southeast Asia. Banana plantain can be severely affected by the devastating pest Odoiporus longicollis that results in severe economic losses in India. Management of weevil pests using chemical methods is harmful to the environment, and cultural methods are also partially successful. Therefore, an alternative approach of plant defense mediated by endophytic fungi to control banana stem borer larvae is necessary, which could affect the extracellular enzyme chitinase and protease. Among four isolates, Beauveria bassiana isolate KH3 is the most virulent entomopathogenic fungus compared with other isolates, and species identification was achieved using molecular phylogenetic characteristics. The B. bassiana isolate KH3 (1 × 108 conidia/mL-1) is more bioeffective against O. longicollis larvae, causing >90% significant mortality in 12 and 18 days. Adult weevils were treated with a higher concentration (1 × 108 conidia/mL-1) of the endophytic B. bassiana isolates KH3 and KH9, which resulted in a maximum mortality of 76.6% at 27 days of post inoculation compared to that with other isolates (p > 0.05). Based on our results, B. bassiana isolate KH3 (MN165867) can be exploited as a potential biocontrol agent for reducing insect population and stem damages in banana by lure and kill method.

Three new Pythium species from rice paddy fields

ABSTRACTIn an investigation of the oomyceteous flora in rice paddy fields of Fars Province, Iran, three new Pythium species were isolated and identified on the basis of morphological features and molecular phylogenetic characteristics. Their unique morphological traits, including sexual and asexual structural characteristics (i.e., sporangial type; oogonial type and ornamentations; type and the number of antheridia per oogonium; and oospore type), cardinal temperatures, and colony morphology on various media, separated them from other known species. Using nuclear and mitochondrial genes, each species formed discrete lineages in phylogenetic analyses based on Bayesian inference and maximum likelihood methods. This paper describes these three new Pythium species, P. heteroogonium, P. longipapillum, and P. oryzicollum, and compares them with their related taxa via morphological features and molecular characteristics. Pathogenicity tests revealed the ability of P. oryzicollum to cause pre- and post-emergence damping-off, seed rot, crown rot, and reduced growth rate on rice.