Partial DNA Sequences Research Articles

A PLIN1 polymorphism is associated with fat production in male emus

The emu (Dromaius novaehollandiae) is a novel poultry species that produces meat, eggs, and fat. Although emus have recently been domesticated, genetic improvements to establish strains have scarcely progressed. In this study, we investigated the relationship between production traits and perilipin 1-encoding gene (PLIN1) polymorphisms in the emus. We determined the partial complementary DNA (cDNA) sequence of the PLIN1, which is involved in lipid droplet formation. We identified four nucleotide substitution sites (c.270C>T, c.321T>C, c.587A>T, and c.639C>T) in the PLIN1 gene of emus. Of these, c.587A>T is a non-synonymous substitution that converts lysine to methionine at the 196th codon (p.K196M). Although p.K196M was predicted to affect the production traits of emus, a large deflection in genotype frequency was observed in this study; thus, we could not investigate the relationship between genotypes and phenotypes. In males, the fat yields of the CC, CT, and TT genotypes in c.270C>T were 0.25 ± 0.06, 0.22 ± 0.06, and 0.21 ± 0.07 kg, respectively, while the meat yields of the CC, CT, and TT genotypes in c.270C>T were 0.15 ± 0.01, 0.16 ± 0.02, and 0.16 ± 0.03 kg, respectively. These results indicate that male emus with the CC genotype had a significantly higher fat content and lower meat productivity than male emus with the other genotypes (P < 0.05). Therefore, c.270C>T in PLIN1 affects fat and meat production in males. Our findings may contribute to the effective genetic improvement of the emus.

Xoconochcothelphusa, a new genus for Ehecatusa chiapensis (Rodríguez &amp; Smalley, 1972), with notes on Spirothelphusa Pretzmann, 1965 (Crustacea: Decapoda: Pseudothelphusidae)

The species of the genus Ehecatusa Ng &amp; Low, 2010, E. chiapensis (Rodríguez &amp; Smalley, 1972) and E. mixtepensis (Rodríguez &amp; Smalley, in Smalley, 1970), were referred to as incertae sedis in the classification system of the tribe Pseudothelphusini Ortmann, 1897, proposed by Villalobos-Hiriart (2005) and Villalobos &amp; Alvarez(2010) and as members of the subfamily Pseudothelphusinae in the recent phylogenetic proposal by Álvarez et al. (2020), supported with morphology and molecular information. The recent discovery of a new specimen of E. chiapensis, from Chiapas, Mexico, comes to expose again the unresolved taxonomic situation of this species in the genus Ehecatusa. New morphologic evidence from the male first gonopod and a phylogenetic analysis based on partial DNA sequences of mitochondrial and nuclear genes (COI, 16S and H3), support the placement of the two species in different genera. Consequently, Xoconochcothelphusa n. gen. is erected to receive X. chiapensis n. comb. The phylogenetic relationships of X. chiapensis n. gen., n. comb. and E. mixtepensis with other genera of the subfamily Pseudothelphusinae Ortmann, 1893 (Ehecatusa, Smalleyus Alvarez, 1989, and Spirothelphusa Pretzmann, 1965) distributed in Mexico, are examined and a key to the identification of the genera of this subfamily is provided.

Human Schistosomiasis due to Schistosoma bovis in Nigeria.

Schistosomiasis is a global public health challenge, especially in sub-Saharan Africa, where Nigeria has the highest burden of disease. Schistosoma hybrids have been discovered in various countries, including Nigeria, where livestock and human beings share common water resources. This study, carried out in three communities, two of which are endemic for urinary schistosomiasis, aimed to identify the urinary schistosomiasis causative agent by using a species-specific molecular technique and their evolutionary relationship. Polymerase chain reaction using primers specific for a partial DNA sequence of the mitochondrial cox1 gene of Schistosoma haematobium was carried out on pooled urine sediments preserved in 70% ethanol. DNA from the high-intensity pooled samples from Onye-Uku camp amplified, and a BLAST search identified the pooled samples as Schistosoma bovis, whereas S. haematobium DNA did not amplify. The phylogenetic relationship of the sequence showed that it clustered with an S. bovis hybrid obtained from a human host in Côte d'Ivoire and had close ancestry with isolates from cattle in Cameroon. This finding revealed the prevalence of S. bovis among some inhabitants in a zone in Nigeria where schistosomiasis is endemic.

Molecular systematics of chinchilla rats and taxonomic assessment of the Abrocoma cinerea species complex

Abstract Chinchilla rats (family Abrocomidae) are hystricomorph rodents primarily inhabiting the central Andes in South America with 8 species in the genus Abrocoma and 2 in Cuscomys. The systematics of this family—relying only on morphological differences—has faced several controversies, particularly in arid-adapted species of Abrocoma (the A. cinerea species complex, or ACC). By using partial DNA sequences of 1 mitochondrial (Cytochrome b) and 2 nuclear genes (GHR and RAG1), we: (i) inferred phylogenetic relationships between 5 species of Abrocoma and 1 of Cuscomys; (ii) performed molecular species-delimitation analyses in 4 species of the ACC (A. cinerea, A. famatina, A. schistacea, and A. uspallata); and (iii) estimated divergence times using stratigraphic ages of extinct taxa. Our phylogenetic analyses recovered 3 main clades: A. bennettii (basal); C. ashaninka; and the ACC. Striking differences in habitat use, morphology, and genetics suggest that each of these main clades might represent a distinct genus. In species-delimitation analyses in the ACC, only 2 species were recognized, and we thus suggest the synonymy of A. famatina with A. cinerea and of A. uspallata with A. schistacea. The origin of extant abrocomids was estimated at the late Miocene (~5.6 million years ago, Ma) with the subsequent origin of arboreal species during the early Pliocene (~4.3 Ma) and the ACC during the late Pliocene (~3.3 Ma). We hypothesize that topographic uplift of the central Andes and associated climatic and ecological changes were the main drivers of diversification in this family.

Origins and taxonomic status of Hemidactylus geckos on the Îles Éparses of the Western Indian Ocean

Distinguishing between anthropogenic introductions and natural colonizations can be complex, especially in groups that are evidently both capable natural colonizers and anthropophilic, such as some geckos of the genus Hemidactylus. However, such information is fundamental both for constructing appropriate conservation approaches, as well as to identify biogeographical patterns. Here we assessed the origins and taxonomic status of the Hemidactylus geckos found on the Îles Éparses, disjunct small islands of the western Indian Ocean located around Madagascar, using partial mitochondrial DNA sequences. Hemidactylus platycephalus was confirmed on Juan de Nova, presumed introduced from continental Africa. The H. mabouia-mercatorius complex was identified on Juan de Nova, Europa, Tromelin, and Grande Glorieuse, with H. mabouia (sensu stricto) on Juan de Nova and H. mercatorius on Europa, from where it is likely to have been introduced to Tromelin. An unnamed lineage within H. mercatorius, previously known only from the Aldabra Islands, was identified on Grande Glorieuse, and further demonstrated the unexpectedly high genetic diversity within this lineage.

Detection and population genetic analysis of Aedes albopictus (Diptera: Culicidae) based on knockdown resistance (kdr) mutations in the Yangtze River basin of China

BackgroundAedes albopictus is an important vector of chikungunya, dengue, yellow fever and Zika viruses. Insecticides are often the most effective tools for rapidly decreasing the density of vector populations, especially during arbovirus disease outbreaks. However, the intense use of insecticides, particularly pyrethroids, has led to the selection of resistant mosquito populations worldwide. Mutations in the voltage-gated sodium channel (VGSC) gene are one of the main drivers of insecticide resistance in Ae. albopictus and are also known as “knockdown resistance” (kdr) mutations. Knowledge about genetic mutations associated with insecticide resistance is a prerequisite for developing techniques for rapid resistance diagnosis. Here, we report studies on the origin and dispersion of kdr haplotypes in samples of Ae. albopictus from the Yangtze River Basin, China; MethodsHere, we report the results of PCR genotyping of kdr mutations in 541 Ae. albopictus specimens from 22 sampling sites in 7 provinces and municipalities in the Yangtze River Basin. Partial DNA sequences of domain II and domain III of the VGSC gene were amplified. These DNA fragments were subsequently sequenced to discover the possible genetic mutations mediating knockdown resistance (kdr) to pyrethroids. The frequency and distribution of kdr mutations were assessed in 22 Ae. albopictus populations. Phylogenetic relationships among the haplotypes were used to infer whether the kdr mutations had a single or multiple origins; ResultsThe kdr mutation at the 1016 locus had 2 alleles with 3 genotypes: V/V (73.38%), V/G (26.43%) and G/G (0.18%). The 1016G homozygous mutation was found in only one case in the CQSL strain in Chongqing, and no 1016G mutations were detected in the SHJD (Shanghai), NJDX (Jiangsu) or HBQN (Hubei) strains. A total of 1532 locus had two alleles and three genotypes, I/I (88.35%), I/T (8.50%) and T/T (3.14%). A total of 1534 locus had four alleles and six genotypes: F/F (49.35%), F/S (19.96%), F/C (1.48%) and F/L (0.18%); S/S (23.66%); and C/C (5.36%). Haplotypes with the F1534C mutation were found only in Ae. albopictus populations in Chongqing and Hubei, and C1534C was found only in three geographic strains in Chongqing. Haplotypes with the 1534S mutation were found only in Ae. albopictus populations in Sichuan and Shanghai. F1534L was found only in HBYC. The Ae. albopictus populations in Shanghai were more genetically differentiated from those in the other regions (except Sichuan), and the genetic differentiation between the populations in Chongqing and those in the middle-lower reaches of the Yangtze River (Huber, Jiangsu, Jiangxi, and Anhui) was lower. Shanghai and Sichuan displayed low haplotype diversity and low nucleotide diversity. Phylogenetic analysis and sequence comparison revealed that the 1016 locus was divided into three branches, with the Clade A and Clade B branches bearing the 1016 mutation occurring mostly in Jiangsu and the Clade C branch bearing the 1016 mutation occurring mostly in Chongqing, suggesting at least two origins for 1016G. IIIS6 phylogenetic analysis and sequence comparison revealed that F1534S, F1534C and I1532T can be divided into two branches, indicating that IIIS6 has two origins; ConclusionsCombined with the distribution of kdr mutations and the analysis of population genetics, we infer that besides the local selection of pyrethroid resistance mutations, dispersal and colonization of Ae. albopictus from other regions may explain why kdr mutations are present in some Ae. albopictus populations in the Yangtze River Basin.

Phylogenetic analysis of Ellipsomyxa species (Myxosporea) and description of Ellipsomyxa gordeyi n. sp. from the gall bladder of mullets (Mugiliformes: Mugilidae) in Nha Trang Bay of the East Sea, Vietnam

The coastal waters of Vietnam are home to a wide diversity of fishes, but the parasite diversity of these potential hosts is much less well characterized. To begin addressing this knowledge gap, we carried out surveys of myxozoan parasites in fishes collected from Nha Trang Bay in Vietnam's East Sea in 2018–2019. Mugilid fishes were collected in March–April 2018, January–February 2019, and November–December 2019, and examined for myxozoans. Myxospores consistent with those of the genus Ellipsomyxa were found in the gall bladder of four mullet species, and we thoroughly characterized those from Planiliza melinoptera. Myxospores were elliptoid and devoid of striation, with a distinct sinuous suture line. Polar capsules were pyriform and oriented toward the poles of the spore. Morphological features were compared to nominal species and this species from Vietnam was distinct. Phylogenetic analysis based on partial small subunit rDNA sequence revealed that broadly, Ellipsomyxa species split into three phylogenetic lineages, and although in some branches there are groupings by host family, habitat or locality, there are no clear phylogenetic patterns. The new species we encountered in P. melinoptera had a close sister relationship with Ellipsomyxa adlardi, with both species part of a larger subclade within the Ellipsomyxa lineage. Despite this phylogenetic similarity, these species were morphologically distinct, and partial large subunit DNA sequences were only 93% similar to each other. A combination of the morphological characteristics and molecular data suggest that this is an undescribed species and we propose the name Ellipsomyxa gordeyi n. sp.

Neurigona susanrocesae, the first Neurigona species from the Philippines

The first species of the genus Neurigona is described from the Philippines. The diagnosis, description and molecular data based on the partial DNA sequence of mitochondrial cytochrome oxidase subunit I gene were documented. The Neurigona species exhibits distinct characteristics, including a sub-apical dorsal stylus resembling an arista, a peduncle on the seventh abdominal segment, a flat dorsal postcranium, elongated legs devoid of prominent setae or bristles, and a distinctive rectangular bend in wing vein M. Based on the DNA sequence analysis, the unknown species is 90.06% similar to Cyrtona sp. and Neurigona zhejiangensis. However, 10% of its DNA sequences are significantly variable making it a totally different species. The specimen was verified by a dipterist as a species belonging to Genus Neurigona under Family Dolichopodidae. These flies are recognized for their lengthy legs and sizable yellow bodies. The species was named as N. susanrocesae as a tribute to the late Susan Roces, the “Queen of Philippine Movies”.

Uncovering further diversity of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) in South American bats

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0–1.2% interspecific divergence in 28S and 3.3–20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans.

New records of the genus Hypogastrura (Collembola: Hypogastruridae) from the Greater Caucasus, with the description of a new species from the monticola-group

A new species of the genus Hypogastrura is described from the North Caucasus: H. metegensis sp. nov. The species shares the unusual absence of lateral sensillum S9 on Ant. IV with only four congeners of the monticola-group which it belongs to, but it can be distinguished from all by the relatively long mucro with lower lateral lamellae. Regional records of and taxonomic notes with partial DNA sequences of their mitochondrial cytochrome c oxidase subunit 1 (COI) gene on two other Caucasian species of the same genus are also given.

First report of Lasiodiplodia pseudotheobromae causing soft rot of plum in China.

Plum (Prunus salicina) is one of the most important fruit tree species worldwide (Valderrama-Soto et al. 2021). In June 2023, the postharvest soft rot symptoms were observed on plum fruits in several fruit markets of Guiyang city, Guizhou province, China. The disease incidence in these markets ranged from 20 to 25% with 70% disease severity. Plum fruits showed rotting, which was characterized by water soaked fruit tissue, softening and presence of whitish mycelia four days post inoculation. In severe conditions, whole fruits become rotted and were covered with white fungal mycelia. Small sections (5 × 3 mm) from 6 diseased plum fruits were surface sterilized by using 75% ethanol for 30 s followed by 0.1% mercuric chloride solution for 5 min, rinsed three times with ddH2O, and then transferred onto potato dextrose agar (PDA) and incubated at 25 ± 2°C for three days. Three pure cultures (GUCC23-0001 to GUCC23-0003) were obtained by transferring a single hyphal tip to new PDA plates. Colonies of these isolates were grayish-white initially, gradually turning to whitish brown with fluffy aerial mycelia and uneven edges and finally turned to a dark gray colony after five days of inoculation. The pseudoparaphyses were hyaline, cylindrical, aseptate, and rounded at apex. Conidia were ellipsoidal, hyaline, unicellular, and 24.2 to 28.6 × 12.3 to 15.5 µm in size (n = 30) (Fig. S1), which were similar to the morphology of Lasiodiplodia pseudotheobromae (Alves et al. 2008). Furthermore, fungal DNA was extracted from fresh mycelia of PDA after seven days by using fungus genomic DNA extraction kit (Biomiga, USA). Partial DNA sequences from four loci including internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), and polymerase II second largest subunit (rpb2) were amplified with ITS1 and ITS4 (White et al. 1990), EF1-688F and EF1-1251R (Alves et al. 2008), Bt2a and Bt2b (Glass and Donaldson 1995), and RPB2-LasF and RPB2-LasR, respectively (Cruywagen et al. 2017). GenBank accession numbers are OR361680, OR361681, OR361682 for ITS, OR423394, OR423395, OR423396 for tef1, OR423397, OR423398, OR423399 for tub2, and OR423391, OR423392, OR423393 for rpb2, and gene sequencing showed 99.6 to 100% identity with ex-type strain of L. pseudotheobromae (CBS 116459). Phylogenetic analysis also placed our isolates in a highly supported clade with the reference isolate of L. pseudotheobromae (Fig. S2). Another experiment was designed to confirm the pathogenicity test for additional confirmation. Five mm mycelial plugs of L. pseudotheobromae from a three day old culture on PDA were placed on five surface-sterilized and non-wounded plum fruits for 12 hours and incubated at 25°C ± 2°C for four days. Sterilized fungus free PDA plugs were used as a negative control. Mycelial plugs were removed after 12 hours following which whole fruits were incubated in plastic boxes at 25°C ± 2°C. The experiment was repeated twice. The pathogenicity was evaluated under control conditions in laboratory (relative humidity, 70 ± 5% and temperature 25 ± 5˚C). Plum fruits showed rotting, which was characterized by water soaked fruit tissue, softening and presence of whitish mycelia four days post inoculation. These symptoms and signs were similar to the initially observed symptoms on plums in the markets. No disease symptoms were observed on the control fruits. The re-isolated fungus obtained from inoculated plum fruits was very similar to those isolated from diseased samples in morphology, fulfilling Koch's postulates. To the best of our knowledge, this is the first report of L. pseudotheobromae causing postharvest fruit rot of plum in China. In 2022, the total planting area of plum was 1946.5 thousand hectares, which produces approximately 6626300 tons of plum (Food and Agriculture Organization of the United Nations, 2022). Based on the disease incidence and severity reported in the current study, soft rot of plum may be responsible for nearly 35% of yield losses under severe. Therefore, our study laid a theoretical foundation for the prevention and control of this post-harvest disease of plum.

A new species of freshwater snail of Fenouilia (Gastropoda, Pomatiopsidae) from northern Guangxi, China, based on morphological and DNA evidence.

A new species of pomatiopsid freshwater snail, Fenouiliaundata Chen & He, sp. nov., is described from Guangxi, China, based on morphological and molecular evidence. The new species can be distinguished from its congeners by the following combination of characters: shell with low, prosocline, rounded axial ribs and fine spiral striae, broader than high; aperture broader than shell height; radula with lateral teeth have only two or three faint, wavy ridges on inner side. A molecular analysis of partial mitochondrial COI and 16S DNA sequences supports the systematic position of the new taxon.

Comparative plastome analysis and taxonomic classification of snow lotus species (Saussurea, Asteraceae) in Central Asia and Southern Siberia

Four species of Saussurea, namely S. involucrata, S. orgaadayi, S. bogedaensis, and S. dorogostaiskii, are known as the “snow lotus,” which are used as traditional medicines in China (Xinjiang), Kyrgyzstan, Mongolia, and Russia (Southern Siberia). These species are threatened globally, because of illegal harvesting and climate change. Furthermore, the taxonomic classification and identification of these threatened species remain unclear owing to limited research. The misidentification of medicinal species can sometimes be harmful to health. Therefore, the phylogenetic and genomic features of these species need to be confirmed. In this study, we sequenced five complete chloroplast genomes and seven nuclear ITS regions of four snow lotus species and other Saussurea species. We further explored their genetic variety, selective pressure at the sequence level, and phylogenetic relationships using the chloroplast genome, nuclear partial DNA sequences, and morphological features. Plastome of the snow lotus species has a conserved structure and gene content similar to most Saussurea species. Two intergenic regions (ndhJ–ndhK and ndhD-psaC) show significantly high diversity among chloroplast regions. Thus, ITS and these markers are suitable for identifying snow lotus species. In addition, we characterized 43 simple sequence repeats that may be useful in future population genetic studies. Analysis of the selection signatures identified three genes (rpoA, ndhB, and ycf2) that underwent positive selection. These genes may play important roles in the adaptation of the snow lotus species to alpine environments. S. dorogostaiskii is close to S. baicalensis and exhibits slightly different adaptation from others. The taxonomic position of the snow lotus species, confirmed by morphological and molecular evidence, is as follows: (i) S. involucrata has been excluded from the Mongolian flora due to misidentification as S. orgaadayi or S. bogedaensis for a long time; (ii) S. dorogostaiskii belongs to section Pycnocephala subgenus Saussurea, whereas other the snow lotus species belong to section Amphilaena subgenus Amphilaena; and (iii) S. krasnoborovii is synonymous of S. dorogostaiskii. This study clarified the speciation and lineage diversification of the snow lotus species in Central Asia and Southern Siberia.

Molecular detection and characterization of Bartonella spp. in small mammals in the Amazonia and Cerrado biomes, midwestern Brazil

Although Bartonella spp. have been worldwide described in rodents and bats, few studies have reported these agents in marsupials. The present work aimed to investigate the occurrence and genetic diversity of Bartonella in small mammals (rodents, marsupials, and bats) and associated ectoparasites in two ecoregions (Amazonia and Cerrado biomes) in midwestern Brazil. For this purpose, DNA samples from 378 specimens of small mammals (128 rodents, 111 marsupials, and 139 bats) and 41 fleas (Siphonaptera) were screened for the Bartonella genus employing a quantitative real-time PCR assay (qPCR) based on the nuoG (nicotinamide adenine dinucleotide dehydrogenase gamma subunit) gene. Then, positive samples in qPCR were submitted to conventional PCR (cPCR) assays targeting the gltA, ftsZ, and rpoB genes. One (0.78 %) rodent, 23 (16.54 %) bats, and 3 (7.31 %) fleas showed positive results in the qPCR for Bartonella sp. After cPCR amplification and sequencing, 13 partial Bartonella DNA sequences of the following genes were obtained only from bats´ blood samples: 9 gltA (citrate synthase), 3 ftsZ (cell division protein), and 1 rpoB (RNA polymerase beta subunit). The maximum likelihood inference based on the gltA gene positioned the obtained sequences in three different clades, closely related to Bartonella genotypes previously detected in other bat species and bat flies sampled in Brazil and other countries from Latin America. Similarly, the ftsZ sequences clustered in two different clades with sequences described in bats from Brazil, other countries from Latin America, and Georgia (eastern Europe). Finally, the Bartonella rpoB from a specimen of Lophostoma silvicolum clustered with a Bartonella sp. sequence obtained from a Noctilio albiventris (KP715475) from French Guiana. The present study provided valuable insights into the diversity of Bartonella genotypes infecting bats from two ecoregions (Amazonia and Cerrado) in midwestern Brazil and emphasized that further studies should be conducted regarding the description and evaluation of different lineages of Bartonella in wild small mammals and their ectoparasites in different Brazilian biomes.

First Report of Geotrichum candidum Causing Rubbery Rot on Potato in Arizona.

In January 2023, stand loss due to seed decay (<3% incidence) was reported in a commercial seed potato field in Cochise County, Arizona. Several rotted 'Ciklamen' tubers with wilting shoots were submitted to the University of Arizona Extension Plant Pathology Laboratory for diagnosis. External symptoms included sunken black-colored lesions on affected tubers; white fungal-like growth was also present on the surface of the more severely affected tubers. Internal tissues were watery, rubbery-textured, and spongy with slight gray discoloration. Isolation from the necrotic tubers on PDA yielded yeast-like fungal colonies. Two derived fungal isolates were subcultured on PDA at room temperature for three days, and colonies had a white, dense, and creamy appearance. Hyphae were hyaline, septate, dichotomously branched, and broken into chains of cylindrical to oval-shaped, single-celled arthrospores ranging in size from 3.8-5.3 μm × 5.4-12.5 μm (n = 20). These morphological characteristics matched those of Geotrichum candidum (Carmichael, 1957). Other common potato pathogens, including Pythium, Phytophthora, and Fusarium, were not detected in the samples. For molecular identification, genomic DNA was extracted from the mycelia of two isolates using the DNeasy Plant Pro Kit (Qiagen Inc., Valencia, CA) according to the manufacturer's instructions. Partial DNA sequences of the internal transcribed spacer (ITS) region of rDNA and elongation factor 1-alpha (EF-1α) gene were amplified and sequenced using primers ITS1/ITS4 (White et al., 1990), EF1-728F/EF1-1567R (Carbone and Kohn 1999) and EF1-1018/EF1-1620R (Stielow et al., 2015). The resulting sequences showed no difference between the isolates and were deposited under the GenBank Accession Numbers OQ690039 and OQ981192. Based on BLASTn analysis, ITS sequence OQ690039 (369-bp) had a 99.46% match with Geotrichum candidum accessions MN861070 and KY103456. EF-1α sequence OQ981192 (843-bp) showed 100% identity with G. candidum accessions MT346368 of the G. candidum isolates responsible for sour rot of strawberries in Florida (Alonzo et al., 2020). A pathogenicity test was conducted twice on ten healthy 'Ciklamen' tubers, surface sterilized using 10% sodium hypochlorite solution, and rinsed copiously in sterilized water. A conidial suspension (106 conidia/ml) of the isolated fungus was sprayed onto the surface of the tubers. The control consisted of ten tubers that were mock-sprayed with sterilized water. All tubers were incubated in separate sterilized plastic bags in the dark at room temperature for four weeks and examined daily for symptom development. Symptoms of rubbery texture and fluid leaking when cut, and a distinctive smell, were produced on the inoculated tubers. In contrast, the controls did not show any symptoms. These symptoms were similar to those observed on naturally infected tubers. The fungus was re-isolated from symptomatic tissues of inoculated tubers and identified by PCR as G. candidum, thus fulfilling Koch's postulates. To our knowledge, this is the first report of G. candidum as a potato pathogen in Arizona. Rubbery rot of potatoes by G. candidum has been reported previously in Idaho (Duellman et al., 2020) and Michigan (Willbur et al., 2021). Arizona annually grows approximately 1,000 hectares of seed potatoes shipped to various states. Therefore, growers must obtain a correct diagnosis to avoid spreading the disease despite the low incidence of disease.

Genetic diversity of diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae) populations in Türkiye.

Diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), is an important worldwide pest of plants belonging to the Brassicaceae family. In this study, we investigated genetic diversity of DBM populations in Brassicaceae production areas in Türkiye using the partial mtDNA CO1 gene region. We determined 43 samples from 11 different populations for haplotype variations using the partial mitochondrial DNA sequences a 684bp fragment of the CO1 gene. The results indicated that, the average haplotype diversity (Hd) was determined as 0.962 and nucleotide diversity (π) was determined as 0.557%. In neutrality tests, negative values were obtained in Tajima's D and Fu' Fs tests (Fu' Fs=-0.40, Tajima's D=-0.01). Tajima's D test was not found significant (p > 0.05). Fst value among DBM population estimates ranged from 0 to 0.631. Barcode gap distance was determined as 1.6%, but the intraspecies of genetic distance were found to be 0.15%. In conclusion, the presented study provided detailed and fundamental information about the genetic diversity of DBM populations in Türkiye. Further studies are needed to develop alternative pest management strategies for DBM populations integrating genetic approaches.

Electronic equipment and appliances in special wards of hospitals as a source of azole-resistant Aspergillus fumigatus: a multi-centre study from Iran

Azole-resistant Aspergillus fumigatus (ARAf), reported as a global public health concern, has been unexpectedly observed in different countries. To identify ARAf and detect azole resistance related to the CYP51A mutation in different hospital environmental samples. In this multi-centre study from Iran, surfaces of electronic equipment and appliances from different hospitals in Iran were sampled using cotton swabs. All samples were cultured using azole-containing agar plates (ACAPs). Recovered Aspergillus isolates were identified at the species level using partial DNA sequencing of the β-tubulin gene. The azole susceptibility testing of A.fumigatus isolates was performed using the Clinical and Laboratory Standards Institute M38-A3 guideline. The sequencing of the CYP51A gene was also performed to detect mutations related to resistance. Out of the 693 collected samples, 89 (12.8%) Aspergillus species were recovered from ACAPs. Aspergillus fumigatus (41.6%) was the most prevalent, followed by A.tubingensis (23.6%) and A.niger (15.6%). Among 37 isolates of A.fumigatus, 19 (51.3%) showed high minimum inhibitory concentration (MIC) values to at least one of the three azoles, voriconazole, itraconazole, and posaconazole. CYP51A polymorphisms were detected in all 19 isolates, of which 52.6% showed the TR34/L98H mutation. Other detected mutations were G432C, G448S, G54E/G138C, F46Y, and Y121F/M220I/D255E. T289F and G432C were the first reported mutations in ARAf. There was a considerable level of azole resistance in hospital environmental samples, a serious warning for patients vulnerable to aspergillosis. Our findings have also revealed a different mutation pattern in the CYP51A gene.

Phylo-geographic analysis of whitefly on the basis of mitochondrial cytochrome oxidase 1 gene

Abstract Bemisia tabaci is a species complex that causes damage to its broad range of plant hosts through serious feeding. It transmits plant viruses of different groups to important agricultural crops. Some important cash crops of Pakistan are sugar cane, rice, tobacco and seed oil. It shows high genetic variability and is differentiated as races or biotypes. Biotypes are, biotype Q, biotype B, biotype B2, biotype M, biotype L, biotype A, biotype H, biotype C, biotype K, biotype N, biotype R, biotype E, biotype P, biotype J, biotype S, biotype AN. Although the current report based on the Bayesian study of mitochondrial cytohrome oxidase gene1 (CO1) DNA sequences has classified the different populations of whiteflies into twelve genetic groups which are Mediterranean, Sub-Saharan Africa silverleafing, Indian Ocean, Asia II, Asia I, Australia, New World, Italy, China, Sub-Saharan Africa non-silverleafing, Mediterranean/Asia Minor/Africa and Uganda sweet potato. Begomoviruses is largest group of viruses transmitted by B. tabaci and cause major diseases of crops such as tomato and chili leaf curl disease, cassava mosaic disease; yellow mosaic disease of legumes and cotton leaf curl disease. The main objective of current study is to inculpate knowledge regarding genetic diversity of whitefly in cotton fields across Pakistan via analysis of partial DNA sequence of mitochondrial gene Cytochrom Oxidase I (mtCO1).

AAV-based gene editing of type 1 collagen mutation to treat osteogenesis imperfecta

Osteogenesis imperfecta (OI) is a genetic disorder characterized by bone fragility, low bone mass, fractures, and extraskeletal manifestations. Since OI is commonly caused by single-nucleotide mutation(s) in the COL1A1 or COL1A2 genes encodingtype I collagens, we developed a genome-editing strategy to correct a Col1a2 mutation in an OIM mouse model resembling a severe dominant form of human type III OI. Using a recombinant adeno-associated virus (rAAV), we delivered CRISPR-Cas9 to bone-forming osteoblast-lineage cells in the skeleton. Homology-directed repair (HDR)-mediated gene editing efficiency in these cells was improved when CRISPR-Cas9 was coupled with a donor AAV vector containing a promoterless partial mouse Col1a2 complementary DNA sequence. This approach effectively reversed the dysregulation of osteogenic differentiation by a Col1a2 mutation invitro. Furthermore, systemic administration of dual rAAVs in OIM mice lowered bone matrix turnover rates by reducing osteoblast and osteoclast development while improving the cellular network of mechano-sensing osteocytes embedded in the bone matrix. This strategy significantly improved bone architecture/mass/mineralization, skeletal deformities, grip strength, and spontaneous fractures. Our study is the first demonstration that HDR-mediated gene editing via AAV-mediated delivery effectively corrects a collagen mutation in OI osteoblasts and reverses skeletal phenotypes in OIM mice.

Morphological and molecular characterization of a new freshwater Ceratomyxa species (Cnidaria: Myxozoa) from the yellow catfish, Trachysurus fulvidraco in China.

Ceratomyxa Thélohan, 1892 is one of the largest genera of the sub-phylum Myxozoa, and has a worldwide geographical distribution. Ceratomyxa species mainly infect the gallbladder of marine fish and rarely infect histozoically or coelozoically freshwater fish. In the present study, yellow catfish, Trachysurus fulvidraco (Siluriformes, Actinopteri) collected from the low reach of Yellow River was first found to be infected with an unknown Ceratomyxa species in their gallbladder which was identified to be new to science and nominated as Ceratomyxa huangheensis n. sp. by an integrative taxonomic approach for myxosporeans. Spores are typical of the genus Ceratomyxa, and matures spores are arcuate, with posterior angle of 139.2±11.6 (137.0-156.0)° and rounded ends, and measures 4.7±0.6 (3.3-5.5) μm in length, and 10.7±1.3 (8.5-13.3) μm in thickness. Two spore valves are slightly unequal and present elongated ovoid in the lateral view. Two equal spherical polar capsules, measuring 2.2±0.4 (1.4-2.8) μm×2.0±0.4 (1.0-2.5) μm in size locates adjacent to the straight suture line. The obtained partial small subunit ribosomal DNA (SSU rDNA) sequence was unique among all myxozoans in GenBank, and the highest similarity is 85.3% with Ceratomyxa sparusaurati, a marine fish-infecting congener. Phylogenetic analysis further indicated that this novel species did not cluster with other freshwater fish-infecting congeners from South America to form an independent clade, but was phylogenetically positioned within the marine fish-infecting clade. The possible presence of different radiation trajectories between Ceratomyxa huangheensis n. sp. and Amazonian freshwater Ceratomyxa lineage was discussed. This is the first report of Ceratomyxa species in the Yellow River basin and the second freshwater Ceraotomyxa species in China, even in Asia.