Sequence Of Subunit Research Articles

BK Channels Function in Nematocyst Discharge from Vibration-Sensitive Cnidocyte Supporting Cell Complexes of the Sea Anemone Diadumene lineata.

AbstractIntegrated chemo- and mechanosensory pathways, along with activated calcium influxes, regulate nematocyst discharge from sea anemone tentacles. Discharge from vibration-sensitive Type A cnidocyte supporting cell complexes use calcium-conducting transient receptor potential V4-like channels. Because calcium influxes often couple with calcium-activated, large-conductance potassium (BK) channels, we hypothesized that BK channels function in nematocyst discharge. To verify this hypothesis, we first tested five selective BK channel blockers on nematocyst-mediated prey killing in Diadumene lineata (aka Haliplanella luciae). All tested BK channel blockers inhibited prey killing at concentrations comparable to their inhibition of vertebrate BK channels. In addition, the BK channel blocker paxilline selectively inhibited prey killing mediated by vibration-sensitive Type A cnidocyte supporting cell complexes. We queried a mammalian BKα amino acid sequence to the Exaiptasia diaphena database, from which we identified a putative anemone, pore-forming BKα subunit sequence. Using the E. diaphena BKα sequence as a template, we assembled a BKα transcript from our assembled D. lineata transcriptome. In addition, the hydra homolog of D. lineata BKα localizes to nematocytes on the hydra single-cell RNA sequencing map. Our findings suggest that D. lineata expresses BK channels that play a role in vibration-sensitive nematocyst discharge from Type A cnidocyte supporting cell complexes. We believe this is the first functional demonstration of BK channels in nonbilaterians. Because stimulated chemoreceptors frequency tune Type A cnidocyte supporting cell complexes to frequencies matching swimming movements of prey via a protein kinase A signaling pathway and protein kinase A generally activates BK channels, we suggest that D. lineata BK channels may participate in protein kinase A-mediated frequency tuning.

First report and molecular identification of Chilo partellus (Swinhoe, 1885) in South-eastern Türkiye: invasion continues (Lepidoptera: Crambidae)

Chilo partellus (Swinhoe, 1885) (Lepidoptera: Crambidae), is an invasive maize pest that can cause high yield losses. The pest has recently been introduced and established in eastern Mediterranean countries such as Türkiye. However, research regarding its expansion towards other regions in Türkiye and other countries is required. This study is the first to report the presence and abundance of the pest in South-eastern Türkiye provinces, Diyarbakır, Mardin, Ranlıurfa, and Rırnak as well as the infestation rate and its contribution to the stemborer-dependent infestation rates. There were significant differences between the abundances of the pest in different counties, while the infestation rate, the contribution to the stemborer-dependent infestation rate, and the contribution to the total stemborer population were not statistically different between counties. The pest was only recorded in three southern counties of Ranlıurfa, during routine September surveys in 2022 and was absent in Diyarbakır, Mardin, and Rırnak. Specimens were morphologically diagnosed, and for molecular identification and phylogenetic analysis, nucleotide sequencing of the mitochondrial cytochrome oxidase subunit I (COI-I) gene was performed. All specimens were confirmed as C. partellus. Further, phylogenetic analysis revealed intraspecific genetic variation and the similarity of the specimens to reference sequences from Africa.

First report of Neoscytalidium dimidiatum as the causal agent of leaf blight on Clivia miniata

In this survey, the symptomatic leaves of Clivia miniata were collected from a greenhouse in Karaj city of Iran. The isolation and morphological investigation showed Scytalidium-like fungus associated with leaf blight symptom. The phylogenetic analysis of the internal transcribed spacer along with partial sequences of rDNA large subunit and translation elongation factor 1-α (tef-1α) genomic regions confirmed the identification of the recovered isolate as Neoscytalidium dimidiatum. The pycnidial morph of the fungus didn’t observe both in vitro and in vivo. The pathogenicity test on C. miniata and C. nobilis was also conducted to fulfill the Koch’s postulates. To our Knowledges, this is the first report of N. dimidiatum causing leaf blight disease on C. miniata and C. nobilis worldwide, as well as these host plants are new for N. dimidiatum in the world.

Carotenoid assembly regulates quinone diffusion and the Roseiflexus castenholzii reaction center-light harvesting complex architecture.

Carotenoid (Car) pigments perform central roles in photosynthesis-related light harvesting (LH), photoprotection, and assembly of functional pigment-protein complexes. However, the relationships between Car depletion in the LH, assembly of the prokaryotic reaction center (RC)-LH complex, and quinone exchange are not fully understood. Here, we analyzed native RC-LH (nRC-LH) and Car-depleted RC-LH (dRC-LH) complexes in Roseiflexus castenholzii, a chlorosome-less filamentous anoxygenic phototroph that forms the deepest branch of photosynthetic bacteria. Newly identified exterior Cars functioned with the bacteriochlorophyll B800 to block the proposed quinone channel between LHαβ subunits in the nRC-LH, forming a sealed LH ring that was disrupted by transmembrane helices from cytochrome c and subunit X to allow quinone shuttling. dRC-LH lacked subunit X, leading to an exposed LH ring with a larger opening, which together accelerated the quinone exchange rate. We also assigned amino acid sequences of subunit X and two hypothetical proteins Y and Z that functioned in forming the quinone channel and stabilizing the RC-LH interactions. This study reveals the structural basis by which Cars assembly regulates the architecture and quinone exchange of bacterial RC-LH complexes. These findings mark an important step forward in understanding the evolution and diversity of prokaryotic photosynthetic apparatus.

First report of Fusarium falciforme causing root rot of pomegranate (Punica granatum L.) in China.

Pomegranate (Punica granatum L.) is a deciduous shrub or small tree that is native to Iran and Afghanistan. It is also a commercially important fruit tree in China and worldwide. In the summer of 2022, a serious root rot disease occurred in some pomegranate orchards in Xichuan County(32º42´ N, 111º48´ E), Henan Province, China, with an incidence of ~30%. Symptoms included leaf yellowing and wilting, root browning and rotting, and stem-base cracking, eventually leading to defoliation and death. To isolate the causal agent, small pieces (5×5 mm) of diseased root from six trees were surface-sterilized by dipping in 2% NaClO for 8 min followed by 70% ethanol for 15 s, rinsed five times with sterile water, and plated on potato dextrose agar (PDA), then incubated at 28°C in the dark for 5 days. Fifteen pure fungal isolates with the same morphological characteristics were obtained from 24 pieces of roots. All isolates produced white fluffy mycelia. Microconidia were hyaline, oval or reniform, with zero to one septa and dimensions of 7.1 to 19.9 (average 14.5 )× 3.8 to 8.0 (average 5.6) μm (n = 100). Macroconidia were sickle-shaped, one to four septate, and 20.1 to 40.8 (average 26.5) × 4.8 to 8.6 (average 6.5) μm (n = 100). Chlamydospores were spherical, single, in pairs or chains, and 5.6 to 9.8 (average 6.8) µm in diameter (n = 100). Based on the above characteristics, the pathogens were identified as Fusarium sp. (Leslie and Summerell 2006). Genomic DNA was extracted from mycelia of two representative isolates Fs1 and Fs3. The internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF-1α) and RNA polymerase II second largest subunit (RPB2) sequences were PCR amplified using primer pairs of ITS1/ITS4, EF1/EF2, and RPB2-5f2/RPB2-7cr, RPB2-7cf/RPB2-11ar (O'Donnell et al., 2022), respectively. BLAST analysis showed that the ITS, TEF-1α and RPB2 sequences of isolates Fs1(GenBank accession nos. OK001765, OQ921726 and OQ928396) and Fs3 (GenBank accession nos. OK001771, OQ921727 and OQ928397) showed 99%-100% identity with multiple GenBank sequences of Fusarium falciforme (KY617066, MN064683, KF255514, OQ933361, KY556711 and ON331935). A phylogenetic tree based on concatenated sequences of ITS, TEF-1α and RPB2 using maximum-likelihood analysis revealed that both isolates Fs1 and Fs3 were in the same clade with F. falciforme strains. Based on the morphological and molecular characteristics, the isolates were identified as members of F. falciforme. For pathogenicity testing, conidial suspensions (1×108 spores /mL) of isolates Fs1 and Fs3 were poured onto the roots of healthy pomegranate that had been planted in pots two months previously. Ten plants were inoculated for each isolate. Control plants were drenched with sterile water. After 3 months, inoculated plants developed leaf yellowing and wilting accompanied by root browning and rotting, much like symptoms observed in field plants. The same fungi re-isolated from the experimental plants were confirmed to be F. falciforme by morphology and sequence analysis. This is the first report of F. falciforme causing root rot on pomegranate. F. falciforme is a ubiquitous soil-borne pathogen that causes root rot on multiple plants around the world (Xu F., et al. 2022; Qiu R., et al. 2023). The results of pathogen identification are essential precursors to development of effective control of the disease.

Three coralloid species of the genus Trechispora (Trechisporales, Basidiomycota) in China: two newly discovered taxa and one reported for the first time.

Two new species of Trechispora indigenous to southern China, T.laxa and T.tongdaoensis, are described and illustrated, and the first record of T.khokpasiensis in China is reported. Molecular phylogenetic analyses of the concatenated nuclear rDNA ITS1-5.8S-ITS2 and nuclear large subunit sequences supported the inclusion of the three species within the Trechispora clade, together with species formerly classified in Scytinopogon. The new species are similar in micromorphology to species of Trechispora (as traditionally circumscribed) but are distinguished by having coralloid basidiomata. A key to the known coralloid Trechispora species in China is provided.

Antihypertensive peptide resources map of ribulose-1,5-bisphosphate carboxylase/oxygenases (RuBisCO) in angiosperms: Revealed by an integrated in silico and in vitro approach

As the most abundant protein on earth, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) has been considered a promising resource of functional foods. This study aimed to explore the full potential of plant RuBisCO proteins as precursors of antihypertensive peptides on a large scale. In total, 12,766 RuBisCO large subunit and 1,020 RuBisCO small subunit sequences of angiosperms were collected for simulated proteolysis and evaluation of antihypertensive potential, revealing a vast reservoir of antihypertensive peptides. Moreover, RuBisCO-derived novel antihypertensive peptides TTVW, TMW, and VPCL were identified with in vitro IC50 of 12.89 ± 0.82, 23.97 ± 1.02, and 339.12 ± 21.64 μM, respectively. Notably, TTVW and TMW are noncompetitive inhibitors predicted to bound adjacent to the catalytic region of ACE, while VPCL is a competitive inhibitor predicted to bound to the central active site inside ACE. Overall, this work provides a powerful theoretical guidance in developing antihypertensive functional foods utilizing plant RuBisCO.

First report of Pseudocerradoa paullula causing aroid leaf rust on Swiss cheese plant Monstera deliciosa in South Carolina, USA.

In February 2023, two Monstera deliciosa Liebm. (Araceae) plants with typical symptoms of leaf rust disease were detected at a grocery store in Oconee Co., South Carolina. Symptoms included chlorotic leaf spots and abundant brownish uredinia, mainly on the adaxial surface of more than 50% of leaves. The same disease was detected on 11 out of 481 M. deliciosa plants in a greenhouse at a plant nursery located in York Co., South Carolina, in March 2023. The first plant sample detected in February was used for morphological characterization, molecular identification, and pathogenicity confirmation of the rust fungus. Urediniospores were densely aggregated, globose, golden to golden brown in color, and measured 22.9 to 27.9 µm (aver. 26.0 ± 1.1 µm; n=50) in diameter with wall thickness at 1.3 to 2.6 µm (aver. 1.8 ± 0.3 µm; n=50). Telia were not observed. These morphological traits aligned with those of Pseudocerradoa paullula (basionym: Puccinia paullula; Ebinghaus et al. 2022; Sakamoto et al. 2023; Sydow and Sydow 1913; Urbina et al. 2023). Genomic DNA was extracted from urediniospores collected from the naturally infected plant sample and used for PCR amplification and DNA sequencing of the large subunit (LSU) genetic marker with primers LRust1R and LR3 (Vilgalys and Hester 1990; Beenken et al. 2012). The LSU sequence of the rust fungus in South Carolina (GenBank accession: OQ746460) is 99.9% identical to that of Ps. paullula voucher BPI 893085 (763/764 nt.; KY764151), 99.4% identical to that of voucher PIGH 17154 in Florida, USA (760/765 nt.; OQ275201), and 99% identical to that of voucher TNS-F-82075 in Japan (715/722 nt.; OK509071). Based on its morphological and molecular characteristics, the causal agent was identified as Ps. paullula. This pathogen identification was also corroborated by the U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Plant Pathogen Confirmatory Diagnostics Laboratory in Laurel, Maryland. To confirm the fungus's pathogenicity on M. deliciosa and M. adansonii Schott (Sakamoto et al. 2023), three plants of each Monstera species were inoculated by spraying with a suspension of urediniospores collected from the original plant sample (1 × 106 spores per ml; approx. 40 ml per plant). Three non-inoculated control plants of each host species were treated with deionized water in the same manner. Plants were placed in a plastic tray with wet paper towels to maintain moisture. The tray was placed at 22C for an 8-h photoperiod and covered for five days to facilitate infection. On 25 days after inoculation, abundant spots bearing urediniospores were produced on all leaves of inoculated M. deliciosa plants. A few uredinia were observed on two of the three inoculated M. adansonii plants. All non-inoculated control plants remained asymptomatic. Morphological features of urediniospores collected from inoculated plants matched those of Ps. paullula used as the inoculum. Aroid leaf rust on Monstera plants was officially reported in Australia, China, Japan, Malaysia, Philippines, and Florida, USA (Shaw 1991; Sakamoto et al. 2023; Urbina et al. 2023). This is the first report of Ps. paullula causing this disease on M. deliciosa in South Carolina, USA. Monstera species are popular indoor and landscape plants. Potential impact and regulatory responses regarding Ps. paullula, a newly introduced and rapidly spreading pathogen in the USA, warrant further evaluation and discussion.

Isolation and identification of green alga Chlorella vulgaris HNUFU001 for environmental selenium enrichment in heterotrophic growth regime

Microalgae, particularly Chlorella, have promising selenium (Se) enrichment capacity and great potential in wastewater treatment and the development of nutraceuticals. To acquire Chlorella strains with strong heterotrophic propensity and robust Se tolerance, this research utilized a heterotrophic isolation strategy along with a high Se concentration medium as selection factors. The combined sequence of the ribosomal small subunit (SSU) and internal transcribed spacer (ITS) was utilized for phylogenetic analysis, and comparative analysis of ITS secondary structures was also integrated to identify the desired strain. A novel subtropical microalgal strain, Chlorella vulgaris HNUFU001 with high Se tolerance, was isolated from water samples in a lake at Hainan University, China. Gaussian process regression was utilized to assess the strain's growth under varying Se concentrations. The intracellular accumulation and extracellular removal of Se were measured using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) and Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES), respectively. The maximum intracellular organic Se (92 % of total Se) was observed as 108.835 μg/g dry weight (DW) for 1 mg/L Se treatment. The highest Se removal rate was 40 %. The high Se tolerance and enrichment traits enable Chlorella vulgaris HNUFU001 to be a promising candidate for the heterotrophic production of organic Se-related nutraceuticals.

Genetic Diversity of the Fall Armyworm Spodoptera frugiperda (J.E. Smith) in the Democratic Republic of the Congo

In 2016, the fall armyworm (FAW), Spodoptera frugiperda, invaded western Africa and rapidly spread in sub-Saharan Africa, causing significant losses in yields of corn, a major food crop in Africa. Although the Democratic Republic of the Congo (DRC) is a large corn-growing country, the impact of FAW has not been investigated. This study was designed to expand investigations on the genetic diversity of FAW populations in the DRC. We collected FAW individuals from eight provinces across the country, for analysis of genetic variation. Based on the partial sequences of both mitochondrial cytochrome oxidase subunit I (COI) and nuclear triosephosphate isomerase (Tpi) genes, we compared polymorphic features of the COI haplotype and Tpi single nucleotide polymorphisms. The results revealed that most (84%) of the analyzed individuals were heterogeneous hybrids Tpi-corn/COI-rice (Tpi-C/COI-R), whereas 16% were homogenous Tpi-corn/COI-corn (Tpi-C/COI-C). Further analysis of the fourth exon/intron sequences of the Tpi gene identified two subgroups, TpiCa1 and TpiCa2, constituting 80% and 20%, respectively, of the collected individuals. Analysis of genetic variation among native and invasive populations indicated significant genetic differences (10.94%) between the native American and DRC populations, whereas both the DRC and African populations were genetically closer to Asian than American populations. This study provides important information on FAW genetic diversity in the DRC, which can be used for effective management of FAW.

Detection and incidence of pineapple heart rot disease caused by Phytophthora nicotianae in commercial farms of Puerto Rico

Pineapple heart rot disease (PHRD) caused by Phytophthora spp. is one of the most important diseases of pineapple. This disease is characterized by soft rot and water-soaked lesions. In severe cases plant mortality is 100%. During a one-year survey, conducted from March 2018 to March 2019, a total of 29 pineapple hectares were evaluated from five commercial pineapple fields located in the Puerto Rican municipalities of Guánica, Lajas, Manatí and Santa Isabel, and at the Agricultural Experiment Substation of the University of Puerto Rico in Isabela. Symptoms of PHRD were observed in all fields evaluated, except at the Agricultural Experiment Substation in Isabela. Diseased plant tissue was surface disinfected and plated on PARPH-V8 selective media for Phytophthora spp. Aerial photography using a DJI Phantom 3 drone was used to corroborate PHRD incidence in the field. Approximately eight hectares were infected by P. nicotianae with an average of PHRD disease incidence of 28.82%. The highest percentage of PHRD incidence was observed in fields located at Guánica, Lajas and Manatí with 40%, 40% and 30%, respectively. Eleven isolates of Phytophthora nicotianae were collected from pineapple fields in Guánica, Lajas, Manatí and Santa Isabel and identified using morphology and phylogeny of sequences of the mitochondrial cytochrome oxidase subunit I region (COI). The estimation of the incidence of P. nicotianae as the causal agent of PHRD is important as a first step in developing specific control measures in the pineapple fields of Puerto Rico.

First report of Curvularia geniculata causing leave spot disease of Bletilla striata in Zhejiang of China.

Bletilla striata, a perennial herbaceous plant belonging to the family Orchidaceae, is native to China and is widely distributed in the Yangtze River basin. In China, B. striata is a popular medicinal plant that is typically used to reduce wound bleeding and inflammation. In September 2021, distinct leaf spot symptoms were observed in more than 50% of B. striata plants in a traditional Chinese medicine plantation (ca. 10 ha) in Xianju City, Zhejiang Province, China. Small, round, pale brown, necrotic spots were first observed on the leaves. Subsequently, these lesions became grayish brown in the center and dark brown with slight protuberances at the margins and eventually enlarged to 5-8 mm on the leaves. Over time, the small spots enlarged and coalesced into necrotic streaks (1-2 cm). Leaves with symptoms of disease were cut, surface-sterilized, and plated on potato dextrose agar (PDA). Fungal colonies (28×28 mm) with grayish-black mycelia from all tissues were produced after 3 days of incubation at 26 °C. The mature colonies eventually turned black in the center, with obvious rings appearing after 10 days of culture. Basal conidia ranged from pale to dark brown, whereas apical ones were pale brown, with central cells being larger and darker than basal cells. Conidia were smooth and either fusiform, cylindrical, or slightly curved with rounded tips. They ranged in length from 22.34 to 36.82 (mean = 28.63) μm with 2-4 septations and slight septal constrictions. Monospore isolation was performed to obtain a pure culture. Strain BJ2Y5 was subsequently stored in the strain Preservation Center of Wuhan University (Wuhan, China) and the strain preservation number CCTCC M 2023123 was obtained. Fresh mycelia and conidia that grew at 26 ℃ for 7 days were collected from PDA plates. DNA was extracted using the Ezup Column Fungi Genomic DNA Purification Kit (Sangon Biotech Co., Shanghai, China). The phylogenetic position of isolate BJ2-Y5 was clarified based on DNA sequence analysis of three loci, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Berbee et al. 1999), the internal transcribed spacer (ITS) region (White et al. 1990), and partial sequences of the second largest subunit of RNA polymerase II (RPB2; O'Donnell et al. 2007). A BLAST search (GenBank accession nos. OP913168, OP743380, and OP913171) showed 99% homology to the reference isolate CBS 220.52. Based on the morphological and molecular information presented in this study these isolates were identified as C. geniculata (Hosokawa et al. 2003). Furthermore, we evaluated the pathogenicity of B. striata leaves by smearing a conidial suspension (106 conidia/mL) on both sides of leaves with and without wounds. Five inoculated and three non-inoculated leaves (smeared with sterile distilled water as a negative control) were kept in a greenhouse at 26 °C under natural sunlight and covered with plastic bags for 72 h to maintain humidity. After 7 days, small round spots appeared on the wounds. Fifteen days later, the symptoms of disease on the wounded inoculated leaves were similar to those observed in the original sample, whereas the control plants remained healthy. No symptoms of infection were observed in unwounded inoculated leaves. C. geniculata was successfully re-isolated from all five inoculated leaves and was confirmed based on Koch's postulates. To the best of our knowledge, C. geniculata infection has not been previously reported in B. striata.

A new species of Cycloporus from the Adriatic Sea, with an updated phylogeny of the families Euryleptidae and Stylostomidae (Polycladida, Platyhelminthes).

We describe Cycloporus pinkipus sp. n., a new polyclad flatworm species from the Adriatic coast of Croatia using live images, histological sections, and a molecular marker. It is the fifteenth described species of Cycloporus Lang, 1884 and the second described congener in the Mediterranean. The genus Cycloporus is characterised by a small oval body, tentacular bumps and the name-giving marginal pores. Cycloporus pinkipus sp. n. has a smooth dorsal surface, which is transparent creamy white with light brown to yellow spots, covered with prominent serial pink spots on the inner rim of the body margin. There is little variation of the genital organs between different species of the genus, therefore we recognise C. pinkipus sp. n. as a new species in particular due to its unique coloration, and a unique partial large nuclear ribosomal subunit (28S) sequence. In recent years the family Euryleptidae Stimpson, 1857, which also contains the genus Cycloporus, was discussed and revised in several molecular studies. In an updated molecular phylogeny of the Polycladida based on partial 18S and 28S rDNA marker genes, C. pinkipus sp. n. was recovered in a clade of many other Cycloporus species within Euryleptidae.

The population genetic structure and phylogeographic dispersal of Nodularia breviconcha in the Korean Peninsula based on COI and 16S rRNA genes.

A freshwater mussel, Nodularia breviconcha (Mollusca: Bivalvia: Unionida) is endemic to Korean Peninsula. It has recently been taxonomically reexamined and elevated from a subspecies of N. douglasiae to an independent species. But population genetic studies for the species have rarely been conducted. To explore the population genetic structure of N. breviconcha, the nucleotide sequences of cytochrome oxidase subunit I(COI) and 16S rRNA genes from 135 N. breviconcha individuals, including 52 from this study and 83 from Choi et al. (2020). We found 23 COI and 11 16S rRNA genes haplotypes. Phylogeny, TCS network, Principal coordinates analysis, and spatial analysis of molecular variance performed with COI gene indicated that there are exist three different genetic lineages in the N. breviconcha populations: West lineage, Southwest lineage, and Southeast lineage. According to the time calibrated phylogeny, they are likely to be diverged during the late Miocene (8-6 Ma). Geographical distribution patterns of the three genetic lineages may be related to the formation of Taebaek and Sobaek-Noryeong mountain ranges in the Korean Peninsula occurred during the Miocene (30-10 Ma). The present results of this study will be helpful not only for the conservation, but also for the exploration of the population genetic structure of endemic freshwater mussels in the Korean Peninsula.

A New Lichenized Fungus, Lendemeriella luteoaurantia, with a Key to the Species of Lendemeriella

Lendemeriella luteoaurantia B.G. Lee is described as a new lichen species from South Korea. The new species is identified by smaller, yellow-orange apothecia, larger ascospores with wider septum width, and the absence of Cinereorufa-green pigment and teloschistin, different from the closest species, L. aureopruinosa I.V. Frolov, Vondrák, Arup, Konoreva, S. Chesnokov, Yakovczenko and Davydov in morphology and chemistry. Molecular phylogeny employing internal transcribed spacer (nuITS), nuclear large subunit ribosomal RNA (nuLSU), and mitochondrial small subunit (mtSSU) sequences strongly supports the new species as nonidentical in the genus Lendemeriella. A preliminary key is provided to assist in the identification of all 10 species of Lendemeriella.

First Report of Rust Disease on Alcea rosea Caused by Puccinia modiolae in Korea.

Alcea rosea, in the family Malvaceae, is a biennial plant native to China and is grown typically for gardening in Korea (Lee 2003). Seven microcyclic Puccinia species have been reported on A. rosea: P. heterogenea, P. heterospora, P. lobata, P. malvacearum, P. platyspora, P. sherardiana, and P. modiolae (Demers et al. 2015; Aime and Abbasi 2018). In early May 2022, characteristic symptoms of rust were observed on four of ten seedlings of A. rosea purchased at a wholesale nursery (36°50'19.8″N, 128°55'28.7″E) in Bonghwa, Korea. Rust spots were present on almost 90% of the 1,000 seedlings of A. rosea in that nursery during our survey in late May. Through a distribution survey from June to July 2022, similar symptomatic leaves were additionally collected from the leaves of A. rosea grown in gardens at five sites in Gimcheon (two sites), Gumi (one), Seongju (one), and Busan (one). Spots were yellow-orange the center surrounded by chlorotic haloes on the adaxial leaf surface, and reddish-brown or dark brown pustules on the abaxial leaf surface. Over time, the spots enlarged and coalesced, causing the decay of large sections of the leaves, and heavily infected leaves fell early. Spermogonia, produced at the center of the chlorotic spot on the adaxial leaf surface, were subepidermal, obovoid, and 113.2-164.5 × 97.6-153.3 μm in size. Telia were reddish-brown to dark brown, round, mostly grouped, 0.28-0.61 mm in diameter, and mainly formed on the abaxial leaf surface but sometimes on the adaxial leaf surface also. Teliospores were two-celled, but rarely one- or three-celled, and were fusoid and 37. 6-110 × 12.4-21.5 μm in size; the wall was yellowish or almost colorless, smooth, 1.2-2.6 μm thick at the sides, and up to 7.4 μm thick at the apex. The morphological characteristics were similar to those of P. modiolae, although the teliospores in our study were longer than those observed by Aime and Abbasi (2018). For phylogenetic analysis, genomic DNA was extracted from the teliospores of each regional specimen. Partial 18S, internal transcribed spacer (ITS), and partial 28S sequences were amplified using primers NS1, ITS4, ITS5, and LR11. The PCR products were sequenced (Celemics, Seoul, Korea) and deposited in GenBank. The ITS-partial large subunit (LSU) sequence and 28S sequences had 100% homology with other P. modiolae sequences deposited in GenBank (accession numbers are shown in Fig. 2). In the phylogenetic trees of the ITS and LSU sequences, the isolates collected in this study were grouped with the reference sequences of P. modiolae, including the Korean isolate (ON631218) recently reported on Malva verticillata by Lee et al. (2022). For the pathogenicity test, the teliospores with germinating basidiospores were suspended in sterile distilled water and smeared on the upper surface of asymptomatic A. rosea leaves in August. The inoculated plants were sprayed with distilled water and kept in the dark with saturated moisture for 24 h in an isolated glass house of the Animal & Plant Quarantine Agent. After 2 weeks, typical rust spots and telia of P. modiolae were observed on the leaves of the inoculated plants, but not in the control plants, which were only sprayed with distilled, sterilized water and otherwise treated similarly to the inoculated plants. The results of this study show that the casual fungus is P. modiolae, which has been commonly found in A. rosea in Korea. To the best of our knowledge, this is the first report of P. modiolae in A. rosea in Korea.

Diptacus rubuscolum Trinidad, Duarte and Navia (Eriophyoidea: Diptilomiopidae) on blackberry (Rubus ulmifolius Schott) in Mexico: occurrence, complementary description, and biology

Blackberry (Rubus ulmifolius Scott) is an economically important crop in Mexico. Eriophyoidea mites are among the most important arthropod pests affecting production of this crop worldwide. Despite this, reports on species diversity of Eriophyoidea on blackberry in Mexico are scarce. We sampled six orchards from two municipalities in the state of Michoacan, the most important region for blackberry production. Sampling was carried out monthly between May and November 2018. Only one species of eriophyoid mite was found, Diptacus rubuscolum Trinidad, Duarte and Navia. We conducted a detailed morphological study of this mite and a complementary description is presented. In addition, we obtained partial sequences of the mitochondrial cytochrome oxidase subunit I (mt COI), the 18S and 28S of the rRNA. The duration of each developmental stage was determined under controlled conditions (25 °C, 60% RH and 12:12 light regime). We confirmed the identity of D. rubuscolum by comparing partial sequences of the D2 region within the 28S rRNA from our samples and sequences retrieved from GenBank. Overall, morphological characteristics were in line with the type specimens. However, some differences on body shape and empodium structure were pointed out. The developmental stages of D. rubuscolum were egg, larva, nymphochrysalis, nymph, imagochrysalis and adult, with a mean duration of 75 (±10), 56 (±9), 21 (±7), 41 (±8), 19 (±6) and 114 (±12) hours respectively. The information produced provides insight and knowledge on the occurrence of this species on blackberry in Mexico as well as on original biological aspects.

Lessons from the deep: mechanisms behind diversification of eukaryotic protein complexes.

Genetic variation is the major mechanism behind adaptation and evolutionary change. As most proteins operate through interactions with other proteins, changes in protein complex composition and subunit sequence provide potentially new functions. Comparative genomics can reveal expansions, losses and sequence divergence within protein-coding genes, but in silico analysis cannot detect subunit substitutions or replacements of entire protein complexes. Insights into these fundamental evolutionary processes require broad and extensive comparative analyses, from both in silico and experimental evidence. Here, we combine data from both approaches and consider the gamut of possible protein complex compositional changes that arise during evolution, citing examples of complete conservation to partial and total replacement by functional analogues. We focus in part on complexes in trypanosomes as they represent one of the better studied non-animal/non-fungal lineages, but extend insights across the eukaryotes by extensive comparative genomic analysis. We argue that gene loss plays an important role in diversification of protein complexes and hence enhancement of eukaryotic diversity.

Taxonomical, functional, and cytopathological characterization of Bacillus spp. from Lake Magadi, Kenya, against Rhizoctonia solani Kühn in Phaseolus vulgaris L.

A decline in common bean production and the ineffectiveness of synthetic chemical products in managing plant pathogens has led to exploiting Kenyan soda lakes as an alternative search for biocontrol agents. This study aimed to identify phylogenetically Bacillus spp. from Lake Magadi and their antagonistic activity against Rhizoctonia solani under in vitro and in vivo conditions. The 16 S ribosomal RNA (rRNA) subunit sequences of six bacterial strains isolated from Lake Magadi showed diversity similar to the Bacillus genus; Bacillus velezensis, Bacillus subtilis, and Bacillus pumilus. In vitro, antagonism showed varied mycelium inhibition rates of fungi in the coculture method. Enzymatic assays showed the varied ability of isolates to produce phosphatase, pectinase, chitinase, protease, indole-3-acetic acid (IAA), and hydrogen cyanide (HCD). The in vivo assay showed M09 (B. velezensis) with the lowest root mortality and incidence of postemergence wilt. Pre-emergence wilt incidence was recorded as lowest in M10 (B. subtilis). Isolate M10 had the highest phenylalanine ammonia-lyase (PAL) for defense enzymes, while polyphenol oxidase (PPO) and peroxidase were recorded as highest in M09. For the phenolic content, M10 recorded the highest phenolic content. In conclusion, Lake Magadi harbors Bacillus spp, which can be used as a potential biocontrol of R. solani.

Taxonomy and Phylogenetic Relationships of Clavulinopsis (Clavariaceae, Agaricales): Description of Six New Species and One Newly Recorded Species from China.

Specimens of Clavulinopsis (Clavariaceae, Agaricales) collected in China were studied using morphological and molecular methods. Six species-C. aspersa, C. bicolor, C. bispora, C. erubescens, C. incarnata, and C. tropicalis-are described as new to science, and C. trigonospora is a newly recorded species in China. Phylogenetic analysis was conducted based on a combined dataset of internal transcribed spacer and nuclear ribosomal RNA large subunit sequences. The phylogenetic reconstruction revealed that the six new species each formed an independent lineage, and the samples of C. trigonospora from China were nested with accessions of C. trigonospora collected from Italy. The morphology of the seven Chinese species is described in detail, and is illustrated with line drawings and photographs. A key to the known Clavulinopsis species in China is provided.