High Support Values Research Articles

Characterization of the plastome of Physaliscordata and comparative analysis of eight species of Physalis sensu stricto.

In this study, we sequenced, assembled, and annotated the plastome of Physaliscordata Mill. and compared it with seven species of the genus Physalis sensu stricto. Sequencing, annotating, and comparing plastomes allow us to understand the evolutionary mechanisms associated with physiological functions, select possible molecular markers, and identify the types of selection that have acted in different regions of the genome. The plastome of P.cordata is 157,000 bp long and presents the typical quadripartite structure with a large single-copy (LSC) region of 87,267 bp and a small single-copy (SSC) region of 18,501 bp, which are separated by two inverted repeat (IRs) regions of 25,616 bp each. These values are similar to those found in the other species, except for P.angulata L. and P.pruinosa L., which presented an expansion of the LSC region and a contraction of the IR regions. The plastome in all Physalis species studied shows variation in the boundary of the regions with three distinct types, the percentage of the sequence identity between coding and non-coding regions, and the number of repetitive regions and microsatellites. Four genes and 10 intergenic regions show promise as molecular markers and eight genes were under positive selection. The maximum likelihood analysis showed that the plastome is a good source of information for phylogenetic inference in the genus, given the high support values and absence of polytomies. In the Physalis plastomes analyzed here, the differences found, the positive selection of genes, and the phylogenetic relationships do not show trends that correspond to the biological or ecological characteristics of the species studied.

The complete chloroplast genome sequence of Zygophyllum kansuense Y. X. Liou (Zygophyllaceae)

Zygophyllum kansuense Y. X. Liou is a localized species and endemic to China. In this study, the complete chloroplast genome of Z. kansuense was sequenced and described. The length of complete chloroplast genome was 105,383 base pairs (bp) for Z. kansuense, the genome with 33.8% GC content, containing a large single-copy (LSC) of 79,594 bp, a small single-copy (SSC) of 17,061 bp, and a pair of inverted repeats (IRs) of 4,364 bp. The genome contains 112 genes, including 75 protein-coding genes, 33 tRNA genes, and four rRNA genes. The phylogenetic analysis indicated that Z. kansuense and others of Zygophyllum clustered into one clade with a high support value.

Mitochondrial DNA barcoding of mosquito species (Diptera: Culicidae) in Thailand

The correct identification of mosquito species is important for effective mosquito vector control. However, the standard morphological identification of mosquito species based on the available keys is not easy with specimens in the field due to missing or damaged morphological features during mosquito collections, often leading to the misidentification of morphologically indistinguishable. To resolve this problem, we collected mosquito species across Thailand to gather genetic information, and evaluated the DNA barcoding efficacy for mosquito species identification in Thailand. A total of 310 mosquito samples, representing 73 mosquito species, were amplified using mitochondrial cytochrome c oxidase subunit I (COI) primers. The average maximum intraspecific genetic variation of the 73 mosquito species was 1% ranged from 0-5.7%. While, average minimum interspecific genetic variation (the distance to the nearest neighbour) of the 73 mosquito species was 7% ranged from 0.3-12.9%. The identification of success rates based on the "Best Match," "Best Close Match," and "All Species Barcodes" methods were 97.7%, 91.6%, and 81%, respectively. Phylogenetic analyses of Anopheles COI sequences demonstrated a clear separation between almost all species (except for those between An. baimaii and An. dirus), with high bootstrap support values (97%-99%). Furthermore, phylogenetic analyses revealed potential sibling species of An. annularis, An. tessellatus, and An. subpictus in Thailand. Our results indicated that DNA barcoding is an effective molecular approach for the accurate identification of mosquitoes in Thailand.

Morphological and phylogenetic characteristics of Phaeomycocentrospora xinjangensis (Pleosporales, Dothideomycetes), a new species from China

The genus Phaeomycocentrospora (Dothidotthiaceae) is a fungal pathogen causing plant leaf spots. A new species, Phaeomycocentrospora xinjangensis sp. nov. is introduced from Xinjiang Uygur Autonomous Region, Northwest China. Multi-gene phylogenetic analyses based on a concatenated ITS, LSU, and ACT sequence dataset were used to confirm the phylogenetic position of the new species. Phaeomycocentrospora xinjangensis can easily be distinguished from the remaining species based on multi-gene phylogenetic analyses coupled with morphological characters. Morphologically, P. xinjangensis differs from other species in the genus by the presence of dark hyphae, dark brown conidiophores, few branched conidiogenous cells. Phylogenetically, our three strains were clustered together and formed a separate subclade with high support values. Moreover, we provided a description, illustrations, and phylogenetic tree for the new species.

Taxonomy, comparative genomics of Mullein (Verbascum, Scrophulariaceae), with implications for the evolution of Verbascum and Lamiales

BackgroundThe genus Verbascum L. (Scrophulariaceae) is distributed in Africa, Europe, and parts of Asia, with the Mediterranean having the most species variety. Several researchers have already worked on the phylogenetic and taxonomic analysis of Verbascum by using ITS data and chloroplast genome fragments and have produced different conclusions. The taxonomy and phylogenetic relationships of this genus are unclear.ResultsThe complete plastomes (cp) lengths for V. chaixii, V. songaricum, V. phoeniceum, V. blattaria, V. sinaiticum, V. thapsus, and V. brevipedicellatum ranged from 153,014 to 153,481 bp. The cp coded 114 unique genes comprising of 80 protein-coding genes, four ribosomal RNA (rRNA), and 30 tRNA genes. We detected variations in the repeat structures, gene expansion on the inverted repeat, and single copy (IR/SC) boundary regions. The substitution rate analysis indicated that some genes were under purifying selection pressure. Phylogenetic analysis supported the sister relationship of (Lentibulariaceae + Acanthaceae + Bignoniaceae + Verbenaceae + Pedaliaceae) and (Lamiaceae + Phyrymaceae + Orobanchaceae + Paulowniaceae + Mazaceae) in Lamiales. Within Scrophulariaceae, Verbascum was sister to Scrophularia, while Buddleja formed a monophyletic clade from (Scrophularia + Verbascum) with high bootstrap support values. The relationship of the nine species within Verbascum was highly supported.ConclusionBased on the phylogenetic results, we proposed to reinstate the species status of V. brevipedicellatum (Engl.) Hub.-Mor. Additionally, three genera (Mazus, Lancea, and Dodartia) placed in the Phyrymaceae family formed a separate clade within Lamiaceae. The classification of the three genera was supported by previous studies. Thus, the current study also suggests the circumscription of these genera as documented previously to be reinstated. The divergence time of Lamiales was approximated to be 86.28 million years ago (Ma) (95% highest posterior density (HPD), 85.12–89.91 Ma). The complete plastomes sequence data of the Verbascum species will be important for understanding the Verbascum phylogenetic relationships and evolution in order Lamiales.

Next-generation sequencing yields the complete mitochondrial genome of Rhodoprasina callantha (Lepidoptera: Sphingidae) and its evolutionary status

In this study, we sequenced and analyzed the complete mitochondrial genome of Rhodoprasina callantha Jordan, 1929. The complete mitochondrial genome sequence of R. callantha was 15,343 bp in size and encoded 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNA), 22 transfer RNA genes (tRNA) and one control region (CR). The nucleotide composition of mitogenome was highly biased to A and T. Most protein-coding genes (PCGs) initiate with the standard start codon of ATN and terminate with the typical stop codon TAA/TAG. The phylogeny of Sphingidae based on nucleotide sequences of PCGs recovered the monophyly of subfamilies of Sphingidae with high support values. Langinae was the first subfamily diverged in Sphingidae, which was in accordance with previous study. R. callantha was the member of subfamily Smerinthinae, and closely related to the genus Marumba.

First Report of Didymella glomerata Causing Didymella Leaf Blight on Maize in China.

Maize (Zea mays L.) is a staple food crop worldwide. In July 2021, gray leaf blight was observed on maize leaves in a field located in Panjin (41°7'11.98" N, 122°4'14.57" E), Liaoning Province, China. Nearly 5% of the maize plants were affected in the field. The leaves of the affected plants showed oval to oblong, gray, sunken lesions with yellow or tan margins. The lesions were scattered all over the leaf surface; however, they were absent on the stalks and other parts of the affected plants. To isolate the pathogen, leaf discs (1.25 mm2) excised from the blight lesions were surface-sterilized with 70% ethanol for 30 seconds, followed by 20% NaOCl for 2 minutes and finally rinsed three times with sterilized water. The discs were cultured on potato dextrose agar (PDA) plates supplemented with streptomycin (100 mg/L) and incubated at 25oC under a 12-h photoperiod for 7 days. Six single spore isolates (two per sampled infected leaf) were purified from the PDA culture plates. The fungal colonies of three selected isolates (one per sampled infected leaf; Pj-1, Pj-2, and Pj-3) were dark brown on the PDA plates and devoid of aerial hyphae; all three isolates grew 11 mm/day on the PDA plates. The number of conidia produced by the isolates on the 6-cm PDA plates 7 days after incubation was ranged from 160 x 108 to 208 x 108 (n = 36). Conidia were hyaline, single-celled and ellipsoidal (3.35-3.56 µm [width] x 6.47-6.70 [length] µm; n = 36). To identify the pathogen, four loci, i.e., 28S subunit (large subunit [LSU]) of the nuclear ribosomal (nr) DNA, internal transcribed spacer (ITS) region (ITS1, 5.8S subunit of nrDNA, and ITS2), the second-largest subunit of RNA polymerase II (rpb2) and β-tubulin (tub2) were amplified using the primer sets described in the study by Chen el al. 2015. BLASTn search against GenBank revealed that the four amplicon sequences originating from Pj-1, Pj-2, and Pj-3 showed 99-100% homology to the type strain CBS 528.66 of D. glomerata. A phylogenetic tree deduced from a maximum likelihood analysis of a concatenated MUSCLE-based alignment of LSU, ITS region, rpb2, and tub2 sequences of 12 isolates/strains showed that the Pj isolates clustered together with CBS 528.66, along with other D. glomerata isolates/strains, with a high bootstrap support value (i.e., 99). Based on both morphological characteristics and molecular phylogeny, Pj-1, Pj-2, and Pj-3 were identified as the D. glomerata isolates. Since the amplicon sequences of the three isolates were identical, only Pj-2 sequences were deposited in GenBank with accession numbers OM372474 (LSU), OK485138 (ITS), OM406188 (rpb2), and OK485135 (tub2). To confirm pathogenicity, 14-day-old plants (V3 growth stage) of a maize cultivar P178 were spray-inoculated with the Pj-2 conidia (1 x 107 conidia/mL) in a growth chamber. The inoculated leaves exhibited typical gray leaf blight lesions (similar to those detected in the maize field) 7 days post-inoculation at 25oC and 95-100% humidity under a 12-h photoperiod, whereas the leaves spray-inoculated with sterilized water remained healthy. The pathogenicity assay was repeated three times; the pathogen was re-isolated from the inoculated leaves each time and confirmed by the morphological characteristics and the molecular phylogeny based on the four loci to be D. glomerata, fulfilling Koch's postulates. This first report of D. glomerata causing Didymella leaf blight on maize will help develop robust disease management strategies against this emerging fungal pathogen.

Old specimens for old branches: Assessing effects of sample age in resolving a rapid Neotropical radiation of squirrels

Ultraconserved Elements (UCEs) have been useful to resolve challenging phylogenies of non-model clades, unpuzzling long-conflicted relationships in key branches of the Tree of Life at both deep and shallow levels. UCEs are often reliably recovered from historical samples, unlocking a vast number of preserved natural history specimens for analysis. However, the extent to which sample age and preservation method impact UCE recovery as well as downstream inferences remains unclear. Furthermore, there is an ongoing debate on how to curate, filter, and properly analyze UCE data when locus recovery is uneven across sample age and quality. In the present study we address these questions with an empirical dataset composed of over 3800 UCE loci from 219 historical and modern samples of Sciuridae, a globally distributed and ecologically important family of rodents. We provide a genome-scale phylogeny of two squirrel subfamilies (Sciurillinae and Sciurinae: Sciurini) and investigate their placement within Sciuridae. For historical specimens, recovery of UCE loci and mean length per locus were inversely related to sample age; deeper sequencing improved the number of UCE loci recovered but not locus length. Most of our phylogenetic inferences—performed on six datasets with alternative data-filtering strategies, and using three distinct optimality criteria—resulted in distinct topologies. Datasets containing more loci (40% and 50% taxa representativeness matrices) yielded more concordant topologies and higher support values than strictly filtered datasets (60% matrices) particularly with IQ-Tree and SVDquartets, while filtering based on information content provided better topological resolution for inferences with the coalescent gene-tree based approach in ASTRAL-III. We resolved deep relationships in Sciuridae (including among the five currently recognized subfamilies) and relationships among the deepest branches of Sciurini, but conflicting relationships remain at both genus- and species-levels for the rapid Neotropical tree squirrel radiation. Our results suggest that phylogenomic consensus can be difficult and heavily influenced by the age of available samples and the filtering steps used to optimize dataset properties.

Global Taxonomy and Phylogeny of Irpicaceae (Polyporales, Basidiomycota) With Descriptions of Seven New Species and Proposals of Two New Combinations.

The phylogenetic analyses of the family Irpicaceae were carried out based on a complete global sampling. The dataset that included concatenated ITS1-5.8S-ITS2 and nrLSU sequences of 67 taxa of Irpicaceae from around the world was subjected to the maximum likelihood analyses and Bayesian inference. In the phylogenetic tree, species from 14 genera were distributed in nine clades, among which five genera—Irpex, Phanerochaetella, Byssomerulius, Cytidiella, and Meruliopsis, received high support values. The genus Efibula was shown to be paraphyletic and four subclades could be recognized, while Phanerochaete allantospora, Leptoporus mollis, and several species from Ceriporia and Candelabrochaete formed a large clade with relatively strong support. Based on the molecular and morphological evidence, seven new corticioid species—Candelabrochaete guangdongensis, Efibula grandinosa, E. hainanensis, E. shenghuae, E. taiwanensis, Irpex alboflavescens, and Phanerochaetella sinensis, were revealed from the materials mostly from East Asia. The monotypic genus Flavodontia, newly described from southwestern China, is regarded as a later synonym of Irpex, and the new combination I. rosea is proposed. In addition, Phanerochaetella queletii is proposed for a taxon first described from Italy and newly recorded from China; Phanerochaete jose-ferreirae from Portugal is determined to be a later synonym. Descriptions and illustrations of the new species and the newly combined taxa are presented, and morphological comparisons for the known species of Efibula and Phanerochaetella are provided.

A phylogenetic study of a tropical sea urchin, Echinometra sp. EZ (Echinoidea: Camarodonta: Echinometridae) from the Persian Gulf

AbstractSea urchins have important effects on marine ecosystems such as rocky shores and coral reefs across the world. However, species diversity and molecular phylogeny of most echinoid taxa are poorly known in Iran. In this study, the phylogenetic relationships of one of the most abundant species of the genus Echinometra in the Persian Gulf were examined. Echinoids were collected from the intertidal zone of Qeshm Island and Lengeh Port on March and December 2017. Morphological criteria based on valid identification keys combined with molecular analysis of a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) protein-coding gene were used to delineate Echinometra species. Our analyses showed that all specimens (N = 15) belong to Echinometra sp. EZ. Tree topologies indicated that our individuals from two sampling sites formed a distinct monophyletic clade with E. sp. EZ, demonstrating high support values. This is the first phylogenetic analysis of E. sp. EZ from Iran.

Two new species of Dugesia (Platyhelminthes, Tricladida, Dugesiidae) from the subtropical monsoon region in Southern China, with a discussion on reproductive modalities

BackgroundFreshwater planarians of the genus Dugesia (Platyhelminthes, Tricladida, Dugesiidae) are distributed in a major part of the Old World and Australia, although until recently only very few species were known from China.ResultsTwo new species of Dugesia from Southern China are described on the basis of an integrative taxonomic approach. BI and ML phylogenetic trees based on the independent genes and on the concatenated dataset had similar topologies, only differing in some nodes that were weakly supported. Phylogenetic trees based on the concatenated dataset revealed that D. adunca Chen & Sluys, sp. nov. and D. tumida Chen & Sluys, sp. nov. are not closely related and belong to different clades. The two new species occupy separate long branches with high support values and, thus, are well-differentiated from their congeners. Separate species status of D. adunca and D. tumida is supported also by the genetic distances between the species included in our analysis, albeit that COI distances varied greatly among species. Dugesia adunca from Guangxi Province is characterized by the following features: living mature animals rather small; asymmetrical openings of the oviducts into the bursal canal; penis papilla with shape of an aquiline bill, albeit with a blunt tip; asymmetrical penis papilla, with a large antero-dorsal lip and a much smaller ventro-posterior lip; very large seminal vesicle, provided with trabeculae; small diaphragm; mixoploid karyotype with diploid complements of 2n = 2x = 16 and triploid complements of 2n = 3x = 24, with all chromosomes being metacentric. Dugesia tumida from Guangdong Province is characterized by a penis papilla provided with a large, symmetrical penial valve from the middle of which arises the small, distal section of the papilla; a duct intercalated between the seminal vesicle and the small diaphragm; ventrally displaced ejaculatory duct curving upwards before opening to the exterior; penis papilla highly asymmetrical, having a slim and long ventral portion and a short and stubby dorsal part; vasa deferentia separately opening into antero-dorsal portion of seminal vesicle; oviducts openings symmetrically into ventral portion of the bursal canal, near its opening into the atrium; mixoploid karyotype, with diploid chromosome portraits of 2n = 2x = 16, and triploid complements of 2n = 3x = 24, with all chromosomes being metacentric. In the context of the various kinds of mixoploidy and the sexualization of specimens, reproductive modalities within the genus Dugesia are shortly discussed.ConclusionMolecular, morphological, and karyological markers show that the two populations examined represent members of the genus Dugesia and constitute two new, distinct species.

Implementation of the FP-Growth Algorithm in Sales Transactions for Menu Package Recommendations at Warung Oemah Tani

Along with the rapid development of the culinary industry, business competition is also getting tougher. Warung Oemah Tani serves a variety of menus and drinks, but to provide satisfying service to customers, business people must try to develop new products. Under these circumstances, the menu recommendations for Warung Oemah Tani need to be analyzed so that the recommendations made are right on target. This study aims to analyze the sales of Warung Oemah Tani using the FP Growth algorithm. This algorithm identifies the data set with the highest frequency of concurrent sales (frequent itemset). The results of the association rules show that the highest support value is 0.520 and the highest confidence value is 0.929, with a minimum support of 30% and a minimum confidence of 80%. Obtained 14 rule associations that meet the minimum support and minimum confidence.

Characterization of four mitochondrial genomes of Crambidae (Lepidoptera, Pyraloidea) and phylogenetic implications.

Loxostege turbidalis, Loxostege aeruginalis, Pyrausta despicata, and Crambus perlellus belong to Crambidae, Pyraloidea. Their mitochondrial genomes (mitogenomes) were successfully sequenced. The mitogenomes of L. turbidalis, L. aeruginalis, P. despicata, and C. perlellus are 15 240 bp, 15 339 bp, 15 389 bp, and 15 440 bp. The four mitogenomes all have a typical insect mitochondrial gene order, including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and one A + T rich region (control region). The PCGs are initiated by the typical ATN codons, except CGA for the cox1 gene. Most PCGs terminate with common codon TAA or TAG, the incomplete codon T is found as the stop codon for cox2, nad4, and nad5. Most tRNA genes exhibit typical cloverleaf structure, except trnS1 (AGN) lacking the dihydrouridine (DHU) arm. The secondary structure of rRNA of four mitogenomes were predicted. Poly-T structure and micro-satellite regions are conserved in control regions. The phylogenetic analyses based on 13 PCGs showed the relationships of subfamilies in Pyraloidea. Pyralidae, and Crambidae are monophyletic, respectively. Pyralidae comprises four subfamilies, which form the following topology with high support values: (Galleriinae + ((Pyralinae + Epipaschiinae)+ Phycitinae)). Crambidae includes seven subfamilies and is divided into two lineages. Pyraustinae and Spilomelinae are sister groups of each other, and form the "PS clade." Other five subfamilies (Crambinae, Acentropinae, Scopariinae, Schoenobiinae, and Glaphyriinae) form the "non-PS clade" in the Bayesian inference tree. However, Schoenobiinae is not grouped with the other four subfamilies and located at the base of Crambidae in two maximum likelihood trees.

Molecular Phylogeny and Historical Biogeography of Goodyera R. Br. (Orchidaceae): A Case of the Vicariance Between East Asia and North America.

Understanding of intercontinental distribution in the Northern Hemisphere has attracted a lot of attention from botanists. However, although Orchidaceae is the largest group of angiosperms, biogeographical studies on the disjunctive pattern have not been sufficient for this family. Goodyera R. Br. (tribe Cranichideae, subfamily Orchidoideae, family Orchidaceae) is widely distributed in temperate and tropical regions. Although the phylogenetic relationship of Goodyera inferred from both morphological and molecular data has been conducted, the sampled taxa were mainly distributed in Asia regions that resulted in non-monophyly of this genus. In this study, the complete plastid genomes of Goodyera, generated by next-generation sequencing (NGS) technique and sampled in East Asia and North America, were used to reconstruct phylogeny and explore the historical biogeography. A total of 18 Goodyera species including seven newly sequenced species were analyzed. Based on 79 protein-coding genes, the phylogenetic analysis revealed that Goodyera could be subdivided into four subclades with high support values. The polyphyletic relationships among Goodyera taxa were confirmed, and the unclear position of G. foliosa was also resolved. The datasets that are composed of the 14 coding sequences (CDS) (matK, atpF, ndhK, accD, cemA, clpP, rpoA, rpl22, ndhF, ccsA, ndhD, ndhI, ndhA, and ycf 1) showed the same topology derived from 79 protein-coding genes. Molecular dating analyses revealed the origin of Goodyera in the mid-Miocene (15.75 Mya). Nearctic clade of Goodyera was diverged at 10.88 Mya from their most recent common ancestor (MRCA). The biogeographical reconstruction suggests that subtropical or tropical Asia is the origin of Goodyera and it has subsequently spread to temperate Asia during the Miocene. In addition, Nearctic clade is derived from East Asian species through Bering Land Bridge (BLB) during the Miocene. The speciation of Goodyera is most likely to have occurred during Miocene, and climatic and geological changes are thought to have had a part in this diversification. Our findings propose both origin and vicariance events of Goodyera for the first time and add an example for the biogeographical history of the Northern Hemisphere.

Comparative Analysis of Four Complete Mitochondrial Genomes of Epinephelidae (Perciformes).

Groupers are commercial, mainly reef-associated fishes, classified in the family Epinephelidae (Perciformes). This study first sequenced the complete mitogenomes of Cephalopholis leopardus, Cephalopholis spiloparaea, Epinephelus amblycephalus, and Epinephelus hexagonatus. The lengths of the four Epinephelidae mitogenomes ranged from 16,585 base pair (bp) to 16,872 bp with the typical gene order. All tRNA genes had a typical cloverleaf structure, except the tRNA-Ser (AGY) gene which was lacking the entire dihydrouridine arm. The ratio of nonsynonymous substitution (Ka) and synonymous substitution (Ks) indicated that four groupers were suffering a purifying selection. Phylogenetic relationships were reconstructed by Bayesian inference (BI) and maximum likelihood (ML) methods based on all mitogenomic data of 41 groupers and 2 outgroups. The identical topologies result with high support values showed that Cephalopholis and Epinephelus are not monophyletic genera. Anyperodon and Cromileptes clustered to Epinephelus. Aethaloperca rogaa and Cephalopholis argus assembled a clad. Cephalopholis leopardus, C. spiloparaea, and Cephalopholis miniata are also in a clade. Epinephelus hexagonatus is close to Epinephelus tauvina and Epinephelus merra, and E. amblycephalus is a sister group with Epinephelus stictus. More mitogenomic data from Epinephelidae species are essential to understand its taxonomic status with the family Serranidae.

A new species of Arthrographis (Eremomycetaceae, Dothideomycetes), from the soil in Guizhou, China

During a survey of keratinolytic fungi in China, a new species, Arthrographis multiformispora was isolated from soil samples. Morphologically, A. multiformispora differs from other species in the genus by the presence of globose or subglobose chlamydospores and cylindrical arthroconidia. Phylogenetically, our four strains were clustered together with high support values and separated from other clades. We provided a description, illustrations, and phylogenetic tree for the new species.

Diversity investigation by application of DNA barcoding: A case study of lepidopteran insects in Xinjiang wild fruit forests, China.

To investigate the species diversity of lepidopteran insects in Xinjiang wild fruit forests, establish insect community monitoring systems, and determine the local species pool, we test the applicability of DNA barcoding based on cytochrome c oxidase subunit I (COI) gene for accurate and rapid identification of insect species. From 2017 to 2019, a total of 212 samples with ambiguous morphological identification were selected for DNA barcoding analysis. Five sequence‐based methods for species delimitation (ABGD, BINs, GMYC, jMOTU, and bPTP) were conducted for comparison to traditional morphology‐based identification. In total, 2,422 samples were recorded, representing 143 species of 110 genera in 17 families in Lepidoptera. The diversity analysis showed that the richness indices for Noctuidae was the highest (54 species), and for Pterophoridae, Cossidae, Limacodidae, Lasiocampidae, Pieridae, and Lycaenidae were the lowest (all with 1 species). The Shannon–Wiener species diversity index (H′) and Pielou's evenness (J′) of lepidopteran insects first increased and then decreased across these 3 years, while the Simpson diversity index showed a trend of subtracted then added. For molecular‐based identification, 67 lepidopteran species within 61 genera in 14 families were identified through DNA barcoding. Neighbor‐joining (NJ) analysis showed that conspecific individuals were clustered together and formed monophyletic groups with a high support value, except for Lacanobia contigua (Denis & Schiffermüller, 1775) (Noctuidae: Hadeninae). Sixty‐seven morphospecies were classified into various numbers of MOTUs based on ABGD, BINs, GMYC, jMOTU, and bPTP (70, 96, 2, 71, and 71, respectively). In Xinjiang wild fruit forests, the family with the largest number of species is Noctuidae, followed by Geometridae, Crambidae, and the remaining families. The highest Shannon diversity index is observed for the family Noctuidae. Our results indicate that the distance‐based methods (ABGD and jMOTU) and character‐based method (bPTP) outperform GMYC. BINs is inclined to overestimate species diversity compared to other methods.

Macrophomina vaccinii Causes a Basal Stem and Root Rot of Patchouli (Pogostemon cablin) in China.

Patchouli (Pogostemon cablin Benth.) is a perennial herb native to South and South East Asia, which widely used as a traditional herbal medicine against indigestion, diarrhea, cold, fever, vomiting and headache in China (Swamy and Sinniah, 2015). In September 2020, a new basal stem and root rot disease of patchouli was observed in three cultivated fields of Zhanjiang City, Guangdong Province. Symptoms included sudden discoloration, chlorosis and wilting of the leaves and severe rot associated with external and internal browning at the basal part of stems and roots that results in the death of approximately 2 to 5% of plants in each field. To determine the causal agent, symptomatic roots with typical lesions were cut into small pieces, then surface sterilized in 2.5% NaClO for 1 min, rinsed three times in distilled water, and then inoculated on potato dextrose agar (PDA) medium. A fungus with same morphological characteristics was consistently isolated from disease tissue. The mycelia initially white to cream, later turning pale brown to black with age on PDA. To induce the sporulation, the isolated fungi were transferred to synthetic nutrient-poor agar (SNA) with autoclaved pine needles and ten pure cultures were obtained by single spores. Masses of black, hard and oblong microsclerotia (av. 171.2×136.5 µm, n=50) were observed on SNA. Conidia hyaline, ellipsoid to obovoid, smooth, enclosed in a mucous sheath, 15.9~32.5×7.7~11.9 µm (av. 23.9 × 9.6 µm, n=100). The internal transcribed spacer (ITS) regions as well as the partial translation elongation factor (EF-1α) and β-tubulin (TUB) genes of two representative isolates GHX-1 and CHX-2 were sequenced, using primer pairs ITS1/ITS4 (White et al. 1990), EF-688F/EF-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The obtained sequences were submitted to GenBank under the following accession numbers: ITS, MZ375360 and MZ375361; EF-1α, MZ393804 and MZ393805; and TUB, MZ393806 and MZ393807. BLASTn searches revealed that 100% identity with the existing sequences of ex-type culture CGMCC3.19503 of Macrophomina vaccinii (ITS, MK687450; EF-1α, MK687426; and TUB, MK687434), respectively. Phylogenetic analysis using Neighbour-Joining method in Mega7.0 with concatenated sequences of ITS, EF-1α and TUB showed that the isolates clustered in the clade of M. vaccinii with high bootstrap support values. Based on both of the morphological and molecular results, the isolates were identified as M. vaccinii (Zhao et al., 2019). To confirm the pathogenicity of the two isolates, 5-mm-diameter mycelial agar plus from the margins of 5-day-old PDA cultures were placed on the wounded basal stem of 2-month-old patchouli seedlings in each pot. Five separate pots were used for each isolate and plants inoculated with sterile agar plus were served as controls. All plants were covered with plastic bags to maintain 90% relative humidity and kept at 26°C with a photoperiod of 12 h in a greenhouse conditions. The patchouli plants developed similar necrotic symptoms on basal stem and root to those observed in the field after 3 weeks after inoculation, whereas the control plants were asymptomatic. M. vaccinii was reisolated from inoculated plants and identify by morphological and molecular characteristics, fulfill the Koch's postulates and identify. Recently, stem blight on blueberry caused by M. vaccinii was reported in China (Zhao et al. 2019). To our knowledge, this is the first report of M. vaccinii causing basal stem and root rot on patchouli in China and worldwide.

Molecular phylogeny of the subgenus Syrrhophus (Amphibia: Anura: Eleutherodactylidae), with the description of a new species from Eastern Mexico

The Eleutherodactylus subgenus Syrrhophus has a complex taxonomic history. For many years the species were classified into series and groups of species based on morphological and allozyme data. However, most of the species have conservative morphology, and their phylogenetic relationships have not been resolved. Furthermore, recent studies employing molecular tools have shown that Syrrhophus diversity is highly underestimated. Here, we present a near-complete phylogenetic hypothesis based on mitochondrial data (16S rRNA, cytochrome c oxidase subunit I, and cytochrome b) and employing maximum likelihood and Bayesian methods. The Bayesian tree has high support values (posterior probabilities ≥0.95) whereas the maximum likelihood hypothesis has generally low values (bootstrap support <70). Both phylogenetic analyses recovered Syrrhophus as monophyletic and composed of four main, allopatric clades (E. symingtoni clade, E. longipes clade, E. modestus clade, and E. nitidus clade) concordant with geography. Based on the molecular and morphological data, we describe a new species from the state of San Luis Potosí, Mexico. The new species is most closely related to E. campi and E. cystignathoides from which it has a genetic distance >7% in 16S rRNA. Finally, based on the phylogenetic results, we resurrect E. rubrimaculatus from synonymy. Our results increase the number of species in the subgenus Syrrhophus to 41. We highlight the importance of efforts to study this important component of Mexican biodiversity. http://zoobank.org/urn:lsid:zoobank.org:pub:3C2B2315-D5C8-420A-938C-A2B3A75B4248

Comparative Mitochondrial Genome Analysis of Mamestra configurata (Lepidoptera: Noctuoidea: Noctuidae) and Other Noctuid Insects Reveals Conserved Genome Organization and Phylogeny

Abstract Mitochondrial genomes (mitogenomes) advance our understanding of molecular evolution and phylogenetic relationships. In this study, the mitogenome of Mamestra configurata was assembled, annotated, and analyzed. The total length of its genome was 15,322 bp; the A+T content accounted for 81.2%. Both the AT and GC skew values (−0.003 and −0.182) in the mitogenome of M. configurata were negative. Compared with their ancestral insects (trnI-trnQ-trnM), the gene sequence showed a typical trnm rearrangement (trnM-trnI-trnQ). Except for cox1, all the protein-coding genes were initiated with ATN. All tRNAs except trnS1 were of the four-leaf grass type. According to phylogenetic analysis, we found that M. configurata and Protegira songi clustered together in one clade with high node support values. Based on Bayesian inference and maximum likelihood methods, the phylogenetic results revealed that M. configurata was closely related to Protegira songi, the Erebidae was sister to (Nolidae + [Euteliidae + Noctuidae]), Doidae and Notodontidae is sister to the other families of Noctuoidea, respectively. Our results may provide a useful reference for the intensive study of the phylogeny of Noctuidae.