Maximum Parsimony Analyses Research Articles

APPLICATION OF THE DNA BARCODING METHOD FOR IDENTIFICATION AND TAXONOMIC ANALYSIS OF THE LAMIACEAE FAMILY

The Lamiaceae family is one of the largest and most important among the aromatic plants, including many species with pronounced pharmacological properties. Plants of this family, such as mint, rosemary and thyme, are widely used in traditional and modern medicine due to their therapeutic and biochemical properties. The importance of correct species identification is emphasized by the need to preserve the biodiversity and sustainable use of the resources of this family. In this work, plants of Lamiaceae family growing in the territory of the Karaganda region were studied by DNA barcoding. The studies were conducted on the basis of nucleotide sequences of rbcL and trnH-psbA chloroplast DNA barcodes using the methods of paired genetic distances, BLASTn and phylogenetic tree based on the analysis of maximum parsimony. The high efficiency of the rbcL locus was demonstrated for the identification of higher taxonomic groups, such as subfamilies and tribes, while the intergenic spacer trnH-psbAhasproved to be effectiveat the level of speciesandgenus level.

Revision and phylogenetic placement of one of the earliest freshwater gobies from the Lower Oligocene of Central Europe

ABSTRACT Gobioid fishes (Teleostei) are renowned for their remarkable species diversity in marine and freshwater environments, making their evolutionary history a fascinating study subject. Among their oldest known fossils is the small-sized freshwater goby ‘Gobius’ gracilis from the Lower Oligocene of Central Europe, which has not undergone a detailed investigation since its first description. Here, we present a comprehensive re-analysis of ‘G.’ gracilis using type specimens and new material from museum collections. We show that this species possesses a unique combination of traits, leading to the establishment of a new genus, †Laubeichthys gen. nov. Phylogenetic analyses using Bayesian Inference (BI) recover †Laubeichthys gen. nov. as a stem member of the extinct family †Pirskeniidae and suggest a sister relationship between the family Thalasseleotrididae (ocean sleepers) and †Laubeichthys gen. nov. + †Pirskeniidae. Maximum parsimony analyses do not contradict such a relationship, but the trees are less resolved. Today, the Thalasseleotrididae includes four marine species that are restricted to the Southwest Pacific. In contrast, †Pirskeniidae and †Laubeichthys gen. nov. inhabited freshwater environments in Central Europe. Our findings provide new insights into the evolutionary history and historical biogeography of gobioid fishes and highlight the dynamic nature of their past diversity and distribution.

Expanded range of Nasutitermes callimorphus Mathews, 1977 (Isoptera: Termitidae: Nasutitermitinae), comparison with N. corniger (Motschulsky, 1855) and N. ephratae (Holmgren, 1910), and synonymy of N. dasyopsis Thorne, 1989 into N. nigriceps (Haldeman, 1854)

The imago of N. callimorphus is described for the first time. Nasutitermes callimorphus occurs from Mexico to Paraguay. Nasutitermes callimorphus is smaller in all measurements but generally resembles its widely distributed sympatric congeners, N. corniger (Motschulsky, 1855) and N. ephratae (Holmgren, 1910). Molecular phylogenetic analysis of a portion of the mitochondrial 16S rRNA gene and the COII mtDNA marker revealed that N. callimorphus forms a distinct clade using both maximum parsimony and maximum likelihood analysis. Unlike N. corniger and N. ephratae, N. callimorphus does not build epigeal or arboreal carton nests. We further regard N. dasyopsis Thorne, 1989 as a junior synonym of N. nigriceps (Haldeman, 1854).

Is the Current Systematic Subdivision of the Subfamily Neanurinae (Collembola, Neanuridae) Still Valid? Testing the Monophyly and Phylogenetic Relationships of Currently Established Tribes of the Neanurinae.

The subfamily Neanurinae is the largest in the family, with almost 800 described species. These springtails differ significantly from all other Collembola in their morphology, behaviour, and natural habitats. A systematic division of the Neanurinae into tribes was proposed more than 30 years ago by Cassagnau (1989), but it has not yet been tested using cladistic methods. Recent studies, both phylogenetic analyses of individual tribes or genera and descriptions of new taxa, suggest that the currently recognised tribes may not be monophyletic. The phylogenetic relationships among major lineages of the Neanurinae were explored by analysing a dataset of 101 discrete morphological characters. Bayesian and maximum parsimony analyses yielded similar tree topologies. The relationships among the Neanurinae were not resolved in any of the analyses, except for the support for the monophyly of the tribe Lobellini. The results indicate that the taxonomic characters used in the classification of Neanurinae are shared among members of the different tribes, which may have resulted in a classification with little phylogenetic basis. The article discusses the phylogenetic significance of morphological characters, including those recognised as key to the evolution and history of Neanurinae.

Phylogeny of the Chinese species groups of the subgenus Homoneura Wulp, 1891 (Diptera, Lauxaniidae, Homoneurinae) based on morphological characters

The subgenus Homoneura Wulp, 1891 (Diptera, Lauxaniidae, Homoneurinae) is highly diverse with more than 220 species known from China, representing more than 80% of the Chinese genus Homoneura Wulp, 1891. These species were assigned into 21 species groups in studies mainly focusing on the classification and description of species. The phylogenetic relationships of each subgenus of Homoneura and the phylogenetic relationship of the species groups are still not well understood. We investigated the male morphology to provide the basis to further revise the species groups of this subgenus. In this survey, 230 species were examined and 117 morphological characters obtained, a phylogenetic analysis was conducted using the maximum parsimony analysis with TNT and WinClada. The analyses yielded 45 most parsimonious trees and one strict consensus tree. A phylogenetic hypothesis is proposed dividing the subgenus Homoneura into 12 species groups: H. (H.) nigrifacies, H. (H.) pallida, H. (H.) patella, H. (H.) beckeri, H. (H.) formosae, H. (H.) henanensis, H. (H.) nigra, H. (H.) notostigma, H. (H.) ornatifrons, and H. (H.) trispina, H. (H.) laticosta, and H. (H.) quinquenotata. This research provides valuable contributions towards a better understanding of the phylogenetic relationships within the subgenus Homoneura. However, the monophyly of the genus and subgenus was not supported.

Confusing boundaries of the Labrador Tea species: dispersal history explains the lack of clear species structure.

The Labrador Teas (genus Rhododendron, subsection Ledum) are a complex of species widely distributed in the Northern Hemisphere. They occupy cold-resistant plant communities from highlands to forest understory and wetland habitats almost circumboreally and they are especially abundant in Northeast Asia (NE Asia) and northern North America (NN Am), still there are no clear species boundaries in this group. The genetic structure of species of the subsect. Ledum from Eurasia and North America as well as the dispersal history of the group require clarification. Phylogeny and biogeography of the subsect. Ledum of the genus Rhododendron were assessed using phylogenetic trees constructed based on the analysis of variation in chloroplast petB-petD, trnV-ndhC, trnH-psbA, K2R-K707, atpB oligo2 - rbcL oligo5 and nuclear (ITS1) markers of four Eurasian and one American species (65 populations, 408 individuals). The data were evaluated with Maximum Parsimony and Bayesian analysis. Molecular dating and ancestral areas reconstruction were obtained. Dense sampling revealed widespread presence of shared haplotypes and ribotypes among Ledum populations and species. Two American, three Eurasian and one mixed lineage diversified during the Neogene climate cooling and then rapidly dispersed during the Pleistocene. The ability to accumulate high genetic diversity and to preserve it across distribution ranges and generations prevented Ledum from lineage sorting. As a result, a species complex with a reserve of genetic variability appeared. Although no clear phylogenetic inference can be obtained at present, the plastid genealogy is consisted with the nuclear genealogy and demonstrates the processes involved in speciation in the Ledum species complex.

NEW INSIGHTS BASED ON MORPHOLOGICAL AND MOLECULAR DATA REVEAL THE TAXONOMIC STATUS OF HALIOTREMA PTEROISI (MONOGENOIDEA: DACTYLOGYRIDAE) INFECTING DEVIL FIREFISH PTEROIS MILES (PERCIFORMES: SCORPAENOIDEI: SCORPAENIDAE) IN THE RED SEA OFF EGYPT.

Paperna, 1972 (Monogenoidea: Dactylogyridae) was found parasitizing the gill lamellae of devil firefish, Pterois miles (Bennet) (Perciformes: Scorpaenidae), in the Red Sea off Safaga (26°44'N, 33°56'E), Egypt. The parasite species was described based on morphological features of available specimens and transferred to PlatycephalotremaKritsky and Nitta, 2019 (Dactylogyridae) as Platycephalotrema pteroisi (Paperna, 1972) n. comb. The occurrence of Pl. pteroisi off Safaga, Egypt, represented a range extension for the helminth of about 160 km to the southwest of the southern end of the Gulf of Aqaba. The transfer of the species to Platycephalotrema based on an evaluation of morphological features was supported by an analysis of molecular sequences of the 28S rDNA gene of Pl. pteroisi and 49 other dactylogyrid species. Maximum-likelihood, Bayesian inference, and maximum parsimony analyses of this dactylogyrid sequence data revealed H. pteroisi to nest with significant support within the clade of Platycephalotrema spp. During the literature review of dactylogyrid species infecting scorpionfishes, it was determined that Ancyrocephalus sp. of Dyer et al. from luna lion fish Pterois lunulata Temminck and Schlegel collected off Okinawa-jima, Japan represented an undescribed species of Platycephalotrema.

Ultraconserved elements from transcriptome and genome dataprovide insight into the phylogenomics of Sternorrhyncha (Insecta: Hemiptera).

Sternorrhyncha, one of the four major suborders of Hemiptera, is a phytophagous taxon inclusive of nearly 18 000 described species. The phylogenetic relationships within the taxon and the earliest-branching lineage of its infraorders remain incompletely understood. This study attempted to illuminate the phylogenetic relationships within Sternorrhyncha through the use of maximum likelihood, Bayesian inference and maximum parsimony analyses, employing ultraconserved element (UCE) data from 39 genomic and 62 transcriptomic datasets and thereby representing most families within the taxon. The probe set Hemiptera 2.7Kv1 was used to recover a total of 2731 UCE loci: from 547 to 1699 (with an average of 1084) across all genomic datasets and from 108 to 849 (with an average of 329) across all transcriptomic datasets. All three types of phylogenetic analyses employed in this study produced robust statistical support for Sternorrhyncha being a monophyletic group. The different methods of phylogenetic analysis produced inconsistent descriptions of topological structure at the infraorder level: while maximum likelihood and Bayesian inference analyses produced strong statistical evidence (100%) indicating the clade Psylloidea + Aleyrodoidea to be a sister of the clade Aphidoidea (Aphidomorpha) + Coccoidea (Coccomorpha), the maximum parsimony analysis failed to recover a similar result. Our results also provide detail on the phylogenetic relationships within each infraorder. This study presents the first use of UCE data to investigate the phylogeny of Sternorrhyncha. It also shows the viability of amalgamating genomic and transcriptomic data in studies of phylogenetic relationships, potentially highlighting a resource-efficient approach for future inquiries into diverse taxa through the integration of varied data sources.

18S rDNA sequence-structure phylogeny of the eukaryotes simultaneously inferred from sequences and their individual secondary structures

ObjectiveThe eukaryotic tree of life has been subject of numerous studies ever since the nineteenth century, with more supergroups and their sister relations being decoded in the last years. In this study, we reconstructed the phylogeny of eukaryotes using complete 18S rDNA sequences and their individual secondary structures simultaneously. After the sequence-structure data was encoded, it was automatically aligned and analyzed using sequence-only as well as sequence-structure approaches. We present overall neighbor-joining trees of 211 eukaryotes as well as the respective profile neighbor-joining trees, which helped to resolve the basal branching pattern. A manually chosen subset was further inspected using neighbor-joining, maximum parsimony, and maximum likelihood analyses. Additionally, the 75 and 100 percent consensus structures of the subset were predicted.ResultsAll sequence-structure approaches show improvements compared to the respective sequence-only approaches: the average bootstrap support per node of the sequence-structure profile neighbor-joining analyses with 90.3, was higher than the average bootstrap support of the sequence-only profile neighbor-joining analysis with 73.9. Also, the subset analyses using sequence-structure data were better supported. Furthermore, more subgroups of the supergroups were recovered as monophyletic and sister group relations were much more comparable to results as obtained by multi-marker analyses.

Evolution of modes of nutrient absorption in entomoparasitic nematodes of the order Tylenchida revealed by structural and phylogenetic analysis

Summary Adult female entomoparasitic nematodes of the order Tylenchida have degenerate digestive organs and body surfaces that lack cuticles and are covered with microvilli. These morphologic features indicate that tylenchids absorb nutrients via the integument. A previous study hypothesised that the nutrient intake of entomoparasitic tylenchids evolved from oral to transdermal in a step-by-step manner, and this evolutionary process is reflected in the development of the integument ultrastructure of these species. However, the evolution of the nutrient absorption of entomoparasitic tylenchids remains unclear. This evolution has not been phylogenetically examined because of a lack of information on the integument structure associated with phylogenetic relationships. In the present study, we examined the integument ultrastructure and molecular sequence of four entomoparasitic tylenchids: Bradynema sp., Contortylenchus sp., C. genitalicola and Deladenus sp. The body surface of Bradynema sp. was covered with microvilli, suggesting that Bradynema sp. absorbs nutrients via the integument. By contrast, the body surfaces of Contortylenchus sp., C. genitalicola and Deladenus sp. were covered with a thin cuticle-like layer and well-developed epidermis without microvilli, suggesting that these species absorb nutrients only partially via the integument. We also classified the nutrient absorption mode based on the integument ultrastructure and molecular phylogenetic analysis of entomoparasitic tylenchids in previous studies. Our maximum parsimony analysis supported the step-by-step evolution hypothesised in the previous study. However, the parsimony analysis also generated new evolutionary processes in which atavism and/or saltation occur instead of stepwise evolution.

Plastid phylogenomics and fossil evidence provide new insights into the evolutionary complexity of the ‘woody clade’ in Saxifragales

BackgroundThe “woody clade” in Saxifragales (WCS), encompassing four woody families (Altingiaceae, Cercidiphyllaceae, Daphniphyllaceae, and Hamamelidaceae), is a phylogenetically recalcitrant node in the angiosperm tree of life, as the interfamilial relationships of the WCS remain contentious. Based on a comprehensive sampling of WCS genera, this study aims to recover a robust maternal backbone phylogeny of the WCS by analyzing plastid genome (plastome) sequence data using Bayesian inference (BI), maximum likelihood (ML), and maximum parsimony (MP) methods, and to explore the possible causes of the phylogenetic recalcitrance with respect to deep relationships within the WCS, in combination with molecular and fossil evidence.ResultsAlthough the four WCS families were identically resolved as monophyletic, the MP analysis recovered different tree topologies for the relationships among Altingiaceae, Cercidiphyllaceae, and Daphniphyllaceae from the ML and BI phylogenies. The fossil-calibrated plastome phylogeny showed that the WCS underwent a rapid divergence of crown groups in the early Cretaceous (between 104.79 and 100.23 Ma), leading to the origin of the stem lineage ancestors of Altingiaceae, Cercidiphyllaceae, Daphniphyllaceae, and Hamamelidaceae within a very short time span (∼4.56 Ma). Compared with the tree topology recovered in a previous study based on nuclear genome data, cytonuclear discordance regarding the interfamilial relationships of the WCS was detected.ConclusionsMolecular and fossil evidence imply that the early divergence of the WCS might have experienced radiative diversification of crown groups, extensive extinctions at the genus and species levels around the Cretaceous/Paleocene boundary, and ancient hybridization. Such evolutionarily complex events may introduce biases in topological estimations within the WCS due to incomplete lineage sorting, cytonuclear discordance, and long-branch attraction, potentially impacting the accurate reconstruction of deep relationships.

Unravelling the mishmash: A new phylogeny for the family Empheriidae (Psocodea, Trogiomorpha) with a new genus and species from Cretaceous Charentese amber

The order Psocodea, including barklice, booklice, and parasitic lice, is diverse and widely distributed since the Cretaceous. That is particularly the case for the speciose extinct family Empheriidae (Psocodea, Trogiomorpha, Atropetae), recently fused with the ‘Archaeatropidae’. Understanding the evolution of barklice is dependent in part on studying this family, as its representatives have been found from the Early Cretaceous to the Eocene, surviving the K/Pg extinction event. The phylogenetic relationships of Empheriidae in relation to other families, such as Lepidopsocidae or Psoquillidae, have been extensively debated. However, distinguishing diagnostic characters for the Empheriidae has proven challenging. In this study, we describe the new empheriid Santonipsocus mimeticusgen. et sp. nov. from Cretaceous Charentese amber (France). It is the third empheriid species found in this locality. The new genus is compared with the other genera in the family, and Proprionoglaris guyoti and Proprionoglaris axioperierga are revised based on the type material and new specimens. We explore the phylogeny of Empheriidae, both the relationships with other families and the inner relationships between the genera, through maximum parsimony analysis and Bayesian inference analysis. Our results suggest that Empheriidae may represent a paraphyletic evolutionary grade to the rest of Atropetae. The phylogenetic relationships between genera align with the biogeography of the family and support previous hypotheses. In addition, we discuss the possible biology of the members of the family, shedding light on the evolutionary history of Empheriidae.

Phylogenetic analysis, complete mitochondrial genome, and redescription of the East Asian genus Aphaenomurus (Collembola: Tomoceridae)

The genus Aphaenomurus Yosii, 1956 includes one species and one subspecies, with a limited distribution in Korea and Japan. The phylogenetic relationships between Aphaenomurus and other genera of the subfamily Tomocerinae have not been resolved. In Korea, Aphaenomurus is generally observed in caves. In this study, specimens collected from the twilight zones of caves were used for an expanded phylogenetic analysis, complete mitochondrial genome analysis, and morphological redescription of Aphaenomurus interpositus Yosii, 1956. Maximum likelihood, Bayesian, and parsimony analyses based on five regions (COI, 16S rDNA, 18S rDNA, 28S rDNA D1–2, and 28S rDNA D7–10) indicated that A. interpositus is a sister group to the genus Plutomurus Yosii, 1956 with strong support.

DNA barcoding of the genus Verbascum (Scrophulariaceae) in the Arabian Peninsula

AbstractVerbascum and Rhabdotosperma are members of the family Scrophulariaceae. The first genus comprises approximately 360 species from almost all parts of the world, while the second contains a total of 8 species from tropical Africa and the Arabian Peninsula. Since 1977, the relationships between Verbascum and Rhabdotosperma continue to be contested. The present study aims to present the phylogenetic relationships and status among Verbascum species in the Arabian Peninsula. For phylogenetic analyses, maximum parsimony and Bayesian inference were performed. In total, 236 DNA sequences from 59 specimens of Arabian Verbascum were analysed. The phylogenetic analysis of one nuclear (ITS) and three chloroplastic (rbcL, matK, trnL) markers confirmed the monophyly of Verbascum, including the genus Rhabdotosperma. In addition to presenting novel phylogenetic relationships among the different Verbascum species in the Arabian Peninsula, our study reduced the species count of Arabian Verbascum to 16. Moreover, the phylogenetic analysis strongly supports the reinstatement of the genus Rhabdotosperma into Verbascum based on the Bayesian and maximum parsimony analyses.

A new gnetalean macrofossil from the Lower Cretaceous of the Laiyang Basin, eastern China

•A new species Laiyangia compacta gen. et sp. nov. with unusual female spikes was described from the Laiyang Formation.•This species shows transitional morphology between early ephedroid and modern gnetoid.•The epidermal microstructural character of this species provides a driver paleoclimate of Laiyang area in Early Cretaceous.

Phylogeny and Fungal Community Structures of Helotiales Associated with Sclerotial Disease of Mulberry Fruits in China.

Mulberry fruit sclerotiniose is a prevalent disease caused by the fungal species Ciboria shiraiana, C. carunculoides, and Scleromitrula shiraiana of the order Helotiales, and severely affects the production of mulberry. However, these species have only been identified using morphological and rDNA-ITS sequence analyses, and their genetic variation is unclear. To address this, morphological and two-locus (ITS and RPB2) phylogenetic analyses were conducted using culture-dependent and independent methods for 49 samples from 31 orchards across four provinces in China. Illumina MiSeq sequencing was used to assess the fungal communities obtained from fruits varying in disease severity and color from an orchard in Wuhan. Conidial suspensions of C. shiraiana and C. carunculoides isolated from diseased fruits, diseased fruits affected with hypertrophy and pellet sorosis sclerotiniose, and mycelia of Sclerotinia sclerotiorum were determined to be pathogenic to the mulberry cultivar YSD10. However, fruits inoculated with S. sclerotiorum mycelia exhibited nontypical disease symptoms, and mycelia and conidia obtained from C. carunculoides and S. shiraiana strains were not pathogenic. Maximum parsimony and Bayesian analyses using the sequences of the assessed loci indicated species variability with no evidence of geographic specialization. Metagenomic analysis revealed that the diversity of fungal communities was reduced with disease progression. Furthermore, within a single fruit, the presence of two Ciboria spp. was detected. These results provide novel insights into Ciboria spp., revealing the secondary infections caused by conidia in diseased fruits, genetic variations of the pathogens, and the occurrence of coinfection. This improved understanding of fungal pathogens will aid in developing effective disease control strategies.

Phylogeny of Prosthechea (Laeliinae, Orchidaceae) based on nrITS and plastid DNA sequences: Reassessing the lumper‐splitter debate and shedding light on the evolution of this Neotropical genus

AbstractAfter a confused taxonomic history, Prosthechea (Laeliinae, Orchidaceae) has been circumscribed in a broad sense to include species previously placed in Encyclia subg. Osmophytum. However, recent alternative classifications resurrecting old names such as Anacheilium and Hormidium, and proposing new genera such as Euchile, Panarica, Pollardia, and Pseudencyclia were presented, splitting Prosthechea into narrower genera. In this study, we provide a molecular phylogeny of Prosthechea to reassess alternative generic classifications and shed light on evolutionary aspects of this highly diverse Neotropical lineage. Eighty species of Prosthechea s.l. and 12 species of related genera were sampled. Bayesian and maximum parsimony analyses of sequences from three plastid regions (rpl32‐trnL, trnD‐trnT, ycf1) and nrITS were included. Possible conflicts between nuclear and plastid data were assessed with partitioned Bremer support. Ancestral state reconstructions of morphological characters relevant to the genus taxonomy are provided. Our results reinforce the monophyly of the lineage corresponding to a broad circumscription of Prosthechea. The small lineages corresponding to Euchile, Panarica, and Hormidium are monophyletic, whereas the larger Anacheilium, Pollardia, and Pseudencyclia are not monophyletic. Splitting Prosthechea into smaller genera is not supported by our phylogeny, and nonmonophyly also hinders the proposal of a comprehensive infrageneric classification. The phylogenetic relationships recovered reflect better the biogeographic patterns than the previously proposed morphological affinities, pointing to the existence of homoplastic floral traits within the genus. Conflicts between nuclear and plastid partitions were detected, suggesting a possible scenario of reticulate evolution.

PHYLOGENETIC RELATIONSHIP BETWEEN RABIES VIRUS (RABIES LYSSAVIRUS) VARIANTS FROM TWO DIFFERENT HOST SPECIES

Rabies is a fatal zoonotic disease that affects several mammals. Bats are recognized hosts of the rabies virus, and their main food source is the blood of other mammals, particularly cattle. During feeding, bats transmit the virus to cattle, which are victims of the disease, contributing to economic losses and increasing the risk of infection for humans. Based on this affinity in the rabies cycle between bats and cattle, the objective of this study was to analyze the phylogenetic relationships of rabies virus samples in cattle and bats. The G gene of the rabies virus was chosen for this study because it is directly related to the infection process. Nucleotide sequences of the viral G gene were selected from GenBank for samples obtained from infected cattle and bats. Maximum parsimony analyses were conducted using the Molecular Evolutionary Genetics Analysis software. The Maxima Parsimony tree indicated a phylogenetic relationship between the G genes of both hosts, indicating that the virus evolved from bats to cattle. Analysis of parsimoniously informative sites revealed that the viral G gene carried specific mutations in each host. Knowledge of the evolutionary relationships between the rabies virus and its hosts is critical for identifying potential new hosts and the possible routes of infection for humans.

Chromosomal rearrangements played an important role in the speciation of rice rats of genus Cerradomys (Rodentia, Sigmodontinae, Oryzomyini)

Rodents of the genus Cerradomys belong to tribe Oryzomyini, one of the most diverse and speciose groups in Sigmodontinae (Rodentia, Cricetidae). The speciation process in Cerradomys is associated with chromosomal rearrangements and biogeographic dynamics in South America during the Pleistocene era. As the morphological, molecular and karyotypic aspects of Myomorpha rodents do not evolve at the same rate, we strategically employed karyotypic characters for the construction of chromosomal phylogeny to investigate whether phylogenetic relationships using chromosomal data corroborate the radiation of Cerradomys taxa recovered by molecular phylogeny. Comparative chromosome painting using Hylaeamys megacephalus (HME) whole chromosome probes in C. langguthi (CLA), Cerradomys scotii (CSC), C. subflavus (CSU) and C. vivoi (CVI) shows that karyotypic variability is due to 16 fusion events, 2 fission events, 10 pericentric inversions and 1 centromeric repositioning, plus amplification of constitutive heterochromatin in the short arms of the X chromosomes of CSC and CLA. The chromosomal phylogeny obtained by Maximum Parsimony analysis retrieved Cerradomys as a monophyletic group with 97% support (bootstrap), with CSC as the sister to the other species, followed by a ramification into two clades (69% of branch support), the first comprising CLA and the other branch including CVI and CSU. We integrated the chromosome painting analysis of Eumuroida rodents investigated by HME and Mus musculus (MMU) probes and identified several syntenic blocks shared among representatives of Cricetidae and Muridae. The Cerradomys genus underwent an extensive karyotypic evolutionary process, with multiple rearrangements that shaped extant karyotypes. The chromosomal phylogeny corroborates the phylogenetic relationships proposed by molecular analysis and indicates that karyotypic diversity is associated with species radiation. Three syntenic blocks were identified as part of the ancestral Eumuroida karyotype (AEK): MMU 7/19 (AEK 1), MMU 14 (AEK 10) and MMU 12 (AEK 11). Besides, MMU 5/10 (HME 18/2/24) and MMU 8/13 (HME 22/5/11) should be considered as signatures for Cricetidae, while MMU 5/9/14, 5/7/19, 5 and 8/17 for Sigmodontinae.

Leaf Spot Caused by Colletotrichum siamense, C. fructicola, and C. aeschynomenes on Ixora chinensis in Guangxi, China.

Ixora chinensis Lam., an important ornamental flower, has become more and more popular in the southwest and southeast regions of China for its bright and abundant flowers (Li et al. 2019). In March 2022, 100% I. chinensis plants showed typical anthracnose symptoms on leaf in Nanning, Guangxi, China (108°22' N, 22°48' E). The central areas of lesions were grayish white with small black particles arranged in a wheel pattern, and the edges of lesions were light red to brown. Three diseased leaf samples were collected from three gardens, respectively. 5×5 mm tissues were cut from infected margins, surface-disinfected in 75% ethanol for 10 s, 2% NaClO for 2 min, rinsed three times in sterilized distilled water, and incubated on PDA at 25°C under 12/12 h light/darkness. Eighty-three morphologically similar colonies were observed on PDA after 5 days, and eight isolates G1-3, G2-1, G3-3, W-1, W-2, LCH2-1, LCH3-3, and LCH4-1 were selected for further study. Genomic DNA of these isolates were extracted from 7-day-old mycelia. Primer pairs ITS1/ITS4, GDF1/GDR1, T1/βt2b, CHS Ⅰ-79F/CHS Ⅰ-354R, CL1/CL2, ACT-512F/ACT-783R, and MAT1-2-1/apn2 were used to amplify ITS loci and GAPDH, CHS-Ⅰ, CAL, ACT, ApMAT genes, respectively (Yang et al. 2009; Silva et al. 2012; Liu et al. 2015). Sequences have been deposited in GenBank (ITS: OQ771884 to OQ771891, GAPDH: OQ759576 to OQ759583, TUB2: OQ759584 to OQ759591, CHS-1: OQ759568 to OQ759575, CAL: OQ759560 to OQ759567, ACT: OQ759552 to OQ759559, ApMat: OQ759544 to OQ759551). Phylogenetic analysis was performed with raxmlGUI v.2.0.0. based on combined sequences of ITS, GAPDH, TUB2, CHS-1, CAL, ACT, and ApMAT using maximum parsimony analysis. The results revealed that isolates G2-1 and W-2 were clustered with Collectrichum fructicola, G3-3, W-1, G1-3, LCH2-1, and LCH3-3 were clustered with C. siamense, and LCH4-1 was clustered with C. aeschynomenes. Three representative isolates W-2, G3-3, and LCH4-1 were selected for morphology and pathogenicity observation. On PDA, the colonies of three isolates presented white to grey cottony mycelia，from the margin to the center, W-2 was white, grey, and light gray, G3-3 showed light gray, white, and grey, LCH4-1 was white and light gray, respectively. Conidia were all hyaline, one-celled, cylindrical, and straight. The conidial sizes of W-2, G3-3, and LCH4-1 were 11.03 to 17.53 × 4.93 to 8.42 μm (n=100), 10.63 to 19.06 × 3.73 to 6.92 μm (n=100), and 11.61 to 20.39 × 3.65 to 6.67 μm (n=100), respectively. Pathogenicity tests of three isolates were conducted on leaves of 1-year-old I. chinensis plants with and without wounds, three plants for each treatment, and five leaves inoculated for each plant. Conidial suspensions (10 µL, 106 conidia/mL in 0.1% sterile Tween 20) were inoculated on each site. Control group was treated with 0.1% sterile Tween 20. All inoculated sites were covered with wet cotton, and all plants were bagged and placed in the greenhouse to maintain humidity at 25℃. After 10 days, all wounded and inoculated leaves showed leaf spot, whereas unwounded and control leaves remained asymptomatic. Koch's postulates were fulfilled by re-isolating the causal agents from diseased leaves. C. siamense and C. aeschynomenes could cause anthracnose of I. chinensis in China (Liu et al. 2016, Li et al. 2021). However, to our knowledge, this is the first report of C. fructicola infecting I. chinensis in China. This study may provide reference for further epidemiological study and prevention of anthracnose on I. chinensis.