Nuclear ITS Research Articles

Low Genetic Diversity of the Only Clade of the Tick Rhipicephalus microplus in the Neotropics.

This study addresses the variability of the mitochondrial cytochrome oxidase subunit I (COI) and 16S rDNA (16S), and nuclear internal transcriber spacer ITS2 (ITS2) genes in a set of field-collected samples of the cattle tick, Rhipicephalus microplus (Canestrini, 1888), and in geo-referenced sequences obtained from GenBank. Since the tick is currently considered to be a complex of cryptic taxa in several regions of the world, the main aims of the study are (i) to provide evidence of the clades of the tick present in the Neotropics, (ii) to explore if there is an effect of climate traits on the divergence rates of the target genes, and (iii) to check for a relationship between geographical and genetic distance among populations (the closest, the most similar, meaning for slow spread). We included published sequences of Rhipicephalus annulatus (Nearctic, Afrotropical, and Mediterranean) and R. microplus (Afrotropical, Indomalayan) to fully characterize the Neotropical populations (total: 74 16S, 44 COI, and 49 ITS2 sequences included in the analysis). Only the clade A of R. microplus spread in the Nearctic-Neotropics. Both the K and Lambda's statistics, two measures of phylogenetic signal, support low divergence rates of the tested genes in populations of R. microplus in the Neotropics. These tests demonstrate that genetic diversity of the continental populations does not correlate either with the geographic distance among samples or with environmental variables. The low variability of these genes may be due to a combination of factors like (i) the recent introduction of the tick in the Neotropics, (ii) a large, effective, and fast exchange of populations, and (iii) a low effect of climate on the evolution rates of the target genes. These results have implications for the ecological studies and control of cattle tick infestations.

DNA Barcoding Based Identification of Rosa x Damascene and Prunus dulcis Herbs Using ITS2 Barcoding Gene Amplification.

The considerable risk of adulteration in the herbs has raised commercial interest in the identification of medicinal herbs globally. DNA barcoding is the primary techniques for identifying the herbs at genetic level. This technique's key benefit is that it can identify the material's purity. This study focuses on the accurate identification of species utilizing Polymerase chain reaction-based nuclear universal internal transcribed spacer region (a barcode region) amplification and sequencing in 2 medically significant plants (Rosa damascene and Prunus dulcis) procured from the local herbal market of Karachi in the year 2022. Results suggest that the ribosomal nuclear ITS2 region of the selected plant species shows the 100 % identity with the reference genome, therefore it has shown a good rate of identification at the species level. The findings of this concludes that ITS2 is the novel standard barcode that involve in species identification, genomic conservation, and secure utilization of the medically significant plant species.

Genetic diversity and occupation strategy of Egeria najas (Hydrocharitaceae) in different habitats along the Upper Paraná River corridor

Egeria najas is a submerged aquatic macrophyte native to South America, with high propagation in reservoirs and natural lakes, whose reproductive strategy is little known. Understanding the genetic diversity of macrophyte populations can provide important information about this species' dispersion and colonization strategies, and support management actions. We aimed to genetically characterize populations of E. najas that colonize reservoirs and natural aquatic habitats (in a floodplain) in the Upper Paraná River basin, using the molecular markers ITS and trnL-trnF. The results showed the absence of genetic variation for the nuclear marker ITS and 13 distinct haplotypes for trnL-trnF. One of these haplotypes occurred in all habitats and 11 are unique haplotypes, of which 5 occurred in the Itaipu Reservoir and 6 in the floodplain. The null genetic diversity for the nuclear marker and the genetic homogeneity of the studied populations indicates that the reproduction of E. najas is mostly vegetative. The source of chloroplast marker haplotype variability may be somatic mutations. The connectivity among aquatic environments associated with river flow favors the transport of aquatic macrophyte propagules to different habitats. In the case of E. najas, whose vegetative propagules regenerate easily, the frequency of migrations supports the low genetic variability observed in populations of the Upper Paraná. In addition, the ability to occupy new habitats and recolonize disturbed ones strongly indicates that E. najas populations follow the metapopulation dynamics.

Molecular Authentication and Phytochemical Evaluation of Indigenous Germplasm of Genus Physalis for Sustainable Utilization

Physalis species are used as an indigenous food and medicine in Kenya. However, species identification and an analysis of the health-promoting bioactive compounds and antioxidant properties are lacking. In this study, we report the molecular identification and mineral and phytochemical profiling of wild Physalis accessions. Leaf samples of 10 Physalis accessions were collected and used for species identification using nuclear ITS2 and plastid rbcL barcodes. Ripe fruits were collected from the same accessions and analyzed for mineral, total phenolic, tannin, and flavonoid contents, and antioxidant activities. The Physalis species were discriminated based on the ITS2 barcode and identified as Physalis purpurea. The genetic diversity, distance, and polymorphism of the ITS2 region of Physalis accessions were high due to the high rate of singleton and parsimony mutations. No genetic diversity, distance, or polymorphism was observed based on the rbcL barcode. The mineral content was significantly different (p < 0.05) for calcium, zinc, nickel, copper, and lithium among the Physalis accessions. No significant variation (p > 0.05) was found for phenolic acids or flavonoids, but the tannic acid content varied significantly (p < 0.05). DPPH free radical scavenging varied significantly (p < 0.05) among Physalis accessions. In conclusion, nuclear ITS2 was used to successfully identify the Physalis species of all the accessions as Physalis purpurea. The present study confirmed that Physalis purpurea has a significantly high mineral and phytochemical content and antioxidant activity. The findings from this study can be used to facilitate exploitation of Physalis purpurea in genetic breeding, their application in pharmaceutical, cosmetic, and nutritional value as well as conservation and sustainable use.

Species identity and phylogeny of Paramphistomoidea Fischoeder, 1901 occurring in cattle and sheep in North Cameroon.

Paramphistomidae and Gastrothylacidae are parasitic flatworms occurring in wild and domestic ruminants in different parts of the world especially in Asia and Africa. In Central Africa, few studies have been done using molecular techniques to resolve taxonomical groupings and understand the epizootiology of these parasites. In this study, we molecularly characterized two hundred adult flukes collected from the fore stomachs of cattle and sheep in the Adamawa region of the northern Cameroon. PCR and sequencing of the nuclear ITS-2 of the ribosomal DNA gene and a portion of the mitochondrial cox-1 locus revealed the presence of at least nine species belonging to the genera of Cotylophoron, Calicophorn, Orthocoelium and Carmyerius. In Zebu cattle, we identified Ca. microbothrium, Ca. clavula, Ca. phillerouxi, Co. cotylophorum, Co. fuelleborni, O. scoliocoelium, Car. gregarius, Car. graberi and Car. mancupatus and one yet unknown Paramphistomoidea sp, whereas in sheep, only Ca. microbothrium was found. The present study also strongly suggests cross-hybridization between the two Cotylophoron species coexisting in cattle. These results have implications for the diagnosis and control of rumen flukes in the region and point to the need for accurate species identification to understand parasite distribution and population genetics.

Molecular phylogenetic study of Utricularia section Oligocista in Australia and a new Cape York endemic species

Utricularia section Oligocista A.DC. (Lentibulariaceae) is distributed across the old and new world tropics. Wepresent a molecular phylogeny using a matrix of the nuclear ITS and two chloroplast markers that includes 19of the 37 recognised species sampled across the range of each species. The phylogenetic study also includesmembers of the closely allied sections Enskide (Raf.) P.Taylor (including section Minutae Lowrie, Cowie &Conran) and Stomoisia (Raf.) Kuntze from across each of their distributions. The phylogeny shows that Australiacontains species from the two major clades of section Oligocista, with each involving ancestors with yellow andpurple/mauve corolla colours. This study provides morphological and molecular evidence for recognition of anew species, Utricularia irwinica R.W.Jobson & Baleeiro, from Cape York Peninsula, Queensland. A lectotypeis designated for the name Utricularia cyanea var. alba Benth. We also recommend changes to the taxonomyof sections Enskide and Oligocista based on the phylogenetic results and morphology.

Chloroplast genome, nuclear ITS regions, mitogenome regions, and Skmer analysis resolved the genetic relationship among Cinnamomum species in Sri Lanka.

Cinnamomum species have gained worldwide attention because of their economic benefits. Among them, C. verum (synonymous with C. zeylanicum Blume), commonly known as Ceylon Cinnamon or True Cinnamon is mainly produced in Sri Lanka. In addition, Sri Lanka is home to seven endemic wild cinnamon species, C. capparu-coronde, C. citriodorum, C. dubium, C. litseifolium, C. ovalifolium, C. rivulorum and C. sinharajaense. Proper identification and genetic characterization are fundamental for the conservation and commercialization of these species. While some species can be identified based on distinct morphological or chemical traits, others cannot be identified easily morphologically or chemically. The DNA barcoding using rbcL, matK, and trnH-psbA regions could not also resolve the identification of Cinnamomum species in Sri Lanka. Therefore, we generated Illumina Hiseq data of about 20x coverage for each identified species and a C. verum sample (India) and assembled the chloroplast genome, nuclear ITS regions, and several mitochondrial genes, and conducted Skmer analysis. Chloroplast genomes of all eight species were assembled using a seed-based method.According to the Bayesian phylogenomic tree constructed with the complete chloroplast genomes, the C. verum (Sri Lanka) is sister to previously sequenced C. verum (NC_035236.1, KY635878.1), C. dubium and C. rivulorum. The C. verum sample from India is sister to C. litseifolium and C. ovalifolium. According to the ITS regions studied, C. verum (Sri Lanka) is sister to C. verum (NC_035236.1), C. dubium and C. rivulorum. Cinnamomum verum (India) shares an identical ITS region with C. ovalifolium, C. litseifolium, C. citriodorum, and C. capparu-coronde. According to the Skmer analysis C. verum (Sri Lanka) is sister to C. dubium and C. rivulorum, whereas C. verum (India) is sister to C. ovalifolium, and C. litseifolium. The chloroplast gene ycf1 was identified as a chloroplast barcode for the identification of Cinnamomum species. We identified an 18 bp indel region in the ycf1 gene, that could differentiate C. verum (India) and C. verum (Sri Lanka) samples tested.

Intersectional polyphyly of the puzzling Scapania obcordata (Marchantiidae) suggests convergent evolution: resurrection and European occurrence of S. jensenii

We present an example of morphological convergence among two species of Scapania that were considered to be conspecific. We resurrect S. jensenii from synonymy with the morphologically and molecularly variable S. obcordata because our molecular data show that they are distantly related. Chloroplast trnL-trnF and nuclear ITS sequences resolved S. obcordata in sect. Curtae and S. jensenii in sect. Apiculatae sister to S. obscura and we detected morphological differences in secondary pigmentation and leaf anatomy. The similarity comprises the absence of a pronounced keel in vegetative leaves and a weakly defined stem cortex. These features are virtually unknown in other species of Scapania and probably result from convergence in adaptation to the same environmental conditions, as both taxa inhabit alluvial plains characterised by regular disturbance by flooding and covering with sand and silt. While S. obcordata is widespread in the northern Holarctic and known from the Antarctic, S. jensenii is known from a few localities in Greenland, mainland Norway, the Chukchi Peninsula and the Swiss Alps. Its rarity, the absence of female plants, and the lack of genetic variability suggest that S. jensenii underwent a bottleneck event.

Rapid cryptic divergence of the Anastrepha fraterculus complex in the Late Pleistocene: a phylogeographical–ecological approach

Abstract The Anastrepha fraterculus complex (AF complex) is a complex of fruit fly species consisting of eight morphotypes distributed across the Americas. In this study, landscape genetics, phylogeographical inferences and coalescence simulations were used to unveil the evolutionary history of the AF complex. Nuclear ribosomal ITS1 sequences from 331 samples distributed in 10 American countries throughout 32 ecoregions and seven biomes were analysed. Results showed high levels of diversity, intraspecific divergence and structuring associated with the different morphotypes of the complex. The analysis of gene flow and genetic diversity showed a clear geographical and environmental pattern. The morphotype–environment association suggested that the cryptic divergence process has involved local adaptation as a mechanism to withstand harsh, novel or variable conditions. The analysis showed a rapid divergence between morphotypes. Haplotypes have been diverging for ~27 000 years before the present, probably enhanced by environmental changes during the Last Glacial Maximum. Diversity estimators and neutrality tests revealed that populations have been experiencing a recent linear growth, starting in the Early Holocene (~11 500 years before the present). The Bayesian phylogeographical and ecological clustering approach detected five spatially separated and ecologically differentiated clusters. Additionally, equatorial Amazonia was indicated to be the most probable ancestral centre of origin for the AF complex.

Central Asia revealed as a key area in evolution of Eremurus (Asphodelaceae)

Eremurus was described at the beginning of the 19th century. However, due to limited sampling and the small number of gene markers to date, its phylogeny and evolution are largely unknown. In this study, we analyzed plastomes from 27 species belonging to 2 subgenera and 3 sections of Eremurus, which are found in Central Asia (its center of diversity) and China. We also analyzed nuclear DNA ITS of 33 species, encompassing all subgenera and sections of the genus in Central Asia, southwest Asia and China. Our findings revealed that the genus was monophyletic, although both subgenera Eremurus and Henningia were found to be paraphyletic. Both plastome and nrDNA-based phylogenetic trees had three clades that did not reflect the current taxonomy of the genus. Our biogeographical and time-calibrated trees suggest that Eremurus originated in the ancient Tethyan area in the second half of the Eocene. Diversification of Eremurus occurred from the early Oligocene to the late Miocene. Paratethys Sea retreat and several orogenetic events, such as the progressive uplift of the Qinghai-Tibet Plateau and surrounding mountain belts (Altai, Pamir, Tian Shan), caused serious topographic and climate (aridification) changes in Central Asia that may have triggered a split of clades and speciation. In this transformed Central Asia, speciation proceeded rapidly driven mainly by vicariance caused by numerous mountain chains and specialization to a variety of climatic, topographic and soil conditions that exist in this region.

Phylogenetic analysis of the endemic Bornmuellera Hausskn. spp. (Brassicaceae) in Türkiye based on nuclear ITS and chloroplast trnL intron, trnL-F, rbcL and trnQ-rps16 DNA sequences

Phylogenetic analysis of the endemic Bornmuellera Hausskn. spp. (Brassicaceae) in Türkiye based on nuclear ITS and chloroplast trnL intron, trnL-F, rbcL and trnQ-rps16 DNA sequences

Monnina (Polygalaceae), a New World monophyletic genus full of contrasts

Abstract Endemic to the Neotropics, Monnina is the second largest genus of Polygalaceae, yet little is known about its phylogenetic history, biogeography, and morphological character evolution. To address these knowledge gaps, we conducted Bayesian and maximum likelihood (ML) analyses of nuclear ITS and plastid trnL–F regions to test the monophyly of Monnina s.l. We used this phylogenetic framework to (i) infer divergence time estimates of lineages within the genus and reconstruct their historical biogeography; (ii) reconstruct the evolution of morphological characters of putative ecological and evolutionary importance in Monnina; and (iii) test for correlations between our phylogenetic hypothesis and environmental data. Our results reveal that Monnina is monophyletic with an indehiscent, 1–2-seeded fruit as a synapomorphy for the genus. We identify six clades within Monnina based on our combined phylogenetic results: Clades A, B, and D are primarily distributed in southern and eastern South America, Clades C and E are primarily Central Andean, and Clade F is chiefly distributed in the Northern Andes and Central America. The ancestor of the Monnina stem lineage dispersed from Australia/Africa to South America during the late Eocene to early Oligocene. The divergences of major lineages within the genus began in the early Miocene. We inferred the most recent common ancestor of Monnina to be an herbaceous plant with one-seeded samaroid fruits. The origins of fleshy fruits and shrubby habits are phylogenetically correlated within Monnina, and their concerted convergent evolution may have promoted increased net diversification rates in the two most species-rich subclades of the genus.

Kiluluma ceratotherii (Nematoda: Strongylida) in a White Rhinoceros (Ceratotherium simum) from the United States: Case Report

Nematodes of the genus Kiluluma (Strongylidae, Cyathostominae) parasitize African rhinoceros. We describe the case of a one-year-old male white rhinoceros calf that presented with colonic inflammation and hemorrhage at necropsy. The animal had died following a neurological episode. We recovered and identified adult nematodes from the colon using morphology and ITS2 gene sequences as Kiluluma ceratotherii. We also generated nuclear ITS1, 5.8S, ITS2, and mitochondrial cox1 sequences for future studies and deposited them in GenBank (OR142644–OR142653). Since the animal was born in the same zoo and never transported, infection likely originated within the herd. This is the first report of this nematode from a white rhinoceros in the United States.

A new species of the genus Aleurodiscus sensu lato (Russulales, Basidiomycota) from Hachijo Island, Japan.

Aleurodiscus sagittisporus sp. nov. is described and illustrated. This species is characterized by producing basidiomata with a monomitic hyphal system, clampless-septate hyphae, arrowhead-shaped, amyloid, finely verrucose basidiospores, gloeocystidia, dendrohyphidium-like branched paraphysoid hyphae, and variously shaped swelling cells in the hymenium. Phylogenetic analyses based on nuclear rDNA LSU and ITS sequences revealed that the species is distinct from the lineage of Aleurodiscus s. str. and related genera in the Aleurodiscus s. lat. clade. Basidiomata of A. sagittisporus have been collected only from dead petioles attached to living trees of Livistona chinensis var. subglobosa on Hachijo Island, Japan.

Resolving relationships despite past hybridization in Aliciella subsection Subnuda (Polemoniaceae)

AbstractPhylogenetics is crucial in the study of evolutionary processes and events transpiring in the course of species diversification. Phylogenetic studies within kingdom Plantae often reveal hybridization and introgression. Here, we study a subsection rife with historic hybridization and discuss the impacts of such processes on evolutionary trajectories. Aliciella subsection Subnuda comprises seven species of herbaceous plants occurring in Utah, the Navajo Nation, and the Four Corners region of North America. Previous molecular and morphological work left relationships in the subsection unresolved. Here, we use comparative DNA sequencing of nuclear ITS and chloroplast DNA regions and genome‐wide RAD‐seq data to clarify phylogenetic relationships and examine the role of hybridization in the subsection. We construct haplotype and nucleotype networks from chloroplast and nuclear ITS sequence matrices and compare nuclear and chloroplast phylogenies to identify multiple putative chloroplast capture events. The RAD‐seq maximum likelihood phylogeny and multispecies coalescent species tree robustly resolve relationships between six species‐level clades. We use STRUCTURE and HyDe on the RAD‐seq data to evaluate the influence of hybridization within the subsection. The HyDe results suggest that hybridization has occurred among all species in the subsection at some point in their history. Cytonuclear discordance reveals historic chloroplast capture, and we discuss potential causes of the observed discordance. Our study robustly resolves relationships in Aliciella subsection Subnuda and provides a framework for discussing its speciation despite a history of hybridization and introgression.

Plastome evolution in the genus Sium (Apiaceae, Oenantheae) inferred from phylogenomic and comparative analyses

BackgroundSium L. (Apiaceae) is a small genus distributed primarily in Eurasia, with one species also occurring in North America. Recently, its circumscription has been revised to include 10 species, however, the phylogenetic relationships within its two inclusive clades were poorly supported or collapsed in previous studies based on nuclear ribosomal DNA ITS or cpDNA sequences. To identify molecular markers suitable for future intraspecific phylogeographic and population genetic studies, and to evaluate the efficacy of plastome in resolving the phylogenetic relationships of the genus, the complete chloroplast (cp) genomes of six Sium species were sequenced.ResultsThe Sium plastomes exhibited typical quadripartite structures of Apiaceae and most other higher plant plastid DNAs, and were relatively conserved in their size (153,029–155,006 bp), gene arrangement and content (with 114 unique genes). A total of 61–67 SSRs, along with 12 highly divergent regions (trnQ, trnG-atpA, trnE-trnT, rps4-trnT, accD-psbI, rpl16, ycf1-ndhF, ndhF-rpl32, rpl32-trnL, ndhE-ndhG, ycf1a and ycf1b) were discovered in the plastomes. No significant IR length variation was detected showing that plastome evolution was conserved within this genus. Phylogenomic analysis based on whole chloroplast genome sequences produced a highly resolved phylogenetic tree, in which the monophyly of Sium, as well as the sister relationship of its two inclusive clades were strongly supported.ConclusionsThe plastome sequences could greatly improve phylogenetic resolution, and will provide genomic resources and potential markers useful for future studies of the genus.

Genetic Markers for Metabarcoding of Freshwater Microalgae: Review.

The metabarcoding methods for studying the diversity of freshwater microalgae and routine biomonitoring are actively used in modern research. A lot of experience has been accumulated already, and many methodological questions have been solved (such as the influence of the methods and time of sample conservation, DNA extraction and bioinformatical processing). The reproducibility of the method has been tested and confirmed. However, one of the main problems-choosing a genetic marker for the study-still lacks a clear answer. We analyzed 70 publications and found out that studies on eukaryotic freshwater microalgae use 12 markers (different nuclear regions 18S and ITS and plastids rbcL, 23S and 16S). Each marker has its peculiarities; they amplify differently and have various levels of efficiency (variability) in different groups of algae. The V4 and V9 18S and rbcL regions are used most often. We concentrated especially on the studies that compare the results of using different markers and microscopy. We summarize the data on the primers for each region and on how the choice of a marker affects the taxonomic composition of a community.

Phylogenetic position of Dorema within Ferula (Apiaceae)

Dorema (Apiaceae) includes 12 species distributed in the Irano-Turanian region. The genus differs from other genera of subfamily Apioideae in having paniculate inflorescences with simple umbels. Recently, molecular phylogenetic analyses placed six species of Dorema, including the generitype, in Ferula, thus subsuming the former into the synonymy of the latter. The remaining six congeners have not been yet included in molecular analyses. Here, we fill this gap by determining the phylogenetic position of these species. Because of a low resolution in some parts of the Ferula tree in earlier studies, alongside standard set of markers—nuclear nrDNA ITS and plastid rps16 and rpoC1 introns and the rpoB-trnC intergenic spacer—we additionally assessed the utility of nrDNA ETS sequences in resolving infrageneric relationships in the genus. We included 43 ingroup species representing major infrageneric divisions recognised in the most recent taxonomic revision. Although we were not successful in obtaining all markers for all newly studied species, they were placed with high support in one clade within section Peucedanoides together with formerly studied congeners in all maximum likelihood and Bayesian analyses of nuclear, plastid, and combined data sets. The addition of the ETS marker to the combined phylogenetic analyses confirmed the most recent classification of Ferula with increased branch support in some parts of the tree. However, many branches still remained unresolved. In particular, the monophyly of former Dorema within section Peucedanoides as suggested by morphology has not been definitely confirmed. A new combination and two nomina nova are proposed.

Molecular analysis separates the Atlantic population of Montagu's stellate barnacle into two cryptic species

AbstractThe distribution of the intertidal barnacle Chthamalus montagui spans the West Mediterranean Sea and the Northeast Atlantic shores of Europe and West Africa. Knowledge of the phylogeography of this species has been limited to the Mediterranean and the European shores of the Atlantic. The present study considers the populations of West Europe, but also focuses on the overlooked populations of West Africa. We performed a molecular analysis using two markers: the mitochondrial COI gene and the nuclear rRNA ITS gene. Whereas ITS proved to be non‐informative, COI has demonstrated that the East Atlantic population of C. montagui comprises two genetically distinct clades: a northern clade that ranges from Mauritania to Scotland and a southern clade that comprises the populations from Senegal. These clades are separated by the Cape Verde Front, which stretches west of the upwelling area off Mauritania. We consider these clades as two cryptic species of the nominal species C. montagui.

Grimmia shevockii and G. insolita, two new species endemic to California

ABSTRACT Introduction California is a biodiversity hotspot for bryophytes, and its northern counties are among the most endemic-rich areas for bryophytes in North America. Having 29 species in California, Grimmia Hedw. is the third richest moss genus in the state, but recent fieldwork suggests that this number is an underestimate. The identity of some specimens endemic to the Klamath Mountains of California and previously identified as G. brevirostris R.S.Williams remains unclear, and they potentially represent a new species. Two further specimens of Grimmia from Siskiyou County, California, do not correspond to any species described so far. Here we present morphological and phylogenetic arguments for recognising these plants as two new moss species in the genus Grimmia subg. Orthogrimmia Schimp. Methods Morphological characters across potential new species were reviewed and compared with those of related species. Phylogenetic analyses based on the plastid trnS–trnF and nuclear ITS regions were conducted. Key results and conclusions The molecular and morphological analyses revealed two new species of Grimmia for California, which are described and illustrated: G. shevockii J.Muñoz, I.Solano & D.Quandt and G. insolita J.Muñoz, I.Solano & D.Quandt. The former is resolved in a monophyletic clade with high support, whereas the latter has incongruent positions in the plastid and nuclear phylogenies, thus it is hypothesised that it could have a hybrid origin. Both of the two new species and G. hamulosa undoubtly pertain to Grimmia. The richness of the bryophyte flora of California provides a clear warrant for future research.