Comparative study of eco-friendly wire mesh configurations to enhance sustainability in reinforced concrete structures.

Recent and past studies mainly focus on reducing the dead weight of structure; therefore, they considered lightweight aggregate concrete (LWAC) which reduces the dead weight but also affects the strength parameters. Therefore, the current study aims to use varied steel wire meshes to investigate the effects of LWAC on mechanical properties. Three types of steel wire mesh are used such as hexagonal (chicken), welded square, and expanded metal mesh, in various layers and orientations in LWAC. Numerous mechanical characteristics were examined, including energy absorption (EA), compressive strength (CS), and flexural strength (FS). A total of ninety prisms and thirty-three cubes were made. For the FS test, forty-five 100 × 100 × 500 mm prism samples were poured, thirty-three 150 × 150 × 150 mm cube samples were made, and forty-five 400 × 300 × 75 mm EA specimens were costed for fourteen days of curing. The experimental findings demonstrate that the FS was enhanced by adding additional forces that spread the forces over the section. One layer of chicken, welded, and expanded metal mesh enhances the FS by 52.96%, 23.76%, and 22.2%, respectively. In comparison to the remaining layers, the FS in a single-layer hexagonal wire mesh has the maximum strength, 29.49 MPa. The hexagonal wire mesh with a single layer had the greatest CS, measuring 36.56 MPa. When all three types of meshes are combined, the CS does not vary in this way and is estimated to be 29.79 MPa. In the combination of three layers, the chicken and expanded wire mesh had the most energy recorded prior to final failure, which was 1425.6 and 1108.7 J, whereas it was found the highest 752.3 J for welded square wire mesh. The energy absorption for the first layer with hexagonal wire mesh increased by 82.81% prior to the crack and by 88.34% prior to the ultimate failure. Overall, it was determined and suggested that hexagonal wire mesh works better than expanded and welded wire meshes.

Soft ground micro TBM jack speed and torque prediction using machine learning models through operator data and micro TBM-log data synchronization.

Tunnel Boring Machines (TBMs) are pivotal in underground projects like subways, highways, and water supply tunnels. Predicting and monitoring jack speed and torque is crucial for optimizing TBM excavation efficiency. Conventionally, skilled operators manually adjust numerous tunnelling parameters to regulate the machine's progress. In contrast, machine learning (ML) algorithms offer a promising avenue where computers learn from operator actions to establish parameter relationships autonomously. This study introduces an innovative approach to enhancing operator monitoring and TBM data comprehension. A robust correlation between TBM operator behaviour and TBM logged data is established by leveraging an Optuna-assisted ML methodology-the research light on the intricate dynamics influencing TBM advance rate parameters. Operational data is collected from micro slurry tunnel boring machine (MSTBM) umbrella support excavations. The proposed framework harnesses Optuna, an advanced hyperparameter optimization platform, to dynamically refine jack speed and torque settings. Through meticulous analysis of the interplay between TBM operator decisions and real-time logged data, the AI model discerns patterns, empowering informed decision-making. Using Optuna, a range of models, including random forest (RF), K-nearest neighbours (kNN), decision tree (DT), XGBoost, Support Vector Machine (SVM), and Artificial Neural Network (ANN) were automatically compared and tuned. The best model's (RF) performance is evaluated through a correlation coefficient (R2) of 96%, mean squared error (MSE) of 119.7, and mean absolute error (MAE) of 4.42 for jack speed decision making while 83% of R2, MSE of 0.62, and MAE of 0.42 for the torque decision making. This intelligent model can assist the TBM operator in making decisions about TBM control.

Pyrrolizidine alkaloids are synthesized and accumulated in flower of Myosotis scorpioides.

Pyrrolizidine alkaloids (PAs) are specialized metabolites that are produced by various plant families that act as defense compounds against herbivores. On the other hand, certain lepidopteran insects uptake and utilize these PAs as defense compounds against their predators and as precursors of their sex pheromones. Adult males of Parantica sita, a danaine butterfly, convert PAs into their sex pheromones. In early summer, P. sita swarms over the flowers of Myosotis scorpioides, which belongs to the family Boraginaceae. M. scorpioides produces PAs, but the organs in which PAs are produced and whether P. sita utilizes PAs in M. scorpioides are largely unknown. In the present study, we clarified that M. scorpioides accumulates retronecine-core PAs in N-oxide form in all organs, including flowers. We also identified two M. scorpioides genes encoding homospermidine synthase (HSS), a key enzyme in the PA biosynthetic pathway, and clarified that these genes are expressed in all organs where PAs accumulate. Phylogenetic analysis suggested that these two HSS genes were originated from gene duplication of deoxyhypusine synthase gene like other HSS genes in PA-producing plants. These results suggest that PAs are synthesized and accumulated in the flower of M. scorpioides and provide a possibility for a PA-mediated interaction between P. sita and M. scorpioides.

Effect of rock properties on wear and cutting performance of multi blade circular saw with iron based multi-layer diamond segments.

This study is an attempt for comprehensive, combining experimental data with advanced analytical techniques and machine learning for a thorough understanding of the factors influencing the wear and cutting performance of multi-blade diamond disc cutters on granite blocks. A series of sawing experiments were performed to evaluate the wear and cutting performance of multi blade diamond disc cutters with varying diameters in the processing of large-sized granite blocks. The multi-layer diamond segments comprising the Iron (Fe) based metal matrix were brazed on the sawing blades. The segment's wear was studied through micrographs and data obtained from the Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive X-ray (EDS). Granite rock samples of nine varieties were tested in the laboratory to determine the quantitative rock parameters. The contribution of individual rock parameters and their combined effects on wear and cutting performance of multi blade saw were correlated using statistical machine learning methods. Moreover, predictive models were developed to estimate the wear and cutting rate based on the most significant rock properties. The point load strength index, uniaxial compressive strength, and deformability, Cerchar abrasivity index, and Cerchar hardness index were found to be the significant variables affecting the sawing performance.

Assessment of the ground vibration during blasting in mining projects using different computational approaches.

The investigation compares the conventional, advanced machine, deep, and hybrid learning models to introduce an optimum computational model to assess the ground vibrations during blasting in mining projects. The long short-term memory (LSTM), artificial neural network (ANN), least square support vector machine (LSSVM), ensemble tree (ET), decision tree (DT), Gaussian process regression (GPR), support vector machine (SVM), and multilinear regression (MLR) models are employed using 162 data points. For the first time, the blackhole-optimized LSTM model has been used to predict the ground vibrations during blasting. Fifteen performance metrics have been implemented to measure the prediction capabilities of computational models. The study concludes that the blackhole optimized-LSTM model PPV11 is highly capable of predicting ground vibration. Model PPV11 has assessed ground vibrations with RMSE = 0.0181 mm/s, MAE = 0.0067 mm/s, R = 0.9951, a20 = 96.88, IOA = 0.9719, IOS = 0.0356 in testing. Furthermore, this study reveals that the prediction accuracy of hybrid models is less affected by multicollinearity because of the optimization algorithm. The external cross-validation and literature validation confirm the prediction capabilities of model PPV11. The ANOVA and Z tests reject the null hypothesis for actual ground vibration, and the Anderson-Darling test rejects the null hypothesis for predicted ground vibration. This study also concludes that the GPR and LSSVM models overfit because of moderate to problematic multicollinearity in assessing ground vibration during blasting.

Virtual issue: phylogeographic studies in the Japanese Archipelago: from geographic patterns of genetic variation to biodiversity in plants.

Phylogeographic studies have investigated genetic variation and structure within species or closely related lineages and are fundamental for understanding factors and processes of genetic divergence as well as speciation. This virtual issue collects 35 papers on phylogeographic studies published in the Journal of Plant Research and focuses on three major topics in biodiversity: (1) biogeography, (2) systematics, and (3) evolution.

Phylogeography of the temperate grassland plant Tephroseris kirilowii (Asteraceae) inferred from multiplexed inter-simple sequence repeat genotyping by sequencing (MIG-seq) data.

A group of temperate grassland plant species termed the "Mansen elements" occurs in Japan and is widely distributed in the grasslands of continental East Asia. It has been hypothesized that these species are continental grassland relicts in Japan that stretch back to a colder age, but their migration history has not been elucidated. To assess the migration history of the Mansen elements, we performed phylogeographic analyses of Tephroseris kirilowii, a member of this group, using single-nucleotide polymorphisms (SNPs) obtained from multiplexed inter-simple sequence repeat genotyping by sequencing (MIG-seq). It was estimated that the Japanese populations of T. kirilowii were divided from those of continental East Asia at 25.2 thousand years ago (ka) with 95% highest probability density interval (HPD) of 15.3-40.0 ka and that Japanese clades first diverged at 20.2 ka with 95% HPD of 10.4-30.1 ka. As the climatically suitable range during the last glacial maximum (LGM) estimated using ecological niche modeling (ENM) was limited in Japan and there was a slight genetic differentiation among Japanese populations, a post-glacial expansion of T. kirilowii in the Japanese Archipelago was indicated.

Decades-long phylogeographic issues: complex historical processes and ecological factors on genetic structure of alpine plants in the Japanese Archipelago.

Mountain regions are important places for biodiversity, where organisms could persist throughout prolonged periods and accumulate genetic divergence as well as promote speciation. Roles of mountains for biodiversity have been exclusively discussed in regions that have specifically diverse species or covered with ice-sheets during the Pleistocene glacial periods, whereas the importance of mountainous regions in East Asia has been less disputed. High mountains in the Japanese Archipelago, located at the eastern edge of the Eurasia continent, have one of southernmost populations of alpine and arctic-alpine plants that are also distributed in the northern Pacific and/or the circumarctic regions. Phylogeographic studies on the Japanese alpine plants have excluded their possible ephemeral occurrence during the current warm period, and rather, suggest persistence of alpine plants throughout several cycles of climate changes in the Pleistocene on high mountains in central Honshu, the main island of the Japanese Archipelago. In this review, I look through decade long phylogeographic studies and show complicated patterns of range dynamics of Japanese alpine plants. In addition, I note recent findings of genetic relationships of Japanese populations of alpine and/or arctic-alpine plants with those in northern regions and their possible ecological divergence in the Japanese Archipelago. Taken together, I provide several issues for understanding historical processes that established distribution of alpine plants following climate changes as well as their diversification and propose importance of Japanese populations of alpine plants on biodiversity in alpine communities across broader range, especially in the northern Pacific region.

Rapid evolution of post-zygotic reproductive isolation is widespread in Arctic plant lineages.

BACKGROUND AND AIMS: The Arctic tundra, with its extreme temperatures and short growing season, is evolutionarily young and harbours one of the most species-poor floras on Earth. Arctic species often show little phenotypic and genetic divergence across circumpolar ranges. However, strong intraspecific post-zygotic reproductive isolation (RI) in terms of hybrid sterility has frequently evolved within selfing Arctic species of the genus Draba. Here we assess whether incipient biological species are common in the Arctic flora. METHODS: We conducted an extensive crossing experiment including six species representing four phylogenetically distant families collected across the circumpolar Arctic. We crossed conspecific parental populations representing different spatial scales, raised 740 F1 hybrids to maturity and measured fertility under laboratory conditions. We examined genetic divergence between populations for two of these species (Cardamine bellidifolia and Ranunculus pygmaeus). KEY RESULTS: In five of the six species, we find extensive reduction in pollen fertility and seed set in F1 hybrids; 219 (46 %) of the 477 F1 hybrids generated between parents separated by ≥427 km had <20 % pollen fertility. Isolation with migration (IM) and *BEAST analyses of sequences of eight nuclear genes in C. bellidifolia suggests that reproductively isolated populations of this species diverged during, or even after, the last glaciation. Likewise, Arctic populations of R. pygmaeus were genetically very similar despite exhibiting strongly reduced fertility in crosses, suggesting that RI evolved recently also in this species. CONCLUSION: We show that post-zygotic RI has developed multiple times within taxonomically recognized Arctic species belonging to several distantly related lineages, and that RI may have developed over just a few millennia. Rapid and widespread evolution of incipient biological species in the Arctic flora might be associated with frequent bottlenecks due to glacial cycles, and/or selfing mating systems, which are common in the harsh Arctic environment where pollinators are scarce.

Phylogeography of the East Asian grassland plant, Viola orientalis (Violaceae), inferred from plastid and nuclear restriction site-associated DNA sequencing data.

To elucidate the origin and migration history of the "Mansen elements," a group of temperate grassland plants mainly distributed in northeastern Asia, phylogeographic analyses based on chloroplast DNA markers and double-digest restriction site-associated DNA sequencing (ddRAD-seq) data were performed on Viola orientalis, one of the representative species of the group. Phylogenetic analyses using ddRAD-seq data revealed that the populations of V. orientalis were clustered into five clades, among which the continental clades made of populations from Russia and Korea diverged more than 100,000 years earlier than the Japanese clades. The Japanese clade likely diverged during the last glacial period, followed by a further post-glacial divergence into the Kyushu and the Honshu subclades. Our study demonstrated that V. orientalis originated in the continental area of northeastern Asia and, during the last glacial period, has spread southward through the Korean Peninsula across the Japanese Islands. This finding supports the previously proposed evolutionary hypothesis regarding the origin and migration routes of the Mansen elements.

Analysis of Secondary Leukemia and Myelodysplastic Syndrome After Chemotherapy for Solid Organ Tumors Using the Food and Drug Administration Adverse Event Reporting System (FAERS).

PURPOSE: As the prognosis of cancer patients deteriorates, secondary carcinogenesis after chemotherapy, especially secondary hematological malignancies, becomes a serious problem. However, information on the frequency and time of onset of secondary hematological malignancies and the risk of hematological malignancy with different drugs is scarce. This study aimed to evaluate the incidence of leukemia and myelodysplastic syndrome in patients with solid tumors, including breast, colon, gastric, pancreatic, small cell lung, non-small cell lung, esophageal, ovarian, cervical, and endometrial cancers. METHODS: Using the United States Food and Drug Administration Adverse Event Reporting System, we analyzed the reporting rates, reporting odds ratios, and the reporting onset times of secondary leukemia and myelodysplastic syndrome for each drug used. RESULTS: The leukemia reporting rates were higher in breast, small cell lung, ovarian, and endometrial cancers than in other cancers, and the myelodysplastic syndrome reporting rates were higher in ovarian and endometrial cancers than in other cancers. For each cancer type, the reporting odds ratios of cytocidal anticancer agents, such as taxanes, anthracyclines, alkylating agents, platinum, and topoisomerase inhibitors, were higher than those of other drugs. Alternatively, the reporting odds ratios of molecular targeted drugs and immune checkpoint inhibitors were not higher than those of other drugs. Approximately half of the cases of leukemia and myelodysplastic syndrome were reported within 1 to 4 years after chemotherapy. CONCLUSIONS: Our study clarified the risks of leukemia and myelodysplastic syndrome for several anticancer drugs in patients with solid tumors. Our data may aid in the assessment of the risks of secondary leukemia and myelodysplastic syndrome when medical oncologists, clinical pharmacists, and patients select chemotherapy regimens.

Divergence in red light responses associated with thermal reversion of phytochrome B between high- and low-latitude species.

Phytochromes play a central role in mediating adaptive responses to light and temperature throughout plant life cycles. Despite evidence for adaptive importance of natural variation in phytochromes, little information is known about molecular mechanisms that modulate physiological responses of phytochromes in nature. We show evolutionary divergence in physiological responses relevant to thermal stability of a physiologically active form of phytochrome (Pfr) between two sister species of Brassicaceae growing at different latitudes. The higher latitude species (Cardamine bellidifolia; Cb) responded more strongly to light-limited conditions compared with its lower latitude sister (C. nipponica; Cn). Moreover, CbPHYB conferred stronger responses to both light-limited and warm conditions in the phyB-deficient mutant of Arabidopsis thaliana than CnPHYB: that is Pfr CbphyB was more stable in nuclei than CnphyB. Our findings suggest that fine tuning Pfr stability is a fundamental mechanism for plants to optimise phytochrome-related traits in their evolution and adapt to spatially varying environments, and open a new avenue to understand molecular mechanisms that fine tune phytochrome responses in nature.

Hemostatic Effect of Palliative Radiation Therapy in Preventing Blood Transfusions from Bleeding Occurring within Advanced Gastric Cancer.

Purpose: To report the hemostatic effects of palliative radiation therapy (RT) for the prevention of blood transfusions (BT) in patients with advanced gastric cancer (AGC). Methods and Materials: Twenty-eight patients who received palliative three-dimensional conformal RT for hemostasis of gastric bleeding were retrospectively assessed in a study conducted in Japan. The median follow-up was 143.5 days. Changes in hemoglobin (Hb) levels were compared at the beginning of RT and four weeks later. Blood transfusion-free survival (BTFS) and overall survival (OS) were measured from the beginning of RT. Treatment toxicity was evaluated within 60 days of RT initiation. Results: No statistically significant decrease in Hb level was observed four weeks after RT. Twenty-eight patients did not receive BT within a month after RT, of whom three died within a month; 6/28 patients (21%) received BT at a median interval of 99.5 days following RT. The one-year BTFS and OS rates for all patients were 69% and 12%, respectively. The one-year BTFS was statistically significantly higher in 17 patients treated with a biologically effective dose (BED)10 of 39 Gy (30 Gy in 10 fractions) (78%) compared with six patients treated with a BED10 of 48 Gy (40 Gy in 20 fractions) (25%). Grade 1 and 2 nausea (n = 11) and a Grade 2 increase in alanine aminotransferase (n = 1) were observed. One patient died of Grade 5 hemorrhage. Conclusions: Palliative RT is an effective treatment to prevent BT for bleeding occurring within AGC. Specifically, a fractionation regimen of 30 Gy in 10 fractions (a BED10 of 39 Gy) has a more durable hemostatic effect and thus should be considered for better prognosis.

Phylogeography of Pulsatilla cernua (Ranunculaceae), a grassland species, in Japan.

The genetic diversity and structure of Pulsatilla cernua, a continental-grassland relict, were investigated using variations in chloroplast DNA (cpDNA) and microsatellites of nuclear DNA. In the analyses of three cpDNA regions, 17 haplotypes were found in 24 populations of P. cernua from Japan, Korea, and Russia. Although the route and time of migration between the continent of Asia and Japan could not be well resolved, the cpDNA haplotype network suggests the existence of several ancient lineages in Japan and a recent secondary migration from Japan to the continent. Microsatellite analyses did not indicate genetic structure among the Japanese populations, indicating the existence of gene flow across the distribution area until recently. These results indicate that the present fragmentation of P. cernua in Japan may reflect a rapid, recent reduction from a previously large, continuous distribution.

Late Pleistocene origin of the entire circumarctic range of the arctic-alpine plant Kalmia procumbens.

The circumarctic ranges of arctic-alpine plants are thought to have been established in the late Pliocene/early Pleistocene, when the modern arctic tundra was formed in response to climate cooling. Previous findings of range-wide genetic structure in arctic-alpine plants have been thought to support this hypothesis, but few studies have explicitly addressed the temporal framework of the genetic structure. Here, we estimated the demographic history of the genetic structure in the circumarctic Kalmia procumbens using sequences of multiple nuclear loci and examined whether its genetic structure reflects prolonged isolation throughout the Pleistocene. Both Bayesian clustering and phylogenetic analyses revealed genetic distinction between alpine and arctic regions, whereas detailed groupings were somewhat discordant between the analyses. By assuming a population grouping based on the phylogenetic analyses, which likely reflects a deeper intraspecific divergence, we conducted model-based analyses and demonstrated that the intraspecific genetic divergence in K. procumbens likely originated during the last glacial period. Thus, there is no need to postulate range separation throughout the Pleistocene to explain the current genetic structure in this species. This study demonstrates that range-wide genetic structure in arctic-alpine plants does not necessarily result from the late Pliocene/early Pleistocene origin of their circumarctic ranges and emphasizes the importance of a temporal framework of the current genetic structure for understanding the biogeographic history of the arctic flora.

Genetic structure of Hepatica nobilis var. japonica, focusing on within population flower color polymorphism.

How phenotypic or genetic diversity is maintained in a natural habitat is a fundamental question in evolutionary biology. Flower color polymorphism in plants is a common polymorphism. Hepatica nobilis var. japonica on the Sea of Japan (SJ) side of the Japanese mainland exhibits within population flower color polymorphism (e.g., white, pink, and purple), while only white flowers are observed on the Pacific Ocean (PO) side. To determine the relationships between flower color polymorphism, within and among populations, and the genetic structure of H. nobilis var. japonica, we estimated the genetic variation using simple sequence repeat (SSR) markers. First, we examined whether cryptic lineages corresponding to distinct flower colors contribute to the flower color polymorphisms in H. nobilis var. japonica. In our field observations, no bias in color frequency was observed among populations on Sado Island, a region with high variation in flower color. Simple sequence repeat (SSR) analyses revealed that 18% of the genetic variance was explained by differences among populations, whereas no genetic variation was explained by flower color hue or intensity (0% for both components). These results indicate that the flower color polymorphism is likely not explained by cryptic lineages that have different flower colors. In contrast, populations in the SJ and PO regions were genetically distinguishable. As with the other plant species in these regions, refugial isolation and subsequent migration history may have caused the genetic structure as well as the spatially heterogeneous patterns of flower color polymorphisms in H. nobilis var. japonica.

Persistent history of the bird-dispersed arctic-alpine plant Vaccinium vitis-idaea L. (Ericaceae) in Japan.

Arctic-alpine plants have expanded and contracted their ranges in response to the Pleistocene climate oscillations. Today, many arctic-alpine plants have vast distributions in the circumarctic region as well as marginal, isolated occurrences in high mountains at lower latitudes. These marginal populations may represent relict, long-standing populations that have persisted for several cycles of cold and warm climate during the Pleistocene, or recent occurrences that either result from southward step-wise migration during the last glacial period or from recent long-distance dispersal. In light of these hypotheses, we investigated the biogeographic history of the marginal Japanese populations of the widespread arctic-alpine plant Vaccinium vitis-idaea (Ericaceae), which is bird-dispersed, potentially over long distances. We sequenced three nuclear loci and one plastid DNA region in 130 individuals from 65 localities covering its entire geographic range, with a focus on its marginal populations in Japan. We found a homogenous genetic pattern across its enormous range based on the loci analysed, in contrast to the geographically structured variation found in a previous study of amplified fragment length polymorphisms in this species. However, we found several unique haplotypes in the Japanese populations, excluding the possibility that these marginal populations result from recent southward migration. Thus, even though V. vitis-idaea is efficiently dispersed via berries, our study suggests that its isolated populations in Japan have persisted during several cycles of cold and warm climate during the Pleistocene.

Polymorphisms of E1 and GIGANTEA in wild populations of Lotus japonicus.

In plants, timing of flowering is an essential factor that controls the survival rates of descendants. The circadian clock genes E1 and GIGANTEA (GI) play a central role in transmitting signals to flowering locus T (FT) in leguminous plants. Lotus japonicus is a wild Japanese species that ranges from northern Hokkaido to the southern Ryukyus and exhibits a wide range in terms of the time between seeding and first flowering. In this study, we first identified LjGI and analyzed polymorphisms of LjE1 and LjGI among wild populations covering the entire distribution range of this species in Japan. LjGI had a coding sequence (CDS) length of 3495 bp and included 14 exons. The homologies of DNA and amino acid sequences between LjGI and GmGI were 89 and 88% (positive rate was 92%), respectively. LjE1 harbored five nucleic acid changes in a 552 bp CDS, all of which were nonsynonymous; four of the changes were located in the core function area. LjE1 alleles exhibited partial north-south differentiation and non-neutrality. In contrast, the LjGI harbored one synonymous and one nonsynonymous change. Thus, our study suggests that LjE1 may be involved in the control of flowering times, whereas LjGI may be under strong purifying selection.

Molecular evidence for ancient relicts of arctic-alpine plants in East Asia.

Following climate cooling at the end of the Tertiary, arctic-alpine plants attained most of their extant species diversity. Because East Asia was not heavily glaciated, the importance of this region as a location for the long-term persistence of these species and their subsequent endemism during the Pleistocene was proposed in early discussions of phytogeography. However, this hypothesis remains to be fully tested. Here, we address this hypothesis by elucidating the phylogenetic history of Phyllodoce (Ericaceae). A phylogenetic tree based on multiple nuclear loci revealed that Phyllodoce nipponica was not derived from widespread species such as the arctic-alpine Phyllodoce caerulea, but rather represented an independent lineage sister to the clade of widespread relatives. Molecular dating indicated a mid-Pleistocene divergence of P. nipponica. These findings exclude the hypothesis that P. nipponica was derived from an arctic-alpine species that extended its range southwards during recent glacial periods. Instead, our results support the hypothesis that P. nipponica is an ancestral species which persisted in the Japanese archipelago during the mid- and late Pleistocene. Our findings demonstrate support for the early proposal and shed light on the importance of the Japanese archipelago for the evolution and persistence of arctic-alpine species.

Lineage diversification and hybridization in the Cayratia japonica-Cayratia tenuifolia species complex.

The Cayratia japonica-Cayratia tenuifolia species complex (Vitaceae) is distributed from temperate to tropical East Asia, Southeast Asia, India, and Australia. The spatiotemporal diversification history of this complex was assessed through phylogenetic and biogeographic analyses. Maximum parsimony, neighbor-joining, and maximum likelihood methods were used to analyze sequences of one nuclear (AS1) and two plastid regions (trnL-F and trnC-petN). Bayesian dating analysis was conducted to estimate the divergence times of clades. The likelihood method LAGRANGE was used to infer ancestral areas. The Asian C. japonica and C. tenuifolia should be treated as an unresolved complex, and Australian C. japonica is distinct from the Asian C. japonica-C. tenuifolia species complex and should be treated as separate taxa. The Asian C. japonica-C. tenuifolia species complex was estimated to have diverged from its closest relatives during the Late Eocene (35.1 million years ago [Ma], 95% highest posterior densities [HPD]=23.3-47.3Ma) and most likely first diverged in mid-continental Asia. This complex was first divided into a northern clade and a southern clade during the middle Oligocene (27.3Ma; 95% HPD=17.4-38.1Ma), which is consistent with a large southeastward extrusion of the Indochina region relative to South China along the Red River. Each of the northern and southern clades then further diverged into multiple subclades through a series of dispersal and divergence events following significant geological and climatic changes in East and Southeast Asia during the Miocene. Multiple inter-lineage hybridizations among four lineages were inferred to have occurred following this diversification process, which caused some Asian lineages to be morphologically cryptic.