Ecological Research最新文献

{"title":"Estimating Distribution Range Shifts of the Japanese Alpine Butterfly Colias palaeno Induced by Historical Climate Fluctuations and the Crucial Role of Its Host Plant","authors":"Daisuke Shimizu,&nbsp;Yuji Yamazaki","doi":"10.1111/1440-1703.70103","DOIUrl":"https://doi.org/10.1111/1440-1703.70103","url":null,"abstract":"<p>The glacial relict butterfly <i>Colias palaeno</i>, which inhabits alpine zones in Japan and is designated a protected species, depends strongly on its host plant <i>Vaccinium uliginosum</i>. Recent environmental changes have raised concerns about shifts in its habitat range; therefore, understanding its distributional dynamics is important for future conservation. However, despite its ecological and conservational importance, temporal changes in the distribution of <i>C. palaeno</i> and the roles of environmental and biological factors have not been quantitatively evaluated across time scales. In this study, we used a landscape ecology approach to examine historical distribution changes and key environmental factors and then estimated the potential future habitat range. Field surveys were conducted in mountainous areas between 2020 and 2023. MaxEnt (Maximum Entropy) niche models indicated that temperature and the presence of <i>V. uliginosum</i> were major factors influencing <i>C. palaeno</i> occurrence. Analyses using paleoclimate reconstructions revealed that both species occupied broader ranges at lower elevations during the Last Glacial Maximum and then shifted upward and became fragmented during the Holocene Thermal Maximum. The degree-day model supported these results by confirming the physiological constraints on the habitat range over time. Based on future projections under the Shared Socioeconomic Pathways (SSPs; SSP2-4.5, 2041–2060), the distributions of both species are expected to be greatly reduced, with habitat fragmentation in the Northern Japanese Alps range and loss of habitat in the Asama mountain range. These findings clarify the vulnerability of <i>C. palaeno</i> to environmental changes and provide essential baseline information for alpine biodiversity conservation.</p>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1440-1703.70103","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148386521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced Nickel Uptake in Mixed Plant Communities of Hyperaccumulator and Non-Hyperaccumulator Species Under Nickel Stress","authors":"Panayiotis G. Dimitrakopoulos,&nbsp;Maria Aloupi,&nbsp;Dimitris Skopianos,&nbsp;Ioanna Chatzelli","doi":"10.1111/1440-1703.70100","DOIUrl":"https://doi.org/10.1111/1440-1703.70100","url":null,"abstract":"<p>Biodiversity loss can negatively affect ecosystem functioning, yet its influence on processes related to metal uptake remains poorly tested. Here, we investigate for the first time how species richness affects biomass production and nickel phytoextraction under serpentine stress using endemic plants occurring in Mediterranean ecosystems. Monocultures and bicultures of <i>Odontarrhena lesbiaca</i> (nickel hyperaccumulator) and <i>Alyssum xiphocarpum</i> (non-hyperaccumulator) were grown on three substrates representing a gradient of increasing stress: peat, a peat-serpentine mixture, and pure serpentine soil. Serpentine soil had the highest total nickel concentration, and aboveground biomass generally declined with increasing soil nickel levels. Across all substrates, bicultures increased soil nickel bioavailability and total aboveground biomass and enhanced the phytoextracted nickel mass by an average of 29% compared to monocultures. The hyperaccumulator had higher tissue nickel concentrations, while the presence of the non-hyperaccumulator enhanced phytoextraction in mixtures, suggesting facilitative species interactions that promote nickel mobilization and uptake in line with the stress-gradient hypothesis. These findings indicate that combining hyperaccumulator and non-hyperaccumulator species can enhance nickel phytoextraction in metal-stressed soils, offering a promising biodiversity-based approach to agromining and remediation of metal-polluted environments.</p>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1440-1703.70100","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148324472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantifying the Radial Profile of Stem Sap Flux Density in a Cool Temperate Forest of Japan: Tree-To-Tree Variability Outweighs Seasonal and Species-Specific Effects","authors":"Qistan Naufal Faryzan,&nbsp;Quan Wang,&nbsp;Mitsuhiko Hanawa,&nbsp;Noviana Budianti,&nbsp;Masaaki Naramoto,&nbsp;Atsuhiro Iio","doi":"10.1111/1440-1703.70096","DOIUrl":"https://doi.org/10.1111/1440-1703.70096","url":null,"abstract":"<div>\u0000 \u0000 <p>Although quantifying the radial profile of stem sap flux density (SFD) is crucial for accurately estimating tree and stand transpiration, limited information exists regarding species-specific and seasonal variations in SFD profiles, particularly in humid forests. To address this knowledge gap, we measured SFD at four specific depths across different tree species (<i>Acer nipponicum</i>, <i>Acer shirasawanum</i>, <i>Betula grossa</i>, and <i>Fagus crenata</i>) over the growing seasons in a cool temperate forest in Japan. Our findings revealed divergent patterns in radial profiles of SFD among trees, with no distinct species-specific profiles due to significant tree-to-tree variation. Environmental responses of SFD were uniform during the full-leaf period (June–September) regardless of species and individuals. However, significant seasonal variations in the radial profile were observed in <i>F. crenata</i>, where post-summer declines in outermost layers exceeded those in inner layers. Ignoring these seasonal variations resulted in a mean error of 7.4 ± 3.3 (standard deviation) % for <i>F. crenata</i> and 3.5% ± 2.0% for the other species in annual transpiration estimates, whereas neglecting tree-to-tree variability caused up to errors of 29.0% ± 13.0%. Therefore, considering seasonal variations in the radial profiles of SFD is important for accurate transpiration estimation, but accounting for tree-to-tree variations is even more critical.</p>\u0000 </div>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148324318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microenvironmental Heterogeneity Driven by Trees in Arid Woodlands: Effects of Neltuma flexuosa Canopy in the Monte Desert","authors":"Brian Leonardo A. Rios,&nbsp;Bertilde E. Rossi,&nbsp;Juan A. Alvarez,&nbsp;Pablo E. Villagra","doi":"10.1111/1440-1703.70099","DOIUrl":"https://doi.org/10.1111/1440-1703.70099","url":null,"abstract":"<div>\u0000 \u0000 <p>Woody plants modify environmental conditions, increase heterogeneity, and affect the spatial distribution of other species. These changes can include differences between under-canopy and exposed areas, microheterogeneity under the canopy, and temporal variability. Data on the mechanisms underlying microenvironmental heterogeneity in drylands remain scarce, yet understanding these mechanisms is crucial for planning conservation, management, and restoration programs. In the Monte Desert (Argentina), <i>Neltuma flexuosa</i> alters the spatial distribution of shrubs and herbs, as well as soil properties. In this work, we analyzed the changes in light, soil, and water conditions produced by <i>N. flexuosa</i> at two different scales in the Ñacuñán Reserve (Mendoza, Argentina). We randomly selected 12 trees, installed a set of pluviometers and evaporimeters, and measured soil water content, temperature, and photosynthetically active radiation (PAR) intensity once per month in two mesohabitats (under the canopy and exposed areas) and two microhabitats (under the northern or southern canopy of <i>N. flexuosa</i>). Our results showed that <i>N. flexuosa</i> plays a significant role in generating diverse microhabitats. At the mesohabitat scale, it intercepted rainfall and reduced evaporation, thereby lowering the water deficit. In addition, it moderated extreme temperatures and reduced solar radiation. This effect was stronger in the southern microhabitat than in the northern one, and in the summer season. These modifications were heterogeneous and promoted microheterogeneity beneath their canopy, resulting in distinct microenvironments. This heterogeneity aligns with species distribution patterns found in previous studies. The varying requirements of species for these factors likely determine each species' capacity to occupy different microhabitats.</p>\u0000 </div>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148324281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Addressing Uncertainties in Climate–Ecosystem Feedbacks in the Arctic: Ten Priority Research Questions for Integrated Observation and Modeling","authors":"Akira S. Mori,&nbsp;Keita Nishizawa,&nbsp;Shota Masumoto,&nbsp;Ülo Niinemets,&nbsp;Vincent Maire,&nbsp;Xinyu Xu,&nbsp;Shu-Kuan Wong,&nbsp;Kureha F. Suzuki,&nbsp;Joël Bêty,&nbsp;J. Hans C. Cornelissen,&nbsp;Masaki Uchida","doi":"10.1111/1440-1703.70089","DOIUrl":"https://doi.org/10.1111/1440-1703.70089","url":null,"abstract":"<p>Positive biosphere–climate feedbacks are likely to amplify the Arctic warming, yet major uncertainties persist regarding their magnitude and underlying mechanisms. Vegetation shifts, permafrost thaw, and microbial responses critically influence greenhouse gas emissions, surface albedo, and energy balance, ultimately feeding back to climate change. However, the capacity of emerging and changing Arctic ecosystems to sustain biogeochemical functions remains unclear. Here, we identify ten priority research questions that address vegetation, soil, and hydrological processes most relevant to climate feedback. We argue that simply expanding field observations is insufficient. Instead, monitoring must be strategically designed—aligned with satellite observations and models across scales, structured for causal inference, and integrated within coordinated international ecological networks to ensure comparability and synthesis. Advancing next-generation, policy-relevant observation systems will support more accurate projections of Arctic feedbacks, improve the ecological realism of Earth system models, and guide policy-making. Given the disproportionate role of Arctic ecosystems in the global climate system, developing coordinated and ecologically grounded monitoring strategies is a crucial step toward reducing uncertainties in biosphere–climate interactions.</p>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1440-1703.70089","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148282158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative Ionomics of Lycopodiaceae and Selaginellaceae Across Diverse Soil Types in Japan","authors":"Akinori Ren,&nbsp;Asuka Saisako,&nbsp;Takato Saito,&nbsp;Akihiro Yamamoto,&nbsp;Soshi Osaki,&nbsp;Akari Matsumoto,&nbsp;Daisuke Aoshima,&nbsp;Hirotsuna Yamada,&nbsp;Takayuki Nakatsubo,&nbsp;Toshihiro Watanabe,&nbsp;Hayato Maruyama,&nbsp;Takuro Shinano,&nbsp;Jun Wasaki","doi":"10.1111/1440-1703.70094","DOIUrl":"https://doi.org/10.1111/1440-1703.70094","url":null,"abstract":"<p>Lycophytes, one of the earliest-diverging vascular plant lineages, occur in diverse and often extreme environments, yet their ecological adaptation strategies remain poorly understood. Ionomics, characterized by elemental composition, offers insights into how evolutionary history and environmental conditions shape plant function. Here, we investigated how lineage and soil type influence ionomic profiles in terrestrial lycophytes by analyzing shoot elemental concentrations in three Lycopodiaceae species and four Selaginellaceae species collected across Japan, including serpentine soils and solfatara fields. Element concentrations were quantified by inductively coupled plasma mass spectrometry, with nitrogen and phosphorus measured colorimetrically. Statistical analyses included heatmap clustering and principal component analysis. Across non-solfatara environments, Lycopodiaceae and Selaginellaceae showed distinct ionomic profiles at the family level within the sampled taxa, with variation appearing to be more strongly affected by lineage differences than by living environment. In Selaginellaceae, individuals from serpentine soils showed higher magnesium, iron, and arsenic, reflecting serpentine soil characteristics. Lycopodiaceae showed higher aluminum, copper, potassium, rubidium, and manganese, whereas Selaginellaceae had higher calcium, magnesium, phosphorus, nitrogen, strontium, and zinc. For <i>Palhinhaea cernua</i> (Lycopodiaceae), which occurred in both solfatara and non-solfatara fields, shoots from the acidic solfatara environment contained higher sulfur, nitrogen, and phosphorus while maintaining consistently high aluminum concentrations, supporting its status as an aluminum hyperaccumulator. Overall, lycophytes demonstrate family-specific elemental homeostasis that is conserved across environments, alongside characteristic shifts in extreme soils. These findings provide a foundation for understanding ecological diversification in the earliest-diverging vascular plants and highlight ionomics as an effective approach for exploring their environmental adaptation strategies.</p>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2026-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1440-1703.70094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148282099","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Slow Succession: A Comparison of the Saxicolous Lichens of a 50-Year-Old Abandoned Chromite Mine and a Nearby Undisturbed Ultramafic Outcrop","authors":"Michael Mulroy,&nbsp;Isabella Suhr,&nbsp;Jason Dart,&nbsp;Elijah Balderas,&nbsp;Zach Raposo,&nbsp;Nishanta Rajakaruna","doi":"10.1111/1440-1703.70095","DOIUrl":"https://doi.org/10.1111/1440-1703.70095","url":null,"abstract":"<div>\u0000 \u0000 <p>Pioneering and succession are well-studied phenomena in the field of ecology but are poorly understood for lithic habitats dominated by cryptogams. Yet pioneering and establishment of epilithic communities constitutes a fundamental ecological process that provides a precursor to the formation of soil terrestrial communities. Abandoned mines offer an exciting opportunity to explore pioneering and succession of lithic habitats. In our study, we quantitatively compared the lichen communities of an approximately 50-year-old abandoned chromite mine to a nearby, undisturbed rock outcrop of similar ultramafic parent material. Our results reveal several differences between these two communities. The early successional community of the mined site had significantly lower lichen cover and species richness, as well as a species composition distinct from the late successional community of the undisturbed rock outcrop. The early successional community was characterized by relatively high cover and frequency of endolithic lichen taxa, which were mostly absent from the late successional community. Our results highlight the slow progress of pioneering and succession in newly exposed areas of rock and emphasize the importance of preserving undisturbed, late successional lithic habitats.</p>\u0000 </div>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.4,"publicationDate":"2026-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148238096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Trophic Dynamics and Organic Carbon Sources in Washington DC, USA Area Seepage Spring Communities","authors":"William Farmer,&nbsp;Daniel Fong,&nbsp;Stephen E. MacAvoy","doi":"10.1111/1440-1703.70093","DOIUrl":"https://doi.org/10.1111/1440-1703.70093","url":null,"abstract":"<div>\u0000 \u0000 <p>Seepage springs are common ephemeral freshwater habitats in the Washington DC area that are home to a variety of invertebrates including the endemic and rare <i>Stygobromus hayi</i>, the official amphipod of Washington DC, USA. However, little is known about the community structure of these habitats. In this study, we analyzed the general food web dynamics of two seepage springs (Pimmit Run and Goldmine Tract) in the Washington DC area that are known to include a congener of <i>S. hayi</i>, <i>Stygobromus tenuis potomacus</i>. Within these two seepage springs, we identified 29 taxa, with varying degrees of abundances, from 18 field excursions over three seasons: Winter, Spring, and Summer. Using dual abundance stable isotope analysis of δ¹³C and δ¹⁵N, we estimated the trophic positions of Lumbriculidae, <i>Conasellus kenki, S. tenuis potomacus</i>, <i>Crangonyx shoemakeri</i>, <i>Tipula</i> sp., <i>Pseudolimnophila</i> sp.<i>,</i> and a species of Platyhelminthes<i>. S. tenuis potomacus</i> was the dominant predator in both locations, with δ¹⁵N and δ¹³C values between 7‰ and 10‰ and −25‰ and −27‰ respectively. <i>C. kenki</i> and <i>C. shoemakeri</i> were consistently lower in the food web, with δ¹⁵N and δ¹³C values between 4‰ and 7‰ and −24‰ and −27‰ respectively and, in some cases, <i>C. kenki</i> may be prey for <i>S. tenuis potomacus</i>. In both seeps leaf material was 4‰ to 5‰ lower in δ¹³C than the shredding/grazing invertebrates suggesting that they derive nutrients from microbes colonizing leaves not from leaf carbon itself.</p>\u0000 </div>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.4,"publicationDate":"2026-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148237752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mineralogical and Chemical Characterization of Clay Minerals in Albania for Sustainable Soil Management","authors":"Bushi Teuta,&nbsp;Bani Aida,&nbsp;Ottner Franz,&nbsp;Sallaku Fatbardh","doi":"10.1111/1440-1703.70091","DOIUrl":"https://doi.org/10.1111/1440-1703.70091","url":null,"abstract":"<div>\u0000 \u0000 <p>Heavy metal contamination in soils poses serious ecological and food-chain risks, particularly in ultramafic landscapes where total and bioavailable nickel (Ni) concentrations are naturally high. This study investigated the mineralogical and chemical properties of clays derived from different parent materials to evaluate their capacity to immobilize Ni and reduce heavy metal toxicity to agricultural plants. Soil samples from two ultramafic regions (Rrajcë and Prrenjas) and one non-ultramafic site (Durres) were analyzed for texture, pH, organic matter, cation exchange capacity (CEC), and mineralogy (XRD). Smectite-dominated Prrenjas clay exhibited abundant interlayer and edge sorption sites, enabling rapid chemisorption and immobilization, with ~89% of Ni within 240 min when added to Rrajcë soil. In comparison, Durrës clay—containing only 30% smectite and dominated by illite, chlorite, kaolinite, and minor serpentine—immobilized only ~64%. These kinetic and mineralogical contrasts demonstrate that expansive smectite clay types (e.g., smectite), rather than total clay content, are the principal drivers of Ni adsorption capacity and reaction rates. A pot experiment further showed that amending Ni-rich Rrajcë soil with smectite-rich clay (1.07%–3.2%) significantly reduced Ni accumulation in alfalfa (<i>Medicago sativa</i>) by 20%–45%, accompanied by a 2.6%–16% decline in the plant-available fraction of soil Ni. These results highlight strong links between clay mineralogy, Ni bioavailability, and plant uptake. Overall, the findings provide new insights into Ni mobility in ultramafic soils and demonstrate that smectite-rich clay amendments represent an effective, low-cost, and environmentally friendly (low-impact) strategy for reducing Ni bioavailability and mitigating risks to agricultural systems and food crops.</p>\u0000 </div>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.4,"publicationDate":"2026-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148237906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MIG-seq2: Multiplexed Inter-Simple Sequence Repeat Genotyping by Sequencing With Dual-Unique Indices","authors":"Yoshihisa Suyama,&nbsp;Naoko Ishikawa","doi":"10.1111/1440-1703.70092","DOIUrl":"https://doi.org/10.1111/1440-1703.70092","url":null,"abstract":"<p>Genome-wide DNA analysis using high-throughput sequencing is a powerful tool for a wide range of genetic studies. Multiplexed inter-simple sequence repeat (ISSR) genotyping by sequencing (MIG-seq) is an effective method that starts with a first polymerase chain reaction (PCR) to amplify genome-wide regions, followed by a second PCR to add sequencing adaptors and indices. Because the original protocol was designed for a relatively low-throughput sequencing platform at the time, the primary limitation was the restricted number of distinguishable samples that could be achieved using unique indices. To overcome this limitation, this study aims to adapt the method to recent advances in ultrahigh-throughput sequencing technology; the second PCR primers were modified to differentiate more than 1344 samples with higher accuracy using dual-unique indices. Additionally, the tail sequences of the first PCR primers were improved to reduce the number of low-quality sequences. By combining these two modifications, we developed a new method, MIG-seq with dual unique indices (MIG-seq2). Moreover, an optional first PCR primer set, the degenerate oligonucleotide primer MIG-seq2 (dpMIG-seq2), was designed to amplify additional regions, mainly in samples with low genetic diversity. To demonstrate the effectiveness of this improved method, a simple phylogenetic analysis was conducted on two closely related species, <i>Plantago hakusanensis</i> and <i>Plantago asiatica</i> var. <i>densiuscula</i>. The resulting phylogenetic trees clearly showed separate clustering of the two species and their intraspecific populations with high reproducibility across analyses using MIG-seq, MIG-seq2, and dpMIG-seq2. This updated version of MIG-seq is expected to support various genetic studies that utilize high-throughput genotyping.</p>","PeriodicalId":11434,"journal":{"name":"Ecological Research","volume":"41 4","pages":""},"PeriodicalIF":1.4,"publicationDate":"2026-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1440-1703.70092","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148237676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"环境科学与生态学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}