Additionally, a previous demographic reduction was suggested. Thus, considering the increased human-promoted impacts across the entire area of distribution of the giant anteater, our results can illustrate the potential effects of these disturbances on the genetic variation, allowing us to request the long-term conservation of this emblematic species.In general, large mammal species with highly specialized feeding behavior and solitary habits are expected to suffer genetic consequences from habitat loss and fragmentation. To test this hypothesis, we analyzed the genetic diversity distribution of the threatened giant anteater inhabiting a human-modified landscape. We used 10 microsatellite loci to assess the genetic diversity and population structure of 107 giant anteaters sampled in the Brazilian Central-Western region. No genetic population structuring was observed in this region suggesting no gene flow restriction within the studied area. On the other hand, the moderate level of genetic diversity (Ho = 0.54), recent bottleneck detected and inbreeding (F During the last decades, anthropogenic impacts have promoted habitat loss and fragmentation by extensive agriculture, urbanization, and highways and thus threaten biodiversity worldwide  includinMyrmecophaga tridactyla, is a charismatic and large Xenarthra that has been suffering from human activities in several regions of its distribution area. Currently categorized as “Vulnerable” and with decreasing populations in the International Union for Conservation of Nature (IUCN) Red List , under the number 53798-4. The research was approved by the Ethics Committee on the Animal Experimentation (CEUA/UFSCar) protocol number 1584280817, and the genetic resource access was registered under SisGen A9F8717.The study was carried out in the Central-Western region of Brazil, comprising the biome Cerrado  and transition areas with two other biomes, the Pantanal wetlands and inland Atlantic forest . In this1. The sample collection was conducted from April 2013 to February 2017. This sampling represents the largest range for a giant anteater population genetically evaluated so far. A detailed information related to each specimen sample is available in We collected a total of 107 tissue samples, comprising 66 samples from roadkill animals in four main roads crossing our study area and 41 samples obtained from captured wild animals . All tisTM .Total genomic DNA was extracted using the conventional phenol-chloroform protocol . The DNAM. tridactyla , 0.8 mg/ml bovine serum albumin (BSA), 2 pmol forward, and 8 pmol of reverse primers, 8 pmol M13 primers, and ∼30 ng of the target DNA. PCRs were conducted in two steps. PCRs were run with an initial denaturing step of 1 min at 94°C, followed by 20 cycles of 1 min at 94°C, 45 s at locus-specific annealing temperature  was tested with 10 replicates and burn-in at 1,000. We tested for K ranging from 1 to 5 because, for K determination based in the highest value of ΔK, following K = 4, in our case) plus 1. For ΔK estimation, we used the algorithm implemented in STRUCTURE HARVESTER , and the microsatellite loci were tested for linkage disequilibrium (LD) and Hardy–Weinberg equilibrium (HWE), using the exact test of p-values using the Markov chain methods with 10,000 dememorization steps, 1,000 batches, and 10,000 iterations per batch. Sequential Bonferroni corrections were applied to correct for all multiple simultaneous comparisons   and deficit (pS) were calculated for each locus using FSTAT v. 2.9.3.2  , effective number of alleles (Ae), observed (Ho), and expected heterozygosity (He) using GenAlex v. 6.5.0 . Allelicis)  with the 2.9.3.2 . To veritors (r)  among alsoftware . We calcsoftware .To assess whether the current effective population size of giant anteater is sufficient to maintain the observed genetic variation over the next 100 years, we simulated future genetic diversity using the program BOTTLESIM v. 2.6  that meaWe measured the contemporary effective population size (Ne) using the linkage disequilibrium (LD) method  and the M-ratio (M-ratio), where a bottleneck depletes the number of alleles faster than reducing allele size range of the microsatellite , using ARLEQUIN v 3.5  implemented in DIYABC . We desiTo obtain the best fit scenario, the posterior probability (PP) for each scenario was estimated by logistic regression on 1% of the simulated dataset closest to the empirical data. For the scenario with high PP, we evaluated the confidence in the scenario choice estimating the posterior predictive global error using 1,000 pseudo-observed dataset for the logistic regression approach. To assess the precision for each estimated parameter, we calculated the relative median of the absolute error (RMAE) . The besM. tridactyla.The PIC values for each locus were higher than 0.5, with a mean value of 0.53 , indicatK = 2 was obtained according to the K = 1). In turn, the best K based on the Ln value revealed K = 1 , even when females and males were separately analyzed  occurred in five loci, with locus 4, A9, and H5 showing heterozygote deficit. Locus H5 also showed high amount (37%) of null alleles . Low relatedness level was found among the individuals  to eight (11 and 20) with a mean of 5.5, and the mean number of effective alleles (Ae) was 3.10 . Mean obThe prediction of future genetic diversity based on BOTTLESIM simulations projected a genetic diversity decrease in the next 100 years in all tested scenarios. Overall, the genetic diversity reduction was directly affected by the bottleneck intensity tested. The observed allele number and effective allele number declined at a faster rate than expected and observed heterozygosity . The prep < 0.05). Signs of population reduction were significant for the TPM model (p = 0.0048) in the bottleneck analysis, and the M-ratio also showed a signal of population reduction (M = 0.39).The effective population size (Ne) estimate was 375.5 , indicating that all parameters estimated were reliable, suggesting a high confidence for scenario 2  were observed in this large and single giant anteater population inhabiting the studied area. Similar values were previously reported for other local populations studied  within the studied population, and it seems not biased either by a kinship effect. Since the r values found were very low, it is an indicative of a low level of relatedness among the individuals.Lower genetic diversity has already been described in a small anteater population inhabiting a protected area , and it was associated with intense population reduction after recurrent fire events, resulting in inbreeding within the remaining individuals . An incrBesides the potential inbreeding detected, we also found a smaller mean number of effective alleles (Ae = 3.10) compared to the mean allele richness (Na = 5.5), suggesting that fewer alleles are contributing to maintain the current genetic diversity. These results can be a consequence of a Ne that is not large enough to retain all alleles in high frequency, since large Ne is necessary to retain more genetic diversity .The effective population size is an important factor that contributes to genetic variability maintenance because both heterozygosity and number of alleles are less impacted in populations with large effective size . It is kAn effective population size Ne ≥ 1,000 was indicated for retaining the evolutionary potential for fitness in perpetuity . Our resOverall, our results showed a single and large population of giant anteaters inhabiting the southern edge of its geographical distribution, therefore already presenting negative genetic signals, as bottleneck and inbreeding, potentially caused by impacts of the increased human activities in the region. Of note, this work represents the study with the largest microsatellite set used in a Myrmecophagidae species, with a high polymorphic information content (PIC > 0.5), and the largest population genetic study thus far carried out in giant anteater, considering both the sampling area and number of individuals analyzed, reinforcing the importance of these results.M. tridactyla in the next 100 years. Despite the wide distribution of the species and the constant reports of threats for this animal, populations of giant anteater have been poorly studied in Brazil. It is important to highlight that conservation strategies should be urgently adopted to guarantee the species persistence. These strategies should be focused on reducing giant anteater mortality, by reducing the impacts such as road kills, hunting, and habitat loss  protocol number 1584280817.CB-R, PG, and AD designed research and data acquisition. CB-R, RS-S, BS, and PG contributed to new reagents or analytical tools. CB-R, RS-S, BS, AD, and PG analyzed the data, wrote, and review the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Recognition of the importance of plant genetic resources started in Germany at the end of the 19th century. Plant research and breeding began to develop in the 1920s. Formal structures of public institutions were founded, long-term conservation facilities were established, private breeding initiatives developed. In 1990, the German reunification required an assessment of the existing research and breeding landscape. This milestone allowed a comprehensive overview of the great number of stakeholders, active in the entire range of tasks related to plant genetic resources. The Federal Ministry of Agriculture then developed a conceptual approach for an efficient governance structure and published its concept of a national programme for the conservation and sustainable use of genetic resources for food and agriculture in 2000. It recognized the sharing of decentral responsibilities among the respective public and private actors and governmental levels with dis-tributed mandates and funding. It also led to the establishment of a central information and coordination center for genetic resources, which facilitates the data sharing, communication, and co-operation among stakeholders, supports public awareness and advises the Federal Ministry on national policies and efficient European and global cooperation. It also supports efficient contributions of German stakeholders into European structures and international bodies. An equivalent conceptual approach and governance structure is recommended to be established at European level. In Germany, traditional, locally adapted landraces were used in agricultural production and by the mid of the 19th century a few individuals began to select and improve such landraces for better yields. Christian Adolf Leberecht Steiger in Leutewitz (fodder beets 1825), Wilhelm von Borries in Eckendorf (fodder beets 1846), Wilfried Paulsen (potato 1846), Mathias Rabbethge, and others (sugar beets 1862) and Wilhelm Rimpau (rye 1867) were a few of them [The importance of landraces as “plant genetic resources” has been recognized in Germany as early as by the end of the 19th century, when Emanuel Ritter von Proskowetz and Franz Schindler acknowledged the “use value” of traditional landraces for agricultural production in general and plant breeding in particular at the International Agricultural and Forestry Congress in Vienna in 1890 .The Russian scientist Nikolai Ivanovich Vavilov started plant explorations in 1916 and collected landraces worldwide . He descA new Kaiser-Wilhelm-Institute of Research on Plant Breeding was founded in Müncheberg in 1927, with Erwin Baur 1875–1933) becoming its first director. Although this institute collected germplasm samples in Turkey and some Latin American countries its main concern had not been the long-term aspect of conservation but rather topics such as crop plant evolution and genetics. The institute and Erwin Baur developed many fundamental impulses for further research related to plant genetic resources. Hence, in 1943, another Kaiser-Wilhelm-Institute of Crop Plant Research was founded at the Tuttenhof farm near Vienna with its director Hans Stubbe (1902–1989), a student of Erwin Baur. Stubbe laid the foundation of the institute’s future crop plant-related research in a wider interpretation, including the collecting and conservation of global crop diversity and their research in the important botanic disciplines such as systematics, ecology, genetics, biochemistry, biophysics, and physiology [–1933 becIt should be noted that with the increased industrialization of German agriculture at the beginning of the 20th century, the appreciation of traditional German landrace diversity was turned upside down. While important plant explorations were organized into other countries and continents , the NAZI regime conducted a “plant variety cleaning”. This cleaning process was meant to protect the German farmers from inferior seed and resulted in a loss of some three quarters of the varieties of the main crops from the seed market .After World War II, Germany was divided into a Western part, the Federal Republic of Germany, and an Eastern part, the German Democratic Republic. Due to the geographical division and the different political systems of the two German parts, the plant genetic resources activities developed separately throughout the various locations. In Eastern Germany important crop collections were located in Gatersleben and Halle and a fruit collection existed in Dresden-Pillnitz. The collection of the Institute for Plant Breeding in Halle was transferred to Gatersleben between 1945 and 1950 . At the In Eastern Germany, a plant breeding and seed service infrastructure was established by the Sowjet military administration already in 1946. While throughout the country 63 seed selection and development farms (seed stations) with a total area of 27,729 ha were set up, a central coordination mandate of these farms was taken over by the German Seed Association (DSG). The DSG was responsible for the coordination, documentation, collection, and distribution of seed and planting material. Seed production plans became obligatory. Already existing small seed initiatives and institutes could still remain independent but had to follow the instructions of the DSG [In addition, crop-oriented breeding research institutes were established or continued such as the Institute Quedlinburg (vegetables), Institute Bernburg , Institute Groß Lüsewitz (potatoes), Institute of Crop Plant Research, Gatersleben, Institute of Phytopathology, Aschersleben, Agricultural Faculty, Martin-Luther-University Halle-Wittenberg and the Institute of Breeding Research Müncheberg. Important breeders also continued their breeding activities in Hadmersleben, Gülzow, and Klein Wanzleben.After the foundation of the German Democratic Republic in 1949, the new German Academy for Agricultural Sciences (DAL) was founded in 1951 with Hans Stubbe becoming the first president. The former DSG breeding research institutes were now integrated in the Academy as DAL-institutes .Institute Quedlinburg Institute Bernburg  and Research Station Hadmersleben Institute Klein Wanzleben (beets)Institute Groß Lüsewitz (potatoes)Institute Gülzow Institute of Fruit Research Dresden-PillnitzInstitute of Forages PaulinenaueEssential research and breeding partners of these DAL-Institutes  were the Institute of Crop Plant Research (Genebank) Gatersleben, the Institute of Phytopathology Aschersleben, the Agricultural Faculty, Martin-Luther-University Halle-Wittenberg, and numerous existing seed stations. Additional nationally owned seed and planting material companies (VVB) were responsible for country-wide seed supply. The “VVB Seed and Planting Material” comprised 110 farms with over 100,000 ha farmland area .The Institute of Crop Plant Research in Gatersleben played a unique role as it was the home of the important genebank collection of plant genetic resources. The institute was placed within the German Academy for Sciences (DAW) in 1948 and, later in 1972, within the scope of the Academy of Sciences (AdW). From 1970 to 1991 this Institute was called Central Institute of Genetics and Crop Plant Research (ZIGuK) before it was renamed in 1992 in Institute of Plant Genetics and Crop Plant Research (IPK). The institute, with its first director Hans Stubbe, conducted numerous collecting and exploration missions and received plant genetic resources from other collection holders. This growing genebank collection of global importance see  was and After 1945, numerous breeding or seed companies left the Eastern part of Germany and tried to settle in the Western part of Germany. As an indication, a list of companies provided by Röbbelen  gives anLaender) with different mandates at the Federal and Laender levels. As there was no central coordination yet, the breeder associations were also divided in three parts, the Association of Plant Breeders (VdP) in Hanover, the Bavarian Breeders’ Association (VBP) in Munich, and the South-Western Breeders’ Association (VSWP) in Stuttgart. In 1962 (VdP and VBP) and 1966 (VSWP) these three associations were merged and formed the new German Plant Breeders’ Association (BDP) in Bonn (1970). Throughout the years, many private breeding companies extended their businesses based on a particular crop focus, diversification across different crops, and/or by international cooperation [While in Eastern Germany the research, breeding, and seed systems were almost entirely based upon public institutions and infrastructure, in Western Germany the systems were continued and built upon private initiatives and companies. In the West, public institutions were active especially in the areas of research, education, and seed quality control. The Western part of Germany was controlled after the Second World War by the USA, France, and Great Britain, leading in 1949 to a constitution as a federal republic of states  was founded in Hanover to maintain and support private breeding initiatives, to assist in knowledge transfer from latest scientific developments, to facilitate the adoption for technical implementation, to support and enable high economic value-addition and to support international cooperation [The growing necessity to also conserve plant genetic resources for future research and breeding in Western Germany was addressed and promoted especially by Hermann Kuckuck, Dieter Bommer, and the GFP. Based on their initiative, in 1970, a new genebank in Western Germany was established by the Federal Ministry of Agriculture at the Institute of Agronomy and Plant Breeding of the Federal Research Centre for Agriculture (FAL) in Braunschweig . The genDuring the 1970s, the Federal Republic of Germany commenced to support international activities to promote the conservation and use of plant genetic resources. In 1971, the Consultative Group on International Agricultural Research (CGIAR) was established. A number of the international agricultural research centers of the CGIAR focused on the improvement of crop plants, collected plant genetic resources, and established genebanks to support their improvement programmes. The Food and Agriculture Organization of the United Nations (FAO) played a key role to support international cooperation and communication related to plant genetic resources activities. In 1974, largely initiated by the FAO, the International Board for Plant Genetic Resources (IBPGR) was created to support the research, collecting, conservation, documentation, evaluation, and use of the genetic diversity of cultivated plants worldwide. It was also active to organize a global network of genebanks holding base collections within and outside the CGIAR .The growing awareness of the importance of plant genetic resources led to an international agreement at the FAO in 1983, the so-called International Undertaking on Plant Genetic Resources  and the “there develops an internationally coordinated network of national, regional, and international centers, including an international network of base collections in gene banks, under the auspices or the jurisdiction of FAO, that have assumed the responsibility to hold, for the benefit of the international community and on the principle of unrestricted exchange, base or active collections of the plant genetic resources of particular plant species”.Inspired by these formal international processes and developments in plant research, the Federal Ministry of Agriculture initiated an assessment of related activities in 1985 .Cultivated varieties (cultivars) in current use and newly developed varieties;Obsolete cultivars;Primitive cultivars (landraces);Wild and weedy species, near relatives of cultivated varieties;Special genetic stocks (including elite and current breeders’ lines and mutants).While the initiation of the assessment of PGR in Western Germany was started in 1985, the results were only published in 1990 . It was Important for breeding of actual and potential crops as well as important from an ecology perspective and for the conservation of the vegetation in Germany;Required for plant breeding and land improvement, considering the scope of agriculture, horticulture, pomiculture, forestry and landscape management;Important in relation to nature conservation, ecosystems and protection of wild species;Laender institutions and non-governmental organizations.Maintained by Federal and Hence, in a broad interpretation, the assessment addressed resources in Western Germany, which were :ImportanThe most important institution in Western Germany at the time of reunification was the Institute of Agronomy and Plant Breeding of the Federal Research Centre for Agriculture (FAL) in Braunschweig, holding a large genebank collection. Since 1980, endangered wild species in Germany were also integrated in this collection. Reference samples of the Federal Plant Variety Office (BSA) and related information were handed over to the FAL-genebank for varieties after the expiry of their variety protection period. Evaluation and documentation of the resources were a particular priority of the institute. Annually, 7000–8000 samples were provided to recipients at their requests; about 1/3 of the requests were received from abroad .Laender institutes, universities, Max-Planck institutes, and private breeding companies.Apart from the registration of test collections maintained at the Federal Plant Variety Office for the testing period, important collections of agricultural crops were held at Laender and county institutes as well as numerous private companies. This diverse situation was similar for genetic resources of ornamental plants, where botanic gardens, universities, and outdoor museums played a significant role as well. The Federal Institute of Grapevine Breeding in Siebeldingen and six Laender institutes maintained the main genetic resources collections for grapevine research and breeding.For horticultural crops, many fruit and vegetable collections were identified at the FAL, the BSA, the Federal Research Institute for Horticultural Plant Breeding in Ahrensburg and more than 20 Laender forestry research institutes, forest administration, arboreta and private forest owner were the key actors involved. These activities were coordinated by the Federal—Laender Working Group “Conservation of Forest Genetic Resources” [The plant genetic resources concept with its assessment  also covsources” .Laender research institutes, universities, botanic gardens, outdoor museums, nature conservation associations and individuals are engaged in these activities. The main conservation responsibilities, however, were located at the Laender level.Activities for the conservation of genetic diversity of wild species were part of the nature and landscape protection measures. Federal and The main collection of microbial genetic resources existed at the German Collection of Microorganisms  in Braunschweig. The DSM was also the International Depositary Authority under the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure in Germany. The collection consisted of some 4200 strains of microorganisms .At the time of the German reunification in 1990, the study by Bommer and Beese  presenteThese studies provided a comprehensive picture and revealed a large number of public institutions being involved in a wide range of activities related to plant genetic resources for research and breeding. The two large genebanks at the IPK in Gatersleben and at the FAL in Braunschweig played a leading role. They also maintained long-term base collections within the international network of base collections supported by the IBPGR . In addiFurthermore, about 100 private breeding and seed companies were operational for the German and, partially, for the international seed markets. A number of non-governmental organizations were engaged in plant genetic resources activities such as on-farm conservation and management. Pioneering examples were the Association for the Conservation of Crop Plant Diversity (VEN) founded in 1986, the Association for the Promotion of Seed Research in Biodynamic Agriculture in 1988, the Association of Pomologists in 1991 or later the Association for the Conservation and Recultivation of Cultivated Plants (VERN) in 1996.Laender levels. The main responsibility for nature protection, including the conservation of genetic resources, as well as for academic institutions such as universities rests with the Laender governments. The Federal government oversees collaborative activities across the Laender, providing a national policy framework, national documentation and monitoring, and providing for the international cooperation. It was evident that there were different levels of governmental mandates at the Federal and The plant genetic resources system was marked by a rich but scattered research landscape, by a few multi-crop and many crop-oriented genebanks and research institutes. A certain degree of duplication of the respective public research centers, in particular between the two large multi-crop genebanks in Gatersleben and Braunschweig, was highlighted. It was recommended to integrate the collection of the genebank in Braunschweig into the genebank in Gatersleben, which was implemented over a certain period of time to avoid any loss of material or knowledge related to the collection and was concluded in 2004.As part of the initial assessment in Western Germany, before the reunification, a plant genetic resources concept was elaborated on how to integrate the multitude of stakeholders, measures, and programmes and better prepare the entire plant genetic resources system for future challenges and opportunities .Background to this so-called “Bommer and Beese” concept  was the Laender levels should remain as it was operating by that time [The proposed plant genetic resources concept covered a wide range of agricultural crops including fruit crops, vegetables, ornamentals, grapevine, forest, and wild species including crop wild relatives. Moreover, microorganisms were considered. It emphasized the promotion of research and conservation efforts and recognized different mandates related to the federal structure of Germany. Core elements of the concept were the following: the Federal Ministry of Agriculture itself, an inter-disciplinary Advisory Board for Plant Genetic Resources, an Information and Coordination Centre for Genetic Resources with a central documentation system, and crop committees for different crops (crop groups). Besides these new bodies, the existing system with the well-functioning conservation, research, and breeding institutions at Federal and hat time .This decentral or distributed system with a central coordination unit was considered advantageous over a combined and centralized system in one large plant genetic resources institution or a completely decentral system of individual institutions. Advantages were seen in securing the necessary continuity of this long-term task, in supporting the inter-disciplinary collaboration, in maintaining the plant genetic resources activities as part of a broader institutional research setting, and in using available infrastructure for new conservation measures .Laender ministries;To provide advice to the Federal and ○To represent Germany in international bodies such as the FAO Commission on Plant Genetic Resources, the European Cooperative Programme for Plant Genetic Resources (ECPGR) and bodies at the European Commission;○To prepare the 4th International Technical Conference of FAO for Plant Genetic Resources (1996) in Leipzig;To support the Federal Ministry of AgricultureTo provide the Secretariat to the National Committee that was asked to prepare the German national report for this 4th International Technical Conference of FAO;To collect, analyze and disseminate information about national and international conservation measures;To undertake public awareness activities;To support collaboration between the formal and informal sectors;To support national conservation measures in line with international activities;To contribute to improved links between conservation and use activities.A first step to implement the new components of the plant genetic resources concept was the establishment of the proposed Information and Coordination Centre for Genetic Resources (IGR) in April 1991. The IGR was located at the German Centre for Agricultural Documentation and Information (ZADI), an institution under the Federal Ministry of Agriculture. The IGR started and evolved in a stepwise manner. Initially, the main task was to develop a national database for plant genetic resources and support the exchange of data with other national and international databases. As from 1993, with its new director Frank Begemann, the following tasks were added: The National Committee that was created for the 4th International Conference of the FAO, consisted of representatives of all stakeholders involved such as different ministries, science and research, the private sector, non-governmental organizations, international cooperation agencies, associations, farmers organizations, and nature conservation agencies . This inIt is worth mentioning that the IGR could not support and coordinate activities through a dedicated budget line under its control. It rather facilitated conservation, documentation and sustainable use of plant genetic resources merely through appropriate information and communication means. This turned out to be effective and supported the collaboration between stakeholders within the large and diverse plant genetic resources system in Germany. Step by step, the usefulness of these information and coordination functions of the IGR in the plant domain was recognized by the Federal Ministry officials in the domains of animal, forest, and aquatic genetic resources.Based on this experience, and considering international processes such as the broadening of the scope of the FAO Commission on Plant Genetic Resources to the new Commission on Genetic Resources for Food and Agriculture (CGRFA) in 1996, the German Federal Ministry of Food, Agriculture and Forestry published a landmark “Concept for the Conservation and Sustainable Use of Genetic Resources for Food, Agriculture and Forestry” in 2000 . This coFederal Ministry of Food and Agriculture (BMEL) with a coordination division related to genetic resources for food and agriculture and additional domain-specific divisions for plant, animal, forest and aquatic genetic resources;Agrobiodiversity strategy and national programmes for plant, animal, forest and aquatic genetic resources;Laender authorities and a wide range of experts and stakeholders.National expert committees for plant, animal, forest and aquatic genetic resources, consisting of Scientific Advisory Board for Biodiversity and Genetic Resources;Information and Coordination Centre for Biological Diversity (IBV) (successor of the former IGR) at the Federal Office for Agriculture and Food (BLE);National inventories for plant, animal, forest and aquatic genetic resources;https://genres.de/en/)National information platform/website  published a so-called “Agrobiodiversity Strategy”, in furtherance of the genetic resources concept from 2000 and as a supplement to the National Biodiversity Strategy . The titIn addition to the overall agrobiodiversity strategy, domain-specific national programmes for genetic resources for plant, animal, forest, and aquatic genetic resources were developed. They are functional instruments to describe the detailed measures to be implemented throughout Germany in each of the domains within a certain time period, to set priorities, to facilitate the monitoring of activities and achievements, and to assist in linking the required stakeholder groups. A potential programme for microbial genetic resources is being considered. The programmes are updated from time to time to remain useful instruments for priority setting and implementation.The implementation of the programmes is based upon the active involvement of a wide range of stakeholders and constituencies involved in all areas such as the identification, collecting, conservation, documentation, characterization, evaluation, and other uses of genetic resources for research and breeding as well as for direct uses for food and agriculture, horticulture, pomiculture, viticulture, forestry, and fisheries. The stakeholders participate in their respective fields of competence and measures based upon their existing respective responsibilities and budgets.Laender authorities and a wide range of experts and stakeholders are in charge of guiding the implementation of the national programmes.National expert committees for plant, animal, forest, and aquatic genetic resources, consisting of Laender ministries;To provide advice to the Federal and To support the BMEL representing Germany in international bodies such as the FAO Commission on Genetic Resources (CGRFA), the European Cooperative Programme for Plant Genetic Resources (ECPGR), the European Regional Focal Point of Animal Genetic Resources (ERFP) and bodies at the European Commission;Provides national coordinator for ECPGR;Support of the development and implementation of the national programmes for plant, animal, forest and aquatic genetic resources;Secretariat for the national expert committees of the Federal Ministry for Agriculture (BMEL) for plant, animal, forest and aquatic genetic resources, as well as for the Scientific Advisory Board for Biodiversity and Genetic Resources;Data collection and documentation of national inventories as well as user-oriented central dissemination of information on occurrences, characteristics and performance of genetic resources for food, agriculture, forestry and fisheries;Monitoring and assessment of agrobiodiversity trends in Germany;Coordination of conservation activities and assistance to conservation networks;Facilitation of national and international support measures and programmes;Knowledge transfer and advisory services for political decision makers and other stakeholders;Biopatent monitoring and access and benefit-sharing (ABS) issues;Public relations and awareness raising.According to the amended scope of the genetic resources concept and the new agrobiodiversity strategy and in recognition of the usefulness of an information and coordination entity, the former Information and Coordination Centre for Genetic Resources (IGR) was renamed into Information and Coordination Centre for Biological Diversity (IBV) and was integrated in the Federal Office for Agriculture and Food (BLE) in 2005. The current tasks of the IBV relate to agrobiodiversity, in particular to genetic resources for food, agriculture, forestry, and fisheries; they include inter alia:Biological and ecological basics;Economic, social and ethical evaluation;Development of science and technology, including genetics and breeding;Land use, landscaping and rural areas;Importance for raw materials, energy, nutrition and health;Promotion of strategies and concepts;Legal, policy and ethical issues;Information and communication, marketing and awareness.The Scientific Advisory Board on Biodiversity and Genetic Resources was constituted in 2003. The Board advises the BMEL on general and fundamental issues relating to the conservation and sustainable use of biological diversity at national, EU, and international level. Members of the Board are scientists from different disciplines appointed by the Federal Ministry of Agriculture, the four chairpersons of the national expert committees on plant, animal, forest, and aquatic genetic resources, as well as the director of the IBV. The main topics to be considered by the board are:Laender institutes, genebanks, research institutions, fisheries, the private sector and non-governmental organizations, including universities, agricultural and horticultural actors, breeders, farmers and foresters, nature conservation, botanic and zoological gardens.While the BMEL is setting the policy framework, the implementation of the national programmes remains under the responsibility of all stakeholders. Due to the broad scope of the demanding programmes a wide range of actors are involved such as Federal and https://genres.de/en/) is dedicated to providing the overall information platform of all programmes and stakeholders as well as the national inventories and a newsletter.The IBV is keeping an oversight and facilitates the implementation of the national programmes through information, documentation, communication, and coordination measures. A specific website  and the particular importance of climate change, loss of biodiversity, and food security, the essential role of plant genetic resources for food and agriculture is evident. These resources are fundamental for further research and plant breeding to support the diversification of the agriculture and food system and contribute to the climate change adaptation processes.Based on its historical developments, the German plant genetic resources system see  is charaThe National Programme for the Conservation and Sustainable Use of Plant Genetic Resources of Agricultural and Horticultural Crops  describeLaender authorities, professional associations and organizations from science and research, the private sector, representatives of genebanks, the in situ conservation and on farm management sector, and non-governmental organizations. The terms of office of members are five years. The BEKO has provided reports about the implementation of the national programme for the periods 2008–2014 and 2015 to 2019. Since the beginning of the current term (2020–2024), also the nature protection sector is represented through the Federal Agency for Nature Protection.The national expert committee for PGR, called the “Advisory and Coordinating Committee for Agricultural and Horticultural Crops (BEKO)” was established by the BMEL in 2002. It consists of up to 17 members representing Federal and Laender, and local level, and even by private individuals. Four of these genebanks are in fact decentralized networks that are specialized in the conservation of certain crops, namely the German Genebank for Fruit Crops, the German Genebank for Grapevine, the German Genebank for Ornamentals, and the Genebank for Crop Wild Relatives.In Germany, there are currently six national genebanks see . These cDespite the differences in the species conserved and the actors involved, all four decentralized genebank networks follow the same structure, which is set out in a cooperation agreement. The coordinating organizations of the two larger networks, i.e., the Genebank for Fruit Crops and the Genebank for Ornamentals, are supported by scientific advisory boards.All genebanks conserve their accessions according to the FAO genebank standards . The IPKThe IBV, as a higher-level coordinating body, is a partner with defined tasks in all decentralized genebank networks. This includes the integration of the respective genebanks in the national and international processes as well as the integration of the data about the respective genebank holdings into the National Inventory of Plant Genetic Resources (PGRDEU) in Germany.All plant genetic resources conserved in the German genebanks are distributed under the terms of the International Treaty on Plant Genetic Resources for Food and Agriculture (ITPGRFA) for the purposes of research, breeding, and training with the Standard Material Transfer Agreement (SMTA), or a special Material Transfer Agreement for ornamentals based on the SMTA. The IPK genebank provided in 2019 a total of 20,069 samples to recipients at their requests, under SMTAs; 11,351 samples were requested from abroad. The current genebank holdings are listed in Lycopersicon and Lupinus, and the former FAL genebank held global collections of Avena, Beta, four Brassica species and Sinapis, as well as a European collection of Phaseolus [Within the network of IBPGR base collections, the IPK held a global base collection of haseolus . GermanyImprovement of priority CWR in situ conservation in their natural habitats, combined with complementary ex situ conservation in genebanks.Provision of a framework for coordination, management and integration of CWR into in situ conservation activities and for raising awareness about the importance of CWR conservation.Promotion of CWR utilization through documentation and the provision of freely available in situ and ex situ characterization and evaluation data in national and international information systems.Supporting the national PGRFA program in international cooperation and the implementation of the CBD, the GPA2, and the International Treaty on PGRFA.Supporting the fulfilment of international reporting obligations regarding the implementation of GPA2, the International Treaty, and the State of the World Report on PGRFA.Attention to in situ conservation of CWR has increased over the past decade. The “German Network of Genetic Reserves” has been established in 2019 as framework for in situ conservation of priority CWR . It consThe first CWR specific network was the wild celery network established in 2019. Currently it is already composed of 17 wild celery genetic reserves; further reserves are in the process of being designated . The coointer alia led, as reported above, to the representation of the Federal Agency for Nature Protection in the BEKO.In situ conservation of wild plant species is a key task of the nature protection sector, while CWR are of particular interest to the agricultural sector given their importance for plant breeding and crop improvement. Hence, their in situ conservation requires collaboration with the nature protection sector, both at the local level, when designating genetic reserves, as well as regional and federal level. This collaboration is currently being established and extended, and has In Germany, there are a large number of NGOs, most of which are organized through an umbrella organization for crop and livestock diversity. This umbrella organization is also a member in the BEKO to advice on issues related to on farm conservation and management.The EU-Regulation on Conservation Varieties adopted in 2009 created the legal prerequisite to permit and market seeds of landraces and varieties of agricultural species and vegetable species, which are relevant for the conservation of genetic resources under facilitated conditions. This was a supportive prerequisite to enable on-farm management of varieties that no longer have seed approval.The Federal Ministry offers project funding to support on farm conservation and management, through which a large number of projects have been carried out in recent years, including some projects that investigated the recultivation of genebank accessions of old landraces from the IPK genebank.Laender governments “Improvement of the Agricultural Structure and Coastal Protection” (GAK) to support agriculture and forestry, the development of rural areas and to improve coastal and flood protection. One of the funding measures specifically serves to promote the cultivation and conservation of old landraces/varieties, that are listed on the Red List of endangered local crops in Germany. The Laender can offer these funding measures and receive co-financing from the Federal ministry. However, due to the priority setting by the individual Laender, the efforts required and the low funding volume so far only one Federal State (Land) offers this funding measure for plant genetic resources.Within the scope of the EU’s European Agricultural Fund for Rural Development (EAFRD) there is also the national cooperative funding instrument of the Federal and The National Inventory of Plant Genetic Resources in Germany (PGRDEU) is the central documentation of the ex situ, in situ, and on-farm conserved plant genetic resources in Germany. This includesThe documentation of the six national genebanks in Germany,The data from the “German Network of Genetic Reserves”,The list of priority CWRs,Extensive data on the historically used vegetable varieties and species from the period 1850–1950,The red list of endangered indigenous crop landraces/varieties in Germany,An inventory of on-farm actors that is currently being developed,Variety descriptions of genebank material from cultivation trials.The national inventory is hosted and managed by the IBV at the BLE. It is regularly updated and, besides being a central resource for national stakeholders, it serves as data source for fulfilling international data reporting obligations to the European Catalogue of plant genetic resources EURISCO managed by the ECPGR and to FAO for SDG Indicator 2.5.1.The European and global collaboration in plant genetic resources for food and agriculture conservation and use is also coordinated by the PGR experts at the IBV. They coordinate the interactions with and the contributions to ECPGR. They advise and represent the Ministry in the collaboration with and sessions of the ITPGRFA and the Intergovernmental Technical Working Group for PGR of the CGRFA and take care of all international reporting obligations to the ITPGRFA, the CGRFA, and GPA. Through IBV’s various functions and roles in the BEKO and the CWR genetic reserve and genebank networks it communicates relevant international information and necessary actions to the national stakeholders and vice versa. The coordination and governance structure is functioning well since more than 20 years now. It is a light structure based on information, communication, and coordination elements but without a centralized funding structure. While this structure brought many advantages, still some challenges remain to be addressed. These issues will be elaborated based upon the plant genetic resources domain as follows.The benefits of the structure—distributed with a central coordination—can be appreciated by the improved national cooperation with a balanced implementation of the national programme across both larger and smaller stakeholders. Additional capacities could be identified and integrated for conservation of plant genetic resources by including very small and also private actors. This is the case, for instance, in the further development of the German genebanks of fruits and ornamentals, where a number of well-qualified and motivated partners are able to contribute to the national endeavor. The comprehensive national programme, the BEKO, as well as the information and communication means allow them to participate in a fair and equitable manner.The comprehensive representation of a wide range of stakeholders in the BEKO facilitates their engagement and contributions toward the implementation of activities in the national programme. This goes hand in hand with the official acknowledgement and recognition by reporting their valuable activities at national and international levels.Monitoring, regular revisions, and priority setting of the national programme is facilitated by the BEKO and the supporting activities of the IBV. This approach allows to continuously integrate innovations from science, observations of the private sector, and findings of non-governmental organizations, as well as new developments from political debates and decisions, and international developments.The capacity to collect information about activities of and contribution from numerous stakeholders, besides the well-known research institutes such as the IPK genebank in Gatersleben, allows enhanced collaboration and facilitates documentation and reporting of the German contributions toward international cooperation. Especially the ECPGR could benefit by coordinated inputs from Germany. At the same time, the active involvement of German members in ECPGR activities facilitates the harmonious implementation of the concepts developed within the ECPGR in Germany.When developing positions for European and global processes, the German Federal Ministry of Food and Agriculture (BMEL) is benefitting from advice of the BEKO and the IBV and national programme assessments. This is especially the case for FAO processes under the CGRFA and the ITPGRFA, as well as for the CBD processes, including the Nagoya Protocol. An additional advantage of the governance structure is an improved coherence in German positions related to plant genetic resources for food and agriculture at the international level.Laender activities or non-governmental organizations related to in situ conservation and on farm activities to the IBV could be improved. IBV also would benefit from more regular information from research institutes about ongoing projects, especially those funded by third parties . Several coordination and conservation activities within the ex situ and in situ conservation networks would benefit from more long-term institutional support to stakeholders carrying out these functions. Besides such advantages, some challenges of this structure remain to be addressed. In particular, the information flow from While policy coherence could be improved with the new governance structure for policy setting at different national agricultural for a related to plant genetic resources, processes to develop joint positions between the agricultural and environmental sectors at national and international level should still be further enhanced.Laender and Federal governments. More than 30 years have passed since. Looking back today from within a stable and well-functioning plant genetic resources system, the approach to (only) centralize information and coordination functions in a permanent dedicated unit, while keeping or developing concrete implementation of conservation and use, research and breeding embedded in functioning local, regional, and federal structures or distributed networks have proven to be very sustainable and effective.A key step in the development of the current coordination and governance structure was the need to thoroughly assess and analyze the national plant genetic resources landscape at the occasion of the German reunification. This assessment has taken into consideration the existing political and administrative structure and the distribution of competencies between the It is conducive to establishing long-term collaboration both at national and international levels, having allowed Germany to engage effectively in continued collaboration with all relevant bodies, i.e., ECPGR, FAO, ITPGRFA, and CGRFA, including respective working groups and subsidiary bodies.This light governance structure provided by a central coordination unit, i.e., the IBV, deserves appropriately staffed offices. The scientists working at the IBV are civil servants, all experts in their respective field of genetic resources, who are entirely dedicated to carrying out the tasks listed in Laender, and relevant competencies and responsibilities distributed between Laender and federal level, similarly Europe presents a highly diverse genetic resources landscape in terms of conservation, management, use, research and breeding, actors and (agro)ecologies, as well as relevant competencies and responsibilities. It is recommended to establish an equivalent concept and governance structure at European level. Like Germany as a Federal state, with a multitude of stakeholders and significant differences among the


Bigheaded carps (bighead carp and silver carp) originated in Southeast and East Asia, and their hybrids were stocked for economic reasons to Hungarian natural waters such as Lake Balaton for decades, while the Tisza River was populated by escaped individuals with farm origins. The presence of these alien species and their hybrids in Hungarian natural water bodies may pose significant ecological risks (connected with their phytoplankton and zooplankton consumption). To be able to deal with the ecological risks and to understand the potential of invasiveness of these species, one must have information on the population-level genetic structures of these alien fish stocks. Ten microsatellite DNA markers and one mitochondrial marker were used to address these questions. The results showed that the two stocks are genetically different; the lake population was genetically more diverse and consisted of hybrid and silver carp individuals, while the river population contained only silver carps. The mitochondrial sequences found in the two populations originated from the Yangtze River. Based on the different genetic structures of the stocks, one can assume that bigheaded carps do not reproduce in Lake Balaton, while the Tisza River stock represents significant reproductive potential and may become invasive in this river.Hypophthalmichthys molitrix, and silver carp, Hypophthalmichthys nobilis) and their hybrids play an important ecological and economic role in their original habitat, while their introduced stocks may pose serious ecological risks. To address questions about the persistence and invasiveness of these fish, we need to better understand their population structures. The genetic structures of bigheaded carp populations inhabiting Lake Balaton and the Tisza River were examined with ten microsatellite markers and a mitochondrial DNA marker (COI). The Lake Balaton stock showed higher genetic diversity compared with the Tisza River stock. Based on hierarchical clustering, the Tisza population was characterized only by only silver carps, while the Balaton stock included hybrid and silver carp individuals. All COI haplotypes originated from the Yangtze River. Based on the high genomic and mitochondrial diversity, along with the significant deviation from H–W equilibrium and the lack of evidence of bottleneck effect, it can be assumed that bigheaded carps do not reproduce in Lake Balaton. The present stock in Balaton may have originated from repeated introductions and escapes from the surrounding fishponds. The Tisza stock consists solely of silver carp individuals. This stock appears to have significant reproductive potential and may become invasive if environmental factors change due to climate change.Bigheaded carps (bighead carp,  Hypophthalmichthys molitrix) and the bighead carp , collectively known as bigheaded carps or filter-feeder Asian carps, have a significant economic role in freshwater fish production. In order to understand the importance of these species, it is sufficient to look at fish harvest reports. The global production of silver carp, for example, was 4,822,794 tons in 2018, putting this species in second place in the world’s aquaculture. The leading producer countries are China, India, Bangladesh, and Iran. Bighead carp ranks fifth among all cultured freshwater fish globally, with a harvested mass of 3,146,466 tons in 2019, accounting for 7.5% of global freshwater aquaculture production [The silver carp  was made up of invasive species, and the biomass of Bigheaded carps was 10.5% of the total biomass [In the European Union, bigheaded carps have not yet been included in the list of invasive species , as thei biomass . This is biomass . No furtThe two main factors responsible for the successful reproduction of bighead carp and silver carp are optimal temperature and water flow. Regarding temperature, the two species require a minimum temperature of 18 °C to reproduce, which is not a limiting factor in most of Europe. Bigheaded carps require intense water currents in the spawning grounds for egg maturation to occur, while silver carp females are less demanding in this regard . In EuroHowever, the situation in Europe is further complicated by the use of the two species (and their hybrids) in aquaculture production as an integral part of the so-called “carp polyculture system” under pond farming conditions, which is a common practice in Central and Eastern Europe . In thisExamination of genetic diversity can provide information on the adaptability, persistence, and invasiveness of a species under the given conditions. High genetic diversity often implies better adaptability, and multiple introductions from different sources also influence the control of the invasive species. The difference in the genetic structure of the native and introduced populations (reviewed by ) suggestThis study aimed to perform a genetic comparison between two Central European stocks of the bigheaded carps in order to explore the genetic structure between and within lacustrine and riverine habitats.n = 20) and bighead carp (n = 21) samples from a gene bank (reference populations at the Northwest Fisheries Resource Development and Management Project (NFRDMP), Parbatipur, Bangladesh) were also used as controls in the study. DNA was extracted using the Qiagen DNeasy® Blood and Tissue Kit  following the extraction protocol outlined by the manufacturer.The sampling locations are shown in ® HCl (pH 9.0) and 50 mM potassium chloride (KCl). The final reaction conditions were as follows: 1× PCR buffer, 1.5–2.5 mM MgCl2, 200 µM of each 2′ deoxynucleotide triphosphate (dNTP), 10 pmol of each of the forward and reverse primer, 1 unit (U) Taq DNA polymerase, and 10–20 ng genomic DNA template. PCR conditions included a denaturation step of 3 min at 94 °C; 5 cycles of 30 s at 94 °C, 30 s at 55 °C, 30 s at 72 °C; 5 cycles of 30 s at 94 °C, 30 s at 52 °C, 30 s at 72 °C; 30 cycles of 30 s at 94 °C, 30 s at 46 °C, 30 s at 72 °C; and a final extension of 30 min at 72 °C. Amplified fragments were analyzed on an ABI3130 Automatic Fragment Analyzer  using POP7 polymer, a 50 cm capillary array, and GS500-LIZ as an internal standard. Fragment length data were analyzed by Genescan and Genotyper 4.0 softwares. .A total of ten cross-species autosomal microsatellite markers  were use4)2SO4 , 0.8 mM dNTP mix, 250 nM for each primer, 2 mM MgCl2, 100 ng template DNA, and 1 U Taq polymerase (Fermentas) in a 25 µL final volume. The PCR reactions were performed in an ABI 2720 Thermal Cycler  with the following cycling conditions: preliminary denaturation at 95 °C for 2 min, then 30 s at 94 °C, 20 s at 52 °C, and 1 min at 72 °C for 35 cycles. Final elongation was 10 min at 72 °C. PCR products were purified by NucleoSpin Gel and PCR Clean-up Kit . The quality of the purified products was assessed on 1.5% agarose gel run in TBE buffer, then sequenced using the Big Dye Terminator v. 3.1 Cycle Sequencing Kit (Applied Biosystem). The sequences were detected on the ABI 3130 sequencer (Applied Biosystems) with a POP7 polymer and a 50 cm long capillary array.For the sequence analyses, the cytochrome oxidase C subunit I gene (COI) was amplified with the universal primers for fish barcoding CO1_FF2d_F (TTCTCCACCAACCACAARGAYATYGG) and CO1_FR1d_R (CACCTCAGGGTGTCCGAARAAYCARAA). The PCR master mix contained 1× PCR buffer with   . The allst of Weir and Cavalli-Sforza and Edwards genetic distance was calculated using FreeNA software  [st and the INA correction for genetic distance. The ENA method corrects for the positive bias induced by the presence of null alleles on Weir’s Fst estimation, and INA gives biased estimates of the Cavalli-Sforza and Edwards (1967) genetic distance. The number of replicates was 10,000 for the computation of the bootstrap 95% confidence intervals.The pairwise F France) , both wiThe effective population size was calculated in NeEstimator 2.1   using th4 and the number of further MCMC runs was set to 105. Calculations were repeated 10 times for each K.The Bayesian algorithm implemented in the software STRUCTURE  ,35 was uDNA sequences were edited and aligned using Mega-X 10.1 software . The revised alignment was 552 bp for the COI gene. The number of polymorphic (segregating) sites, the total number of mutations, haplotype diversity, and nucleotide diversity were estimated by DnaSP 5.10. software  . A haploThere was no evidence for large allelic dropout, and the presence of null alleles was assumed in loci Hmo 13, 33, 36, 37, 39, and 40 in Balaton; Hmo 02, 13, 33, and 39 in Tisza; locus Hmo 36 in silver carp; and loci 33 and 37 in bighead carp stocks because of the general excess of homozygotes. The following loci showed deviation from the HW equilibrium: all loci in the Balaton stock; Hmo33 in the stock of inflow in Balaton; Hmo 01, 02, 13, 33, and 39 in the Tisza stock; Hmo 34, 36, and 40 in the native silver carp population; and Hmo 02, 13, 33, 37, 39, and 40 in the native bighead carp population.Regarding the diversity data in the two studied stocks, the Balaton stock showed significantly higher values for the number of alleles (Na) and the effective number of alleles (Neff). Allelic richness was also higher in the Balaton stock, but the difference was not significant. In the case of private allelic richness (ARp), the Tisza stock showed a significantly higher value. Heterozygosity values were similar in all stocks/populations. The diversity of individuals collected in the inflow area of Lake Balaton was lower compared with both the Balaton stock and the Tisza stock; however, a significant difference could only be detected in the Na compared with Balaton and in ARp compared with the Tisza.The gene bank stocks of both species showed significant discrepancies from the Balaton stock in the case of NA values and from the bighead stock in the case of Ne values. The ARp showed a difference between the Balaton inflow and bighead stocks .The values of the stock sizes (Ne) in the three Hungarian stocks estimated by the linkage disequilibrium method were 49.7 (CI 95%: 45.5–54.6), 129.0 (CI 95%: 9.9-infinite), and 98.4 (CI 95%: 50.1–570.4) individuals in the Balaton, Balaton inflow, and Tisza stocks, respectively. Unexpectedly, the Wilcoxon signed-rank test for heterozygosity excess performed by BOTTLENECK did not support a recent population bottleneck in any of these stocks.st was 0.142 (95% CI: 0.105–0.187) and 0.139 (95% CI: 0.100–0.186) without and with ENA correction, indicating only moderate genetic distances. The pairwise Fst and Cavalli-Sforza and Edwards genetic distance values between stock pairs are shown in The global FHierarchical clustering by STRUCTURE software  resultedIn total, 138 mtDNA sequences were analyzed, resulting in 52 haplotypes .Within the haplotype groups, the number of polymorph sites was 175 , and 43 Based on the STRUCTURE analysis of the microsatellite data, 31 individuals were clustered as hybrids, of which 25 individuals were confirmed by the allele length of the Hmo1 and Hmo3 microsatellites that have different allele size ranges in the bighead carp and silver carp . The mtDBigheaded carps show different genetic and stock/population characteristics in their native range and in their introduced habitats. The loss of genetic diversity results in population decline in the native habitat , but in the Mississippi River Basin, the two introduced species became invasive parallel with the ability of hybridization in natural waters . The genLepomis gibbosus) was introduced at the end of the 19th century in several countries in Europe. The species has spread from Germany to Central Europe through the Danube River [Pseudorasbora parva) was introduced to Europe along with the bigheaded carp [Thus, the genetic diversity of silver carp stocks in the two stocks is the same or slightly lower than that of native populations. Silver carp also show similar or higher diversity compared with other invasive species that also occur in the region. The pumpkinseed sunfish  have been introduced and settled in Hungary via the Danube River and extended further upstream [Pseudorasbora parva), which settled simultaneously with bigheaded carps, has now become an invasive species in Europe [The Danube River is a part of the Southern invasion corridor in Europe and can be considered an “invasion gateway” . Severalupstream . The topn Europe . The sprn Europe . The repn Europe . In the n Europe . Accordin Europe , successn Europe . PredictOur investigation highlights that depending on the habitat quality, the reproduction of the silver carp varies in riverine habitats. The genetic information suggests that the species may become invasive similarly in the North American populations. It is important to develop detailed management plans and monitoring activities to control the spread of the silver carp. It seems that the distinct Balaton stock does not pose a significant hazard because of the lack of regular reproduction. The elimination of the stock could be implemented by the rigorous monitoring of the inflow waters and the continuous selective fishing of the adult individuals. However, the Tisza River represents a different situation, where management coupled with the effects of climate change could not control the gradation of the bigheaded carp species.


Plasmodium vivax, a major contributor to the malaria burden in India, has the broadest geographic distribution and shows higher genetic diversity than P. falciparum. Here, we investigated the genetic diversity of two leading P. vivax vaccine candidate antigens, at three geographically diverse malaria-endemic regions in India. Pvama1 and Pvmsp119 partial coding sequences were generated from one hundred P. vivax isolates in India  and ~1100 published sequences from Asia, South America, North America, and Oceania regions included. These data were used to assess the genetic diversity and potential for vaccine candidacy of both antigens on a global scale. A total of 44 single nucleotide polymorphism (SNPs) were identified among 100 Indian Pvama1 sequences, including 10 synonymous and 34 nonsynonymous mutations. Nucleotide diversity was higher in Rourkela and Nadiad as compared to Chennai. Nucleotide diversity measures showed a strong balancing selection in Indian and global population for domain I of Pvama1, which suggests that it is a dominant target of the protective immune response. In contrast, the Pvmsp119 region showed highly conserved sequences in India and across the Oceania, South America, North America and Asia, demonstrating low genetic diversity in the global population when compared to Pvama1. Results suggest the possibility of including Pvmsp119 in a multivalent vaccine formulation against P. vivax infections. However, the high genetic diversity seen in Pvama1 would be more challenging for vaccine development. Plasmodium vivax is the dominant malaria species in several countries, and a safe and effective vaccine against the species is needed if progress is to be made towards malaria control and elimination. The P. vivax Apical Membrane Antigen 1 (AMA1) ectodomain and the C-terminal region (19 kDa) of the Merozoite Surface Protein 1 (MSP1) are important malaria vaccine antigens. However, antigenic diversity remains a critical factor for acquisition of immunity and also a major barrier for development of an effective vaccine. Here, investigations on the genetic diversity of two blood stage vaccine candidate antigens, Merozoite surface protein 119 and Apical membrane antigen 1, of P. vivax from three malaria endemic regions of the India was attempted and compared with global population. High genetic variation was observed in Pvama1 domain I, which has been reported as a dominant target of host immune response. In contrast, Pvmsp119 showed highly conserved sequences from three geographically diverse populations of India and globally, which further suggests Pvmsp119 as an extremely conserved protein. Collectively, the diversity data generated in this study provides a baseline data towards development of a broadly efficacious Pvama1 and Pvmsp119 based vaccine. Plasmodium vivax is a major malaria species which affects a large part of the world’s population [P. vivax malaria transmission areas and its burden has been increased as compared to P. falciparum over the last few years [P. vivax has a higher genetic diversity than the most common species Plasmodium falciparum indicating better fitness and potential to escape the immune response [P. vivax resistance to the antimalarial drug chloroquine in various countries including India has raised a threat on malaria elimination [Plasmodium species and have augmented the malaria burden in India [P. vivax and P. falciparum are unevenly distributed but also co-circulate in several Indian states. In the southern states (e.g. Tamil Nadu), P. vivax is the dominant malaria parasite species [P. falciparum cases and state like Gujarat (from the western part of India) showed the prevalence of both species, including mixed species infections [pulation . Worldwiew years . India rew years . P. vivaresponse . In addimination . India’sin India . The two species . Easternfections . To contP. falciparum and P. vivax [P. vivax Apical membrane antigen 1 (Pvama1), required for both merozoite invasion of erythrocytes and sporozoite invasion of hepatocytes [Pvama1 codes for a cysteine-rich ectodomain, a conserved cytoplasmic region, and a transmembrane region [PvAMA1 ectodomain is divided into three subdomains by disulfide bridges referred to as Domain I (DI), Domain II (DII) and Domain III (DIII) [PvAMA1 is highly immunogenic and high antibody titers against the ectodomain have been reported in natural infections, which effectively inhibit erythrocyte invasion [Hundreds of malaria vaccine candidates have been characterized, of which Apical membrane antigen 1(AMA1) and Merozoite surface protein 1(MSP1) are the most promising blood-stage vaccine candidates for P. vivax , 8. P. vatocytes , is an ae region . The PvAI (DIII) . The ectinvasion , 10. DI invasion .Plasmodium vivax merozoite surface protein 1 (PvMSP1) is a highly abundant protein present on the surface of merozoites. Initially it is synthesized as a high molecular weight precursor, which is further processed to various low molecular weight fragments [19 fragment contains two epidermal growth factor-like domains that have been shown to be efficient blood-stage vaccine candidates [19 remains on the surface of the merozoite and participates in the initial adhesion to the reticulocyte [19 can inhibit parasite growth [ragments . The C tndidates . At the culocyte . Severale growth –19 and ae growth –23.P. vivax population [P. vivax malaria burden from Southeast Asia region, however, genetic diversity in PvAMA1 and PvMSP119 vaccine candidate antigens from malaria endemic region of India has rarely been done [PvAMA1 and PvMSP119 kDa region from three geographically diverse malaria endemic regions of India. Published Pvama1 and Pvmsp119 sequences from the global parasite population and sequences generated in this study was analyzed to identify genetic polymorphism and selective forces acting on these genes. The results of present study thus have important implications in Pvama1 and Pvmsp119 based vaccine designing.To develop an effective malaria vaccine which can work in diverse regions of the world, it is important to study genetic diversity of vaccine candidates from geographically diverse regions to include alleles that can induce the immune response and cover the antigenic diversity of pulation . India ceen done –27. TherThe clinical samples collected in this study received ethical approval from the Institutional ethic committee of ICMR-NIMR in New Delhi, India and Institutional Review Board at New York University, School of Medicine in New York, New York, USA. Samples were collected after written consent was obtained from all participants.P. vivax as the dominant species during the malaria season in July and October. Samples were collected at Besant Nagar Malaria Clinic, and in cross-sectional surveys conducted in the coastal fishing community, including a few slums and middle and upper-class urban dwellings. Nadiad is a small town located in the central part of Gujarat state, which is considered as hypo-endemic for P. vivax and P. falciparum, with a slightly higher prevalence of P. vivax. Samples were collected at the Malaria Clinic, located in the Civil Hospital of Nadiad and in cross-sectional surveys conducted in the rural area of Nadiad town. The city of Rourkela located in the Sundargarh district of the eastern state of Odisha, where P. falciparum is the major infecting malaria species, displays meso to hyper endemic transmission in this region. Samples were collected at a malaria clinic in a suburb of Rourkela and cross-sectional surveys were conducted in a forest area of Sundargarh.Samples were collected as part of a U.S. National Institutes of Health Center for the Study of Complex Malaria in India (CSCMi) project  from thrP. vivax infection was determined by RDT and microscopic examination of thick and thin blood smear. From each individual 2–3 ml of blood was collected in EDTA vacutainers . Samples were centrifuged at room temperature at 1500 g for 15 min. Plasma and platelets were removed and stored at -80°C. DNA was extracted from infected red blood cells using DNA Blood MIDI kit . A total of 100 P. vivax samples collected from infected individuals in Chennai , Nadiad  and Rourkela  between January 2013 and May 2015 were used in this study (P. vivax infections were confirmed by species-specific nested-polymerase chain reaction (nPCR) at each study site [is study . P. vivaudy site , and recPvama1 (PVX_092275) and Pvmsp119 (PVX_099980) with Primer3 (version0.4.0) using the P. vivax Sal-1 genome reference from the malaria database PlasmoDB (www.PlasmoDB.org). Two primer sets were designed for Pvama1 and one primer set was designed for C-terminal, 19kDa region of the Pvmsp1gene  was amplified using two sets of primers .MSP-119C terminal region was PCR amplified . PCR products were purified using the Exonuclease-I and Shrimp Alkaline Phosphatase  followed by sequencing reactions in both forward and reverse directions (2x coverage) by a commercial sequencing facility using ABI BigDye Terminator Cycle Sequencing kit on an ABI 3730XL automatic DNA Analyzer .The  primers . The ampmplified  as abovePvama1 sequences (n = 100) and Pvmsp119 sequences (n = 100) were aligned to the P. vivax Sal-I reference strain sequence. A 1335 bp region encompasses the complete ectodomain  of Pvama1, and 266 bp region of Pvmsp119 was assembled excluding primer regions.Raw sequence data were assembled and edited using the SeqMan . The DNA sequence chromatograms were carefully inspected to identify the SNP’s. All Pvama1 and Pvmsp119, published sequences were retrieved from GenBank  were retrieved from GenBank and added into the analysis. A total of 582 Pvmsp119 sequences (full-length Pvmsp119 gene sequence and Pvmsp142 region sequence) from 14 distinct P. vivax subpopulations and two reference strains  were retrieved for the analysis. Combined, these represent Pvama1 and Pvmsp119 sequences from seven global regions: East Asia , South Asia , Southeast Asia , West Asia , South America , North America (Mexico), and Oceania (PNG. Vanuatu).To investigate the global genetic diversity of  GenBank . A totalPvama1 and Pvmsp119, which includes total number of polymorphic sites (S), nucleotide diversity (π), the number of haplotypes (h), haplotype diversity (Hd), the number of synonymous mutation (SP), non-synonymous mutation (NS), using a 100 bp sliding window with a 25 bp step size in DnaSP.All sequences were aligned by the CLUSTAL W program in MEGA6.0  and nuclPvama1 and Pvmsp119, Tajima’s D, Fu and Li’s D and F were calculated in DnaSP. Further, to evaluate natural selection the McDonald-Kreitman (MK) test was performed [Plasmodium cynomolgi AMA1 was used as the interspecies outgroup (GenBank accession no. XM_004222507.1) [FST by DnaSP was calculated to determine the degree of genetic variance in Pvama1 and Pvmsp119 kDa region among global isolates. The minimum number of recombination events (Rm), and linkage disequilibrium (LD) was calculated. The relationship between LD and nucleotide sites distance was plotted by using indices D’ and R2. To investigate the genetic relatedness among Indian and global Pvama1 haplotypes, network analysis was performed using a haplotype frequency > 1. For Pvmsp119 all Indian and global haplotypes were used and analysis were performed using Network software version 5.0.0.3 [PvAMA1 ectodomain were visualized as a three dimensional structure of PvAMA1 (PDB ID: 1W81) by Pymol [To test the neutral theory of evolution in erformed .The sequ22507.1) . Wright’ 5.0.0.3 . Mutatioby Pymol .Pvama1 sequences MH657021—MH657120, and 100 Pvmsp119 sequences MH657121—MH657220Sequences generated in this study have been deposited in GenBank under the accession numbers: 100 Pvama1 was amplified from 100 P. vivax isolates collected from Chennai , Nadiad  and Rourkela . The average number of nucleotide differences (K) were higher in Rourkela and Nadiad population when compared to Chennai  for Nadiad and Rourkela populations indicates deviation from neutral evolution and the tendency for diversifying selection in the corresponding regions. The negative Tajima’s value, Fu and Li (F and D) value for Chennai population indicates purifying selection. McDonald-Kreitman test showed that the entire Pvama1 ectodomain, particularly DI, had more nonsynonymous substitution as compared to synonymous substitution.Natural selection was assessed across the region encoding the Pvama1. The average number of nucleotide differences (K) for the domain I was higher in the Nadiad and Rourkela population and lower in the Chennai population  as well as seven geographically diverse P. vivax populations, including from Thailand (n = 231), Sri Lanka (n = 23), Iran (n = 29), Korea (n = 66), China-Myanmar border (n = 73), Venezuela (n = 73), PNG (n = 102), and a set of monkey adapted reference isolates (n = 8) (To study  (n = 8) .Pvama1 sequences here onwards referred as the “global” (including the 100 Indian sequences obtained in this study and 616 published sequences retrieved from GenBank described above) were used to determine nucleotide diversity and genetic differentiation. A total of 126 SNP’s was identified amongst the 716 global isolates, including 41 synonymous mutations and 85 nonsynonymous mutations. The average number of nucleotide difference (k) for the Pvama1 ectodomain across global isolates was 13.568. The higher number of nucleotide differences was found in Iran and India and the least number of differences were seen in isolates from South Korea. The haplotype diversity for the entire Pvama1 ectodomain of global isolates was (Hd = 0.9898). This value was higher in Iran, PNG, and India when compared to the other populations. The nucleotide diversity (π) was higher in Domain-I (DI) of the Pvama1 among global isolates  estimated was 23  sequences revealed that this region is highly conserved, and it didn’t show any polymorphic site in three geographically diverse malaria endemic regions of India.Nucleotide sequence analysis of the 100 Pvmsp119 kDa region is a potential vaccine candidate, its global genetic diversity has not been carefully evaluated. To study the genetic diversity of Pvmsp119, sequence data generated in the present study from India were compared to previously published sequences from India (n = 28), Thailand (n = 209), Sri Lanka (n = 106), Mexico (n = 35), Singapore (n = 50), Brazil (N = 11), Cambodia (n = 44), Myanmar (n = 28), Turkey (n = 40), South Korea (n = 22), China (n = 2), Bangladesh (n = 5), Vanuatu (n = 2), and two reference strains .Although the Pvmsp119 sequences here onwards referred as “global” (including 100 Indian sequences obtained in this study and 582 published sequences retrieved from GenBank described above) were used to determine nucleotide diversity and genetic differentiation. There was no genetic or nucleotide diversity found in Indian, South American (Brazil), and Oceanian (Vanuatu) isolates.A total 682 Pvmsp119 kDa region across global isolates was 0.229. The highest number of nucleotide differences were found in Turkey (K = 0.508) and Bangladesh (0.4) while the least was seen in South Korea (K = 0.091) and Thailand (K = 0.181). The haplotype diversity (Hd) for the entire Pvmsp119 region of global isolates was Hd = 0.2. This value was higher in Turkey (Hd = 0.508) than the other populations. The nucleotide diversity (π) of the entire Pvmsp119 kDa region for global isolates was π = 0.00937  for the  0.00937 . Overall9 region .Pvmsp119 is under purifying selection. The Fu and Li’s D and F values for Pvmsp119 global isolates -5.13595 (P < 0.02) and -4.74044 (P < 0.02) were also negative. For the global Pvmsp119 isolates, the minimum number of recombination events (Rm), estimated was 1. The LD index (R2) was also very low suggesting the absence of recombination events and low LD may have contributed to the low diversity in Pvmsp119 , indicating that Pvmsp119 .Sequence analysis of the global data resulted in 14 different haplotypes with amino acid changes at 11 codons as aligned to Sal I. All 11 amino acid changes were dimorphic and region specific. Amino acid change 1706E was found only in Turkey . We observed 8 amino acid changes specific to isolates from Singapore S1643P, D1649N, R1669G, C1681R, C1689R, A1697V, N1708D and P1721Q and one amino acid change specific to South Korea N1692K. Only one amino acid change (K1709E) was found in three different populations from Thailand, Cambodia and Myanmar.Pvama1 due to genetic variation among the three India populations, we calculated the FST value using Pvama1 sequences from the Chennai, Nadiad and Rourkela. A moderate level of genetic differentiation (FST = 0. 143) was detected between Nadiad and Chennai isolates, a low level of genetic differentiation (FST = 0.075) was detected between the Rourkela and Chennai population, and the FST value observed between Nadiad and Rourkela (FST = -0.003) demonstrated a high degree of genetic similarity between these two populations.To understand population differences in FST values for the eight worldwide Pvama1 populations with full-length ectodomain sequence were also calculated  was also detected for South Korea and other population. Moderate FST values (0.07–0.13) were detected among the India, China Myanmar, Thailand, Venezuela, PNG, and Sri Lanka populations. A negative FST value (-0.00687) was observed between Indian and Iranian isolates, showing a high degree of genetic similarity between these two populations.lculated . A high FST values of the thirteen worldwide Pvmsp119 populations were evaluated  when compared to India, China, Myanmar, Thailand, Singapore, Cambodia, Brazil, Mexico and Vanuatu populations, which represents a high degree of genetic distance. Moderate FST values (0.02–0.18) were detected when the Indian population was compared with population from Cambodia, Thailand, and Myanmar.valuated . FST valPvama1, demonstrating an extremely high level of haplotype diversity across the three sites (Hd 0.968). The Pvama1 data showed only 4 haplotypes  shared among these three populations. One haplotype (Hap6) is shared between the Chennai and Rourkela populations, one haplotype (Hap8) is shared between the Chennai and Nadiad populations, and three haplotypes  are shared between the Nadiad and Rourkela isolates. We observed region specific haplotypes in Chennai (n = 6), Nadiad (n = 27) and Rourkela (n = 12)  .Pvama1 ectodomain, a total of 352 haplotypes were identified in 716 global isolates with no matches to the Sal1 reference strain; only one isolate from Thailand matched to the Belem reference strain. A haplotype network formed using haplotypes with a frequency > 1 revealed a clearer network. Haplotype 2, haplotype 4 and haplotype 22 were shared among the Indian, Sri Lankan and Iranian populations with frequencies of 2.3, 1.9 and 0.69 respectively. Haplotypes 6, 7, 11 and 20 were shared among the Indian and Iranian population with a frequency of 1.1, 0.5, 0.5, and 0.2 respectively  in Rourkela and Nadiad as previously reported for both P. vivax and P. falciparum from other regions [P. vivax from Chennai, Nadiad and Rourkela as analyzed by Tajima’s D and Fu and Li (D and F) test which is in agreement with other studies from Venezuela, China-Myanmar Border, and PNG [P. vivax population from Sri Lanka showed evidence of positive selection in domain II, reported as highly immunogenic [Pvama1 is different in geographically diverse regions of the world.In ia cases . The preia cases  which pose areas , 40, 41. history , higher P. vivax . In Pvam regions , 45, sug regions . Results regions , 48, 49. and PNG , 45, 49.unogenic , 51. ThePvAMA1 ectodomain. However, domain I was the most variable, suggesting that this portion of the protein is potentially more exposed and accessible to host immune responses [P. vivax isolates from Rajasthan [PvAMA1 ectodomain 23 residues were located on one side, suggesting that this side might be targeted by host immune responses. As reported previously [P. vivax isolates at all three study sites in India. In DII of PvAMA1 highly antigenic region (aa 290–307 SASDQPTQYEEEMTDYQK) during natural infection [P. vivax malaria [Pvama1 haplotypes were observed in present study and only few haplotypes were shared between the three Indian subpopulations. In 716 global dataset, 352 haplotypes were identified with no matches to the Sal1 reference strain, which has been used for immune-epidemiological studies and vaccine development. Malaria vaccine based on only reference strains may result in poor clinical efficacy due to high genetic diversity and strain specific immunity in vaccine candidates [FST estimation between India and the closest geographical site Iran showed little genetic differentiation, which may be due to human population movement between the two countries. Only few haplotypes were shared between India, Sri Lanka and Iran indicating that similar allelic forms of Pvama1 circulating in these regions. The complex network of Pvama1 connected by rare admixed haplotypes and evidence of geographically restricted haplotypes suggest that covering Pvama1 diversity would be more challenging [Pvama1 gene may contribute to antibody binding and escape, which needs further investigation to understand weather Pvama1 haplotypes are antigenically distinct as previously reported for PfAMA1 [Polymorphic sites were distributed throughout esponses , which cesponses , 27. At esponses , 27. A teviously , 49, resnfection  was foun malaria . An effe malaria , 53. In ndidates , which fllenging . Geneticr PfAMA1 . GeneticPvMSP1 is a merozoite surface antigen, The 42 kDa region breaks into two fragments 33 kDa and 19 kDa. The predicted structure shows that 33 kDa fragment covers the 19 kDa fragment. This observation could explain the diversity and balancing selection present in the 33 kDa fragment but not in 19 kDa fragment [Pvmsp119 kDa fragment demonstrated that this region is highly conserved in P. vivax isolates from Chennai, Nadiad and Rourkela. A study conducted at same study sites reported high antibody response against PvMSP119 antigen as compared to PvAMA1 in people living in these areas [Pvmsp119 region was found to be conserved in many parts of India i.e. New Delhi (Delhi), Aligarh (Uttar Pradesh), Panjim (Goa), Panna (Madhya Pradesh) Sonapur (Assam), and Car Nicobar (Andaman & Nicobar Islands) [Pvmsp119 region were observed among India, China, Vanuatu and Mexico and found to be identical at the nucleotide level to the Belem and Salvador-1 types. Low genetic diversity and geographic clustering was observed in Pvmsp119, among the parasite populations from Turkey, Thailand, Bangladesh, South Korea, Sri Lanka, Singapore, Cambodia, and Myanmar suggesting that immune selection maintains similar Pvmsp119 alleles around the globe.fragment . Sequencse areas . The PvmIslands) . SimilarPvmsp119 haplotypes was identified in 682 global isolates. Since negative Tajima’s D was observed in multiple populations, and only Turkey and Cambodia showed positive Tajima’s D values, which indicates that Pvmsp119 region has undergone a recent bottleneck and lacks evidence of balancing selection. The level of selection pressure appears to vary on different parts of the protein in different Plasmodium species. In P. falciparum, msp119 is under positive selection pressure and Pvmsp133 is under purifying selection [Pvmsp119 region maintains low genetic diversity in geographically diverse malaria endemic regions shows that Pvmsp119 based vaccines may work effectively worldwide. Multiple-allele immunogens appear to be protective against heterologous infection and vaccine constructs based on two or more representative strains and antigens may offer an effective solution to the problem of polymorphism [PvMSP119 is advantageous, because of the improved cellular immune response.A total of 14 election . As repoelection  Pvmsp119morphism , 58. A rPvMSP119 and PvAMA1 vaccine candidate antigens in three geographically diverse malaria endemic regions of India and global dataset available in public domain. This is the first extensive study from India which contributes towards understanding the genetic diversity of PvAMA1 and PvMSP119 vaccine candidates. Highly conserved Pvmsp119 region is observed in present study and previously reported high antibody response against PvMSP119 antigen in malaria endemic populations of India confirm it as one of the most promising malaria vaccine candidates. In contrast, exceptionally high genetic diversity is observed in PvAMA1 among Indian and global population. Here, domain I of PvAMA1 is observed to be a dominant target of balancing selection. Overall, it seems that both the studied malaria antigens are under distinct selective pressure by the human immune system and thus genetic diversity in vaccine candidate antigens must be considered while developing a broadly efficacious malaria vaccine.In summary, this study presents the genetic diversity in S1 Fig(PPTX)Click here for additional data file.S2 FigPvama1 ectodomain sequences from three geographical diverse malaria-endemic regions of India were used to create a median-joining network. This network represents the mutational paths connecting Pvama1 haplotypes that may explain the observed sequence diversity. Each node represents one haplotype, node size indicates haplotype frequency and node color corresponds to the country of origin. Line length is proportional to genetic distance.(PPTX)Click here for additional data file.S3 FigPvama1 ectodomain sequences from India. Chennai sequences are in yellow, Nadiad sequences are in red and Rourkela sequences are in blue. The Sal-1 reference sequence was also included and indicated as a triangle. The unrooted neighbor-joining tree was constructed using 10,000 bootstrap replicates and the branch lengths are in the same units as evolutionary distance used to infer the tree.Neighbor-joining tree constructed using 100 (PPTX)Click here for additional data file.S1 Table(PPTX)Click here for additional data file.S2 Table(PPTX)Click here for additional data file.


Tetrahymena thermophila. We test the hypothesis that at temperatures closer to the species’ thermal optimum , genetic diversity should have a weaker effect on population growth compared to temperatures away from the thermal optimum. To do so, we grew populations of T. thermophila with varying levels of genetic diversity at increasingly warmer temperatures and quantified their intrinsic population growth rate, r. We found that genetic diversity increases population growth at cooler temperatures, but that as temperature increases, this effect weakens. We also show that a combination of changes in the amount of expressed genetic diversity (G) in r, plastic changes in population growth across temperatures (E), and strong G × E interactions underlie this temperature effect. Our results uncover important but largely overlooked temperature effects that have implications for the management of small populations with depauperate genetic stocks in an increasingly warming world.Genetic diversity and temperature increases associated with global climate change are known to independently influence population growth and extinction risk. Whether increasing temperature may influence the effect of genetic diversity on population growth, however, is not known. We address this issue in the model protist system  Understanding how temperature influences population growth is important in a warming world. Here, we show that temperature weakens the positive effect of genetic diversity on population growth in a microbial system. These results have consequences for conservation of genetically depauperate populations in a changing world. We hypothesize that lower genetic diversity may depress population growth , as observed in many other organisms, while higher genetic diversity may increase growth .In particular, we address (1) whether genetic diversity affects population growth in y Figure ; blue. Ct Figure ; red. An22.1Tetrahymena thermophila from the Cornell University Tetrahymena Stock Center from across three putatively different genetic backgrounds . The lines were reared in Carolina Biological protist medium®  in 200 ml autoclaved borosilicate jars, and a 16–8 day/night cycle at 22ºC within Percival growth chambers .We sourced five clonal lines  of the protist To determine whether temperature alters the effects of genetic diversity on population growth, we manipulated the temperature and initial genetic diversity of microcosm populations. To manipulate genetic diversity, we started populations with a varying number of clonal lines . Monoclonal cultures were initialized with 50 individual protists. For all other treatments, the initial abundance of each clone depended on the total number of clones present, to control for possible effects of initial density: two‐clone populations started with 25 individuals/clone, three‐clone populations started with ~16 individuals/clone, four‐clone populations started with ~12 individuals/clone, and five‐clone populations started with 10 individuals/clone. Each monoclonal population, and each combination of four and five clones, was replicated four times. Each combination of two and three clones was replicated twice, for a total of 84 experimental populations per temperature. All experimental microcosms were reared in 3 ml of growth media in 35 mm petri dishes.r, of a well‐mixed population , at seven different temperatures . Experimental microcosms were grown in Percival growth chambers with all other environmental variables mimicking rearing conditions.We crossed the five genetic diversity treatments with four temperature treatments  along the rising portion of the thermal performance curve (TPC) of the organism Figure , for a tr, which has units of t−1) of each microcosm population was calculated as [log(fN)–log(iN)]/time, with time = 1 day, fN being the final abundance, and iN the initial density .After a 24‐h incubation period, we estimated final population size by sub‐sampling each microcosm and counting individual cells under a stereomicroscope . Assuming exponential growth, the intrinsic growth rate , environmental variation in r (E), or G × E interactions, could explain observed changes in r across genetic diversity and temperature treatments. To do so, we used Analysis of Covariance (ANCOVA) in R packages “rstatix” v0.7  increased with temperature  are at least in part responsible for the lower levels of variation in r observed at higher temperatures, compared to those observed at lower temperatures  in es Table . Indeed,es Table , while ces Table . MoreoveBecause the presence of strong G × E effectively shifts which genotypes grow better at different temperatures, there is a possibility for temperature‐mediated clonal sorting in these microbial populations. Rapid evolutionary change has been suggested as a possible mechanism through which organisms may fend off the negative impacts of climate change  as was hypothesized in the introduction and then confirmed empirically , which represents a clear next step. We hypothesize that as temperature rises past Topt, we should see a strengthening of the effect of genetic diversity on r , for the same reasons that we observe a weakening of the effect of genetic diversity on r as temperature increases toward Topt. If that is the case, increasing genetic diversity could “rescue” populations ; Data curation (lead); Formal analysis (lead); Investigation (lead); Writing‐original draft (lead); Writing‐review & editing (lead). Megan Liu: Investigation (supporting). Samantha Votzke: Investigation (supporting); Writing‐review & editing (supporting). Andrea Yammine: Conceptualization (supporting); Investigation (supporting); Writing‐review & editing (supporting). Jean P. Gibert: Conceptualization ; Data curation (supporting); Formal analysis (supporting); Funding acquisition (lead); Resources (lead); Supervision ; Writing‐original draft (supporting); Writing‐review & editing .https://doi.org/10.5061/dryad.sqv9s4n52; https://github.com/JPGibert/Genetic_Diversity_And_TemperatureThis article has been awarded <Open Data, Open Materials>Badges. All materials and data are publicly accessible via the Open Science Framework at Appendix S1Click here for additional data file.