Prunus avium L.) is a temperate fruit species whose production might be highly impacted by climate change in the near future. Diversity of plant material could be an option to mitigate these climate risks by enabling producers to have new cultivars well adapted to new environmental conditions. In this study, subsets of sweet cherry collections of 19 European countries were genotyped using 14 SSR. The objectives of this study were (i) to assess genetic diversity parameters, (ii) to estimate the levels of population structure, and (iii) to identify germplasm redundancies. A total of 314 accessions, including landraces, early selections, and modern cultivars, were monitored, and 220 unique SSR genotypes were identified. All 14 loci were confirmed to be polymorphic, and a total of 137 alleles were detected with a mean of 9.8 alleles per locus. The average number of alleles (N = 9.8), PIC value (0.658), observed heterozygosity (Ho = 0.71), and expected heterozygosity (He = 0.70) were higher in this study compared to values reported so far. Four ancestral populations were detected using STRUCTURE software and confirmed by Principal Coordinate Analysis (PCoA), and two of them (K1 and K4) could be attributed to the geographical origin of the accessions. A N-J tree grouped the 220 sweet cherry accessions within three main clusters and six subgroups. Accessions belonging to the four STRUCTURE populations roughly clustered together. Clustering confirmed known genealogical data for several accessions. The large genetic diversity of the collection was demonstrated, in particular within the landrace pool, justifying the efforts made over decades for their conservation. New sources of diversity will allow producers to face challenges, such as climate change and the need to develop more sustainable production systems.Sweet cherry ( The ECPGR network is organized in thematic and crop Working Groups, including the Prunus WG (https://www.ecpgr.cgiar.org/working-groups/prunus accessed on 9 September 2021). From 2003, the European Search Catalogue for Plant Genetic Resources , coordinated and funded by ECPGR, provides information at the accession level of PGRs conserved ex situ and in situ [Prunus collections, in 2012, the Prunus WG developed the European Prunus DataBase . More recently, in the frame of the EU.Cherry ECPGR project (https://www.ecpgr.cgiar.org/working-groups/prunus/eucherry accessed on 9 September 2021), a collaborative action focused on sweet cherry, the WG has embarked on updating, documenting, and communicating the patrimonial richness of cultivated Prunus avium L. in Europe, aiming to connect information of the EURISCO and EPDB databases.Agriculture will face crucial challenges in the near future, including the need for substantially increased food production in order to satisfy 8 billion people. The impact of climate change is already causing a reduction in agricultural production at a global scale while food demand increases steadily. Experts from the Intergovernmental Panel on Climate Change predict that this situation will worsen in the coming years . Diversihttps://cerezadeljerte.org/en/the-picota-of-jerte accessed on 9 September 2021), and the ‘Itxassou’ village in Southwest France (http://www.cerise-itxassou.com/ accessed on 9 September 2021). European public institutes and non-governmental organizations have also been working for decades to collect and preserve ex situ cherry landraces and modern cultivars . The beneficial health effects have attracted attention as sweet cherry is rich in polyphenols ,5 in addhttps://www.bordeaux.inra.fr/cherry/ accessed on 9 September 2021) recently joined forces to analyze a large set of European sweet cherry landraces together with some popular old and modern cultivars. One of the long-term aims of this initiative is to define a first European core-collection, highly representative of sweet cherry diversity still available in Europe. Here we investigated accessions from 19 countries using 18 microsatellite loci, with the following objectives: (i) assess genetic diversity; (ii) analyze patterns of population structure; (iii) and identify germplasm redundancies.Currently, molecular markers  are being used to analyze the genetic diversity of European landraces and identify putative synonyms in a whole range of fruit tree species and other vegetatively propagated crops . StudiesP. cerasus) instead of sweet cherry. These were discarded from further analysis. Among the 314 diploid accessions, a total of 180 accessions represented genotypes that were only identified once. The remaining 134 accessions could be allocated to 40 groups based on having identical genotypes. These groups varied in size from groups with only two accessions each (19 groups) to a group containing 12 accessions. Some redundancies were found within samples from the same national collection, and some were found between collections  showed inconsistent allele patterns, and so a subset of 14 loci was used for further analysis. Three hundred and fourteen accessions presented allelic patterns corresponding to diploid genotypes, whereas 10 accessions presented additional alleles in various loci suggesting that they were tetraploid and most likely belonging to sour cherry  to 0.838 (EMPaS06) with an average of 0.658. Observed heterozygosity (Ho) varied from 0.350 (EMPa017) to 0.950 (BPPCT034) with a mean value of 0.709. Expected heterozygosity (He) varied from 0.380 (EMPa017) to 0.850 (EMPaS06) with a mean value of 0.699. The inbreeding coefficient Fis ranged from −0.17 (BPPCT034) to 0.08 (EMPa017), six loci showed a slight excess of heterozygotes, but only BPPCT034 showed a statistically significant deviation from the Hardy–Weinberg equilibrium. Thirty-nine percent of the alleles occurred in less than 1% of the accessions ; from 8 alleles in EMPa018 to only 1 in CPSCT038 and EMPaS02 .e were slightly higher in L, whereas Ho was slightly lower. , while early selections (11%) and modern cultivars (20%) reported fewer . The Sloy lower. . In ES-My lower. .The most likely number of populations determined by Bayesian analysis was K = 4 . Using aClusters 1, 2, and 4 contained 14, 14, and 12 accessions, respectively, while cluster 3 was smaller (containing 6 accessions). The number of admixed genotypes was 174 . However, by lowering the membership threshold to 55%, 13% of the previously admixed accessions could be assigned to cluster 1, 13 % to cluster 2, 21% to cluster 3, and 12% to cluster 4. More than a third, i.e., 78 accessions, still remained as admixed since they had a membership coefficient below 0.55 . When usThe results obtained with STRUCTURE were confirmed by the representation of accessions in PCoA analysis. Accessions could be divided into four groups corresponding to the populations previously identified, with the admixed accessions dispersed among the rest a,b.In both PCoA plots, cluster K3 separated clearly from the three others. In terms of allelic diversity variation, cluster K2 appeared as the largest.A Neighbor-joining (N-J) tree based on a simple matching dissimilarity matrix between the 220 accessions of the European collection separated the sweet cherry accessions into three main clusters and six subclusters . NeverthCluster I consisted of a group of 36 accessions, which were mainly early selections and modern breeding cultivars . All the accessions forming the STRUCTURE K1 population were present in this cluster, including most of the Italian accessions, which were mainly landraces. Among the latter, ‘Duroncino di Cesena’ showed the greatest diversity. ‘Mora di Cazzano’, another Italian landrace maintained in the French collection, appeared related to ‘Adriana’, which is one of its offspring. Associated with them was ‘Bigarreau Camus de Venasque’, which is a French landrace from Southwest France near the Italian border. The rest of the cluster consisted mainly of modern cultivars, as well a popular old cultivars, the early selection ‘Emperor Francis’. Four Romanian cultivars  clustered relatively close to each other; indeed, they all derive from crosses with ‘Boambe de Cotnari’ as parent. ‘Ludovic’ and ‘Bucium’ are offspring of the well-known Canadian cultivar ‘Van’, which also clustered in the same region of the dendrogram. The remaining cultivars were modern ones from different Northeast European countries, most of them having at least a German accession as parent.Cluster II grouped 85 accessions; among them, 62% were landraces and 38% early selection and modern breeding cultivars. Accessions in cluster II were mainly from North, Northwest, and Central Europe. Cluster II comprised most of the admixed accessions identified by Bayesian analysis, as well as all the accessions found in the K3 population. In the upper part of this cluster, a group of ‘guignes’, including ‘Ham Green Black’, ‘Guigne du Champ de l’air’, ‘Karting Fran Djupekas’, ‘Goodnestone Black’, and ‘Guigne Presgaux 2’ were closely related. All the Belgian landraces in this study except ‘Bigarreau Ghijssens’ and ‘Rouge dorée’ were found in this cluster, more or less close to ‘Cerise Proces’. All the accessions from the K3 STRUCTURE population were grouped in this part of the dendrogram, and they were in majority landraces from distant countries, such as Belgium (‘Cerise Proces’), Hungary (‘Torbágyi Késői’ and ‘Solymári Gömbölyű’), and Norway (‘Osabaer’). The French early selection (‘Bigarreau Marmotte’) was grouped close to several well-known early selections and modern cultivars, such as ‘Napoleon’ (the accession maintained in the French collection), ‘Hedelfinger’, ‘Bing’, its offspring ‘Sue’, and the Japanese cultivar ‘Nanyo’, which results from ‘Napoleon’ open pollination. Other well-known European modern breeding cultivars, such as the Italian ‘Ferrovia’, the Romanian ‘Uriase de Bistrita’, and the Hungarian ‘Kavics’ clustered closely, and the two latter having ‘Gemersdorfer’ as parent. The lower part of cluster II included numerous landraces from central European countries, such as one landrace from Switzerland maintained in the French collection, ‘Hâtive de Bâle’, four landraces from Austria, all Bosnian landraces and the only studied Croatian landrace, ‘Okićka’, but also landraces from North and Northwest Europe (Norway and Great Britain). Several early selections and modern cultivars could, though, be found in these subclusters. For instance, German accessions ‘Knauffs Schwarze’, ‘Kassins Frühe’, ‘Nafrina’, and ‘Werder’ were clearly grouped, the last one belonging to the Norwegian collection and supposed to be one of the parents of ‘Nafrina’. The other early selections or modern cultivars were mostly originated from Germany, Romania, or Sweden. Among them, we can cite another accession named ‘Napoleon’, maintained in the Norwegian collection, and the well-known German cultivars ‘Schmidt’ and ‘Regina’.Cluster III was the largest one with 99 accessions. It included all the accessions of the K4 and K2 populations identified by Bayesian analysis. The upper part was constituted mainly by early selection and modern breeding cultivars. It comprised all accessions of the K4 STRUCTURE population, which are mainly maintained in the Estonian and Latvian collections and originated from or were released in Northeastern Europe . Many of these cultivars share the genitor ‘Leningradskaya Chornaya’ in their pedigree. All the Ukrainian modern bred cultivars were also found in this cluster which has the German early selection cultivar ‘Drogans Gelbe Knorpelkirsche’ as common parent. In the lower part of cluster III all accessions of the K2 STRUCTURE population were grouped, with the exception of the Romanian cultivar ‘Galata’, present in cluster II. Among these, the early selection cultivars ‘Early Birchenhayes’ and ‘Burcombe’ from Great Britain, the Greek landrace ‘Tragana Edessis’, and the Romanian modern cultivar ‘Maxut’ were clearly separated from a large group of accessions which were all landraces. There were several groupings of accessions from the same geographical regions, e.g., early selections from Great Britain, Greek landraces along with the modern cultivar ‘O.T.E.A.’, a large group of French landraces integrating the K2 STRUCTURE population and three cultivars from Latvia and Lithuania. The last accession from STRUCTURE population K2, ‘Lisboeta’, was grouped in the dendrogram along with landraces from Bosnia and Herzegovina and France, as well as with the well-known French early selections ‘Bigarreau Hâtif Burlat’ and ‘Bigarreau Moreau’. The American modern breeding cultivar ‘Glacier’, resulting from the hybridization of cultivars ‘Stella’ and ‘Bigarreau Hâtif Burlat’, was closely clustered. Finally, the accessions grouped within the lower part of cluster III presented a high geographical diversity (9 countries involved) and a predominance of landraces.Overall, considering the different clusters, in terms of genetic diversity, the most original accessions were ‘Duroncino di Cesena’, ‘Bigarreau Camus de Venasque’, ‘Hedemora’, ‘Techlovicka’, ‘Alica’, ‘Galata’, ‘Draganele de Pitesti’, ‘Malizia’, ‘Durette’, ‘Dvijla Cerna’, ‘Maardu Maguskirss’, and ‘Radica’. The most distant accession of the whole study was the Italian landrace ‘Malizia’ within Cluster III.One of the goals of this study was to contribute to the development of a joint European germplasm collection, which is easy to use by sweet cherry breeders. Among the 314 accessions initially collected, 40 groups with shared genotypes were found. Redundancies occurred both within and between samples of accessions from the national collections. A range of errors was indicated, including incorrect passport data due to spelling mistakes in accession names translation, in particular when transcribing accessions names from the Cyrillic to Latin alphabet: e.g., ‘Iput’ (Estonian collection) and ‘Iputj’ (Latvian collection) or to mislabeling, e.g., ‘Fryksås’ (Swedish collection) which was deemed to match ‘Gårdebo’ and is believed to be a mislabeled accession. This latter finding agrees with the alignment against the Swedish national dataset, which also associated these two last accessions with accessions of the name ‘Wils Frühe’ in both Germany and Great Britain collections . Accessio = 0.71), and expected heterozygosity (He = 0.70) were higher in this study compared to values reported in most other sweet cherry studies [e = 0.56 and 6.5 alleles per locus [e = 0.59 and 4.3 alleles per locus [o and He, respectively (0.68 and 0.66) [o = 0.66 and He = 0.64 [o = 0. 62; He = 0.63) [o = 0.61; He = 0.65) [o = 0.63; He = 0.63) were obtained when characterizing 207 accessions (landraces and modern bred cultivars) from a French germplasm collection [o and He values obtained in this work possibly reflect the large geographical sampling range. In addition, they reflect the high genetic diversity among the European accessions forming the collection, and similar values were found from the analysis of a larger group of accessions aligned across more complete samples from a subset of the national collections [Average number of alleles N = 9.8), PIC value (0.658), observed heterozygosity  . A studye = 0.64 . All thee = 0.64 , 0.50 [2e = 0.64 , 0.66 [2e = 0.64 , 0.59 [2e = 0.64 , 0.61 [2e = 0.64 . More re = 0.63) . Similar = 0.65) . Likewisllection . With th.8, PIC vo, He, % of rare alleles, and inbreeding coefficient (Fis) between landraces and either early selection or modern breeding cultivars agreed with the loss of diversity associated with breeding [The differences for Hbreeding ,15,29. Tbreeding , a highePrunus avium L. in Europe and our findings in relation to the numbers of rare alleles in Greek samples, the genetic resources of some essential regions including Greece, Croatia, Serbia, and Slovakia were underrepresented in our study. Moreover, geographical regions, including Bulgaria and Spain, were completely absent, and it would be valuable to consider these in further analyses.The two different approaches (STRUCTURE and PCoA) used here to analyze the structure of the European sweet cherry accessions provided similar results. This clustering reflected, to some extent, the geographical origin of the accessions. Thus, population K1 contained most of the Italian accessions, and some of these  clustered together also in a previous study . Other aUnderstanding the genetic relationships among accessions from different countries’ collections is crucial to define a European core collection. The N-J tree grouped the 220 sweet cherry accessions within three main clusters not necessarily related to the accession geographical origin. However, at least some clusters were associated with accession country of origin, i.e., Italian accessions in cluster I, German early selection cultivars in cluster II, and accessions from Northeast countries  in cluster III. So far, few studies have investigated the genetic relationships of such a large number of accessions from so many countries. Our study confirmed previous results obtained in a study by the INRAE germplasm collection analyzed with the RosBREED cherry 6K SNP array v1 , such asOur findings demonstrated that a group of sweet cherry landraces were more diverse than a similar-sized group of early selections and modern cultivars. The bottleneck observed in modern cultivars has been well documented for the North American breeding programs . AlthougAn interesting finding of this study was that landraces, early selections, and modern cultivars co-occurred in most of the main groups determined by STRUCTURE except for K4, which was composed mainly of Northeastern European modern cultivars. Despite a lower genetic diversity within the pool of early selections and modern cultivars, the genetic distances between accessions belonging to these groups appeared to be relatively important. It is highly likely that by carefully selecting popular modern cultivars, including key founders , breeders could maximize genetic diversity in order to avoid the negative effects of inbreeding. However, it is also recommended to incorporate landraces with particularly interesting agronomic traits  and/or with a particularly deviating genetic composition .https://www.bordeaux.inra.fr/cherry/ accessed on 9 September 2021). Accessions were chosen to meet the following criteria: (i) they should, as far as possible, represent original landraces from the country of provenance; (ii) they should exhibit at least one trait of potential interest to sweet cherry breeders, and (iii) they should ideally be accompanied by characterization data. Some popular old cultivars (resulting from early selection in previous centuries), such as ‘Bigarreau Hâtif Burlat’, ‘Hedelfinger’, and ‘Napoleon’, were included as were some modern cultivars released in breeding programs, such as ‘Bing’, ‘Leningraskaja Chornaya’, ‘Sue’, ‘Valerij Tchkalov’, and ‘Van’. Accessions were noted to classify as either: landraces, early selections, or modern cultivars, according to Campoy et al. [Leaf samples from 324 cherry accessions were obtained from a total of 19 countries, including partners in the EUCherry project and collaborators in the COST Cherry network . Finally, only 14 SSRs were kept for the study; they were distributed across the eight cherry linkage groups and were not closely linked  following a slightly modified protocol . Eighteeping set , 4 loci ping set ,37, 2 loping set , and 1 gping set . Even thTwo approaches were followed to identify accessions that shared the same multilocus genotype: visual checking and comparison of profiles of the accessions and the ‘Multilocus Matches’ function available in GenAlex 6.503 software . Followio), expected heterozygosity (He), and inbreeding coefficient  were estimated using Cervus 3.0 software [Parameters of genetic diversity, including the average number of observed alleles, polymorphism information content (PIC), observed heterozygosity  was performed using DARwin 6.0.010 software  . Second,To our knowledge, the present study, although preliminary, is the largest genetic diversity and structure analysis of combined sweet cherry germplasm from across Europe. Among the samples selected from collections, some new supposed redundancies were identified, and others were confirmed. However, mislabeling and synonymy were not sufficient to explain redundancies concerning some accessions which differ in the country (and sometimes continent) of origin, status, and period of release. A large genetic diversity was found in our collection, in particular within the landrace pool, justifying the efforts made over decades for their conservation. The availability of new sources of diversity should help to address the new challenges that producers face, such as the changing climate and the need to develop more sustainable production systems. However, we would suggest that the picture we present can be further improved with more exhaustive sampling in some of the countries studied and in some of the countries not included, particularly those important in the evolutionary history of the species, including the supposed area of origin of sweet cherry, within the Caucasus region.


It has been well established that bushfire/wildfire smoke can taint grapes (and therefore wine), depending on the timing and duration of exposure, but the risk of smoke contamination from stubble burning (a practice employed by some grain growers to prepare farmland for sowing) has not yet been established. This study exposed excised bunches of grapes to smoke from combustion of barley straw and pea stubble windrows to investigate the potential for stubble burning to elicit smoke taint. Increased levels of volatile phenols  were detected in grapes exposed to barley straw smoke (relative to control grapes), with smoke density and the duration of smoke exposure influencing grape volatile phenols. However, the sensory panel did not perceive wine made from grapes exposed to low-density smoke to be tainted, despite the presence of low levels of syringol providing compositional evidence of smoke exposure. During the pea stubble burn, grapes positioned amongst the burning windrows or on the edge of the pea paddock were exposed to smoke for ~15–20 and 30–45 min, respectively, but this only resulted in 1 µg/kg differences in the cresol and/or syringol concentrations of smoke-affected grapes (and 1 µg/L differences for wine), relative to controls. A small, but significant increase in the intensity of smoke aroma and burnt rubber flavor of wine made from the grapes positioned amongst the burning pea stubble windrows provided the only sensory evidence of any smoke taint. As such, had vineyards been located immediately downwind from the pea stubble burn, it is unlikely that there would have been any smoke contamination of unharvested grapes. Smosyringol ,6,7. Sevsyringol ,12,13,14syringol ,16,17. Much of the smoke taint research undertaken to date has been in response to recurring incidents of vineyard exposure to bushfire/wildfire smoke and has investigated: the physiological responses of grapevines to smoke exposure ,18,19; fStubble is the term given to the cut stalks and straw residue remaining in the field after the grains of cereal crops such as wheat, canola, barley, lupins and oats are harvested ,29,30. WThe smoke and particulates released from stubble burning can have adverse effects on human health and the environment if not properly managed ,30. The In regions where grains and wine grapes are grown in close proximity, there is increasing concern that smoke from burning stubble could taint unharvested grapes, especially where burn programs overlap the grape growing season. Although the risk of grapes being contaminated by stubble smoke is likely to be lower than that arising from exposure to bushfire smoke (due to the relative durations of smoke exposure), the potential for stubble burning to taint grapes and wine has not previously been investigated, and was therefore the primary aim of this study. A preliminary trial was first undertaken to confirm that excised grape bunches could be used as ‘sinks’ to monitor smoke exposure during a stubble burn and to establish to what extent smoke density affects temporal changes in grape volatile phenols during smoke exposure, and their carry over into wine. Environmental data were collected before, during and after the (5 h) preliminary field trial . The air10 > PM2.5 > PM1.0, in agreement with particle size distributions previously reported for smoke from domestic wood fires [3 after 270 min, which was attributed to the corresponding drop in wind (Particulate matter (PM) was monitored as a measure of smoke emission b, and asod fires . PM was  in wind a. Neverto-cresol were detected, at concentrations of 6.0 and 1.0 µg/kg, respectively, as natural constituents of control grapes , but only after 2 or more hours of smoke exposure. Collectively, these results confirm the accumulation of smoke-derived volatile phenols in grapes depends on both the density of smoke (being a function of the proximity of grapes to the source of smoke) and the duration of smoke exposure. Only guaiacol and l grapes . Smoke-ao-cresol and syringol were detected in the control wine , with similar levels of guaiacol and o-cresol being detected in Smoke C wine. However, syringol was present at a significantly higher concentration, i.e., 9.7 µg/L. While the guaiacol content of Smoke B wine was also the same as the control, significantly higher cresol and syringol levels were observed. The highest levels of volatile phenols were found in Smoke A wine, with guaiacol and syringol again the most abundant. Interestingly, whereas the guaiacol concentration of Smoke A wine was almost two-fold higher than the levels detected in grapes (at t = 5), only a 5 µg/L (~15%) difference in syringol was observed between grapes (at t = 5) and wine. The volatile phenol levels observed in this wine were comparable to, or slightly higher than, concentrations reported in several smoke-tainted red wines resulting from field trials involving the application of smoke to grapevines  of different cultivars [The differences in the volatile phenol concentrations observed for grapes were also seen amongst wines . GuaiacoThe volatile phenol glycoconjugate profiles of wines were also determined, but of the glycoconjugates measured, few were detected, and none were found at concentrations above 14 µg/L . This waStatistically significant differences were observed amongst wines for measurements of residual sugar, pH, malic acid, color and phenolics , but couThe sensory profiles of control and Smoke C wines were very similar, with these wines exhibiting the most intense fruit aromas and flavors, and least intense smoke-related attributes . This suEnvironmental data were again collected before, during and after the (~2 h) stubble burn trial . During 3  also detected the smoke plume detected by sensor A, but the smoke had dispersed substantially, such that the PM10 concentration peaked at only ~450 µg/m3, while PM2.5 and PM1.0 remained below 50 µg/m3  seemingly did not detect any smoke; with the exception of one time point, PM concentrations remained below 50 µg/m3 throughout the stubble burn  in the adjacent paddock, were unlikely to have been exposed to any smoke, due to the combined vertical and lateral dispersion of smoke plumes.o- and m-cresols were detected in grapes from the excised bunches deployed during the stubble burn trial, nominally ‘smoke-affected grapes’  were continuously captured  and uploaded to the manufacturer’s network via an internal Wi-Fi connection. Data were subsequently exported from the network as Excel files. After 5 h of smoke exposure, the remaining fruit was collected and stored overnight (at 0 °C), together with control fruit , prior to small-scale winemaking. Control fruit was also sampled  and frozen (−20 °C) for chemical analysis.Shiraz grapes were hand harvested at maturity  were 27 °Brix) from a commercial vineyard located in the Padthaway wine region of South Australia , and stored in a 0 °C cold room prior to use. The preliminary trial, which involved exposure of grape bunches to smoke over 5 h, was conducted at the University of Adelaide’s Waite Campus. Bunches (60 per treatment) were suspended on wire frames positioned at three distances  downwind from the smoke source, hereafter positions A, B and C, respectively . Barley 1.0, PM2.5 and PM10 data , while the sensor at position A also recorded temperature, relative humidity, wind speed and direction , with data again uploaded to the manufacturer’s network via Wi-Fi. On completion of the stubble burn , grape samples  were collected and frozen (at −20 °C) for chemical analysis. The remaining fruit was collected and stored overnight (at ambient temperature), together with control fruit , prior to transportation to the University of Adelaide’s Waite Campus for small-scale winemaking. The environmental sensors remained in the field overnight and continued to collect data, which were subsequently exported from the manufacturer’s network as Excel files. Cabernet Sauvignon grapes were hand harvested (when TSS were 24 °Brix) from a commercial vineyard located in the Coonawarra wine region of South Australia , approximately 3 h prior to use. The trial, which involved exposure of grape bunches to smoke from prescribed burning of pea stubble, was conducted in a commercial pea field located in Glenroy , north-west of the Coonawarra. The pea stubble was raked into windrows prior to burning, with the property manager commencing the burn when the prevailing weather conditions  were in accordance with the local Council’s prescribed burning permit conditions. Bunches (100 per treatment) were suspended on wire frames positioned (i) amongst stubble windrows in the pea field, (ii) on the edge of the pea field (upwind) and (iii) in an adjacent field (approximately 500 m upwind), hereafter positions A, B and C, respectively . PortablGrape bunches  were destemmed by hand and berries divided (by mass) into three fermentation replicates . Berries were then crushed by hand, with 50 mg/L additions of sulphur dioxide (added as an 8% solution of potassium metabisulphite) and pH adjustment (to 3.5 with a 10% solution of tartaric acid) made to must prior to inoculation with 300 mg/L Maurivin PDM  and addition of 200 mg/L of diammonium phosphate. Musts were fermented on skins at 18–20 °C, with the cap plunged twice daily. Wines were pressed when residual sugar was <1 g/L and cold stabilized at 0 °C for 8 weeks. Following cold stabilization, samples  were taken for chemical analysis, before wine replicates were assessed by four sensory experts from the University of Adelaide  to establish there were no off-flavors or obvious differences between replicates. Replicate wines were then blended for sensory analysis .d3-guaiacol, d3-4-methylguaiacol, d7-o-cresol, and d3-syringol as internal standards, with a limit of detection of 1–2 µg/L; volatile phenol glycoconjugates were measured using an Agilent 1200 high performance liquid chromatograph equipped with a 1290 binary pump coupled to an AB SCIEX Triple QuadTM 4500 tandem mass spectrometer, with a Turbo VTM ion source , using d3-syringol gentiobioside as an internal standard, with a limit of detection of 1 µg/L.Basic wine chemistry parameters were determined by the Australian Wine Research Institute’s (AWRI) Commercial Services Laboratory  and comprised: alcohol, residual sugar, pH, titratable acidity  and volatile acidity  measured using a Foss WineScan analyzer ; and red wine color and total phenolics, measured using the modified Somers method and methyl cellulose precipitable tannin assay , respectThe sensory profiles of wines were determined using the Rate-All-That-Apply (RATA) method  with panp ≤ 0.05), to determine significant differences between wines, using XLSTAT .Chemical data were analyzed by one-way analysis of variance (ANOVA) using GenStat . Sensory data were analyzed by two-way ANOVA using participants as a random factor and wines as a fixed factor, with Fischer’s LSD post hoc test  can taint grapes, with both the density of smoke and the duration of smoke exposure affecting the uptake of smoke-derived volatile phenols by grapes and the perceived intensity of smoke taint in resulting wines. Interestingly, the sensory panel did not detect smoke taint in a wine made from grapes exposed to low-density smoke, despite the presence of low levels of syringol providing compositional evidence of smoke exposure. The use of excised bunches of grapes as ‘sinks’ for monitoring smoke exposure was also demonstrated and this approach was employed in the subsequent stubble burn trial, along with environmental sensors that measured smoke density and the duration of smoke exposure based on the concentration of particulate matter. Results from the stubble burn trial strongly suggest that the risk of grapes being contaminated by stubble smoke are low, especially when guidelines for managing emissions are followed, e.g., consideration of prevailing weather conditions  and not scheduling burns after rain (when stubble is more likely to smolder than burn), and/or where unharvested vineyards are not immediately downwind from stubble burns. Nevertheless, there are alternative stubble management practices that landowners or operators could consider instead of burning, so as to minimize emissions and their potential impacts on human health and the environment.


Our article aims to provide a comprehensive portrayal of how seven Asian jurisdictions have sought to address the challenge of genetic discrimination (GD) by presenting an analysis of the relevant legislation, policies, and practices. Based on our findings, policy discussion and action on preventing or mitigating GD have been narrowly framed in terms of employment, insurance, disability, marriage, and family planning. Except for South Korea, none of the jurisdictions we examined has adopted specific legislation to prevent GD. However, for Asia to truly benefit from its recent scientific and technological progress in genomics, we highlight the need for these jurisdictions to engage more proactively with the challenges of GD through a coordinated regulatory and governance mechanism. Genomic technologies have generated a wealth of new knowledge regarding health and disease. A wide spectrum of genetic tests—once only a figment of a distant reality—is becoming widely available, from whole-genome or -exome sequencing to multi-omics and epigenetic testing. These tests are being progressively integrated within healthcare systems, taken up in clinical practices, and offered as direct-to-consumer genetic testing (DTC-GT) services worldwide.1, and BioBank Japan gathered human DNA and serum samples from 291,274 participants in 20172. In Taiwan, a National Biobank Consortium was established in 2020 to integrate existing 33 biobanks for a large and comprehensive biomedical big data network3 and the Hong Kong government has in 2020 established the Hong Kong Genome Institute to conduct large-scale genome sequencing4. Asia also has a vibrant private sector, which markets technologies and genetic services to consumers both within the region and across the globe.Asia in particular has taken its place at the forefront of the genomic and precision medicine revolution. Asian countries are active in large international collaborative projects such as the International Cancer Genome Consortium, the Global Alliance for Genomics and Health, and the International Human Epigenome Consortium. They have actively set up public biobanks with the big data extracted from their populations. For example, the Korean Genome and Epidemiology Study included ~245,000 participants’ samples with omics data in 20177. The recent processing of biological materials and genetics data biobanks and data repositories, along with the exponential growth of DTC-GT, across national borders, have signaled the need to re-initiate discussion of the ELSI on a global scale8.While genetic testing may improve disease prediction, diagnosis, and treatment, the rapid uptake and application of genetics and genomics raise numerous ethical, legal, and social issues (ELSI). One of the most prominent among these is the growing number of possibilities of using genetic information to justify treating individuals differently or profiling specific population groups that may lead to genetic discrimination (GD). GD usually refers to the unfair and differential treatment of an individual based on actual or suspected genetic characteristics, and is recognized to arise in a number of contexts such as insurance, employment, and health care9, life insurance11, and employment12. More recent studies vary considerably, ranging from those that present comparative data on the use of genetic testing results by private life insurance companies13 to the systematic reviews of international public policies and studies concerning GD7, and analyses of the perceptions and concerns regarding GD14. For instance, a public survey was conducted across Europe to examine apprehensions surrounding GD in the case of population-based genetic databases15. Another study examined public perception of GD associated with genetic testing in primary care, finding that perceptions substantially varied by nationality among US and Canadian residents16.By the first decade of the twenty-first century, several transatlantic studies documented GD in clinical settings17. Another study regarding genomic technologies and consent practices in Japan, Malaysia, and Taiwan highlighted the need to inform genomic research participants about the risk of GD18. In the light of growing collaborations in genomic research and the use of gene technologies across the Asian regions and beyond, our paper seeks to reinvigorate the use of comparative law and policy approaches to identify existing and emerging forms of GD and to harmonize legal and regulatory responses in ways that facilitate international collaboration.In contrast, there are only a few published studies on GD in Asia in which comparative law and policy approaches have been applied. Yoshizawa et al. discussed how genetic technologies might be used in discriminatory ways in six Asian jurisdictions, documenting a case of pre-employment GD in ChinaThis article begins with a review of legislation, policies, and practices related to GD in seven Asian jurisdictions—China ), India, Japan, Philippines, South Korea, Singapore, and Taiwan. These jurisdictions were selected primarily due to the legal and bioethical expertise available in the Asian Bioethics Network on Genomics, which was formed in 2016 to promote ethical dialog and regulatory harmonization across jurisdictions in Asia that have committed substantial resources to advance gene technologies and genomic medicine. All the contributing authors from these jurisdictions are members of the Network and have been involved in active and continuing dialog through in-person meeting in Seoul on April 16 and 17, 2018 and further electronic communication. These have in turn given rise to a degree of consensus on identifying and discussing GD across these jurisdictions that are highly varied in demographic composition, socio-political constitution, and level of technological development. The authors of the seven Asian jurisdictions conducted an in-country analysis of GD through policy and regulatory review using Google and official legal databases in the jurisdictions between January to May 2019. Further data searches were subsequently conducted by Y.J. and H.Kim during the period from May to July 2020.19. Because this type of use of genetic information does not create GD per se, but rather exacerbate existing types of discrimination based on other factors such as population group, citizenship status, wealth level, etc., it warrants a distinct investigation in future research.This review is presented in two stages. In the first stage, an overview of the regulatory governance of GD in each jurisdiction is presented, particularly in relation to insurance, where concerns over prejudicial exclusion are widely recognized. The second stage is focused on outlining existing privacy frameworks that protect people against GD in these jurisdictions. Subsequently, this article highlights the progress of GD policies and the identified GD policy gaps to provide interpretations specific to the Asian context, before providing further comparisons with their European and North American counterparts. An emerging area of concern that we decided not to include in our manuscript is that of GD caused by the forensic use of genetic information and databases in AsiaBioethics and Safety Act (BSA). In addition, measures protecting people against GD have been included in research oversight frameworks, such as national ethics guidelines and research funding policies.The brief comparative review presented in this part illustrates the emerging legal and political trends developed to address GD in the selected Asian jurisdictions Table . India a20. Under the Insurance Act (2015), the CBIRC mandates that business licenses be issued to both state-owned or private insurers, including foreign-funded insurers20. The MAHI applies to those types of insurance companies and their employees20. Specifically, the MAHI prohibits insurance companies from “mak[ing] differentiated pricing” of an insured person based on genetic information and genetic testing results (art. 17). The MAHI also prohibits insurance companies from collecting and requesting genetic testing results of an insured person “when an insurance company sells health insurance products” (art. 38). However, the MAHI allows the use of the “family genetic medical history” of the insured person for purposes of differentiated pricing of the insured person or collecting genetic information when selling health insurance products (arts. 17 and 38).The China Banking and Insurance Regulatory Commission (CBIRC) adopted new government guidelines—the Measures for the Administration of Health Insurance (MAHI)—in 2019, to prohibit GD in health insuranceDisability Discrimination Ordinance and the Personal Data (Privacy) Ordinance provide some protection against GD, although neither statutes are specific to genetics or gene technology. Under this statute, a court has ruled that it is unlawful for the civil service to discriminate against potential or actual employees based only on a family history of mental illness related to genetic testing results21. In a recently published blueprint to drive the local development of genomic medicine, an expert group appointed by the SAR government indicated that more specific regulatory measures may be needed to prevent GD in life, critical illness, and health insurance22. Considerable emphasis has also been placed on public engagement in the blueprint, which may support better identification of GD and means to address related concerns.In Hong Kong SAR, the M/S United India Insurance Co. v. Jai Parkash Tayal—highlights the pervasive effect of GD as it relates to private health insurance and employment23. The Court concluded that such discrimination did not adhere to proper standards of genetic testing and failed to comply with the test of intelligible differentia, a standard of natural justice and equity (The Constitution of India). However, the Supreme Court of India stayed this judgment and prevented the operative part of the Delhi High Court’s judgment from transforming India’s laws regarding GD24. Indeed, the majority of employers and insurance providers in the country are in the private sector, and impleading the insurance regulator, the Insurance Regulatory Development Authority of India, is the right step to address potential GD in the insurance sector.A 2018 High Court of Delhi case—National Ethical Guidelines for Biomedical and Health Research Involving Human Participants in 201725. The guidelines affirm that social stigmatization and discrimination in education, employment, health care, and general insurance are probable consequences of the irresponsible use and sharing of genetic research. The guidelines also specify the need to obtain informed consent for secondary uses of genetic information and prescribe norms relating to the maintenance of data confidentiality, as well as pre- and post-test counseling. However, these guidelines do not legally bind the scientific community, serving instead as ethical guidelines.The Indian Council of Medical Research developed the Fundamental Principles of Research on the Human Genome issued in 2000 by the Bioethics Committee of the Council for Science and Technology addresses this topic, its normative strength is limited. The Principles state that “…regardless of their genetic characteristics, all individuals or groups of individuals are respected for their dignity and human rights, and that they are equal to one another and are not subjected to any form of discrimination.”26 The subsequently adopted government guidelines for human genome research—Ethical Guidelines for Human Genome/Gene Analysis Research (2001)—do not mention GD27.Japan does not have a law regulating discrimination based on personal genetic traits. While the 29. In 2019, a non-partisan group of lawmakers approved the draft outline of a Bill to promote responsible use of patients’ genetic information in disease treatment and to prevent GD30.However, recently, more active initiatives have emerged in Japan. In 2019, the Japanese insurance industry announced that guidelines banning life insurance companies from collecting or using personal genetic information for purposes related to coverage eligibility or premium calculations are being planned to prevent GDBill 1875 in 2013, entitled An Act Prohibiting Discrimination because of Genetic Information in Health Insurance Coverage31. The Bill was meant to prevent insurers from discriminating based on predictive genetic data in the context of both individual and group health insurance contracts. The National Ethical Guidelines for Health and Health-Related Research (2017) expressed concerns that “there is potential harm to research participants arising from the use of genetic information, such as stigmatization or discrimination.” This is why “researchers shall take special care to protect the privacy of participants and confidentiality of such information.”32The latest unsuccessful attempt to directly address GD in health insurance was the Senate Code of Practice for the provision of clinical and laboratory genetic services that have regulatory force from 2020 onwards33. In the Code, all licensed clinical laboratories providing laboratory genetic/genomic testing services are prohibited from disclosing genetic test results without consent to any third parties, including family members, although some limited exceptions may apply. However, insurance companies usually require such consent to be provided in application forms to purchase personal insurance . In biomedical research (such as the SG10K pilot study), the use of genetic technologies and information is mainly governed under the Personal Data Protection Act (PDPA) and Human Biomedical Research Act, which implements recommendations of the Bioethics Advisory Committee34.The Ministry of Health issued the Constitution of the Republic of Korea, the National Human Rights Commission of Korea Act proscribes discriminatory acts violating equality rights, including those based on disability or medical history (art. 23)35. However, genetic features are not explicitly included under the terms “disability” and “medical history” in the provision.Under the 36. Furthermore, the BSA contains a criminal sanction for those who discriminate based on genetic information with imprisonment up to 2 years or fines of up to 30 million KRW (art. 67-1-4)36.However, the BSA (2004), the first legislation to explicitly prohibit GD in Asia, seems to alleviate the above concerns. The Act stipulates that discrimination “on the grounds of genetic information in education, employment, promotion, insurance, or any other social activity” (art. 46-1) is illegal, and prohibits “compelling any person to undergo a genetic test or to submit the results of a genetic test” (art. 46-2)Human Subjects Research Act37 and the Basic Law of Indigenous Peoples38, Regulations of Consulting and Obtaining the Consent and Participation of Indigenous Peoples and Tribes have recently been implemented to avoid ethnically related stigmatization of Taiwanese aboriginal groups in the context of genetics studies and biomedical research39.GD, to some extent, is linked with the recognition of the indigenous rights of the aboriginal groups in Taiwan. Under the Constitution (art. 7) . However, when encountering certain cases, it is not appropriate to apply the Constitution directly without invoking a specific law. Protection against GD in the workplace may be available through a similarly broad interpretation of the Employment Services Act, which prohibits discrimination in employment based on several characteristics such as race, religion, and disability (art. 5)40. Nevertheless, GD is not explicitly addressed under the Act, and there are yet to be judicial decisions regarding GD under this legislative scheme.Otherwise, GD in Taiwan could be proscribed according to a broad interpretation of the equality right stipulated in the Enforcement Decree of the Personal Information Protection Act (PIPA) in South Korea, the PDPA in Taiwan, and the Data Privacy Act (DPA) of 2012 in the Philippines stipulate genetic data as sensitive personal data (Enforcement Decree of the PIPA art. 18(1), PDPA art. 2(1), DPA Sec. 3(t))43. In addition, in the reformed 2017 Act on the Protection of Personal Information in Japan, genetic information is interpreted as a personal identification code similar to sensitive information44. These are similar to the General Data Protection Regulation in the European Union that specifies special categories of personal data, including genetic data45. Although personal genetic data that can cause an infringement of the rights of the subject through data leakage is categorized as sensitive information or personal identification code in those laws, the laws require additional safeguards to process sensitive data. For instance, the laws provide specific security and safeguard measures for personal data controllers who process such sensitive personal data. They make it more difficult for personal genetic data to be used in a way that renders them vulnerable to stigmatization and discrimination, but these safeguards do not directly refer to GD. As the other example, there also has been no case to test the illicit use of genetic or health information presented to the Personal Information Protection Commission, which performs privacy impact assessment under the PIPA in South Korea46.Among the general privacy legislation, the Personal Information Security Specification (PISS)—the major digital privacy governmental guidelines in China—also protects individual genetic information, to some extent47. If personal biometric and health information can be a source of personal reputational damage or discrimination, it is categorized as personal sensitive information (PISS art. 3(2)). Compared to the data protection laws in South Korea and Taiwan that explicitly specify genetic data as sensitive personal data, PISS perceives genetic information as personal biometric and health information. PISS also includes more exceptions to confidentiality besides authorized consent, such as national security and national defense, public safety, public health, a criminal investigation, prosecution, trial, and judgment enforcement (PISS art. 5(4)).The 48. This is similarly the case in Hong Kong SAR under the Personal Data (Privacy) Ordinance49.Another type of genetic privacy governance can be seen in Japan and Singapore. In the PDPA in Singapore, genetic information is perceived as personal information only if it identifies an individual on its own or when combined with other informationRegulation of the People’s Republic of China on the Administration of Human Genetic Resources in 2019 provides for the respect of the privacy rights of human genetic resources providers in collecting, preserving, utilizing, and providing the resources (art. 9)50. Further, it prohibits the collection of human genetic resources of genetically related families without ethical approval issued by the national authorities (art. 11). If anyone violates the provision of the Regulation, the authorities and the Ministry of Science and Technology of China can order that the illegal act be stopped and illegally collected human genetic resources be confiscated, with the rule breakers paying a fine of up to five million yuan (art. 36).In the limited contexts of clinical or research testing, including biobanking, some legislation and regulations provide rules to protect the confidential nature of genetic information; however, those laws are also silent on the topic of GD. For instance, China’s Medical Service Act of 1962 (2019) provides a rule that prevents any disclosure or transmission of a patient’s data to a third party without the patient’s consent (art. 21-2(1))51. However, this does not apply to genetic data held outside clinical records51. The BSA protects the confidentiality of genetic information in medical records, preventing disclosure to anyone other than the patient (art. 46-3)36.In South Korea, the Human Biobank Management Act enacted in 201052. The Act establishes a privacy protection framework for the storage of, use of, and access to genetic samples and data contained therein. However, this legislative effort has not extended its protection to GD.The biobanks in Taiwan, which hold genetic data and samples from Taiwanese citizens, benefit from a governance structure set by the Constitution, which may afford its citizens some protection against GD, but this interpretation of the right to equality in relation to GD has not yet been tested in a court of law. Based on our findings, the protection against GD provided by policies and laws in the selected Asian jurisdictions varies significantly, from rather weak, partial, and uncertain, to relatively high (only in South Korea).The results of our review showcase the existing GD policy framework in selected Asian jurisdictions, raising important questions about their comprehensiveness and effectiveness. South Korea enforces the legislative prohibition of GD with criminal sanctions, but these have never been tested in a court of law. Case law in India invalidated a discriminatory clause about genetic disorders in health insurance, but the persisting ambiguity of parameters of deniability in insurance plans, adequate regulation of the use, and sharing of health data, including genetic information, remains challenging. The recent government guidelines in China prohibit some discriminatory practices involving the collection and use of genetic information only in the context of health insurance. Other surveyed jurisdictions have chosen instead to address the GD topic in nonbinding ethical guidelines in research . Taiwan has broadly formulated the right to equality in its This review also tested whether existing policies and laws focusing on data protection provides some protection against GD. Surveyed jurisdictions show different coverage of genetic privacy to some extent Table . The genHowever, these legal and regulatory data privacy laws have not succeeded, so far, in providing a sufficiently strong degree of genetic nondiscrimination protection to individuals who use genetic testing services in Asia. These frameworks usually include standard exemption clauses, such as “with the consent of the individual to whom the information belongs,” that limit the extent of protection they provide. Specifically, no surveyed jurisdiction explicitly delimited the lawful use of genetic information or alleviated public concerns surrounding compelled, unexpected, or unrecognized disclosures of genetic data that can result in discrimination.53, South Korea is the only Asian jurisdiction where GD in employment is punishable through a criminal sanction36. Specifically, there has been a reported case in pre-employment, Xie v. Human Resources and Social Security Bureau in Foshan City, adjudicated in mainland China. In 2009, 31 applicants to the Foshan local government were denied employment for being thalassemia gene carriers and three of them filed a lawsuit alleging discrimination54. However, in 2010, the Foshan Intermediate People’s Court ruled that it is legal to deny applicants who carry the thalassemia gene jobs in civil service55. Until now, there have been no guidelines regulating thalassemia genetic testing in pre-employment physical examination in the civil service as well as in state-owned companies. This situation has created some apprehension among job applicants, especially in southern China, where a high prevalence rate of α and β thalassemia carriers is reported (11%)56.The lack of laws or policies against discriminatory use of genetic information may raise substantial concerns beyond clinical or research contexts. Discrimination in the workplace owing to genetic traits would be one area of concern. Although the International Labour Organization already argued for the protection of people with a genetic predisposition to developing diseases in the workplace back in 2007Genetic Information Nondiscrimination Act (2008)57 in the US and the Genetic Nondiscrimination Act (2017)58 in Canada currently prohibit the acquisition and use of genetic information for employment with criminal sanctions. Amendments to the Canadian Labour Code and the Human Right Code appoint the filing of GD claims to designated governmental agencies59. In the United Kingdom, the Information Commissioner’s Office, which oversees the Data Protection Act, has issued the Employment Practices Code60. This code prohibits the use of genetic testing information to predict a worker’s health condition, along with requests for disclosure of a worker’s genetic test results (4.5.1 and 4.5.2.)60. The code allows for exceptions where the genetic condition of a worker is likely to pose a serious safety risk to other people, or where a specific working environment or practice might pose risks to workers who have certain genetic traits (4.5.3)60.Compared to this, both the 61. The Selective Benefit Coverage System under the National Health Insurance Act in South Korea has also covered the gene panel testing based on the next-genome sequencing for genetic disorders and cancers from 201962. Since 2019, Japan has covered 70–90% of fees of the newly introduced cancer gene panel tests for intractable cancer patients, who have not responded to surgery and anticancer drug treatment63. PhilHealth—the National Health Insurance Corporation of the Philippines—extended its coverage of neonatal screening from six disorders to 28 additional disorders in the expanded program, and the confirmatory tests have included genetic testing since 201864.In contrast to North America, concerns regarding GD in health insurance appear to be connected with universal health coverage in Asia. Japan, the Philippines, Hong Kong SAR, Singapore, South Korea, and Taiwan have adopted national health insurance systems in the form of compulsory social insurance since the latter half of the twentieth century. The systems have recently expanded their coverage range to reduce the financial burden of genetically at-risk individuals. In Taiwan, the tests subsidized by the national health insurance system are mainly for clinical diagnosis rather than early detection, which means only those with pre-existing symptoms of a genetic disease can receive a free diagnosis65. In particular, anxiety over GD in health insurance was considered low in the study, while GD in employment was a matter of concern for young participants.However, there remains limited evidence to identify the effectiveness of the public health coverage system against GD. From a 2019 national survey of public perception and knowledge of genomic research and medicine in Japan, 22% of respondents considered that the avoidance of discrimination in employment or insurance is important when personal genetic information is used in research and hospitals66. In Singapore, as the possibility of being denied insurance coverage is a common concern with genetic testing, the Life Insurance Association of Singapore declared in 2006 that its members would not require applicants to undergo genetic testing to obtain insurance coverage67. However, an important caveat to this policy statement is that individuals who are already aware of genetic test results relevant to their health must disclose this information to their insurer. There is currently no regulatory control over private insurers’ decision to extend insurance coverage, or not, to genetically at-risk individuals in the country. The recent move by the Japanese insurance industry to adopt GD guidelines is laudable, but independent monitoring and review of their implementation would provide added credibility to this initiative. A 2020 survey on the attitudes of insurance employees toward adverse selection and GD shows that it is necessary to improve understanding of GD through communication and consultation to prepare the guidelines for Japanese insurers68. As an example of a successful model, a 2008 agreement between the Association of Insurers, patient associations, and professional associations in the Netherlands led to the adoption of guidelines on mortality and morbidity risk determination owing to familial hypercholesterolemia. A 2012 tracking study confirmed that the guidelines facilitated access to life insurance, helped to reduce the negative perception about GD, and had been appropriately applied by insurers69.Furthermore, little is discussed of the need to develop national policies and agreements to prohibit GD in the life insurance or private health insurance in Asia. A 2013 survey of 99 Korean physicians in genomics provided some evidence that the legal protection against GD for people with genetic risks was insufficient in the private health insurance at that time (50.5%)74. A 1998 experts’ survey in China also showed that 64% of Chinese geneticists were aware of the lack of legal prohibition on discrimination based on disability75. Compared to this, Canada shows how the use of evidence of GD in a specific context, for example, that of Huntington’s disease76, as well as dynamic advocacy from patient organizations, can spark constructive debates surrounding the necessity of implementing legislation to prevent GD. In Canada, such debates led to the creation of the Canadian Coalition for Genetic Fairness and the adoption of Act S-201 (GNDA) to prevent GD, even outside the context of Huntington’s disease. Information presented in this context—in addition to the Huntington studies—included privacy surveys78, economic modeling of anti-selection risks in the event of a prohibition on the use of genetic results79, and research related to the life insurance industry practices80.Disability discrimination related to genetic traits remains a great concern in some Asian jurisdictions. When associated with the negative social perception identified in the Philippines and India toward patients with disabilities and rare diseases, such findings could reflect a trend toward stigmatization and marginalization of carriers of genetic diseases, which could be interpreted as GD81. In a 2012 public survey of 1000 participants, 71.9% expressed that they would refuse to marry someone with genetic factors rendering them “high risk” for developing cancer82. In the aforementioned 1998 Chinese experts’ survey, a majority of Chinese geneticists agreed on the following: partners should know each other’s genetic status before marriage (92%); carriers of the same defective gene should not marry one another (91%); women should have prenatal diagnosis if medically indicated (91%); and governments should require premarital carrier tests (86%)75. Conversely, GD does not appear to be a significant concern when considering marriage or starting a family in North America and Europe, outside of specific population groups that are genetically at risk of developing a serious illness85.In addition, this review found some noticeable differences between the perception of GD in Asia and that in North America and Europe. In Asia, serious concerns exist regarding the possible use of genetic information when considering potential spouses for marriage and starting a family. In the 2017 Japanese public survey, the respondents were largely concerned about suffering potential disadvantage in the context of marriage and pregnancy (41.0%), employment (37.6%), and insurance (43.8%) owing to their genetic characteristics86. We encourage researchers to further document GD concerns and incidents in Asia using consistent, recognized, quantitative, and qualitative research methodologies. Third, we limited our comments on laws, policies, and practices by focusing on seven Asian jurisdictions. GD seems to have become a concern that merits policymakers’ attention in these jurisdictions, but this phenomenon could develop differently in other Asian regions.This GD review is limited by several factors. First, our comparative assessment used only two approaches—the GD-specific frameworks and genetic privacy—so might not adequately identify more general concerns about GD in this region. A more broadly framed assessment would enable researchers and policymakers to understand and recognize how their nations provide some degree of protection from GD to their citizens with or without having to specifically regulate GD. Second, this review cannot be used to support or refute the view that incidents and concerns about GD are substantial in Asia. The reports on GD that we reviewed generally did not have a robust methodology or conveyed little evidence. This situation is not unique to Asia, as documenting GD has also been a struggle globallyOur article aims to provide a comprehensive portrayal of how seven Asian jurisdictions have sought to tackle the challenge of GD. It provides an overview of laws, regulations, and policies adopted in these jurisdictions to address this complex issue. Unlike North America and Europe, where the GD debate was in full force by the late 1990s, in Asia, GD has generated fewer discussions to date reflecting both similar and distinct concerns from those observed in the western hemisphere.We found that GD has called forth a variety of legal and political responses in Asia. However, this topic requires further investigation in order for countries in this region to be able to develop the kind of multilevel, nuanced, anti-GD policies required to account for different social, ethical, and economic factors. More specifically, the jurisdictions we reviewed have not yet succeeded in specifying the lawful use of genetic information or alleviating public concerns surrounding compelled, unexpected, or unrecognized disclosures of genetic data that can lead to discrimination.For Asia to truly benefit from its innovative surge in genomics and precision medicine, such progress needs to be matched by a corresponding investment in addressing ELSI raised by genomics, predominant among which is GD. Our transnational collaboration could be the start of a promising regional discussion to ensure that issues are sufficiently presented and addressed in Asia. This would in turn drive the debate forward at the national level, thus promoting greater solidarity and fostering much-needed trust in science in this region.


This research empirically analyzes the psychological impact of smog pollution on investors. Results indicate that smog pollution has negative impact on investor sentiment which is weakened by the positive tone in media reporting. Empirical evidence for the impact of smog pollution on investor sentiment and the related moderating role of media tone is presented in this study. Smog pollution not only affects people’s physical health , as wellThe original contributions of this study include three aspects. First, instead of previous studies that focus on changes in investor sentiment as climatic factors that influence financial markets, or studies that use air quality as an indirect proxy variable for investor sentiment, this study focuses on the influence of smog pollution on investor emotion on the fluctuate of stock markets. Second, while prior studies prove that markets react to pollution events , this stTo define investor sentiment, some studies used stock market mispricing  and the In the external environment context of our fucus topic, some scholars suggested macroeconomic development expectations such as macroeconomic conditions and monetary policy expectations affected investor sentiment . AnotherAir pollution increases the negative sentiments among investors, thus triggering more conservative investment-related activities . This stFirstly, smog pollution affects the development of companies. For instance, some studies found that smog pollution affects the debt financing ability , obtainiSecondly, information disclosed about smog pollution affects investors’ perceptions. According to the attentional resources theory, investor attention tends to be limited. For instance, investors may not fully respond to all market information due to intervening variables  such as H1: Smog pollution in the areas that companies located in is negatively influent investor sentiment.For many small and medium investors in China’s capital market, media concerning is important channels for obtaining market information. For instance, media reports reveal a company’s operating status, improving the efficiency of company information distribution and dissemination, and help investors access “soft information” to capture a company’s fundamentals . When coH2: Media tone has inhibitory effects on the negative relationships between smog pollution and investor sentiment.1 is expected to be significantly negative, supporting Hypothesis 1. In Model (2), the intersection items of smog pollution and media tone  is added to test the moderating effects of media tone on the relationships between smog pollution and investor sentiment. β1 is expected to be significantly negative and β3 is expected to be significantly positive, thus verifying Hypothesis 2.Base on the related literature , this stISi,t). Following f), the residual, Qr = Q–Qf, is used to measure mispricing due to investor sentiment (IS).The dependent variable is investor sentiment , which is based on the air quality monitoring data released by the China National Environmental Monitoring Centre (CNEMC). Considering the hysteretic effect of air pollution on the human body, this paper uses the first-order lag term of PM2.5 as the independent variable.The independent variable is smog pollution (TONES) is delayed by one period. The measure of media tone is derived from the overall sentiment score indicator in the news and the social media quantitative public opinion database. Using text analysis technology, the media tone indicator is scored after analyzing the news published by 327 important newspapers. To provide continuous and comparable sentiment scores for media reports, text with a positive tone are assigned positive scores while negative scores are assigned to texts with a negative tone. Finally, the aggregated sentiment scores of media report items are matched with the most relevant listed companies mentioned in the reports. The daily data of the news sentiment tendency score in the database is summarized by company, and the media tone annual index is obtained.Because of the lag in media news reports, media tone (SOE), enterprise scale (SIZE), years of establishment (AGE); company profitability and management activities, including profit margin on net assets (ROE), asset-liability ratio (LEV), future development opportunities (GROW), operating net cash flow (CRAA), capital investment (INV), management expense ratio (ADM), audit costs (DCOST), and proportion of independent directors (IDRATIO). In this scenario, major shareholders, institutional investors, and management staff can have information advantages outside and inside the enterprise. Thus, their shareholding ratio may affect the judgment of investors. Understandably, control variables also include the shareholding ratio of the largest shareholder (SHRCRL), the institutional shareholding ratio (INHOLDS), and the executive shareholding ratio (MAHARE). Moreover, the characteristics and performance of company stocks can also affect investor sentiment. Thus, four typical control variables and included: earnings per share (EPS), book value per share (BPS), book to market value (BTM), and stock liquidity (TRADE). In addition, the control variables include year and industry dummy variables. From related studies , a serieThe financial data for all listed companies in this study is obtained from the China Stock Markets and Accounting Research (CSMAR) database, while the urban smog concentration monitoring data of 74 key cities in China from 2013 to 2017 is obtained from the CNEMC. In this dataset, the data between 2013 and 2017 have higher internal consistency because the Science and Technology Innovation Board was established in China in 2018, The data on news reporting tone is derived from the News and Social Media Quantitative Public Opinion Data published by the Chinese University of Hong Kong. To calculate relevant variables, we exclude companies with missing data and companies in the financial industry, as well as companies located in cities that do not disclose smog monitoring data, and. The final sample contains 5704 observations.IS) is -0.192, indicating that investors have a negative view of a company. The average value of PM2.5 is 52.21 g/μ3, which is on the second level for air quality (35–75 g/μ3) as stipulated in China, while the maximum value of 130 g/μ3 is on the fourth level (115–150 g/μ3), which indicates moderate smog pollution. The average value of media tone (TONES) is 23.61, indicating that the media’s news reporting tone of all the companies in the sample is generally positive.PM2.5 is -8.9972 10–3, which is significant at the 5% level and indicates a negative impact of smog pollution on investor sentiment. In Model (2), the coefficient of TONES is calculated as 1.703×10––51, which is significant at the 1% level, indicating that a more positive media tone can create a more positive investor sentiment. The regression results of the interaction of PM2.5 and TONES in Model (2) show that the coefficient of PM2.5×TONES is 0.287×10–4, which is significant at the 5% level. Because the coefficient of PM2.5 in Model (1) is significantly negative and the coefficient of PM2.5×TONES in Model (2) is significantly positive, it can be concluded that media tone inhibits the negative influence of smog pollution on investor sentiment. Therefore, Hypotheses 1 and 2 are supported.Empirical results indicate that investors have certain negative expectations for listed companies and underestimate the value of listed company located in smog- polluted areas. However, a positive tone from media can inhibit the negative investor expectations caused by smog pollution. These results are consistent with the literature on smog pollution in terms of firm value loss  from negVarious methods were used to ensure robustness in the conclusions of this study.10 to replace PM2.5 as the alternative proxy variable for smog pollution to test Hypothesis 1 and Hypothesis 2. Resulting estimates show the robustness of Hypotheses 1 and 2 is supported.To prove that the selection of the independent variable index does not affect the research conclusion, we apply PMTo prove that the selection of the dependent variable index does not affect the research conclusion, we apply the method of To prove that the selection of the moderating variable index does not affect the research conclusion, we apply a weighted adjustment of the original probability of the overall sentiment orientation of the news as the alternative proxy variable for media tone. The estimation results indicate that the two hypotheses in this study are robust.This study on the psychological impact of smog pollution on investors finds that smog pollution in a company’s location can negatively affect investor sentiment, and that the tone of related news reportage can weaken this negative impact. This study highlights the effects of changes in the external natural environment on investor sentiment and is significant in maintaining the sustainable development of financial markets.The findings point toward the following policy implications: (1) As the main governing body in smog pollution regulation and as the micro-subjects affected by smog pollution, companies should take the initiative in bearing the social responsibilities of pollution prevention and ecological protection. (2) Investors should take a rational, guiding role in the tone of related news reporting, and (3) regulatory authorities should guard against the expansion of the negative tone in news reports.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.This study was conducted in accordance with the recommendations of the Ethics Committee of the Beijing University of Chemical Technology.BL and YW performed the initial analyses and wrote the manuscript. YL and TZ assisted in the data collection, data analysis, and textual modifications. All authors revised and approved the submitted version of the manuscript.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Mexico is a rich source for anthropological and population genetic studies with high diversity in ethnic and linguistic groups. The country witnessed the rise and fall of major civilizations, including the Maya and Aztec, but resulting from European colonization, the population landscape has dramatically changed. Today, the majority of Mexicans do not identify themselves as Indigenous but as admixed, and appear to have very little in common with their pre-Columbian predecessors. However, when the maternally inherited mitochondrial (mt)DNA is investigated in the modern Mexican population, this is not the case. Control region sequences of 2021 samples deriving from all over the country revealed an overwhelming Indigenous American legacy, with almost 90% of mtDNAs belonging to the four major pan-American haplogroups A2, B2, C1, and D1. This finding supports a very low European contribution to the Mexican gene pool by female colonizers and confirms the effectiveness of employing uniparental markers as a tool to reconstruct a country’s history. In addition, the distinct frequency and dispersal patterns of Indigenous American and West Eurasian clades highlight the benefit such large and country-wide databases provide for studying the impact of colonialism from a female perspective and population stratification. The importance of geographical database subsets not only for forensic application is clearly demonstrated. In cultural terms, Mexico is part of Mesoamerica in its center and South and the Greater Southwest and Aridoamerica in its North [Mexico, officially the United Mexican States, is the northernmost Central and southernmost North American country. It covers more than 1.9 million kmts North ,4,5,6,7.ts North ,9. Mesoats North ,11,12. Tts North ,5,13,14.ts North . Populatts North . While its North ,17. Mixets North . The Victs North ,19. The ts North . The meats North . Since cts North ,13,18,20ts North . Howeverts North  and undets North ,14. Recets North . Considets North . One clats North . For thets North ,27,28,29ts North ,31,32, tts North ,34. Prevts North ,36), mtDts North  , ancts North ,39, datats North ,40) wherts North , US Mexits North ), did nots North ), or a cts North . Furtherts North . To addrn = 567), Center , and South  , except for the South . The country-wide Western and Eastern datasets contained 1377 and 521 individuals, respectively. A further 123 Mexican individuals were included in the study whose TMA could not be assigned to a specific state for the lack of information or recurrent toponyms . All experimental procedures and individual written informed consent, obtained from all donors, were reviewed and approved by the Western Institutional Review Board, Olympia, Washington (USA). Ethno-linguistic affiliation of donors was not recorded. Only individuals where genealogical investigation revealed a terminal maternal ancestor (TMA) from Mexico and unrelatedness were included. The sample set covers all 32 administrative units of Mexico , except the easternmost state of Quintana Roo. Sample numbers range from 3 to 511, with a mean and median of 61 and 36, respectively . For intn = 709) . The mactoponyms .https://empop.online (accessed on 16 August 2021)) [mt, build 17 [Total DNA was extracted using the QIAamp DNA Blood Maxi Kit  according to the manufacturer’s recommendations and stored at −20 °C. The entire control region  was amplified and sequenced as described in . Sequenct 2021)) . The SAMt 2021))  implement 2021))  accordinbuild 17 . SAM2 dobuild 17 . The exaForensic and population genetic molecular statistics and diversity indices were calculated from the complete haplotypes using Arlequin v3.5.1.2  disregarAll 2021 haplotypes passed quality control and will be uploaded on the forensic online mtDNA population database EMPOP with reference numbers EMP000849-852 . They coThe dataset contained 799 different haplotypes in the range nps 16000–16569 1–580, 502 of which (62.8%) were unique, comprising 26.1% of the included individuals. An RMP of 0.5% was calculated, PD reached 99.6%, and DC was 41.5%. The mean number of pairwise differences between two randomly chosen haplotypes  was 14.995 ± 6.709, resulting in a nucleotide diversity (π) of 0.013 ± 0.006 over the 1168 nps. The most frequent haplotype was 16223T 16290T 16319A 16362C 64T 73G 146C 153G 182T 235G 263G 315.1C 523del 524del, relative to the rCRS , belongeThe dataset revealed a wide-ranging spectrum of mtDNA haplogroups present in the general country-wide Mexican population .mt build 17 [n = 103; 5.1%) and C1b*   of PhyloTreebuild 17  accordin9; 4.9%) . Differe9; 4.9%) .The high number of sub-lineages  likely rThe observed proportions of lineages in this dataset of modern cosmopolitan Mexicans are well in line with reports of self-estimated ancestry as well as mitogenetic data from various admixed population samples that revealed an extremely high Indigenous American prevalence  with soThe spatial distribution of both Indigenous American and West Eurasian haplogroup clusters within Mexico was found to be strikingly heterogeneous. The Indigenous lineages showed peculiar and specific patterns along the country. Haplogroup A2 (41.8%) was most frequent in the South, where it comprised 57.0% of individuals, and showed a gradient via the Center (43.6%) to the North (22.2%). Always, the Eastern regions had a higher percentage than the Western parts. Haplogroup C1 (23.7%) presented the opposite pattern: it was particularly frequent in the Northwest (49.5%) and revealed a decline from the North (38.8%) to the Center (16.1%) and South (17.8%) .Both clines were also observed in Indigenous and admixed populations when analyzed at a country-wide scale ,17,18,62It has been postulated for Indigenous populations of Mexico and beyond that geography rather than linguistic classification predicts mtDNA structure and diversity, suggesting that genetic divergence predates linguistic diversification ,10,36,63n = 545), Central (n = 589), and Southern (n = 679) haplotypes were analyzed, all surpassing the minimum size considered necessary for a forensic mtDNA database [ST revealed highest differentiation between North and South (1.48|0.094) and lowest between Center and South (0.19|0.014), while the results for the comparison Center-North were intermediate (0.79|0.049), respectively  at the other pole. Analysis of the contribution of variables to the first and second PC revealed clear haplogroup structuring. In particular, the second PC separated the Indigenous American from the West Eurasian clades, causing the outgroup position of the Northeastern state group of Nuevo León-Tamaulipas in the PCA . The findatabase  . This study provides for the first time a comprehensive overview on the mtDNA variation in the modern general population of Mexico using a large sample covering the entire country. The findings confirm that the genetic impact of European conquest was small in terms of maternal lineage introgression, and the changes in population structure that followed have likely not substantially changed the ample pre-Columbian pattern of mtDNA variation in Mexico. The mitogenetic structure of the general population is still mainly Indigenous. The proportion of West Eurasian mtDNA lineages in the population was found to be low, but with some exceptions, mainly restricted to the Northeast, where the West Eurasian component yielded high proportions. More detailed analyses argued pro regional databases at least for forensic genetic investigations. MtDNA is just one genetic marker and, obviously, only a synopsis of all genomic data will fully open the window into the past. This study has also provided insights of a possible sex-differentiated mobility and mixture that impacted cultural as well as biological survival in Mexico, as well as in other countries. The specific insights from the maternal contribution have proven their importance for genetic and anthropological studies aiming to reconstruct and date human settlement. No mitogenome is inherited independently from the nuclear genome. Therefore, this dataset can also help to provide general ancestry estimates for the cosmopolitan population to avoid bias. Understanding population stratification, genetic contributions, and their association with medical traits in admixed populations and individuals is highly relevant to biomedical research and personalized medicine ,74,77,78


Microtus oeconomus, a species preferring moist habitats. We intended to check what barriers and what distances affected its genetic isolation on a local scale. The study was conducted in the area of Kampinoski National Park in central Poland (Europe). DNA variability of 218 root vole individuals was assessed by genotyping nine microsatellite loci. Despite its spatial fragmentation, the studied population did not seem to be highly structured, and isolation through distance was the main differentiating factor. Even a distance of several kilometres of unfavourable natural habitats and unfavourable terrain did not exclude the exchange of genes between subpopulations. Our results suggest that the genetic effects of the fragmentation of wetlands have been considerably compensated (delayed) due to the migratory abilities of this species. Our study does not provide clear results on the impact of anthropogenic barriers but suggests that such barriers may have a much stronger effect than natural barriers.Climate-related changes have a severe impact on wetland ecosystems and pose a serious challenge for wetland-dependent animals as their preferred habitats decline, lose spatial continuity, and appear as isolated islands in the landscape. In this paper, we studied the effects of long-term habitat changes (drying out and fragmentation of wet non-forest habitats) on the genetic structure of the population of the root vole  Climate-related changes have a severe impact on natural ecosystems, including wetlands. Wetlands have already suffered greatly in recent decades due to widespread drainage and other anthropogenic changes; therefore, they are more susceptible to climate warming ,2. As thWarming-related changes pose a serious challenge for wetland-dependent animals, as their preferred habitats decline, lose spatial continuity, and appear as isolated islands in the landscape ,3. In a Microtus oeconomus   has ofteconomus ,16,17,18f Poland . Geograpf Poland ,21. It hf Poland ,23. HoweIn order to study the local effects of the drying up, fragmentation, and isolation of wetlands on the genetic structure of species associated with these habitats, it is important to select appropriate study sites and methods. Nowadays, molecular methods are valuable tools for the analysis of the impact of habitat fragmentation on populations. The most widely used molecular markers are microsatellites  due to their high polymorphism and possibility of amplification from a small amount of tissue ,24,25. AThe aim of this paper was to assess the effects of the decline and fragmentation of wetland habitats on the genetic structure of the root vole population. Using microsatellites as molecular markers, we expected to define genetically distinct subpopulations on a local scale, determine the distances between patches of wetlands that restrict gene flow, and indicate the main barriers  hindering the migration of the studied species.2. There is a mosaic of forest and non-forest habitats in this area, and wet habitats (required for the root vole) are separated by contrasting dry habitats formed on belts of postglacial sand dunes covered mainly by pine forests (Peucedano-Pinetum and Leucobryo-Pinetum). In addition, there are anthropogenic elements , but their share is small in this area. Samples were collected in the area of Kampinoski National Park (coordinates: 52°19′ N 20°34′ E) situated in central Poland near Warsaw. The park covers an area of 385.44 kmThe habitat changes and the basic abiotic parameters of this area have been documented over the last few decades, especially since the National Park was established in 1959 . One of Samples were collected from 10 sites: 5 from the northern swampy belt and 5 from the southern swampy belt. Two sampling sites in the western and eastern part and one site in the central part of each swampy belt were chosen . The disThe northern swampy belt with the W, B, Z, K, and M study sites is bordered by wide dune belts covered by coniferous forest in the north and south. It adjoins the Vistula River valley and an urbanised area in the east and reaches an agricultural area and the Bzura River valley in the west. The southern swampy belt with the A, P, G, L, and X study sites is bordered by a dune belt with a coniferous forest in the north and by a forest and infrastructure of the city of Warsaw in the east. The other parts of the belt adjoin agricultural areas and scattered buildings. In the southern swampy belt, the relative continuity of sedge habitats is maintained only in the western part, and there are several small and isolated open areas covered by sedges in the central and eastern part. Root voles were captured in patches of sedge communities. A transect of 50 wooden live traps spaced at 20-m intervals was set within each patch. The trapping lasted for 2–4 consecutive days and all captured rodents were identified to the species, sexed, and weighed. When the rodent was identified as a root vole, a tissue sample for DNA analysis was taken by toe clipping (permission for this procedure was obtained from the ethics committee). The clipped toes were transferred into sterile Eppendorf tubes and dried using silica gel. Captured individuals were released in the site of capture. Trapping sessions were carried out in the autumn (from October to November) of 2008 and 2009. Based on previous studies , it is kDNA from dried toes was isolated with chelex resin pursuant to the protocol proposed by Larbi . The cona), allelic richness (AR), observed (Ho) and expected (He) heterozygosity, and the inbreeding coefficient (Gis) for all subpopulations. Pairwise differentiation based on G’st [Standard genetic diversity indices were estimated with Genodive v. 3.02  and FSTA on G’st  between Deviations from the Hardy–Weinberg equilibrium (HWE) were examined for each population calculating the inbreeding coefficient (Gis) with GenoDive and GenAlEx v. 6.5 . The sigst/(1 − Gst) between the groups. A genetic tree based on G’st matrices [To test the proportion of genetic variance between individuals and subpopulations, we performed an Analysis of Molecular Variance (AMOVA) for standardized data  implemenmatrices  and an Umatrices  was consmatrices .6 Markov chain Monte Carlo (MCMC) iterations and a burn-in period of 105. Admixture and correlation models were implemented. Structure Harvester was used to assess the optimal value of K for this study . The ray Evanno .The number of alleles per locus ranged from 8 (MM5) to 13 (Ma54), with an average of 9.78. The average allelic richness and effective number of alleles was 6.47 and 4.16, respectively. Allelic richness (AR) across subpopulations ranged from 5.40 to 6.47, with the highest in deme B and the lowest in deme K. Two private alleles were found: in deme M  and in deme P . We found a significant heterozygosity deficit in 8 out of 9 loci (with the Bonferroni correction for multiple tests). Due to a lack of evidence for null alleles or linkage disequilibrium, all loci were kept for further analyses. The PI analysis showed that using two markers  and fiver markers  reduces the likelihood of finding two individuals with the same genotype for the certain loci below 1% . p-value < 0.001). Pairwise genetic differentiation was significant in most of the comparisons, except in pairs B-Z and G-Z . The highest G’st values were observed for P and K  of the total variance. Most of the variability occurred within the sites. The AMOVA indicated that the variance among subpopulations accounted for only 4%  decline in the groundwater level and the increasing forest cover (described in detail in ). In genIn contrast to our predictions, the main division of the vole population in the study area did not run along the north–south line (between the two swamp belts) but along the east–west line. This means that the belts of dry glacial dunes covered by pine forest do not constitute a significant barrier to gene exchange between populations. It should be noted here that the differences in the relative height between the tops of the dunes and the almost flat areas of the swamp belts are up to 30 m, and the width of these belts is usually more than two kilometres. Therefore, it was assumed that migration, at least theoretically, should be limited not only by the unfavourable habitat (dry pine forest) but also by the topography of the area. The ecological literature provides no data indicating natural terrestrial ecosystems that may constitute significant barriers to the migration of voles. Kelemen et al.  showed tThe isolating effect of anthropogenic barriers on the genetic structure of the root voles cannot be unequivocally demonstrated, because the Kampinoski National Park has only a few elements of this type. Only one large asphalt road  runs through the park, with an average traffic of about 6000 vehicles per day. This road divides the park into the eastern and western parts, and a similar division was exhibited by the genetic structure of the studied vole population. It has been emphasised in many studies that roads can be perceived as a barrier to rodent movement ,64,65,66Rural buildings are another type of a potential anthropogenic barrier in the park. Our research has shown that subpopulations A and P show clear genetic differences from subpopulation W, and these differences cannot be explained by geographic distance. These subpopulations are divided by a line of rural buildings, but the infrastructure  is scattered and does not constitute a tight barrier. The role of the barrier effect of such rural infrastructure can be reinforced by companion animals, i.e., cats and dogs, which can prey on voles migrating through the building line ,68. Can The above-described ability of the root vole to overcome even wide and unfavourable terrestrial natural barriers certainly does not mean that the drying out of the park’s wetlands will not have any negative consequences for this species. Changes in the surface of the wet habitats and their fragmentation will certainly affect the range and abundance of the species, which was previously pointed out by the author of . HabitatDespite the spatial fragmentation, the studied population turned out to be poorly genetically diverse, and the isolation through distance was the main differentiating factor. Natural habitats and topography  did not constitute a significant barrier to gene exchange between root vole subpopulations. As a result, the genetic effects of the fragmentation of wetlands in the Kampinoski National Park were substantially compensated by the migratory abilities of this species. Our study does not provide clear results on the impact of anthropogenic barriers but suggests that such barriers may have a much stronger effect than natural barriers.