Skip to main content
NCBI home page
PLOS One logoLink to PLOS One
. 2020 Jan 31;15(1):e0228420. doi: 10.1371/journal.pone.0228420

Is the lady's-slipper orchid (Cypripedium calceolus) likely to shortly become extinct in Europe?—Insights based on ecological niche modelling

Marta Kolanowska 1,2,*, Anna Jakubska-Busse 3
Editor: Jana Müllerová4
PMCID: PMC6993984  PMID: 32004339

Abstract

Lady's-slipper orchid (Cypripedium calceolus) is considered an endangered species in most countries within its geographical range. The main reason for the decline in the number of populations of this species in Europe is habitat destruction. In this paper the ecological niche modelling approach was used to estimate the effect of future climate change on the area of niches suitable for C. calceolus. Predictions of the extent of the potential range of this species in 2070 were made using climate projections obtained from the Community Climate System Model for four representative concentration pathways: rcp2.6, rcp4.5, rcp6.0 and rcp8.5. According to these analyses all the scenarios of future climate change will result in the total area of niches suitable for C. calceolus decreasing. Considering areas characterized by a suitability of at least 0.4 the loss of habitat will vary between ca. 30% and 63%. The highest habitat loss of ca. 63% is predicted to occur in scenario rcp 8.5. Surprisingly, in the most damaging rcp 8.5 prediction the highest overlap between potential range of C. calceolus and its pollinators will be observed and in all other scenarios some pollinators will be available for this species in various geographical regions. Based on these results at least two approaches should be implemented to improve the chances of survival of C. calceolus. In view of the unavoidable loss of suitable habitats in numerous European regions, conservation activities should be intensified in areas where this species will still have suitable niches in the next 50 years. In addition, for C. calceolus ex-situ activities should be greatly increased so that it can be re-introduced in the remaining suitable areas.

Introduction

Various statistical models are used to predict the spatial distribution of plant and animal species based on presence-only data [1–4]. This approach is also used in many conservation studies [5–6] such as evaluating the distributions or areas suitable for conservation [6–7] and identifying priority areas for conservation [8]. Unfortunately, species distribution models are rarely used for research on the largest angiosperm plant family, the Orchidaceae. Orchids are one of the most threatened groups as their complex life history make them particularly vulnerable to the effects of global environmental change [9–10].

Cypripedium calceolus is one of the most intensively studied European plants [11–20]. It is the only slipper orchid in Europe—just one additional species, Cypripedium macranthos, is found in Belarus. The geographical range of C. calceolus is relatively broad and includes Europe (except the extreme north and south), the Crimea, Mediterranean, Asia Minor, western and eastern Siberia, Far East of Russia and south of Sakhalin Island [21–23]. Lady's-slipper orchid used to be more widespread in Europe, but the number of its populations declined in the 19th century due to the over-collection for horticulture and habitat degradation [24].

Nowadays C. calceolus is considered as endangered in most countries within its range [21] is listed in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) and also in Annex II of the Habitats Directive and under Appendix I of the Convention on the Conservation of European Wildlife and Natural Habitats (Bern Convention) [18] [23] [25].

Natural populations of this slipper orchid are included in Natura 2000 sites and other types of protected areas. This plant is also included on several national red lists and red data books as threatened [23]. In many countries this taxon is extremely rare, critically endangered and/or regionally extinct [19] [21] [23] and in others it is classified as Endangered (e.g. Croatia, Czech Republic, Hungary, Russia and Spain) or Vulnerable (e.g. Austria, Belarus, Denmark, France, Germany, Lithuania, Slovakia and Switzerland [23] [26]. Noteworthy, C. calceolus is a differential species for the unique Polish Kashubian region plant community—Fagus sylvatica-Cypripedium calceolus [27].

In the last century a significant decline in the number of populations of this species was recorded in almost all of Europe. This is due to many reasons, above all habitat destruction, especially expansion of agriculture, inappropriate forest management such as clearcutting, widespread use of herbicides and pesticides, equipment that can severely compact the soil, road and trail construction and collecting [23]. In addition, according to Rankou, & Bilz [23] browsing and grazing can pose a threat in two different ways: overgrazing affects individuals whereas the abandonment of traditional grazing leads to natural succession and therefore an increase in competition for this orchid. The replacement of natural forest with spruce plantations has caused habitat degradation as the soil is de-calcified and this species is linked to calcareous soils [23].

Climate change, especially the lack of rainfall and dry seasons, as well as the fires recorded in recent years in almost all regions of Europe may be responsible for the decline in the number of specimens in natural populations of C. calceolus. Currently, numerous (sub)populations of this species in various regions of Europe are fragmented remnants and genetically isolated. This raises the question—what is the future of this orchid? Is this species becoming extinct before our eyes? For instance, the dramatic decline of C. calceolus populations in Lower Silesia (SW Poland) was recorded and documented for over 100 years [28–30]. Among the 30 localities of this species, 12 were listed after 1945, in 2012 only 9 of them were confirmed [31], however, in 2019 only 7 were confirmed.

Furthermore, the occurrence of C. calceolus may be limited in the future by extinction or modification of the geographical ranges or ecology of its pollinators. While the lady's-slipper orchid is self-compatible, insects are required to transfer pollen to the stigma [32] as the position of the stigma and anthers prevent self-pollination [33]. Recent studies indicated that global warming can disturb the pollination of other European orchid, Ophrys sphegodes, which is pollinated by Andrena nigroaenea [34]. Noteworthy, Andrena bees are also one of the most important pollinators of C. calceolus. Undoubtedly, reproduction success is crucial for the long-term existence of the surviving populations [35].

The aim of this study was to evaluate the predicted effect of global warming on the distribution and coverage of the ecological niches that are currently suitable for C. calceolus in Europe as well as to estimate the impact of the climate changes on the availability of its pollinators.

Materials & methods

List of localities

The database of C. calceolus localities was compiled based on information in public facilities (e.g. GBIF, ukrbin.com, tela-botanica.org, iNaturalist, redbook.minpriroda.gov.by, WildSlovenia, Portale della Flora d'Italia, naturamediterraneo.com), published articles and books [21] [24] [36–49], conservation reports (FAO, Berne Convention Resolution 6, Krajowy plan ochrony gatunku obuwik pospolity; [50]) and field observations made by Jakubska-Busse. While identification of numerous orchid species requires taxonomic skills and experience and for such taxa using information derived from public databases is not recommended, C. calceolus is the most spectacular terrestrial orchid in Europe which can be easily recognized even by amateur naturalists.

The list of C. calceolus pollinators was compiled based on available literature data [15] [18] [24] [51–55]. Information about distribution of 21 from a total of 24 reported insect species was gathered from GBIF. Due to the lack of sufficient, precise information about distribution of Musca autumnalis, Andrena fulvicrus and Andrena ovina, these species were not included in the analyses. Pollinators of C. calceolus belong mostly to Hymenoptera, but two, Chrysotoxum festivum and Syrphus ribesii, represent Syrphidae, Diptera. Nomada panzeri is classified within Nomadinae, and Colletes cunicularius within Colletidae. Seven species, Halictus tumularum, Lasioglossum albipes, L. calceatum, L. fratellum, L. fulvicorne, L. morio, and L. quadrinotatum belong to Halictidae. The highest number of pollinators represent genus Andena (Andrenidae)–Andrena carantonica, A. cineraria, A. flavipes, A. fucata, A. haemorrhoa, A. helvola, A. nigroaenea, A. praecox, A. scotica, A. tibialis.

Ecological niche modelling

The ecological niche modelling was done using the maximum entropy method in MaxEnt version 3.3.2 [56–58] based on presence-only observations of this species. From the total of 932 locations of C. calceolus gathered during the study (Fig 1, S1 Table) the duplicate presence records (records within the same grid cell) were removed using MaxEnt. Considering pollinators input data, due to the various coordinate precision used in public databases only records georeferenced with the precision of at least 2 km were used to guarantee correct location of the observation in the grid cell. For data thinning and to minimize geographical overrepresentation of the samples, the initial catalogue was then reduced to include only records distanced one from another for at least 10 km and again the duplicate presence records (records within the same grid cell) were removed using MaxEnt. The final database included 519 localities of Chrysotoxum festivum, 2040 of Syrphus ribesii, 739 of Nomada panzeri, 621 of Colletes cunicularius, 1004 of Halictus tumularum, 1151 of Lasioglossum albipes, 1122 of L. calceatum, 940 of L. fratellum, 469 of L. fulvicorne, 699 of L. morio, 123 of L. quadrinotatum, 273 of Andrena carantonica, 1201 of A. cineraria, 325 of A. flavipes, 731 of A. fucata, 1477 of A. haemorrhoa, 1258 of A. helvola, 694 of A. nigroaenea, 486 of A. praecox, 421 of A. scotica, and 189 of A. tibialis (Figs 2–4; S2 Table).

Fig 1. Localities of C. calceolus georeferenced in this study.

Fig 1

Open in a new tab

Map was generated in ArcGis 9.3 (ESRI, 2006).

Fig 2. Localities of pollinators georeferenced in this study.

Fig 2

Open in a new tab

Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), and Andrena tibialis (J) georeferenced in this study. Map was generated in ArcGis 9.3 (ESRI, 2006).

Fig 4. Localities of pollinators georeferenced in this study.

Fig 4

Open in a new tab

Chrysotoxum festivum (A), Colletes cunicularius (B), Halictus tumulorum (C), Nomada panzeri (D), Syrphus ribesii (E) georeferenced in this study. Map was generated in ArcGis 9.3 (ESRI, 2006).

Fig 3. Localities of pollinators georeferenced in this study.

Fig 3

Open in a new tab

Lasioglossum albipes (A), Lasioglossum calceatum (B), Lasioglossum fratellum (C), Lasioglossum fulvicorne (D), Lasioglossum morio (E), Lasioglossum quadrinotatum (F) georeferenced in this study. Map was generated in ArcGis 9.3 (ESRI, 2006).

For the modelling bioclimatic variables in 2.5 arc-minutes (± 21.62 km2 at the equator) of interpolated climate surface were used. This approach was justified considering the precision of georeferenced records of both, orchid and its pollinators–the information available in public databases was not sufficient to conduct further analyses using more detailed maps (in 30 arc-seconds). Because some previous studies [59] indicated that usage of a restricted area in ENM analysis is more reliable than calculating habitat suitability on the global scale, the area of the analysis was restricted to 78.83˚N-34.08˚N– 13.12˚W-77.29˚E.

In this study the most widespread source of data for ecological studies was used. WorldClim [60] is commonly applied to produce species distribution models (> 15000 citations). Of 19 climatic variables (“bioclims”, Table 1) available in WorldClim (version 1.4, www.worldclim.org) seven were removed as they were significantly correlated with one another (above 0.9) as evaluated by Pearsons’ correlation coefficient computed using ENMTools v1.3 [61]. As a result of the reduction of multi-collinearity the following variables were excluded from further analyses: bio6, bio7, bio9, bio10, bio11, bio16 and bio17. Because MaxEnt is relatively robust against collinear variables [62] we decided not to remove other data from the analyses. The most recent research results suggested that the strategy of excluding highly correlated variables has little influence on models derived from MaxEnt [63].

Table 1. Codes of climatic variables developed by Hijmans et al. [60].

Code Description
bio1 Annual Mean Temperature
bio2 Mean Diurnal Range = Mean of monthly (max temp − min temp)
bio3 Isothermality (bio2/bio7) * 100
bio4 Temperature Seasonality (standard deviation * 100)
bio5 Max Temperature of Warmest Month
bio6 Min Temperature of Coldest Month
bio7 Temperature Annual Range (bio5—bio6)
bio8 Mean Temperature of Wettest Quarter
bio9 Mean Temperature of Driest Quarter
bio10 Mean Temperature of Warmest Quarter
bio11 Mean Temperature of Coldest Quarter
bio12 Annual Precipitation
bio13 Precipitation of Wettest Month
bio14 Precipitation of Driest Month
bio15 Precipitation Seasonality (Coefficient of Variation)
bio16 Precipitation of Wettest Quarter
bio17 Precipitation of Driest Quarter
bio18 Precipitation of Warmest Quarter
bio19 Precipitation of Coldest Quarter
Open in a new tab

Predictions of the future extent of the climatic niches of C. calceolus in 2070 were made using climate projections obtained from the Community Climate System Model (CCSM4) which was commonly used in previous studies on orchids (e.g. [64–65]). Four representative concentration pathways (RCPs: rcp2.6, rcp4.5, rcp6.0, rcp8.5) were analyzed. These pathways are trajectories adopted by the Intergovernmental Panel on Climate Change (IPCC) for its fifth Assessment Report in 2014. These four scenarios describe potential future climate of the world assuming various amounts of greenhouse gases will be emitted. The RCPs are named after a possible range of radiative forcing values in 2100, relative to pre-industrial values (+2.6, +4.5, +6.0 and +8.5 W/m2 respectively; [66–67]). These climate projections were used in several previous studies on threatened plants (e.g. [68–69]) and endangered animals (e.g. [70–71]).

In all analyses the maximum number of iterations was set to 10000 and convergence threshold to 0.00001. The neutral (= 1) regularization multipler value and auto features were used. All samples were added to the backgroud. The “random seed" option which provided a random test partition and background subset for each run was applied. 10% of the samples were used as test points. While often larger test samples are used in species distribution models [72], we followed Oraie et al. [73], Ashraf et al. [74], and Tobeña et al. [75] in our analyses. The run was performed as a bootstrap with 100 replicates, and the output was set to logistic. While bootstrap is also recommended for small-sample analyses we followed Slater & Michael [76] in our modelling. All operations on GIS data were carried out on ArcGis 10.6 (Esri, Redlands, CA, USA). The evaluation of the created models was made using the most common metric—the area under the curve (AUC; [77–79].

To visualize the climatic preferences of C. calceolus the predicted niche occupancy profiles (PNOs) were created using the Phyloclim package [80]. SDMtoolbox 2.3 for ArcGIS [81–82] was used to visualize changes in the distribution of suitable niches of studied orchid and its pollinators caused by the global warming [81]. To compare distribution model created for current climatic conditions with future models all SDMs were converted into binary rasters and projected using Albers EAC (as implemented in SDMtoolbox 2.3) as projection. The presence threshold was estimated individually for each species based on the values of grids in which studied species occur in models created using present-time. For C. calceolus, Chrysotoxum festivum, Syrphus ribesii, Nomada panzeri, Colletes cunicularius, Halictus tumularum, Lasioglossum albipes, L. calceatum, L. fratellum, L. fulvicorne, L. morio, Andrena carantonica, A. cineraria, A. fucata, A. haemorrhoa, A. helvola, A. nigroaenea, A. praecox, and A. scotica the threshold for presence was set as 0.4. The habitat suitability of at least 0.3 was considered as sufficient for the occurrence of Andrena flavipes, A. tibialis, and Lasioglossum quadrinotatum. Furthermore, to estimate the pollinator availability, the binary models of predicted range of C. calceolus were compared with future distribution of its pollinators to calculate the number of grid cells in which both orchid and insect could occur.

Results

Predicted distribution of C. calceolus

The average training AUC for the replicate runs received scores of 0.912–0.914, which indicates that the MaxEnt models are very reliable (Table 2).

Table 2. The average training AUC for the replicate runs for created models of C. calceolus.

Scenario AUC score
present 0.914
rcp2.6 0.913
rcp4.5 0.914
rcp6.0 0.912
rcp8.5 0.914
Open in a new tab

Created map of the potential distribution of C. calceolus (Fig 5) is consistent with the known location of populations of this species. However, some additional regions (e.g. Eastern Carpathians and western valley of the Danube river) were indicated by the ENM analysis as potentially suitable for this orchid.

Fig 5. Present distribution of suitable niches of C. calceolus.

Fig 5

Open in a new tab

Map was generated in ArcGis 9.3 (ESRI, 2006).

The total area of niches suitable for C. calceolus will decrease in 2070 according to three of four scenarios of future climate change analyzed (Fig 6). Considering areas characterized by a suitability of at least 0.4 the loss of habitat will vary between ca. 30% and 63%. Surprisingly scenario rcp 6.0 will be slightly less harmful than rcp 4.5. The highest habitat loss of ca. 63% is predicted in rcp 8.5. In this scenario relatively large suitable areas will still be available in Scandinavia but the niche coverage in the Pyrenees and the Alps will be significantly smaller than currently. C. calceolus will almost disappear from the Carpathians and there will be no suitable niches for this orchid in the Apennines, Balkans, lowlands of Baltic countries and valleys of the major European rivers. The changes in the distribution of the coverage of suitable niches of C. calceolus are presented in Fig 7 and Table 3.

Fig 6. Future distribution of suitable niches of C. calceolus.

Fig 6

Open in a new tab

Estimations based on rcp2.6 scenario (A), rcp4.5 scenario (B), rcp6.0 scenario (C) and rcp8.5 scenario (D). Maps were generated in ArcGis 9.3 (ESRI 2006).

Fig 7. Changes in the distribution of the coverage of suitable niches of C. calceolus in various climate change scenarios.

Fig 7

Open in a new tab

Rcp2.6 (A), rcp4.5 (B), rcp6.0 (C) and rcp8.5 (D). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction. Maps were generated in ArcGis 10.6 (ESRI). Albers EAC projection.

Table 3. Changes in the coverage of suitable niches of C. calceolus.

Scenario Number of grid cells ≥ 0.4 Range expansion [km2] Range contraction [km2]
present 125382 - -
rcp2.6 88063 135057.6446 690919.2031
rcp4.5 67337 107792.1356 933432.8332
rcp6.0 70680 136271.5200 939595.5852
rcp8.5 46517 131135.8933 1236528.1833
Open in a new tab

Of the bioclimatic factors analyzed the most important variables influencing the distribution of C. calceolus are temperature seasonality (bio4) and precipitation in the warmest quarter (bio18; Table 4). Somewhat less significant for the occurrence of this species is precipitation in the driest month (bio14). The PNO profiles of C. calceolus for these three vital variables are presented in Fig 8.

Table 4. The estimates of relative contributions of the environmental variables to the Maxent models.

scenario variable 1 variable 2 variable 3
present bio4 (40.8) bio18 (30.9) bio14 (15.1)
rcp2.6 bio4 (43.5) bio18 (33.3) bio14 (12.8)
rcp4.5 bio4 (43.4) bio18 (31.1) bio14 (14.8)
rcp6.0 bio4 (43.5) bio18 (31.9) bio14 (11.9)
rcp8.5 bio4 (44.6) bio18 (30.4) bio14 (13.5)
Open in a new tab

Fig 8.

Fig 8

Open in a new tab

Predicted niche occupancy profiles created for present models (A-C), and future climate change scenarios (D-F). Diagrams generated in RStudio using the Phyloclim package (Heibl & Calenge, 2013). Albers EAC projection.

Predicted availability of C. calceolus pollinators

The average training AUC for the replicate runs received scores of 0.878–0.989, which indicates that the MaxEnt models are very reliable (Table 5). The predicted potential ranges of all studied insect species are presented as S1–S4 Figs.

Table 5. The average training AUC for the replicate runs for created models of C. calceolus pollinators [SD–standard deviation].

Species Scenario
present rcp2.6 rcp4.5 rcp6.0 rcp8.5
Andrena carantonica 0.979, SD = 0.001 0.980, SD = 0.001 0.980, SD = 0.001 0.980, SD = 0.001 0.980, SD = 0.001
Andrena cineraria 0.921, SD = 0.001 0.922, SD = 0.001 0.922, SD = 0.001 0.921, SD = 0.001 0.920, SD = 0.001
Andrena flavipes 0.969, SD = 0.001 0.969, SD = 0.001 0.969, SD = 0.002 0.968, SD = 0.001 0.969, SD = 0.001
Andrena fucata 0.947, SD = 0.001 0.946, SD = 0.001 0.947, SD = 0.001 0.948, SD = 0.001 0.947, SD = 0.001
Andrena haemorrhoa 0.914, SD = 0.001 0.914, SD = 0.001 0.912, SD = 0.001 0.914, SD = 0.001 0.916, SD = 0.001
Andrena helvola 0.955, SD = 0.001 0.955, SD = 0.001 0.950, SD = 0.001 0.956, SD = 0.001 0.955, SD = 0.001
Andrena nigroaenea 0.956, SD = 0.001 0.956, SD = 0.001 0.956, SD = 0.001 0.955, SD = 0.001 0.956, SD = 0.001
Andrena praecox 0.963, SD = 0.001 0.963, SD = 0.001 0.962, SD = 0.001 0.963, SD = 0.001 0.961, SD = 0.001
Andrena scotica 0.968, SD = 0.001 0.969, SD = 0.001 0.970, SD = 0.001 0.969, SD = 0.001 0.970, SD = 0.001
Andrena tibialis 0.979, SD = 0.002 0.978, SD = 0.002 0.976, SD = 0.002 0.979, SD = 0.002 0.979, SD = 0.002
Chrysotoxum festivum 0.955, SD = 0.001 0.953, SD = 0.001 0.956, SD = 0.001 0.955, SD = 0.002 0.956, SD = 0.001
Colletes cunicularius 0.954, SD = 0.001 0.954, SD = 0.001 0.955, SD = 0.001 0.953, SD = 0.001 0.955, SD = 0.001
Halictus tumulorum 0.935, SD = 0.001 0.935, SD = 0.001 0.935, SD = 0.001 0.937, SD = 0.001 0.936, SD = 0.001
Lasioglossum albipes 0.931, SD = 0.001 0.932, SD = 0.001 0.930, SD = 0.001 0.930, SD = 0.001 0.930, SD = 0.001
Lasioglossum calceatum 0.925, SD = 0.001 0.926, SD = 0.001 0.928, SD = 0.001 0.927, SD = 0.001 0.926, SD = 0.001
Lasioglossum fratellum 0.933, SD = 0.001 0.934, SD = 0.001 0.935, SD = 0.001 0.934, SD = 0.001 0.936, SD = 0.001
Lasioglossum fulvicorne 0.960, SD = 0.001 0.961, SD = 0.001 0.960, SD = 0.001 0.960, SD = 0.001 0.960, SD = 0.001
Lasioglossum morio 0.955, SD = 0.001 0.954, SD = 0.001 0.955, SD = 0.001 0.954, SD = 0.001 0.955, SD = 0.001
Lasioglossum quadrinotatum 0.988, SD = 0.001 0.988, SD = 0.001 0.989, SD = 0.001 0.989, SD = 0.001 0.988, SD = 0.001
Nomada panzeri 0.935, SD = 0.002 0.937, SD = 0.002 0.938, SD = 0.001 0.939, SD = 0.001 0.937, SD = 0.002
Syrphus ribesii 0.878, SD = 0.001 0.878, SD = 0.001 0.879, SD = 0.001 0.881, SD = 0.001 0.879, SD = 0.001
Open in a new tab

Except of a single case of Andrena helvola (only rcp4.5 scenario) all pollinators of C. calceolus will face habitat loss caused by the climate changes (Table 6). The highest decrease of 410327–786796 km2 in the coverage of the suitable niches will be observed in Syrphus ribesii. Generally, the rcp8.5 scenario will cause the most significant damages in the available habitats of studied species. In this scenario Diptera representatives, Chrysotoxum festivum Syrphus ribesii will lose respectively 78995 km2 and 786796 km2 of their current niche coverage. The potential range of the only Nomadinae species, Nomada panzeri, will be smaller for 479387 km2. The decrease of 387973 km2 will be observed in Colletes cunicularius. Within Halictidae the most significant habitat loss is predicted for Lasioglossum calceatum (474419 km2) and within Andrena representatives the highest range contraction will be observed in Andrena cineraria (624529 km2).

Table 6. Loss of suitable niches [km2] of studied pollinators of C. calceolus in various climate change scenarios.

Species rcp2.6 rcp4.5 rcp6.0 rcp8.5
Andrena carantonica–total habitat loss 106690.3 183650 183463.3 162565.9
Andrena cineraria–total habitat loss 270283.4 523591.1 458826.2 624529.6
Andrena flavipes–total habitat loss 23885.33 39871.14 130351.5 350735.3
Andrena fucata–total habitat loss 196535.8 287893.9 294411.5 301601.3
Andrena haemorrhoa–total habitat loss 315719.7 514010.9 470610.2 545384.9
Andrena helvola–total habitat loss 92721.41 -226677 198814.1 123030.9
Andrena nigroaenea–total habitat loss 130706.4 289014.4 256669.3 416247.2
Andrena praecox–total habitat loss 26163.68 194388.1 193622.5 314188.3
Andrena scotica–total habitat loss 34175.26 83589.33 77314.53 141892.7
Andrena tibialis–total habitat loss 42579.01 68966.8 89845.45 134123.9
Chrysotoxum festivum–total habitat loss 8833.278 51094.82 56753.34 78995.28
Colletes cunicularius–total habitat loss 155955 379625.5 345879.8 387973.3
Halictus tumulorum–total habitat loss 101311.9 184285 154442.3 219468.7
Lasioglossum albipes–total habitat loss 237676.8 302703.2 320911.3 351575.7
Lasioglossum calceatum–total habitat loss 226695.9 369765.1 396395.7 474419.9
Lasioglossum fratellum–total habitat loss 248825.8 373257.3 350660.6 407059.1
Lasioglossum fulvicorne–total habitat loss 46295.34 167047.9 176497.5 234800.9
Lasioglossum morio–total habitat loss 107698.8 166357 172407.7 227162.8
Lasioglossum quadrinotatum–total habitat loss 105812.6 123890 105289.7 118567.6
Nomada panzeri–total habitat loss 246640.8 376450.8 357140.8 479387.4
Syrphus ribesii–total habitat loss 410327.2 657883.1 710023.7 786796.7
Open in a new tab

Considering the predicted range overlap of C. calceolus and studied insects (Fig 9, Table 7), the highest pollination potential in the future will be attributed to Syrphus ribesii which will occur in 45.85–66.81% of C. calceolus range. The global warming will almost exclude the possibility of pollen transfer by Lasioglossum quadrinotatum which will overlap with the lady's-slipper orchid in just 0.27–2.59% of the orchid niches coverage. Similar situation will be observed in Andrena carantonica (0.42–7.31%) and A. scotica (2.06–7.86%). Surprisingly, the highest overlap between C. calcolus and its pollinators is expected in rcp8.5 scenario. In this generally unsuitable climatic conditions seven of the studied insects will be available for the lady's-slipper orchid in more than 40% of its range—Andrena cineraria, A. fucata, A. haemorrhoa, Lasioglossum albipes, L. fratellum, Nomada panzeri, and Syrphus ribesii. Nomada panzeri, the only representative of Nomadinae, will occur in 25.30–52.96% of the predicted range of C. calceolus, while Colletes cunicularius (Colletidae) will be able to pollinate orchid populations in 10.10–21.46% of the range. Lasioglossum fratellum will be the most important pollinator of C. calceolus within Halictidae–this species will be available in 22.74–51.64% of the orchid range. Considering Andrena species, the most significant contribution to the orchid propagation will be attributed to Andrena fucata which can occur in 18.36–44.60% of the lady's-slipper orchid range.

Fig 9. Predicted niche overlap between C. calceolus (red diagonal shade) and its pollinators (green shade) in various climate change scenarios.

Fig 9

Open in a new tab

Present time (A), rcp2.6 scenario (B), rcp4.5 scenario (C), rcp6.0 scenario (D) and rcp8.5 scenario (E). Maps were generated in ArcGis 10.6 (ESRI).

Table 7. The number of grid cells where both C. calceolus and specific pollinator can occur in various climate change scenarios.

present rcp26 rcp45 rcp60 rcp85
number of common grid cells part of C. calceolus range number of common grid cells part of C. calceolus range number of common grid cells part of C. calceolus range number of common grid cells part of C. calceolus range number of common grid cells part of C. calceolus range
Andrena carantonica 9163 0.073 4132 0.047 281 0.004 467 0.007 628 0.014
Andrena cineraria 40077 0.320 27903 0.317 14583 0.217 19745 0.279 20514 0.441
Andrena flavipes 20249 0.161 11444 0.130 9442 0.140 6442 0.091 1229 0.026
Andrena fucata 23024 0.184 21360 0.243 14047 0.209 17928 0.254 20748 0.446
Andrena haemorrhoa 32401 0.258 23890 0.271 13568 0.201 18670 0.264 18969 0.408
Andrena helvola 15286 0.122 10581 0.120 20474 0.304 5472 0.077 7354 0.158
Andrena nigroaenea 15130 0.121 10503 0.119 3167 0.047 6195 0.088 3073 0.066
Andrena praecox 19039 0.152 19895 0.226 9360 0.139 11974 0.169 7646 0.164
Andrena scotica 2586 0.021 2680 0.030 1206 0.018 2646 0.037 3657 0.079
Andrena tibialis 16841 0.134 13401 0.152 8424 0.125 8745 0.124 7687 0.165
Chrysotoxum festivum 19656 0.157 18478 0.210 11385 0.169 13988 0.198 16490 0.354
Colletes cunicularius 26466 0.211 18899 0.215 6803 0.101 9200 0.130 8711 0.187
Halictus tumulorum 22195 0.177 19054 0.216 9620 0.143 13448 0.190 13636 0.293
Lasioglossum albipes 23108 0.184 20308 0.231 14743 0.219 19327 0.273 20080 0.432
Lasioglossum calceatum 28482 0.227 23254 0.264 14971 0.222 17002 0.241 18402 0.396
Lasioglossum fratellum 28508 0.227 26083 0.296 18417 0.274 21548 0.305 24021 0.516
Lasioglossum fulvicorne 18992 0.151 20538 0.233 10093 0.150 10924 0.155 8576 0.184
Lasioglossum morio 17897 0.143 12864 0.146 6254 0.093 7365 0.104 6848 0.147
Lasioglossum quadrinotatum 2013 0.016 241 0.003 711 0.011 1084 0.015 1203 0.026
Nomada panzeri 31722 0.253 27968 0.318 21341 0.317 23132 0.327 24635 0.530
Syrphus ribesii 58772 0.469 41718 0.474 30877 0.459 34297 0.485 31078 0.668
Open in a new tab

Discussion

Based on the results of this research the area of niches suitable for C. calceolus will significantly decrease under all the currently available scenarios of climate change. The ENM was used previously in a very few studies on the effect of climate change on orchids. While global warming is predicted to negatively affect European species of Dactylorhiza [64] suitable niches for holomycoheterotrophic Neottia nidus-avis and Epipogium aphyllum are predicted to become more widespread [83]. In the recent regional study Kaye et al. [84] evaluated the probability of extinction of American Cypripedium fasciculatum based on population size, time between surveys, and elevation. This research revealed that 39–52% of C. fasciculatum populations are likely extinct. In our study we investigated exclusively the impact of climate changes on European C. calceolus but the similar loss of habitats was predicted (30–63%).

As evaluated in this study temperature seasonality and precipitation in the warmest quarter are crucial climatic factors limiting the distribution of C. calceolus. Therefore, it is not surprising that global warming will cause a decrease in the availability of suitable niches for this species in Europe. According to the National Aeronautics and Space Administration (NASA) rising temperatures will intensify the world’s water cycle and increase evaporation. As a result, storm-affected regions will experience increases in precipitation, while the increased risk of drought is predicted for areas located far away from storm tracks. As a result of global warming the heat capacity of the surface layer will increase due to loss of sea ice. Dwyer, Biasutti & Sobel [85] reported that when seasonality of surface temperature is considered, the phase delay and amplitude decrease are strongest at high latitudes and will drive the global response.

Noteworthy, while our analyses included the evaluation of possible effects of the global warming on the distribution of suitable niches of both, studied orchid and its pollinators, there are other factors that can increase the extinction rate of the lady's-slipper orchid. Like all other Orchidaceae representatives, C. calceolus requires mycorrhizal fungi for germination and seedling nutrition. Distribution of Cypripedium can be hereby limited by mycorrhizal specificity [86] and while this relationship is not primarily limited by fungal distribution but by genetically controlled specialization [87], the further studies could be improved and include also analysis of the possible changes of European mycobiota.

Based on our results at least two approaches should be implemented to improve the chances of survival of C. calceolus. In view of the unavoidable loss of suitable habitats in numerous European regions, conservation activities over the next 50 years should be concentrated in areas where there are still suitable niches for this species. The prioritization of preservation zones is suggested by Seaton et al. [88] but their proposal is mainly for the most biodiverse, tropical regions of the world.

In addition, for C. calceolus ex-situ activities should be carried out at a large scale. Seed storage will enable the cultivation of this rare orchid in the future and successful reintroduction into the wild. This method is already effectively being used to re-establish the lady's slipper orchid in Britain [89] and could be used in the future to introduce C. calceolus in the remaining suitable areas.

Conclusions

Our research results indicated significant loss (30%-63%) of suitable habitat of C. calceolus in 2070, but the pollinator availability should not further limit the chance of survival of this species. The highest decrease of niches coverage was predicted in rcp 8.5 scenario of future climate change. Temperature seasonality and precipitation in the warmest quarter are crucial climatic factors limiting the distribution of C. calceolus, therefore, it is not surprising that global warming will cause a decrease in the availability of suitable niches for the studied species in Europe. Based on our results at least two approaches should be implemented to improve the chances of survival of the lady's-slipper orchid in Europe. In view of the unavoidable loss of suitable habitats in numerous European regions, conservation activities over the next 50 years should be concentrated in areas where there are still suitable niches for this species. Furthermore, for C. calceolus ex-situ activities, e.g. steed storage, should be carried out at a large scale. Noteworthy, while both orchid and its pollinators were included in our analyses, the extinction of the lady's-slipper orchid may be further driven by the modification of local mycobiota.

Supporting information

S1 Table. Localities of C. calceolus gathered in this study.

(XLS)

Click here for additional data file. (122KB, xls)
S2 Table. Localities of pollinators of C. calceolus gathered in this study.

(XLSX)

Click here for additional data file. (483.6KB, xlsx)
S1 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp2.6 climate change scenario.

Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

(TIF)

Click here for additional data file. (20.4MB, tif)
S2 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp4.5 climate change scenario.

Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

(TIF)

Click here for additional data file. (18.5MB, tif)
S3 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp6.0 climate change scenario.

Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

(TIF)

Click here for additional data file. (19.3MB, tif)
S4 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp8.5 climate change scenario.

Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

(TIF)

Click here for additional data file. (21.5MB, tif)

Acknowledgments

We would like to thank prof. Maxim A. Dzhus (Belarusian State University), Peter Efimov, PhD (Komarov Botanical Institute of the Russia Academy of Sciences), Myroslav Shevera, PhD (National Academy of Sciences of Ukraine), Mindaugas Lapele, PhD (Dzukija National Park, Lithuania) and Spyros Tsiftsis, PhD (Eastern Macedonia and Thrace Institute of Technology) for providing data on the distribution of C. calceolus. We are grateful to Prof. Anthony F.G. Dixon (University of East Anglia) for the valuable comments on the manuscript. Thanks are also due to Mr Jacek Stefaniak, MSc (University of Wrocław) for helpful discussions and to Giuseppe Brundu for valuable comments on the manuscript.

Data Availability

All relevant data, including list of samples used in this study are provided as supplementary material.

Funding Statement

The research described here was supported by the Education Youth and Sports (MSMT) grant nr LO1415 (MK). https://app.dimensions.ai/details/grant/grant.6884710 The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

References

  • 1.Guisan A, Zimmermann NE. Predictive habitat distribution models in ecology. Ecol Modell. 2000; 135: 147–186. [Google Scholar]
  • 2.Peterson AT, Soberón J, Pearson RG, Anderson RP, Martínez-Meyer E, Nakamura M, et al. Ecological Niches and Geographic Distributions. Princenton: Princenton University Press; 2011. [Google Scholar]
  • 3.Brambilla M, Saporetti F. Modelling distribution of habitats required for different uses by the same species: Implications for conservation at the regional scale. Biol Conserv. 2014; 174: 39–46. [Google Scholar]
  • 4.Yañez-Arenas C, Martínez-Meyer E, Mandujano S, Rojas-Soto O. Modelling geographic patterns of population density of the white-tailed deer in central Mexico by implementing ecological niche theory. Oikos. 2012; 121: 2081–2089. [Google Scholar]
  • 5.Micchi de Barros Ferraz KMP, Frosini de Barros Ferraz S, de Paula RC, Beisiegel B, Breitenmoser C. Species Distribution Modelling for Conservation Purposes. Nat Conserv. 2012; 10(2): 214–220. [Google Scholar]
  • 6.Micchi de Barros Ferraz PMB, Beisiegel BM, de Paula RC, Sana DA, de Campos CB, de Oliveira TG, et al. How species distribution models can improve cat conservation—jaguars in Brazil. CatNews. 2012; 7: 38–42. [Google Scholar]
  • 7.Chefaoui RM, Hortal J, Lobo JM. Potential distribution modelling, niche characterization and conservation status assessment using GIS tools: a case study of Iberian Copris species. Biol Conserv. 2005; 122(2): 327–338. [Google Scholar]
  • 8.Thorn JS, Nijman V, Smith D, Nekaris KAI. Ecological niche modelling as a technique for assessing threats and setting conservation priorities for Asian slow lorises (Primates: Nycticebus). Diver Distr. 2009; 15(2): 289–298. [Google Scholar]
  • 9.Swarts ND, Dixon KW. Terrestrial orchid conservation in the age of extinction. Ann. Bot. 2009; 104(3): 543–556. 10.1093/aob/mcp025 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Gale SW, Fischer GA, Cribb PJ, Fay MF. Orchid conservation: bridging the gap between science and practice. Bot J Linn Soc. 2018; 186: 425–434. [Google Scholar]
  • 11.Kull T. Identification of clones in Cypripedium calceolus (Orchidaceae). P Est Acad Sci. 1988; 37: 195–199. [Google Scholar]
  • 12.Kull T. Fruit-set and recruitment in populations of Cypripedium calceolus L. in Estonia. Bot J Linn Soc. 1998; 126: 27–38. [Google Scholar]
  • 13.Bergström G, Birgersson G, Groth I, Nilsson LA. Floral fragrance disparity between three taxa of Lady’s slipper Cypripedium calceolus (Orchidaceae). Phytochemistry. 1992; 31: 2315–2319. [Google Scholar]
  • 14.Case MA. High levels of allozyme variation within Cypripedium calceolus (Orchidaceae) and low levels of divergence among its varieties. Syst Bot. 1993; 18(4): 663–677. [Google Scholar]
  • 15.Erneberg M, Holm B. Bee size and pollen transfer in Cypripedium calceolus (Orchidaceae). Nord J Bot. 1999; 19: 363–367. [Google Scholar]
  • 16.Brzosko E. Dynamics of island populations of Cypripedium calceolus in the Biebrza river valley (northeast Poland). Bot J Linn Soc. 2002; 139: 67–77. [Google Scholar]
  • 17.Blinova IV. A northernmost population of Cypripedium calceolus L. (Orchidaceae): demography, flowering, pollination. Selbyana 2002; 23: 111–120. [Google Scholar]
  • 18.Antonelli A, Dahlberg CJ, Carlgren KHI, Appelqvis T. Pollination of the Lady’s slipper orchid (Cypripedium calceolus) in Scandinavia. Nord. J. Bot. 2009; 27: 1–8. [Google Scholar]
  • 19.Fay MF, Bone R, Cook P, Kahandawala I, Greensmith J, Harris S, et al. Genetic diversity in Cypripedium calceolus (Orchidaceae) with a focus on northwestern Europe, as revealed by plastid DNA length polymorphisms. Ann Bot 2009; 104: 517–525. 10.1093/aob/mcp116 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Rasmussen HN, Pedersen HÆ. Cypripedium calceolus germination in situ: seed longevity, and dormancy breakage by long incubation and cold winters Eur J Environ Sci. 2011; 1(2): 69–70. [Google Scholar]
  • 21.Devilliers-Terschuren J. Action Plan for Cypripedium calceolus in Europe. Strasbourg: Council of Europe Pub; 1999. [Google Scholar]
  • 22.Delforge P. Orchids of Europe, North Africa and the Middle East. Portland: Timber Press; 2006. [Google Scholar]
  • 23.Rankou H, Bilz M. Cypripedium calceolus. The IUCN Red List of Threatened Species 2014, e.T162021A43316125; 2014. Available at: 10.2305/IUCN.UK.2014-1.RLTS.T162021A43316125.en [DOI] [Google Scholar]
  • 24.Kull T. Biological flora of the British Isles. Cypripedium calceolus L. J Ecol. 1999; 87: 913–924. [Google Scholar]
  • 25.Govaerts R, Dransfield J, Zona SF, Hodel DR, Henderson A. World checklist of selected plant families. Kew: The Board of Trustees of the Royal Botanic Gardens; 2009. [Google Scholar]
  • 26.eřovský J. Endangered plants. London: Sunburst Books; 1995 [Google Scholar]
  • 27.Matuszkiewicz W. Przewodnik do oznaczania zbiorowisk roślinnych Polski. Warszawa: Wydawnictwo Naukowe PWN; 2006. [Google Scholar]
  • 28.Limpricht W. Kalkpflanzen der östlichen Grafschaft Glatz. Feddes Repert Spec Nov Regni Veg. 1942; 131: 126–141. [Google Scholar]
  • 29.Limpricht W. Kalkpflanzen der westlichen Grafschaft Glatz. Englers Bot Jahr. 1943; 3: 151–174. [Google Scholar]
  • 30.Jakubska-Busse A, Szczęśniak E, Śliwiński M, Narkiewicz C. Zanikanie stanowisk obuwika pospolitego Cypripedium calceolus L., 1753 (Orchidaceae) w Sudetach. Przyroda Sudetów. 2010; 13: 43–52. [Google Scholar]
  • 31.Szczęśniak E, Jakubska-Busse A, Śliwiński M. Zróżnicowanie i rozmieszczenie zbiorowisk z udziałem Cypripedium calceolus L. (Orchidaceae) na Dolnym Śląsku. Acta Bot Siles. 2012; 8: 97–128. [Google Scholar]
  • 32.Edens-Meier R, Arduser ME, Westhus E, Bernhardt P. Pollination ecology of Cypripedium reginae Walter (Orchidaceae): size matters. Telopia. 2011; 13: 327–340. [Google Scholar]
  • 33.Pemberton RW. Pollination of slipper orchids (Cypripedioideae): a review. Lankesteriana. 2013; 13(1–2): 65–73. [Google Scholar]
  • 34.Robbirt KM, Roberts DL, Hutchings MJ, Davy AJ. Potential Disruption of Pollination in a Sexually Deceptive Orchid by Climatic Change. Curr Biol. 2014; 24(23): 2845–2849. 10.1016/j.cub.2014.10.033 [DOI] [PubMed] [Google Scholar]
  • 35.Roberts DL. Pollination biology: the role of sexual reproduction in orchid conservation In: Dixon KW, Kell SP, Barrett RL, Cribb PJ, editors. Orchid Conservation. London: Natural History Publications; 2003. pp. 113–136. [Google Scholar]
  • 36.Lazare J-J, Miralles J, Villar L. Cypripedium calceolus L. (Orchidaceae) en el Pirineo. Anales Jard Bot. Madrid. 1986; 43: 375–382. [Google Scholar]
  • 37.Perazza G. Cartografia della orchidee (Orchidaceae) spontanee in Trentino-Alto Adige (Italia). Ricerca Sull'erbario dell'Universita di firence (FI). Ann Mus Civ Rovereto. 1995; 11: 231–256. [Google Scholar]
  • 38.Perezza G, Decarli Perezza M. Cartogrfia orchidee trdentine (COT): Cypripedium calceolus L. e Liparis loeselii (L.) Rich., specie citate nella directiva habitat della CEE. Atti Acc Rov Agiati. 2002; 252: 129–210. [Google Scholar]
  • 39.Isaja A, Dotti L. Le orchidee spontanee della Val di Susa (Piemonte-Italia) primi dati sulla distribuzione di tre orchidee rare: Cypripedium calceolus L. (1735), Corallorhiza trifida Chatelain (1760) e Aceras anthropophorum R.Br ex Aiton fil. (1814). Riv Piem St Nat. 2003; 24: 205–215. [Google Scholar]
  • 40.Gudžinskas Z, Ryla M. Lietuvos gegužraibiniai (Orchidaceae) Orchids (Orchidaceae) of Lithuania. Vilnius: Botanikos instituto leidykla; 2006. [Google Scholar]
  • 41.Raðomavièius V. Lietuvos Raudonoji Knyga Red Data Book of Lithuania. Vilnius: Publishing Company; 2007. [Google Scholar]
  • 42.Petrović D, Stešević D, Vuksanović S. Materials for the Red Book of Montenegro. Natura Montenegrina. Podgorica. 2008; 7(2): 605–631. [Google Scholar]
  • 43.Didukh YP. Red data book of Ukraine Vegetable kingdom. Kyiv: Globalconsalting; 2009. [Google Scholar]
  • 44.Teteryuk LV, Kirillova IA. Rare and protected Orchids of the Komi Republic. Arbeitskreis Heimische Orchideen. 2011; 28(1): 133–179. [Google Scholar]
  • 45.Balázs ZR, Roman A, Balázs HE, Căpraş D, Podar D. Rediscovery of Cypripedium calceolus L. In the vicinity of Cluj-Napoca (Romania) after 80 years. Contrib. Bot. 2016; 51: 43–53. [Google Scholar]
  • 46.Tsiftsis S, Tsiripidis I. Threat categories of the Greek orchids (Orchidaceae). Chronia Botanica. 2016; 21: 43–74. [Google Scholar]
  • 47.Khapugin AA, Chugunov G, Vargot EV. Cypripedium calceolus (Orchidaceae) in central Russia: A case study for its populations in two protected areas in the Republic of Mordovia (Russia). Lankesteriana. 2017; 17(3): 417–431. [Google Scholar]
  • 48.Melnyk V, Shynder OI, Nesyn Y. Distribution of Cypripedium calceolus (Orchidaceae) in Ukraine. Ukr Bot J. 2018; 75(1), 20–32 (2018). [Google Scholar]
  • 49.Melnyk V, Shynder OI, Nesyn Y. Habitats and the current state of populations of Cypripedium calceolus (Orchidaceae) in Ukraine. Ukr Bot J. 2018; 75(2): 160–168. [Google Scholar]
  • 50.Pedrini P, Brambilla M, Bertolli A, Prosse F. Definizione di "linee guida provinciali" per l’attuazione dei monitoraggi nei siti trentinidella rete Natura 2000. LIFE+T.E.N—Azione A5; 2014. [Google Scholar]
  • 51.Müller H. Beobachtungen an westphälischen Orchideen. Verh Naturh Ver Rheinl. 1868; 25: 1–62. [Google Scholar]
  • 52.Müller H. Die Befruchtung der Blumen durch Insekten und die gegenseitigen Anpassungen beider. Leipzig: Kessinger Publishingl; 1873. [Google Scholar]
  • 53.Daumann E. Zur Bestäubungsbiologie von Cypripedium calceolus L. Österreichische Botanische Zeitung. 1969; 115: 434–446. [Google Scholar]
  • 54.Ishmuratova MM, Zhimova TV, Ishbirdin AR, Sujundukov IV, Magafurov AM. Ant ecology, phenology and consorts of Cypripedium calceolus L. and Cypripedium guttatum Sw. in the southern Ural. Bull Moscow Soc Nat. 2006; 110: 40–46. [Google Scholar]
  • 55.Claessens J, Kleynen J. The flower of the European orchid. Form and function. Voerendaal, Netherlands: Jean Claessens & Jacques Kleynen; 2011. [Google Scholar]
  • 56.Phillips SJ, Dudík M, Schapire RE. A maximum entropy approach to species distribution modeling. ICML '04. Proceedings of the twenty-first international conference on Machine learning 655–662. New York: ACM; 2004.
  • 57.Phillips SJ, Anderson R, Schapire RE. Maximum entropy modeling of species geographic distributions. Ecol Modell. 2006; 190: 231–259. [Google Scholar]
  • 58.Elith J, Phillips SJ, Hastie T, Dudík M, Chee YE, Yates CJ. A statistical explanation of MaxEnt for ecologists. Divers Distrib. 2011; 17: 43–57. [Google Scholar]
  • 59.Barve N, Barve V, Jimenez-Valverde A, Lira-Noriega A, Maher SP, Peterson AT, et al. The crucial role of the accessible area in ecological niche modeling and species distribution modeling. Ecol Mod. 2011; 222: 1810–1819. [Google Scholar]
  • 60.Hijmans RJ, Cameron SE, Parra JL, Jones PG, Jarvis A. Very high resolution interpolated climate surfaces for global land areas. Int J Climatol. 2005; 25: 1965–1978. [Google Scholar]
  • 61.Warren DL, Glor RE, Turelli M. ENMTools: a toolbox for comparative studies of environmental niche models. Ecography. 2010; 33: 607–611. [Google Scholar]
  • 62.Braunisch V, Coppes J, Arlettaz R, Suchant R, Schmid H, Bollmann K. Selecting from correlated climate variables: a major source of uncertainty for predicting species distributions under climate change. Ecography. 2013; 36: 971–983. [Google Scholar]
  • 63.Feng X, Park DS, Liang Y, Pandey R, Papeş M. Collinearity in ecological niche modeling: Confusions and challenges. Ecol Evol. 2019; 9: 10.1002/ece3.5555 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 64.Naczk A, Kolanowska M. Glacial Refugia and Future Habitat Coverage of Selected Dactylorhiza Representatives (Orchidaceae). PLoS ONE. 2015; 10(11): e0143478 10.1371/journal.pone.0143478 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 65.Gamisch A, Comes HP. Clade-age-dependent diversification under high species turnover shapes species richness disparities among tropical rainforest lineages of Bulbophyllum (Orchidaceae). BMC Evol Biol. 2019; 19: 93 10.1186/s12862-019-1416-1 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 66.Moss R, Babiker M, Brinkman S, Calvo E, Carter T, Edmonds J, et al. Towards New Scenarios for Analysis of Emissions, Climate Change, Impacts, and Response Strategies. Geneva: Intergovernmental Panel on Climate Change; 2008. [Google Scholar]
  • 67.Weyant J, Azar C, Kainuma M, Kejun J, Nakicenovic N, Shukla PR, et al. Report of 2.6 Versus 2.9 Watts/m2 RCPP Evaluation Panel. Geneva: IPCC Secretariat; 2009. [Google Scholar]
  • 68.Sohel SI, Akhter S, Ullah H, Haque E, Rana P. Predicting impacts of climate change on forest tree species of Bangladesh: evidence from threatened Dysoxylum binectariferum (Roxb.) Hook.f. ex Bedd. (Meliaceae). iForest. 2016; 10(1): 154–160. [Google Scholar]
  • 69.Xu X, Zhang H, Xie T, Xu Y, Zhao L, Tian W. Effects of Climate Change on the Potentially Suitable Climatic Geographical Range of Liriodendron chinense. Forests. 2017; 8(399): 1–14. [Google Scholar]
  • 70.Lamsal P, Kumar L, Aryal A, Atreya K. Future climate and habitat distribution of Himalayan Musk Deer (Moschus chrysogaster). Ecol Inform. 2018; 44: 101–108. [Google Scholar]
  • 71.Sony RK, Sen S, Kumar S, Sen M, Jayahari KM. Niche models inform the effects of climate change on the endangered Nilgiri Tahr (Nilgiritragus hylocrius) populations in the southern Western Ghats, India. Ecol Eng. 2018; 120: 355–363. [Google Scholar]
  • 72.Briscoe DK, Hiatt S, Lewison R, Hines E. Modeling habitat and bycatch risk for dugongs in Sabah, Malaysia. Endanger Species Res. 2014; 24: 237–247. [Google Scholar]
  • 73.Oraie H, Rahimian H, Rastegar-Pouyani N, Rastegar-Pouyani E, Ficetola GF, Hosseinian Yousefkhani SS, et al. Distribution pattern of the Snake-eyed Lizard, Ophisops elegans Ménétriés, 1832 (Squamata: Lacertidae), in Iran. Zool Middle East. 2014; 60: 125–132. [Google Scholar]
  • 74.Ashraf U, Ali H, Nawaz Chaudry M, Ashraf I, Batool A, et al. Predicting the Potential Distribution of Olea ferruginea in Pakistan incorporating Climate Change by Using Maxent Model. Sustainability. 2016; 8: 722. [Google Scholar]
  • 75.Tobeña M, Prieto R, Machete M, Silva MA. Modeling the Potential Distribution and Richness of Cetaceans in the Azores from Fisheries Observer Program Data. Front Mar Sci. 2016; 3: 202. [Google Scholar]
  • 76.Slater H, Michael E. Predicting the current and future potential distributions of lymphatic filariasis in Africa using maximum entropy ecological niche modelling. PLoS One. 2012; 7: e32202 10.1371/journal.pone.0032202 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 77.Hosmer DW, Lemeshow S. Applied Logistic Regression. New York: John Wiley and Sons; 2000. [Google Scholar]
  • 78.Mason SJ, Graham NE. Areas beneath the relative operating characteristics (ROC) and relative operating levels (ROL) curves statistical significance and interpretation. Q J R Meteorol Soc. 2002; 128: 2145–2166. [Google Scholar]
  • 79.Evangelista PH, Kumar S, Stohlgren TJ, Jarnevich CS, Crall AW, Norman III JB, et al. Modelling invasion for a habitat generalist and a specialist plant species. Divers Distrib. 2008; 14: 808–817. [Google Scholar]
  • 80.Heibl C, Calenge C. Phyloclim: Integrating Phylogenetics and Climatic Niche Modelling. R package version 0.9–4; 2013. Available from: http://CRAN.R-project.org/package=phyloclim [Google Scholar]
  • 81.Kremen C, Cameron A, Moilanen A, Phillips SJ, Thomas CD, Beentje H, et al. Aligning conservation priorities across taxa in Madagascar with high-resolution planning tools. Science. 2008; 320: 222–226. 10.1126/science.1155193 [DOI] [PubMed] [Google Scholar]
  • 82.Brown JL. SDMtoolbox: a python-based GIS toolkit for landscape genetic, biogeographic and species distribution model analyses. Methods Ecol Evol. 2014; 5: 694–700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 83.Kolanowska M, Kras M, Lipińska M, Mystkowska K, Szlachetko DL, Naczk A. Global warming not so harmful for all plants—response of holomycotrophic orchid species for the future climate change. Sc Rep. 2017; 7: 12704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 84.Kaye TN, Bahm MA, Thorpe AS, Gray EC, Pfingsten I, Waddell C. Population extinctions driven by climate change, population size, and time since observation may make rare species databases inaccurate. PLoS ONE. 2019; 14(10): e0210378 10.1371/journal.pone.0210378 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 85.Dwyer JD, Biasutti M, Sobel AH. Projected Changes in the Seasonal Cycle of Surface Temperature. American Meteor Society. 2012; 25: 6359–6374. [Google Scholar]
  • 86.Shefferson RP, Weiss M, Kull T, Taylor DL. High specificity generally characterizes mycorrhizal association in rare lady's slipper orchids, genus Cypripedium. Mol Ecol. 2005; 14(2): 613–626. 10.1111/j.1365-294X.2005.02424.x [DOI] [PubMed] [Google Scholar]
  • 87.Shefferson RP, Taylor DL, Weiß M, Garnica S, McCormick ML, Adams S, et al. The evolutionary history of mycorrhizal specificity among lady’s slipper orchids. Evolution. 2007; 61: 1380–1390. 10.1111/j.1558-5646.2007.00112.x [DOI] [PubMed] [Google Scholar]
  • 88.Seaton P, Kendon JP, Pritchard HW, Puspitaningtyas DW, Marks TR. Orchid conservation: the next ten years. Lankesteriana. 2013; 13(1–2): 93–101. [Google Scholar]
  • 89.Ramsay MR, Stewart J. Re-establishment of the lady's slipper orchid (Cypripedium calceolus L.) in Britain. Bot J Linn Soc. 2008; 126(1–2): 173–181. [Google Scholar]

Decision Letter 0

Jana Müllerová

Transfer Alert

This paper was transferred from another journal. As a result, its full editorial history (including decision letters, peer reviews and author responses) may not be present.

10 Dec 2019

PONE-D-19-27369

Is the lady's-slipper orchid (Cypripedium calceolus) likely to shortly become extinct in Europe? - insights based on ecological niche modelling

PLOS ONE

Dear Dr. Kolanowska,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

We would appreciate receiving your revised manuscript by Jan 24 2020 11:59PM. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter.

To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'.

  • A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'.

  • An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'.

Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

We look forward to receiving your revised manuscript.

Kind regards,

Jana Müllerová, Ph.D

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We noticed you have some minor occurrence of overlapping text with the following previous publication(s), which needs to be addressed:

https://www.iucnredlist.org/species/162021/43316125

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed.

3. Thank you for including your competing interests statement; "no"

Please complete your Competing Interests on the online submission form to state any Competing Interests. If you have no competing interests, please state "The authors have declared that no competing interests exist.", as detailed online in our guide for authors at http://journals.plos.org/plosone/s/submit-now

 This information should be included in your cover letter; we will change the online submission form on your behalf.

Please know it is PLOS ONE policy for corresponding authors to declare, on behalf of all authors, all potential competing interests for the purposes of transparency. PLOS defines a competing interest as anything that interferes with, or could reasonably be perceived as interfering with, the full and objective presentation, peer review, editorial decision-making, or publication of research or non-research articles submitted to one of the journals. Competing interests can be financial or non-financial, professional, or personal. Competing interests can arise in relationship to an organization or another person. Please follow this link to our website for more details on competing interests: http://journals.plos.org/plosone/s/competing-interests

4. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information.

Additional Editor Comments (if provided):

Dear Author, your manuscript is interesting and worth publishing but needs more work, make sure in your re-submission to adequately address all issues raised by the reviewer, especially trying to enlarge the data collection following the suggestions.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: General comments

An interesting Ms that, in my opinion, is it worth to be published after major revision.

I would strongly suggest to address more carefully the distribution of Cypripedium calceolus and all the references and data-bases that could be used. I had the impression the total the number of presence records could be increased after a more careful verification of all the available sources (for a number of Countries). A number of additional potential databases and refences is provided as an example. I think it is worth to improve the data collection and re-run the model.

I think it is very important to start with the better available distribution dataset, if not the level of uncertainly in the model rises too much. In fact, there are already a number of uncertainties areas due to, e.g.: (1) using a single model and not, for example, ensemble modelling techniques; (2) excluding from the model the future changes in land use and the future distribution of beech forest. A number of studies consider the Fagus sylvatica s.l. as a species sensitive to climatic extremes, especially drought and water deficit, which reduces its competitive advantage over less drought-sensitive species, and this will ultimately result in forest vegetation transformation, (3) including in the model the areas that are not and will not be suitable in the future (this could be achieved with masking le land use such as the urban areas that are not of interest).

In the discussion session I would suggest to discuss the results of the present research also in comparison with the modelling of Cypripedium fasciculatum in US done by ThomasN.Kaye (Population extinctions driven by climate change, population size, and time since observation may make rare species databases inaccurate, - PLoSONE 14(10):e0210378.https://doi.org/10.1371/journal.pone.0210378).

Minor comments

LL 63-64: “Appendix I of the Convention on the Conservation of European Wildlife and 64 Natural Habitats of Bern Convention” – please rephrase: Convention on the Conservation of European Wildlife and Natural Habitats (Bern Convention);

L 67: “forms” – types;

L 161: “Figure 1. Localities of C. calceolus georeferenced in this study” -. This map seems not to take into account the distribution of the species in Italy, in particular in the NW, probably due to the fact that Italy is not enough included in GBIF. I would suggest to have a look at: http://dryades.units.it/floritaly/index.php?procedure=taxon_page&tipo=all&id=8099 and http://www.naturachevale.it/wp-content/uploads/2016/06/Cypripedium-calceolus-L_new.pdf

http://fll-italia.it/UploadDocs/6103_G_Perazza___M__Decarli_Perazza_p_129.pdf

http://www.storianaturale.org/anp/PDF%20ANP/24_2003_Isaja%20Dotti_Le%20Orchidee%20spontanee%20della%20Val%20di%20Susa%20Primi%20dati%20sulla%20distribuzione%20di%20tre.pdf

https://www.naturamediterraneo.com/forum/topic.asp?TOPIC_ID=118207

http://www.fondazionemcr.it/UploadDocs/15_art08.pdf

http://www.isprambiente.gov.it/public_files/direttiva-habitat/Manuale-140-2016.pdf (page 128)

I also would strongly suggest to take into consideration the distribution and the reference in the EuroMed PlantBase at:

http://ww2.bgbm.org/euroPlusMed/PTaxonDetailOccurrence.asp?NameId=48350&PTRefFk=8000000

In addition, detailed distribution records are included in the “Action plan for Cypripedium Calceolus in Europe (Nature and Environment No. 100) (1999), Council of Europe” and in TIIU KULL, Journal of Ecology, 1999, 87, 913-924 (BIOLOGICAL FLORA OF THE BRITISH ISLES) and: http://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:320700-2#distribution-map

- LAZARE, J.-J., J. MIRAIXES & L. VILLAR (1987). Cypripedium calceolus (Orchidaceae) en el Pirineo. Anales Jard. Bol. Madrid43(2): 375-382.;

http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.602.3796&rep=rep1&type=pdf

https://www.conservacionvegetal.org/wp-content/uploads/publicaciones/Catalogo%20de%20especies%20amenazadas%20en%20Aragon.pdf

L 368: “suitable habitat” – The modelling do not consider habitats but climate, so I would not use the word “habitat” here; in fact in L 333 the term “niches suitable” is used.

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Giuseppe Brundu

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step.

PLoS One. 2020 Jan 31;15(1):e0228420. doi: 10.1371/journal.pone.0228420.r002

Author response to Decision Letter 0

16 Dec 2019

Additional Editor Comments (if provided):

Dear Author, your manuscript is interesting and worth publishing but needs more work, make sure in your re-submission to adequately address all issues raised by the reviewer, especially trying to enlarge the data collection following the suggestions.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: General comments

An interesting Ms that, in my opinion, is it worth to be published after major revision.

I would strongly suggest to address more carefully the distribution of Cypripedium calceolus and all the references and data-bases that could be used. I had the impression the total the number of presence records could be increased after a more careful verification of all the available sources (for a number of Countries). A number of additional potential databases and refences is provided as an example. I think it is worth to improve the data collection and re-run the model.

I think it is very important to start with the better available distribution dataset, if not the level of uncertainly in the model rises too much. In fact, there are already a number of uncertainties areas due to, e.g.: (1) using a single model and not, for example, ensemble modelling techniques; (2) excluding from the model the future changes in land use and the future distribution of beech forest. A number of studies consider the Fagus sylvatica s.l. as a species sensitive to climatic extremes, especially drought and water deficit, which reduces its competitive advantage over less drought-sensitive species, and this will ultimately result in forest vegetation transformation, (3) including in the model the areas that are not and will not be suitable in the future (this could be achieved with masking le land use such as the urban areas that are not of interest).

In the discussion session I would suggest to discuss the results of the present research also in comparison with the modelling of Cypripedium fasciculatum in US done by ThomasN.Kaye (Population extinctions driven by climate change, population size, and time since observation may make rare species databases inaccurate, - PLoSONE 14(10):e0210378.https://doi.org/10.1371/journal.pone.0210378).

Authors: Corrected.

Minor comments

LL 63-64: “Appendix I of the Convention on the Conservation of European Wildlife and 64 Natural Habitats of Bern Convention” – please rephrase: Convention on the Conservation of European Wildlife and Natural Habitats (Bern Convention);

Authors: Corrected.

L 67: “forms” – types;

Authors: Corrected.

L 161: “Figure 1. Localities of C. calceolus georeferenced in this study” -. This map seems not to take into account the distribution of the species in Italy, in particular in the NW, probably due to the fact that Italy is not enough included in GBIF. I would suggest to have a look at: http://dryades.units.it/floritaly/index.php?procedure=taxon_page&tipo=all&id=8099 and http://www.naturachevale.it/wp-content/uploads/2016/06/Cypripedium-calceolus-L_new.pdf

http://fll-italia.it/UploadDocs/6103_G_Perazza___M__Decarli_Perazza_p_129.pdf

http://www.storianaturale.org/anp/PDF%20ANP/24_2003_Isaja%20Dotti_Le%20Orchidee%20spontanee%20della%20Val%20di%20Susa%20Primi%20dati%20sulla%20distribuzione%20di%20tre.pdf

https://www.naturamediterraneo.com/forum/topic.asp?TOPIC_ID=118207

http://www.fondazionemcr.it/UploadDocs/15_art08.pdf

http://www.isprambiente.gov.it/public_files/direttiva-habitat/Manuale-140-2016.pdf (page 128)

I also would strongly suggest to take into consideration the distribution and the reference in the EuroMed PlantBase at:

http://ww2.bgbm.org/euroPlusMed/PTaxonDetailOccurrence.asp?NameId=48350&PTRefFk=8000000

In addition, detailed distribution records are included in the “Action plan for Cypripedium Calceolus in Europe (Nature and Environment No. 100) (1999), Council of Europe” and in TIIU KULL, Journal of Ecology, 1999, 87, 913-924 (BIOLOGICAL FLORA OF THE BRITISH ISLES) and: http://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:320700-2#distribution-map

- LAZARE, J.-J., J. MIRAIXES & L. VILLAR (1987). Cypripedium calceolus (Orchidaceae) en el Pirineo. Anales Jard. Bol. Madrid43(2): 375-382.;

http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.602.3796&rep=rep1&type=pdf

https://www.conservacionvegetal.org/wp-content/uploads/publicaciones/Catalogo%20de%20especies%20amenazadas%20en%20Aragon.pdf

Authors: We have already used several sources provided by the Reviewer and these were cited in the previous version of ms (e.g. Lazare et al. 1986, Kull 1999, Devilliers-Terschuren 1999 – Action plan, etc.). We could not use all data suggested by the Reviewer due to the lack of possibility of precise georeferencing. Not all links provided by the Reviewer included specific locations but just general information about occurrence of C. calceolus (e.g. http://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:320700-2#distribution-map). However, we included additional 43 Italian records based on:

- Perezza G., Decarli Perezza M. 2002. Cartogrfia orchidee trdentine (COT): Cypripedium calceolus L. e Liparis loeselii (L.) Rich., specie citate nella directiva habitat della CEE.

- https://www.naturamediterraneo.com

- Perazza G. 1995. Cartografia della orchidee (Orchidaceae) spontanee in Trentino-Alto Adige (Italia). Ricerca Sull'erbario dell'Universita di firence (FI). Ann. Mus. Civ. Rovereto 11: 231-256.

- Pedrini P., Brambilla M., Bertolli A., Prosse F. 2014. Definizione di "linee guida provinciali" per l’attuazione dei monitoraggi nei siti trentinidella rete Natura 2000. LIFE+T.E.N - Azione A5

- Isaja A., Dotti L. 2003. Le orchidee spontanee della Val di Susa (Piemonte-Italia) primi dati sulla distribuzione di tre orchidee rare: Cypripedium Calceolus L. (1735), Corallorhiza Trifida Chatelain (1760) e Aceras anthropophorum R.Br ex Aiton fil. (1814). Riv. Piem. St. Nat. 24: 205-215.

All models for C. calceolus were run again and all statistics were calculated using new outcomes.

L 368: “suitable habitat” – The modelling do not consider habitats but climate, so I would not use the word “habitat” here; in fact in L 333 the term “niches suitable” is used.

Authors: Corrected.

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Giuseppe Brundu

Attachment

Submitted filename: Response to Reviewers.docx

Click here for additional data file. (19.9KB, docx)

Decision Letter 1

Jana Müllerová

15 Jan 2020

Is the lady's-slipper orchid (Cypripedium calceolus) likely to shortly become extinct in Europe? - insights based on ecological niche modelling

PONE-D-19-27369R1

Dear Dr. Kolanowska,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

With kind regards,

Jana Müllerová, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

All issues of previous version were addressed and by my opinion, the manuscript is now ready for publication. I have no further comments.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: I am not native English speaker, so I do not feel enough qualified to evaluate point 5 above. However, I had no problems in reading and understanding the Ms and I have not found any major mistake.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: Yes: Giuseppe Brundu

Acceptance letter

Jana Müllerová

21 Jan 2020

PONE-D-19-27369R1

Is the lady's-slipper orchid (Cypripedium calceolus) likely to shortly become extinct in Europe? - insights based on ecological niche modelling

Dear Dr. Kolanowska:

I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

For any other questions or concerns, please email plosone@plos.org.

Thank you for submitting your work to PLOS ONE.

With kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Jana Müllerová

Academic Editor

PLOS ONE

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 Table. Localities of C. calceolus gathered in this study.

    (XLS)

    Click here for additional data file. (122KB, xls)
    S2 Table. Localities of pollinators of C. calceolus gathered in this study.

    (XLSX)

    Click here for additional data file. (483.6KB, xlsx)
    S1 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp2.6 climate change scenario.

    Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

    (TIF)

    Click here for additional data file. (20.4MB, tif)
    S2 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp4.5 climate change scenario.

    Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

    (TIF)

    Click here for additional data file. (18.5MB, tif)
    S3 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp6.0 climate change scenario.

    Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

    (TIF)

    Click here for additional data file. (19.3MB, tif)
    S4 Fig. Changes in the distribution of the coverage of suitable niches of C. calceolus pollinators in 2070 based on rcp8.5 climate change scenario.

    Andrena carantonica (A), Andrena cineraria (B), Andrena flavipes (C), Andrena fucata (D), Andrena haemorrhoa (E), Andrena helvola (F), Andrena nigroaenea (G), Andrena praecox (H), Andrena scotica (I), Andrena tibialis (J), Chrysotoxum festivum (K), Colletes cunicularius (L), Halictus tumulorum (M), Lasioglossum albipes (N), Lasioglossum calceatum (O), Lasioglossum fratellum (Q), Lasioglossum fulvicorne (P), Lasioglossum morio (R), Lasioglossum quadrinotatum (S), Nomada panzeri (T), Syrphus ribesii (U). -1 = range expansion, 0 = no occupancy (absence in both), 1 = no change (presence in both), 2 = range contraction.

    (TIF)

    Click here for additional data file. (21.5MB, tif)
    Attachment

    Submitted filename: Response to Reviewers.docx

    Click here for additional data file. (19.9KB, docx)

    Data Availability Statement

    All relevant data, including list of samples used in this study are provided as supplementary material.

    Articles from PLoS ONE are provided here courtesy of PLOS

    RESOURCES