Nutrient Analysis and Species Diversity of Alpine Grasslands: A Comparative Analysis of Less Studied Biodiversity Hotspots

: The alpine grasslands of Kashmir Himalaya act as a treasure house of ﬂoristic biodiversity. They have remained largely unstudied because of their remoteness and inaccessibility. It is imperative to have quantitative studies of these areas to allow the long-term monitoring of ﬂora in these fragile ecosystems. During the present study, nutrient analysis and species diversity of some alpine grasslands were investigated. Electroconductivity (EC) of the soils ranged between 0.12 and 0.33 (dSm − 1 ). With an increase in altitude and precipitation and a decrease in temperature, soil pH and available macro-nutrients (OC, N, P, K) show a considerable decrease. Sixty-six plant species belonging to twenty-nine families and ﬁfty-one genera were reported with members predominantly from the Asteraceae, Rosaceae and Plantaginaceae families. Seven species were common to all study areas and Renyi diversity proﬁles showed that Kongwattan was the most diverse followed by Poshpathri and Yousmarg. The results of the Sorensen β diversity index showed a relatively lower dissimilarity index among the three studied alpine sites. In the majority of the growth forms, growth initiation was recorded in April, whereas senescence occurred in September. The highest bloom was seen in June-July. The plant species exhibited a greater variability in their phenophases under different environmental conditions and altitudinal gradients. Plants were more vigorous at lower altitudes and showed rapid response to the prevailing conditions. Stoloniferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense Poa compressa Poa angustifolia, and Plantago lanceolata showed a greater importance value index (IVI). The sedentary system of livestock rearing at Yousmarg resulted in the decreased density of the palatable species. This study allowed us to conclude that direct knowledge of soil nutrient composition and species diversity in alpine ecosystems can enhance conservation and ensure better management practices over a period of time. G.N.; formal analysis, M.M.G. and S.V.; investigation, I.A.W.; M.M.G. and R.G.; data curation, S.V.; writing — original preparation, I.A.W. and S.V.; — review and I.A.W. and R.G.; visualization, G.N. S.V., G.N. Author Contributions: Conceptualization, S.V., M.M.G. and G.N. methodology, I.A.W. and R.G.; software, I.A.W.; validation, S.V., I.A.W., M.M.G. and G.N.; formal analysis, M.M.G. and S.V.; investi-gation, I.A.W.; resources, M.M.G. and R.G.; data curation, S.V.; writing—original draft preparation, I.A.W. and S.V.; writing—review and editing, I.A.W. and R.G.; visualization, G.N. and M.M.G.; super-vision, S.V. and G.N.; project administration, H.M.S. and F.A.A.-M.; funding acquisition, F.A.A.-M.


Introduction
The Himalaya is home to one of the most diverse and unique ecosystems on the planet [1]. Himalayan alpine plant communities are ecologically significant because they govern soil stability, play an important role in ecosystem functioning, and are important in cultural, ethical, and aesthetic aspects [2]. These regions show low productivity due lands, (2)study the floristic composition and phenology of different alpine grasslands , (3) understand the variation in the phenology and growth characteristics of plant species at different study areas, and (4) to study the dominance pattern and impact of grazing on species composition at different alpine grasslands. Statistical analysis was performed using different statistical analyses with R software.

Study Sites
The present study was carried out at the alpine regions of Yousmarg, Poshpathri and Kongwattan in Kashmir, India. Yousmarg lies in the Pir Panjal range of Kashmir Himalaya and is renowned for harboring a large number of alpine grasslands. The study was conducted in the upper reaches at 2831 m above mean sea level (m.a.s.l.). These areas remain free from snow from April to late October. In early growing months (April and May), the weather is cloudy and foggy, however, in June, July and August, itis clear with bright and longer durations of sunlight. The alpine grasslands of Poshpathri-Tral lie in the Zanskar mountainous range of Kashmir Himalaya. The grassland is located at 3103 m.a.s.l. The grassland of the Kongwattan lies in the Pir Panjal range of Kashmir Himalaya. It is located in the Kulgam district at an altitude of about 3347 m.a.s.l. The study was carried out on a gentle slope at all three sites. Poshpathri is situated on the southern slope of the mountain range, while Yousmarg and Kongwattan are on the northern slope. The meteorological information of all the sites during different months was provided by the Meteorological Department, Rambagh, Srinagar (J and K) and is given in Figure 1.

Soil, pH, Nutrient Analysis,and Bulk Density
Hydrogen ion concentration (pH) of the soil samples was determined in 1:2.5 soil: water ratio (w/v) with the help of a glass electrode pH meter [45]. Electrical conductivity was estimated in 1:2.5 soil: water suspension with EC meter [45]. The texture class of the

Soil, pH, Nutrient Analysis, and Bulk Density
Hydrogen ion concentration (pH) of the soil samples was determined in 1:2.5 soil:water ratio (w/v) with the help of a glass electrode pH meter [45]. Electrical conductivity was estimated in 1:2.5 soil:water suspension with EC meter [45]. The texture class of the soil was determined by the hydrometer method [46]. Organic carbon was estimated by the rapid titration method [47]. Available nitrogen was determined by using alkaline permanganate as per the modified Kjeldahl method [48]. Available phosphorus was extracted from the soil with 0.5 M NaHCO 3 (pH 8.5) and determined by the ammonium molybdate blue color method using a spectrophotometer [49]. A total of 1 N ammonium acetate was used as an extractant and the available potassium content was determined by feeding the extract to a flame photometer [45]. Exchangeable Ca was analyzed using ammonium acetate extract through the EDTA method. Available micronutrients (Zn, Cu, Mn, and Fe) were analyzed through the DTPA extractable method [50]. The bulk density of the study areas was determined by the soil core sampling method [51].

Floristic Composition and Phenological Studies
Regular field visits were made from April to October at all three study sites to determine their floristic composition. Flora analysis was performed by placing random quadrats (1 m 2 ), following Dad and Khan, [52]. Unknown plant species were identified with the help of regional floras of Dhar and Kachroo [53]. The phenology of the species was recorded throughout the study period from April to October. Data on each of the six phenophases viz. germination, vegetative growth, flowering, fruiting, seed maturation, and senescence were recorded [54,55]. The appearance of the first leaf in case of dicots was considered as initiation of germination [54,55], while in the case of graminoids seedlings upto 2 cm length were considered in the germination phase [55].
The impact of different environmental conditions (temperature and precipitation) and edaphic factors (altitude) was studied on the phenology (germination, vegetative phase, flowering, fruiting, seed maturation, and senescence) of the plant species.

Species Dominance Pattern
The quadrat method was used for the collection of such data following the method outlined by Curtis and Cottam [56]. Appropriate numbers of quadrats (1 m 2 ) were laid randomly in the study area. Density and abundance were calculated on a per meter square basis. Frequency was determined by dividing the total number of quadrants in which species were present by the total number of quadrants laid, and multiplying the result by 100. The seasonal changes in the flora were studied through tiller analysis [57]. Other structural parameters of different grasslands were reported using the following formulae: Relative density = number of individuals of particular species total number of individuals in area × 100 Relative frequency = frequency value of particular species total of all frequency values for all species × 100 Area = C 2 /4π Relative area = Basal cover of particular species/Total basal area IVI = Relative density + Relative frequency + Relative area 2.5. Statistical Analysis 2.5.1. Ecological Indices All analyses were conducted in R software v4.0.2 [58]. We used the Renyi diversity profile approach to calculate the plant species diversity of the three studied alpine sites using the vegan (v2.5-7) package [59]. Renyi diversity profiles are a function dependent parametric family of diversity indices that reflect sensitivity to rare and common species and display a graphical ordering of community diversity [60][61][62]. Renyi diversity profile values (H α ) were calculated from the frequencies of each contributing species and a scaling parameter (α) that ranges from zero to infinity [63] according to the following formula: where p i is the proportion abundance of each species and α is a scaling parameter [47,49,51]. The values of the Renyi profile at the given scales of 0, 1, 2, and ∞ correspond to species richness (S), the Shannon diversity index (H ), the Simpson diversity index (D −1 ) and the Berger-Parker diversity index (d −1 ), respectively [62,63]. According to the Renyi diversity profile, a community can be regarded as more diverse if all of its Renyi diversities are higher than that of the other community [60,62]. We calculated the diversity values in the current study at the α scale of 0, 0.25, 0.5, 1, 2,4,8,16,32,64, and infinity (∞) and plotted the diversity profiles for each site separately.

β-Diversity
To gain an in-depth understanding of composition change between the studied alpine sites, we calculated the turnover (i.e., species replacement between sites) and nestedness (i.e., species gain or loss between sites) components of beta diversity to study spatial patterns of turnover and nestedness-resultant dissimilarity among the three sites using a betapart package [64]. More specifically, it partitions the pairwise Sorenson dissimilarity between the two sites (βsor) into two additive components which in turn account for species spatial turnover (βsim) and nestedness-resultant dissimilarities (βsne) [65].

Predictors of Phenological Stages
We evaluated the relative effects of altitude, temperature, and precipitation on different phenological events with the generalized linear mixed models (GLMMs) and a Poisson error distribution using the lme4package [66]. Model performance was evaluated with the help of the Akaike information criterion corrected for small sample sizes (AICc) and the marginal (mR 2 ) and conditional (cR 2 ) R 2 values, using the 'dredge' function in the Mu Min package [67] to select the simplest model with ∆AICc < 2.

Pearson's Correlation Coefficient
We also performed Pearson's multiple correlation between community indices (i.e., plant species richness, diversity, and evenness measures) and environmental variables (temperature, precipitation, and altitude). The statistical significance associated with the resulting correlation was detected at the 5% level.

Nutrient Analysis
The soil pH showed a considerable decrease with an increase in altitude and precipitation. pH at Yousmarg ranged between 7.11 and 8.3, while at Kongwattan the pH of the soil ranged between 6.3 and 6.5. Electroconductivity of the soil ranged from 0.18 ± 0.03 dSm −1 (at Yousmarg) to 0.19 ± 0.04 (at Kongwattan). Soil organic carbon and macro soil nutrients (nitrogen, phosphorus, and potassium) showed a significant decrease with an increase in altitude and precipitation, and a decrease in the temperature. The highest organic carbon was reported at Yousmarg (2.33 ± 0.31) followed by Poshpathri (1.84 ± 0.29) and Kongwattan (1.45 ±0.36). The concentration of the micronutrients (zinc, iron, calcium, and copper) did not show any significant change with respect to change in altitude, temperature, and precipitation. There were no significant statistical differences in the bulk density and electroconductivity of soils at different study areas. Bulk density ranged between 1.23 ± 0.9 (at Kongwattan) and 1.34 ± 1 Mgm 3 (at Yousmarg). Detailed results are provided in Table 1.

Floristic Composition
During the entire study period, 23 plant species from17 genera and 17 families were documented at Yousmarg Budgam, 32 plant species from 25 genera and 18 families were reported at Poshpathri Tral, and 36 plant species from 32 genera and 21 families were reported at Kongwattan Kulgam. Only 7 plant species were common at all the three study sites, while Yousmarg had 9 exclusive plant species, Tral had 15, and Kongwattan had 22. A total of 9 plant species were common between the Yousmarg and Poshpathri grasslands, 10 between Poshpathri and Kongwattan, and 5 common species were reported from the grasslands of Kongwattan and Yousmarg (Table 2).   Environmental (temperature, precipitation) and edaphic (altitude) factors at different grasslands show a profound effect on the dominance pattern of the plant families. Asteraceae followed by Plantaginaceae and Polygonaceae were the dominant families with the highest genera and species at Kongwattan. Rosaceae followed by Asteraceae, Brassicaceae, and Fabaceae dominated the flora at Poshpathri, while Rosaceae followed by Polygonaceae, Plantaginaceae, Caryophylliaceae and Poaceae with a maximum number of genera and species dominated the flora of the alpine grassland at Yousmarg (Figure 2).

Phenology
The phenological spectrum as analyzed for the various plant species documented during the survey in the alpine grasslands of Kashmir Himalaya revealed that all the plants exhibited a distinct phenological pattern corresponding to their response to the harsh climatic conditions of the alpine grasslands. In alpine conditions the climate is not favorable for plant growth throughout the year, thus the plants have got a fixed period of 7-8 months to complete their life cycle. Germination of most plant species started in April following the melting of the snow. The melting of the snow makes moisture available which is necessary for germination. This period also witnessed a temperature increase favoring plant germination. The highest percentage of germination was recorded in April, followed by May and June. The highest number of species in the vegetative phase was found in May followed by June and July. The flowering of the plant species was observed from May to August. The highest number of plants in the flowering phase was seen in July followed by June, May, and August. Fruiting started in June and ended in September. The highest percentage of plants in the fruiting phase was seen in August followed by July and September. The signs of seed set were seen from the month of July. The highest number of species in the seed maturation phase was observed in September followed by August. By the end of September almost all of the species entered their senescence phase. Though the majority of the plant species completed their life cycle by the end of September, some plant species were found to be in the final stage of their life cycle (senescence) in October (Supplementary Tables S1-S3).

Impact of Environmental and Edaphic Factors on the Phenology of Plant Species
The climate and edaphic factors provide a significant impact on the germination and senescence of the plant species, whereas flowering, fruiting, and seed maturation were least affected. Lower altitude and precipitation and higher temperature at Yousmarg resulted in the early germination of Taraxicum officinale, Ranunculus hirtellus, Iris hookeriana, Sibbaldia cuneata, Poa compressa, Plantago major, and Trifolium repens as compared to the grassland sites at Poshpathri and Kongwattan where germination started in May. Similar phenological shifts were seen for plant species that were common between different study areas (Kongwattan and Poshpathri, Kongwattan and Yousmarg, Poshpathri and Yousmarg). Senescence of the plant species was initiated earlier at Kongwattan where the majority of the plant species entered their senescence phase in the month of September. Detailed results are provided in Figure 3.

Phenology
The phenological spectrum as analyzed for the various plant species documented during the survey in the alpine grasslands of Kashmir Himalaya revealed that all the plants exhibited a distinct phenological pattern corresponding to their response to the harsh climatic conditions of the alpine grasslands. In alpine conditions the climate is not favorable for plant growth throughout the year, thus the plants have got a fixed period of 7-8 months to complete their life cycle. Germination of most plant species started in April following the melting of the snow. The melting of the snow makes moisture available which is necessary for germination. This period also witnessed a temperature increase favoring plant germination. The highest percentage of germination was recorded in April, followed by May and June. The highest number of species in the vegetative phase was found in May followed by June and July. The flowering of the plant species was observed from May to August. The highest number of plants in the flowering phase was seen in July followed by June, May, and August. Fruiting started in June and ended in September. The highest percentage of plants in the fruiting phase was seen in August followed by July and September. The signs of seed set were seen from the month of July. The highest number of species in the seed maturation phase was observed in September followed by August. By the end of September almost all of the species entered their senescence phase. Though the majority of the plant species completed their life cycle by the end of September, some plant species were found to be in the final stage of their life cycle (senescence) in October (Supplementary Tables S1-S3).

Impact of Environmental and Edaphic Factors on the Phenology of Plant Species
The climate and edaphic factors provide a significant impact on the germination and senescence of the plant species, whereas flowering, fruiting, and seed maturation were least affected. Lower altitude and precipitation and higher temperature at Yousmarg resulted in the early germination of Taraxicum officinale, Ranunculus hirtellus, Iris hookeriana, Sibbaldia cuneata, Poa compressa, Plantago major, and Trifolium repens as compared to the grassland sites at Poshpathri and Kongwattan where germination started in May. Similar phenological shifts were seen for plant species that were common between different study areas (Kongwattan and Poshpathri, Kongwattan and Yousmarg, Poshpathri and Yousmarg). Senescence of the plant species was initiated earlier at Kongwattan where the majority of the plant species entered their senescence phase in the month of September. Detailed results are provided in Figure 3.

Environmental Correlates of Phenological Stages
The results of the generalized linear mixed models (GLMMs) are shown in Figure 4 and Table 3. The results indicate that temperature and elevation were the most influential environmental variables which significantly affected all of the studied phenological stages except vegetative growth and reaching mR 2 value from 0.34 to 0.94 ( Figure 4; Table  3). In contrast, precipitation had a significant effect on germination, vegetative growth, seed maturation, and senescence only ( Figure 4; Table 3). Table 3. Results of generalized linear mixed models (GLMMs) testing the effect of best performing environmental variables on different phenological stages. Numbers for variables show the z statistic of model significance. mR 2 and cR 2 indicate the fit of the models without (marginal) and with(conditional) random effect, respectively. *** p < 0.001, ** p < 0.01 and * p < 0.05.

Environmental Correlates of Phenological Stages
The results of the generalized linear mixed models (GLMMs) are shown in Figure 4 and Table 3. The results indicate that temperature and elevation were the most influential environmental variables which significantly affected all of the studied phenological stages except vegetative growth and reaching mR 2 value from 0.34 to 0.94 ( Figure 4; Table 3). In contrast, precipitation had a significant effect on germination, vegetative growth, seed maturation, and senescence only ( Figure 4; Table 3).

Species Dominance Pattern
Density, frequency, and abundance were reported on a monthly basis from April to October to determine the effect of grazing, temperature, and precipitation on plant species. Relative density, relative frequency, cover, and relative cover was analyzed to determine the importance value index (IVI). A definite trend in the increase of temperature, favorable precipitation, least pressure of grazing, and a considerable increase in the density of plant species was reported from April to July at all three grasslands. However, the decrease in temperature and movement of shepherds to these areas to feed their livestock resulted in a drastic decrease in the density of plant species from July onwards. While analyzing the density of the plant species, it was calculated that Trifolium repens, Poa angustifolia, Poa compressa, Sibbaldia cunenata, Mazus reptans, Trifolium pratense, Primula denticulata, and Myosotis stricta showed the highest density, while Impatiens brachycentra, Geum vernum, Viola biflora, Arabis alpina, Arenaria stricta, Crepis sancta, and Fimbristylis dichotoma were the least dense plant species at their respective sites. The highest total tiller density was reported at Kongwattan followed by Poshpathri and Yousmarg. Grazing intensity was highest at Yousmarg, while Kongwattan showed the lowest pressure of grazers and tramplers. The grassland of Yousmarg is located at an altitude of below 3000 m.a.s.l. and receives a sedentary system of livestock rearing. To feed their livestock, shepherds and the Bakharwal tribe show early and prolonged movement to these regions. This results in greater anthropogenic stress causing the decreased diversity of palatable species and the increased density of non-palatable species, such as C. falconeri, C. wallichii, etc. Stoloniferous forbs and tussock forming graminoids were the dominant groups at these alpine grasslands. Based on importance value index (IVI), Trifolium repens, Sibbaldia cuneata, Poa compressa, Plantago major, Plantago lanceolata, Trifolium pratense, Poa angustifolia, Primula denticulata, and Sambucus wightiana dominated the flora composition on these alpine grasslands (Table 4). Plant species such as Trifolium repens, Trifolium pratense, Sibbaldia cuneata, Plantago major, Poa compressa, Poa annua, and Plantago lanceolata were reported to be the most frequent plant species with a frequency percentage greater than 50%. Plant species such as Viola biflora, Thymus serpyllum, Rheum webbianum, Podophyllum hexandrum, Fimbristylis dichotoma, Epilobium hirsutum, Cardamine impatiens, and Capsella bursa-pastoris were the least frequent plant species with a frequency percentage less than 5%.

Species Dominance Pattern
Density, frequency, and abundance were reported on a monthly basis from April to October to determine the effect of grazing, temperature, and precipitation on plant species. Relative density, relative frequency, cover, and relative cover was analyzed to determine the importance value index (IVI). A definite trend in the increase of temperature, favorable precipitation, least pressure of grazing, and a considerable increase in the density of plant species was reported from April to July at all three grasslands. However, the decrease in temperature and movement of shepherds to these areas to feed their livestock resulted in a drastic decrease in the density of plant species from July onwards. While analyzing the density of the plant species, it was calculated that Trifolium repens, Poa angustifolia, Poa compressa, Sibbaldia cunenata, Mazus reptans, Trifolium pratense, Primula denticulata, and Myosotis stricta showed the highest density, while Impatiens brachycentra, Geum vernum, Viola biflora, Arabis alpina, Arenaria stricta, Crepis sancta ,and Fimbristylis dichotoma were the least dense plant species at their respective sites. The highest total tiller density was reported at Kongwattan followed by Poshpathri and Yousmarg. Grazing intensity was highest at Yousmarg, while Kongwattan showed the lowest pressure of grazers and tramplers. The grassland of Yousmarg is located at an altitude of below 3000 m.a.sl. and receives a sedentary system of livestock rearing. To feed their livestock, shepherds and the Bakharwal tribe show early and prolonged movement to these regions. This results in greater anthropogenic stress causing the decreased diversity of  Table 3. Results of generalized linear mixed models (GLMMs) testing the effect of best performing environmental variables on different phenological stages. Numbers for variables show the z statistic of model significance. mR 2 and cR 2 indicate the fit of the models without (marginal) and with(conditional) random effect, respectively. *** p < 0.001, ** p < 0.01 and * p < 0.05.

Species Diversity
The Renyi diversity profiles of the three studied alpine sites are presented in Figure 5. The results showed that among all three sites Kongwattan is the most diverse followed by Poshpathri, and Yousmarg. In addition, this pattern of Kongwattan being the most diverse is consistent with all the α values used ( Figure 5).

Species Diversity
The Renyi diversity profiles of the three studied alpine sites are presented in Figure  5. The results showed that among all three sites Kongwattan is the most diverse followed by Poshpathri, and Yousmarg. In addition, this pattern of Kongwattan being the most diverse is consistent with all the α values used ( Figure 5).

β-Diversity
The results of the multiple-site dissimilarities are shown in Figure 6. The results indicate that the Sorensen dissimilarity among the three studied alpine sites was relatively low. Furthermore, the nestedness component (βsne) was found to contribute largest to the overall dissimilarity among the studied sites, as shown by the higher peak of βsne (nestedness-resultant dissimilarity) ( Figure 6A). This in turn reflects that the observed dissimilarity among the sites is most likely a consequence of richness difference (i.e., nestedness component) and to a lesser extent because of species replacement (i.e., turnover component). Moreover, the clustering obtained from the dissimilarity matrices of the turnover component showed that Poshpathri was highly dissimilar to the other two sites (Yousmarg and Kongwattan), which were similar to a greater extent as they formed the same sub-cluster ( Figure 6B). Contrary to this, the clustering obtained from the dissimilarity matrices of nestedness showed a quite different pattern in which the Yousmarg site was highly dissimilar to the other two sites (PP and KW), which were similar to a greater extent as they formed the same sub-cluster ( Figure 6C).

Correlation
The results of the multiple correlation are presented in Figure 7. The results indicate that species richness is positively correlated with altitude, precipitation, Shannon diversity, and Simpson diversity (r = 0.96, 0.72, 1, and 1, respectively), but the resultant correlation is significant only between species richness and Shannon diversity index (p < 0.05). Similarly, species richness is negatively correlated with temperature and Pielou's evenness (r = −0.9 and −1, respectively), but the resultant correlation is significant between species richness and Pielou's evenness only (p < 0.05). In addition, there was a significant positive correlation between Shannon diversity index and Simpson diversity index (r = 1; p < 0.05) and a significant negative correlation between Shannon diversity index and Pielou's evenness (r = −1; p < 0.05) and between Simpson diversity index and Pielou's evenness (r = −1; p < 0.05) (Figure 7).
The results of the multiple-site dissimilarities are shown in Figure 6. The results indicate that the Sorensen dissimilarity among the three studied alpine sites was relatively low. Furthermore, the nestedness component (βsne) was found to contribute largest to the overall dissimilarity among the studied sites, as shown by the higher peak of βsne (nestedness-resultant dissimilarity) ( Figure 6A). This in turn reflects that the observed dissimilarity among the sites is most likely a consequence of richness difference (i.e., nestedness component) and to a lesser extent because of species replacement (i.e., turnover component). Moreover, the clustering obtained from the dissimilarity matrices of the turnover component showed that Poshpathri was highly dissimilar to the other two sites (Yousmarg and Kongwattan), which were similar to a greater extent as they formed the same sub-cluster ( Figure 6B). Contrary to this, the clustering obtained from the dissimilarity matrices of nestedness showed a quite different pattern in which the Yousmarg site was highly dissimilar to the other two sites (PP and KW), which were similar to a greater extent as they formed the same sub-cluster ( Figure 6C).

Correlation
The results of the multiple correlation are presented in Figure 7. The results indicate that species richness is positively correlated with altitude, precipitation, Shannon diversity, and Simpson diversity (r = 0.96, 0.72, 1, and 1, respectively), but the resultant correlation is significant only between species richness and Shannon diversity index (p < 0.05). Similarly, species richness is negatively correlated with temperature and Pielou's evenness (r = −0.9 and −1, respectively), but the resultant correlation is significant between species richness and Pielou's evenness only (p < 0.05). In addition, there was a significant

Discussion
Grasslands account for about 40% of total land area and thus act as a significant ecosystem on a global scale [68]. Grasslands are a key component of Alpine and pre-Alpine landscapes, with a wide range of appearances ranging from heavily exploited grasslands in the lower areas to very diverse seasonal alpine pastures and specialized

Discussion
Grasslands account for about 40% of total land area and thus act as a significant ecosystem on a global scale [68]. Grasslands are a key component of Alpine and pre-Alpine landscapes, with a wide range of appearances ranging from heavily exploited grasslands in the lower areas to very diverse seasonal alpine pastures and specialized natural ecosystems at the higher elevations [69,70]. Besides providing significant economic value, alpine pastures deliver a wide range of ecosystem services, such as storage and cycling of different nutrients (prominently nitrogen and carbon) and water retention [71][72][73]. Previous studies have shown that species diversity decreases with increasing altitude [74]. In our study, species diversity increased with an increase in altitude. This is possibly due to the sedentary livestock grazing at the lower alpine sites, resulting in decreased species diversity as compared to higher diversity at upper alpine reaches where the pressure of grazing and trampling is less [75,76].
Soil pH values increased with the increase in altitude and the decrease in temperature at different grassland sites. These results are consistent with the findings of Yimer et al. [77], Oyonarte et al. [78], and Roukos et al. [79]. At warmer temperatures, decomposition results in greater soil organic carbon density [80] which tends to increase the concentration of H + ions [81], resulting in lower pH at lower altitudes with higher temperatures. Nitrogen, phosphorus, and potassium get significantly reduced with increased altitude. These findings are supported by the results of Bhandari and Zhang [82] who also reported similar findings on the grasslands of Tibet, China, and Manang District, Nepal.
The flora of alpine regions face demanding environmental conditions with cold soils having low moisture and low winter temperatures with extended permafrost and a brief growing season [83,84]. To cope with such adverse conditions, plants of alpine regions show various morphological and physiological adaptations and tend to complete their life cycle within a short duration during favorable climatic conditions [85][86][87]. The survival of most species in the grasslands can also be attributed to a modest degree of species competition during early regeneration, which has resulted in the dominance of only a few species [88]. In the present study, plant species from the Asteraceae, Fabaceae, Plantaginaceae, and Rosaceae families were dominant and this can be attributed to their adaptation to the alpine environment by producing large amount of seeds and showing diverse reproductive strategies.
Prolonged snow cover, reduced photoperiod, low temperature, and frost (hence, the lack of moisture) during the early months of the spring prevent the plant species from overcoming their dormancy [89][90][91]. The weather shift from late March results in a change in environmental conditions on the alpine grasslands. The increase in temperature not only results in the melting of snow, which makes optimum soil moisture available for the floral evocation of plant species, but also helps in breaking the dormancy of seeds and buds under the alpine conditions. During this study, species specific responses to environmental conditions were reported while analyzing the germination at these grasslands. About 76.26% of the plant species started germination in the month of April, while the remaining 23.74% of the plant species germinated in the month of May. While analyzing the effect of climate change on plant species at different alpine grasslands, various investigators reported that the change in the weather conditions (increase in temperature, melting of snow, change in photoperiod, melting of permafrost) during the early days of spring played a vital role in the germination of the plant species [92][93][94].
Observations made to assess the floral evocation of plants revealed that flowering attains its peak in July, exploiting the period of most favorable climatic conditions. Such observations are supported by the findings of different researchers who also reported direct proportionality of peak flowering with peak temperatures [89,[95][96][97][98]. Plant species tend to complete their life cycle before they face a drastic decline in temperature and precipitation in the alpine regions. Completing their life cycle within the stipulated time of 7-8 months was observed to be an important adaptation by these alpine plants to survive the severe conditions of the alpine regions. Vashistha [98], while studying the phenology of plant species at an alpine region of north-west Himalaya (India), also reported initiation of senescence from September onwards and reported that the plants of alpine regions exploit the favorable weather conditions in such a way that they complete their whole life cycle within this period (April-October).
IVI analysis gives information about a species' social interactions and may be identified as a pattern of dominating species in a population [99]. IVI analysis revealed distinct combinations of species with different dominants and co-dominants at different study sites. Based on the importance value index (IVI), dominance of the plant species reflects a considerable degree of variation. Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia, and Plantago lanceolata depicted wide amplitude and showed greater IVI (more dominant) as compared to other plant species, reflecting their adaptation to the conditions of alpine regions. The species found to be dominant were rhizomatous and grow as tussocks. Such observations are supported by the results of different workers [100,101] who also reported these plant species to be dominant on other alpine grasslands of the western Himalayas. Changing environmental conditions and pressure caused by grazers and tramplers exert an immense impact on the community characteristics of plants in alpine regions. A considerable increase in the values of density and frequency of plant species upto July was observed to be in accordance with the increase of favorable climatic conditions and minimal grazing pressure. A decline in such values from July onwards is attributed to the change in weather conditions (temperature, precipitation, and photoperiod) and the increase of anthropogenic pressure caused by the grazers and tramplers. Such results are in accordance with the findings of other researchers [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.

Supplementary Materials:
The following supporting information can be downloaded at: www.mdpi.com/xxx/s1, Table S1: Phenology of plant species at Yousmarg.  environmental conditions and pressure caused by grazers and tramplers exert an immense impact on the community characteristics of plants in alpine regions. A considerable increase in the values of density and frequency of plant species upto July was observed to be in accordance with the increase of favorable climatic conditions and minimal grazing pressure. A decline in such values from July onwards is attributed to the change in weather conditions (temperature, precipitation, and photoperiod) and the increase of anthropogenic pressure caused by the grazers and tramplers. Such results are in accordance with the findings of other researchers [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.

Supplementary Materials:
The following supporting information can be downloaded at: www.mdpi.com/xxx/s1, Table S1: Phenology of plant species at Yousmarg.   [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.

Supplementary Materials:
The following supporting information can be downloaded at: www.mdpi.com/xxx/s1, Table S1: Phenology of plant species at Yousmarg.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions. environmental conditions and pressure caused by grazers and tramplers exert an immense impact on the community characteristics of plants in alpine regions. A considerable increase in the values of density and frequency of plant species upto July was observed to be in accordance with the increase of favorable climatic conditions and minimal grazing pressure. A decline in such values from July onwards is attributed to the change in weather conditions (temperature, precipitation, and photoperiod) and the increase of anthropogenic pressure caused by the grazers and tramplers. Such results are in accordance with the findings of other researchers [102,103] who also reported a decline in the density and frequency values due to climate change and an increase in grazing.

Conclusions
The present work on the nutrient analysis, species diversity, and ecological analysis allowed us to conclude that:  Nutrient composition (N, P, K) of the alpine soil show negative correlation with altitude and precipitation and positive correlation with temperature.  The altitudinal gradients and grazing intensity had a significant impact on the floristic diversity of alpine grasslands in Kashmir Himalaya.  The phenology of the plant species is significantly affected by the altitude, temperature, and precipitation at different study areas.  Observations related to the phenology have radically proved that certain factors such as low temperature, heavy snowfall, frost, and high altitude interfere with the vegetative growth of plant species, and are essential for the morphogenesis of flowering.  Renyi diversity profiles show that the alpine grassland of Yousmarg has the least floristic diversity due to a sedentary system of livestock rearing.  The Sorensen β diversity index reveals a relatively great similarity index among the studied sites which reflects the adaptability of the plant species in the higher altitudes of alpine regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.  Stonliferous forbs and tussock forming graminoids such as Sibbaldia cuneata, Trifolium repens, Plantago major, Trifolium pratense, Poa compressa, Poa angustifolia and Plantago lanceolata showed greater importance value index (IVI) and dominated the floristic composition of these regions.