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    Extensive range contraction predicted under climate warming for two endangered mountaintop frogs from the rainforests of subtropical Australia

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    Response of Canola productivity to integration between mineral nitrogen with yeast extract under poor fertility sandy soil condition

    Photosynthetic pigmentsBased on the analysis of variance, data of Photosynthetic pigments as presented in Table 1 indicate that photosynthetic pigments as chlorophyll a (Chl. a) had non-significant for three Canola genotypes AD201 (G1), Topaz (G2) and SemuDNK 234/84 (G3), but chlorophyll b (Chl. b) and chlorophyll a/b ratio (Chl. a/b) had significant difference for three genotypes. Chl. a, Chl. b and Chl. a/b were positively responded to different N application i.e. without nitrogen fertilization (control F0), 95 kg N ha−1 (F1), 120 kg N ha−1 (F2) and 142 kg N ha−1 (F3) (without yeast); and integrated between nitrogen fertilization and yeast extract (YE) treatments as follows: 95 kg N ha−1 + YE (F4), 120 kg N ha−1 + YE (F5) and 142 kg N ha−1 (F6) (with yeast), data indicated that F5 and F6 gave the highest values of Chl. a and Chl. a/b ratio and lowest values of Chl. b Table 1. Interaction data showed that three Canola genotypes that were fertilized with N without yeast or with yeast had a slight difference with statistically significant in chl. a. The highest values of Chl. an obtained by G2 under F5 treatment followed by G1 under F6 treatments. In respect to Chl. a/b ratio, statistical analysis showed that Interaction between Canola genotypes treated with N applications without or with yeast had a significant difference whereas the highest values were recorded when Canola genotypes G3 and G2 fertilized with F6 and F5 with slight differences. While the interaction was significant between N treatments and Canola genotypes for Chl. b. and Canola genotype (G1) gave the highest value when treated with F1. Generally, F6 and F5 improve the contents of chl. a and chl. a/b ratio for three Canola genotypes Table 1. Chl. contents were increased in plants grown under middle and high N conditions as compared with plants grown under low N conditions, which significantly affected photochemical processes20. N is a fundamental element for leaf plants, insufficient N supply lead to decreased photosynthetic rate in plants21, this occurs to many factors such as a decrease in pigment degradation22, reduction in stomatal conductance23 and a decline in the light and dark reaction of photosynthesis. Canola is a nitrophilous plant, wherein a high concentration of NO3 in the culture media results in higher Chl. contents in the plant leave compared with controls20. The Chl. a/b ratio can be a valuable indicator of N element within a leaf because this ratio must be positively related to the ratio of PSII cores to light-harvesting chlorophyll-protein complex (LHCII)24. LHCII contains the majority of Chl. b, consequently it has a lower Chl a/b ratio than other Chl. binding proteins associated with PSII25. Thus, Chl. a/b ratios should increase with decreasing N availability, especially under high light conditions26, the Chl. a/b ratio and the ratio of PSII to Chl. are independent of N availability for spinach27, and lower Chl. a/b ratios were noticed when plants were subjected to low N28, while Kitajima and Hogan29 revealed that the Chl. a/b ratio increased when Chl. content decreased in response to N restriction in photosynthetic cotyledons in leaves of seedlings of four tropical woody species in the Bignoniaceae, and Bungard et al.30 demonstrated that there is a tiny response in Chl. a/b ratios to light or N. The yeast includes bio-regulators i.e. plant growth regulators and endogenous plant hormones, which enhance photosynthesis, also it produces 5-Aminolevulinic acid which is vital to tetrapyrrole biosynthesis and biochemical processes in plants, including heme and Chl. biosynthesis25.Table 1 Photosynthetic pigments for the three Canola genotypes under different N applications without and with yeast extract.Full size tableYield and its attributesComparing of mean data through the Duncan Multiple Range Test in the probability level of 5%, data showed significant differences among the Canola genotypes for the highest plant (cm), branches number/plant, and pods number/plant. On contrary, there wasn’t a significant difference for seed number/pods, seed yield (t ha−1), biological yield (t ha−1), and harvest index, wherein G2 gave the highest value for the highest plant (cm). In the same trend, G2 gave the highest values of branches No./plant and pods No./plant followed by G3 for the previous two treats Table 2. All examined N without or with yeast caused a significant difference in yield and its attributes, wherein F6 positively affected on abovementioned traits and gave the highest values on the highest plant (cm), branches No./plant, pods No./plant, seed No./pods, seed yield (t ha−1), and harvest index. While the highest values of biological yield (t ha−1) were obtained with F3, F6, and F5, respectively Table 2.Table 2 Growth, yield and its attributes for the three Canola genotypes under different N applications without and with yeast extract.Full size tableThe interaction between the Canola genotype and different N rates without or with yeast extract as shown in Table 2, demonstrated a significant difference. Data showed that the highest values of plant height and pods No./plant were recorded by G2 under F6 and the highest values of branches No./plant, seed No./pods, and seed yield (t ha−1) got by G3 and G2 under F6. There was a slight difference with statistically significant biological yield (t ha−1) and highest values established by G1 under F3 and F6; and G2 and G3 under F3, F5, and F6 respectively; and the highest values of harvest index recorded by G1, G2 and G3. under F6. Generally, data proved that 142 kg N/ h−1 + YE (F6) was enhanced the yield and its components of three Canola genotypes i.e. AD201 (G1), Topaz (G2), and SemuDNK 234/84 (G3). Many researchers reported that there are significant differences among Canola varieties and growth and yield traits are significantly increased by increasing N rates11. Increasing N fertilizer rates significantly increased most of the yield and its components31, N enhances metabolites synthesized by the plant which leads to more transformation of photosynthesis to reproductive parts, and induces different physiological mechanisms to access the nutrient32. Yeast extract as bio-fertilizer had a significant and positive effect on plant height and yield traits of Canola. The role of bread yeast in increasing the growth and yield traits; may be due to the content of yeast to many important nutrients elements i.e. N, Mg, Ca, Zn, Cu, and Fe, and the production of some growth regulators such as Auxin and Gibberellin and cytokinin which is necessary for plant biological processers especially photosynthesis and cell division and elongation33. Also, Yeast extract had stimulatory effects on cell division and enlargement, protein and nucleic acid synthesis, and chlorophyll formation34, in addition to its content of cryoprotective agent, i.e. sugars, protein, amino acids, and also several vitamins35. Consequently, it improves growth, flowering, and fruit set and formation and increases yield34.Correlation of Canola seed yield and chlorophyll a/b ratioPartial correlation coefficients of Canola seed yield and Chl. a/b ratio is given in Fig. 1. This result showed that seed yield was positively correlated with Chl. a/b ratio when the amount of N applied without or with yeast extract is increased. Chl. a/b ratio can be an important indicator of N within a leaf, this ratio must be positively related to photosynthesis and biological processers which reflect on seed yield.Figure 1Correlation of Canola seed yield (t/h) and chlorophyll a/b ratio as affected by different nitrogen rates without and with yeast extract.Full size imageCorrelation of Canola seed yield and its attributesCorrelations of seed yield and yield components of Canola are a function of the plant height, number of branches/plant, number of pods/plant, and number of seeds/pod as shown in Fig. 2a–d. These results proved that grain yield was strongly positively correlated with some of the abovementioned traits when N fertilization increased without or with yeast extract. Sufficient N contributes to enhance physiological processes, improves growth, flowering, seed formation, and the seed yield finally.Figure 2(a) Correlation of Canola seed yield (t/h) and plant height (cm) as affected by different nitrogen rates without and with yeast extract, (b) Correlation of Canola seed yield (t/h) and branch No/plant as affected by different nitrogen rates without and with yeast extract, (c) Correlation of Canola seed yield (t/h) and pods No/ plant as affected by different nitrogen rates without and with yeast extract, and (d) Correlation of Canola seed yield (t/h) and seeds No/ pod as affected by different nitrogen rates without and with yeast extract.Full size imageChemical propertiesRegarding results of the oil yield (t ha−1), seed oil %, protein %, N % in seed, and N% in straw as presented in Table 3, data showed significant differences among three Canola genotypes; AD201 (G1), Topaz (G2) and SemuDNK 234/84 (G3), excepted oil yield had non-significant difference. G1 was surpassed in oil %; G2, G3 surpassed in protein % and N % in seed, and G3 surpassed in N% in straw. Different N fertilization applies without or with yeast extract had a significant effect on the abovementioned traits, wherein F6 treatment gave the highest oil yield, protein %, N % in seed, and N% in straw, while seed oil % significantly increased with F1 and F4 treatments. There was significant interaction concerning with abovementioned traits, Table 3, as well as the highest values of seed oil yield (t ha−1), protein % in seeds, and nitrogen % in seeds were obtained with G1, G2, and G3 when treated with F6. Wherein the highest values of oil % were obtained by G1 under F1 and F4 treatments. Concerning N% in straw was increased by increasing the rate of N fertilizer application and the highest value was recorded by adding F6 to G336. Seed oil percentage was decreased by increasing nitrogen rates; the effect of interaction between Canola cultivars and nitrogen fertilization treatments was significant on seed oil. % High rates of N led to decreases in seed oil % and increase in protein concentrations in Canola seed37, the increase in seed protein % because N is an integral part of protein and the protein of Canola.Table 3 Effect of different N applications without and with yeast extract on oil yield, oil %, protein %, N % in seed and N% in straw for the three Canola genotypes.Full size tableCorrelation of Canola seed yield and seed oil percentageA strong negative correlation was detected between seed oil percentage as shown in Fig. 3. The result indicates that seed oil percentage decreases with increasing in different N fertilization rates without or with yeast extract. That’s a negative correlation between seed yield and seed oil %; it might be due to N application which results in delaying maturity leading to poor seed filling and a greater proportion of green seed38.Figure 3Correlation of Canola seed yield (t h−1) and oil % as affected by different nitrogen rates without and with yeast extract.Full size imagePhysico-chemical properties of Canola oilThe effects of different N application rates without or with yeast extract on Canola genotypes on physico-chemical properties i.e. Acid value (mg g−1), saponification number (mg g−1) and peroxide value (mg kg−1) were shown in Table 4. Data of chemical properties of Canola oil showed significant differences among Canola genotypes, the highest acid value and peroxide value were obtained from G2 followed by G1 and G3, respectively, while the highest saponification number was obtained by G3 followed by G1 and G2, respectively.Table 4 Oil properties for three Canola genotypes under different N applications without and with yeast extract.Full size tableData had significant differences among different N application rates without or with yeast extract, by increasing the N rated from F0 to F6 caused decreases in Acid value, Saponification number, and peroxide value. Also, data showed a significant interaction between Canola genotypes and different N application rates without or with yeast extract for all abovementioned traits, wherein the highest values of saponification number were obtained by G1 and G3 under F0 treatment. In addition, the highest values of peroxide value and the acid value were obtained by G2 with F0. The acid value is a physicochemical indicator38, wherein oils which have higher acid value posse poor quality39, on another hand, Low acid value of Canola genotype shows their higher oil quality. The peroxide value varied between 7.1 and 9.06 meq. O2/kg indicates that the tested vegetable oils are fresh, and the lowest initial peroxide value is suitable for consumption40. High saponification value indicated that Canola oil possesses normal triglycerides and may be useful in the production of liquid soap and shampoo41. Saponification number was significantly different among genotypes and a higher nitrogen rate resulted in an increase in the unsaponifiable matter and led to a decrease in oil acid value and saponification value42.Fatty acids composition percentages in Canola oilThe main values of fatty acids composition percentages in Canola oil were determined and calculated in the second season Table 5. Gas–liquid chromatographic analysis showed that, saturated fatty acids (Palmitic, 16:0, Stearic, 18:0, Arachidic, 20:0, and Behenic, 22:0) represent about 9.1 of the total fatty acids. Palmitic was the dominant acid among the saturated ones. In respect of unsaturated fatty acids i.e., Oleic acid (18:1), Linoleic (18:2), Linolenic (18:3), and Erucic (22:1), they all represent about 90.9% of total fatty acids. Therefore, Oleic acid (18:1) was the major fatty acid in Canola oil (59.43%) followed by Linoleic (20.80%) and Linolenic (9.02%). Erucic acid was less than 2%.Table 5 Saturated and unsaturated fatty acids (%) in seeds of the three Canola genotypes and different N applications without and with yeast extract.Full size tableData in Table 5, showed slight differences in saturated fatty acids between Canola varieties. AD201(G1) variety contained more amount of Palmitic (4.78%) and Stearic (1.52%) acids followed by Topaz (G2) for Palmitic and SemuDNK 234/84 (V3) for Stearic. However, Behenic acid (1.20%) was higher in G3 than G2 (1.17%), while G2 was the highest in Arachidic acid than G3 variety. These results are in line with those obtained by El Habbasha et al.43. They reported that AD 201, Silvo, and Topas (G2) were different in their oil contents of saturated and unsaturated fatty acids. Canola varieties were also slightly differed in their content of the unsaturated fatty acids Table 5, G3 variety contained more amounts of Oleic (60.36%) acid followed by the G2 variety. G1 recorded the lowest amount of Oleic acid (58.36%) in comparison with the other two varieties. On the other hand, G1 showed a high increment in Linoleic and Linolenic acids followed by G3 for Linoleic and Linolenic acids. The second oil quality breeding objective is to reduce the percentage of Linolenic acid from the percent 8–10% to less than 3% while maintaining or increasing the level of Linoleic acid44. Lower Linolenic acid is desired to improve the storage characteristics of the oil, while higher Linolenic acid content may be nutritionally desirable. Similar observations were reported by Ref.45. Topaz variety recorded the highest value for Erucic acid (1.77%) followed by AD201 variety, whereas Semu DNK gave the lowest value (1.45%). The increase in Erucic acid content in the Topaz variety may be due to the decrease in Oleic acid content46. Stated that the concentrations of Oleic and Erucic acids were negatively correlated and a high Oleic acid concentration ( > 50%) was always associated with a low Erucic acid concentration ( More

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    Community succession and functional prediction of microbial consortium with straw degradation during subculture at low temperature

    Changes of straw degradation characteristics at different culture stagesCorn straw degradation ratioCorn straw weight loss in M44 at F1 reached 35.90% at 15 ℃ for 21 days, which was greater than that at F5, F8, and F11 by 2.33%, 3.01%, and 3.35%, respectively. There were no significant differences between F8 and F11(Fig. 1).Figure 1Corn straw degradation ratio was measured at different culture stages. The same small letter means there was no significant difference, and different small letters indicate significant differences at p  More

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    Extinction magnitude of animals in the near future

    Selection of environmental-biotic events to be studiedIn global warming events associated with mass extinctions, the current environmental changes are similar to those recorded during the end-Ordovician, end-Guadalupian, and end-Permian mass extinctions. Therefore, I analyzed global surface temperature anomalies, mercury pollution concentrations, and deforestation percentages in these three mass extinctions and in the current crisis. The asteroid impact at the K–Pg boundary and nuclear war cause the formation of stratospheric soot aerosols distributed globally, thus inducing sunlight reductions and global cooling (impact winter and nuclear winter). I also analyzed stratospheric soot aerosols as a possible cause of future extinctions.Most likely case and worst caseThe most likely case corresponds to the reduction of CO2 emissions resulting from human conduct, the protection of forests, and the introduction of anti-pollution measures in the future under the Paris Agreement on Climate change and Sustainable Development Goals (SDGs). The worst case corresponds to the scenario in which humans fail to stop increasing global surface temperatures, pollution, and deforestation until 2100–2200 CE.I use the average of the RCP4.5 and RCP6.0 cases in the Intergovernmental Panel on Climate Change (IPCC)8 as the most likely case of GHG emissions, representing the middle of the four potential GHG emissions cases (RCP2.6, 4.5, 6.0, and 8.5) in Fifth Assessment Report of the IPCC8, approximately corresponding to the middle of SSP2-4.5 and SSP3-7.0 in Sixth Assessment Report of the IPCC9. The timing of decreased global GHG emissions is 2060–2080 CE. Therefore, I use the average GHG emissions and global surface temperature anomalies of the RCP4.5 and RCP6.0 cases as the most likely values and those of the RCP8.5 case as the worst-case scenario, marked by stopping GHG emissions from 2090 to 2100 CE8,9, as this case corresponds to the highest GHG emissions8,9.Surface temperature anomaly, environment, and extinction magnitude dataData on surface temperature anomalies and extinction percentages are from Kaiho4. Changes in industrial GHG emissions and global surface temperature anomalies are sourced from the Fifth and Sixth Assessment Report of the IPCC8,9.Pollution can be represented by mercury concentrations measured in sedimentary rocks recording mass extinctions8 and in recent sediments deposited in seas and lakes25,26 because mercury is toxic to plants and animals and because its sources include volcanic eruptions, meteorite impacts, and the combustion of fossil fuels10,33, which are common sources of pollutants, and because it can be commonly measured from sedimentary rocks recording mass extinctions33. The mercury concentration is related to the CO2 emission amount during global warming because of the common sources of mercury and CO2 (volcanism and fossil fuel combustion influencing global warming). Thus, the future mercury concentrations are estimated based on the CO2 emission amounts estimated by the IPCC8,9. Since mercury and the other pollutants mainly come from oil, coal, and vegetation33, the amount of mercury released should change in parallel with industrial CO2 emissions because there is a good correlation between mercury and CO2 emissions11.Deforestation occurs by the expansion of agricultural areas and urban areas, which are strongly related to human populations13,28. Thus, future deforestation percentages are estimated based on estimated future population data27 (Supplementary Table S2). The severity of deforestation in each event is expressed by the occupancy % of the deforested area in the pre-event forest area in (i) the Permian–Triassic transition marked by the largest mass extinction based on plant fossil records24 and (ii) 2005–2015 CE as a representative of the Anthropocene epoch12,13,28 based on the actual forest area relative to the pre-agriculture phase before 4000 BP. Deforestation is related to the human population because agriculture and urbanization have caused deforestation13,28. I estimate the past and future deforestation percentage using human population data in the past and future21 based on the parallel growth of the human population and deforestation13,28.Amount of stratospheric soot was calculated using a method of Kaiho and Oshima34 (Supplementary Table S1). I obtained global surface temperature anomaly caused by stratospheric soot using Fig. 5 of Kaiho and Oshima34.I then use those data to estimate the future extinction magnitude based on the assumption that the Earth and contemporary life at the time of each crisis are more or less mutually comparable throughout time and to the present day.I estimate the magnitude of the species animal extinction crisis between 2000 and 2500 CE using Figs. 1, 2 and Supplementary Tables S1 and S2 in each cause under the most likely case and worst case under three nuclear war scenarios (zero, minor, and major; Fig. 2d)15 in the PETM and mass extinction cases, respectively (Supplementary Tables S3, S4; Fig. 3). Finally, I estimate the magnitude of current animal extinction crisis by the four causes as an average of the species extinction magnitude by the four causes in Fig. 3. I use two different contribution rates of temperature anomalies, pollution, deforestation, and stratospheric soot by nuclear wars, 1:0.2:0.1:1 for marine animals and 1:0.5:1:1 for terrestrial tetrapods (different contribution case considering lower influence of pollution and deforestation to marine animals rather than terrestrial animals) and 1:1:1:1 for marine animals and 1:1:1:1 for terrestrial tetrapods (equal contribution case considering high influence of pollution and deforestation to marine animals via rain and soil erosion) (Supplementary Tables S5–S9). These contribution rates are estimated as end-members to show ranges of animal species extinction magnitude (%). More

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    Ecologists should create space for a wide range of expertise

    Madhusudan Katti says ecology would benefit from including perspectives from all of Earth’s inhabitants.Credit: Marc Hall

    Decolonizing science

    Science is steeped in injustice and exploitation. Scientific insights from marginalized people have been erased, natural history specimens have been taken without consent and genetics data have been manipulated to back eugenics movements. Without acknowledgement and redress of this legacy, many people from minority ethnic groups have little trust in science and certainly don’t feel welcome in academia — an ongoing barrier to the levels of diversity that many universities claim to pursue.
    In the next of a short series of articles about decolonizing the biosciences, Madhusudan Katti suggests five shifts that ecologists need to make to unravel the effects of colonization on their field. Katti, an evolutionary ecologist at North Carolina State University in Raleigh, would also like to see stronger inclusion of uncredentialed experts and Indigenous communities in research.

    Last year, my colleagues and I wrote a paper highlighting five shifts that would help to decolonize ecology (C. H. Trisos et al. Nature Ecol. Evol. 5, 1205–1212; 2021). Ecologists need to improve how they incorporate varied perspectives, approaches and interpretations from the diverse peoples inhabiting Earth’s natural environments. The five shifts are: the individual need to decolonize one’s mind; understand the history of colonization and how it shaped Western ecology; facilitate access to and dissemination of data; recognize diverse scientific expertise; and establish inclusive research groups. Although it can be difficult to make reforms given how resistant institutions are to change, we are optimistic because we have received invitations to speak on these issues. People are ready for these conversations.
    Decolonizing science toolkit
    My colleagues and I developed a workshop around the five shifts. We have conducted the workshop at my institution, and at the annual conference of the Society for Integrative and Comparative Biology. For each of the shifts, I have participants brainstorm and write down challenges and solutions that might lead to progress in these areas for their own research departments or institutions. We address them, shuffle groups and suggest policy changes and future action.Some organizations are already moving forward with some low-hanging fruit, such as making data and published results more accessible. However, open-access publishing models put an even greater burden of publication costs on authors and perpetuate inequalities, because early-career researchers and those in the global south often can’t afford them.The most contentious area tends to be the reluctance of academia to accept non-credentialed expertise such as traditional knowledge. Universities are in the business of giving out credentials in the form of degrees. If academia no longer requires a PhD, that can be a challenge to that model. There are also few, if any, incentives or rewards to spend time working towards decolonizing academia, even though it takes time and effort away from furthering individual careers.As an Indian American, I would like to see institutions expand antiracism conversations rather than introduce new checklists of things to do. For example, at annual meetings, it would be great to see scientific societies make more connections with the Indigenous communities where we work and invite them to share their perspectives.
    This interview has been edited for length and clarity. More

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    Wildflower phenological escape differs by continent and spring temperature

    We used a hierarchical Bayesian modeling approach to evaluate the relationship between the spring phenology of tree and wildflower species and various climate drivers (see Methods). Following model selection, our final model structure included fixed effects of average spring (March–April) temperature and elevation, as well as species-level random effects. We show continental distributions of spring temperature values in Fig. 1b (means and standard deviations are listed in Table S2). We report estimates for spring temperature sensitivities from the final model structure in the main text. Parameter estimates for elevation sensitivities as well as the model performance of other potential drivers and combinations of drivers are reported in Tables S3 and S4. An extended discussion of model assumptions and limitations is included in the Supplementary Information.Sensitivity differences by strataTree leaf out phenology (LOD) was substantially more sensitive to average spring temperature in North America (mean = −3.62 days °C−1; 95% credible interval (CI) = [−3.76, −3.49]) than in Europe (mean = −2.79; CI = [−3.27, −2.30]) and Asia (mean = −2.62; CI = [−2.97, −2.26]; Fig. 2). These values are consistent with previously reported phenological sensitivities in North America7 (−5.5 to −3.3 days °C−1) and Europe8 (−4.1 to −3.0 days °C−1), as the credible intervals from our results overlap with the reported credible intervals of prior studies. However, the Asian LOD sensitivity was less sensitive than previously reported27 (−3.50 to −3.03 days °C−1), potentially owing to differences in species selection28 or model structure. Previously reported sensitivities were determined in separate studies using either observational data7,8 or long-term observation-based weather station data27. The general consistency between our findings suggests that phenology data from herbarium collections are good indicators of patterns in natural systems29,30,31, a point supported by a recent study of phenological sensitivity derived from herbaria and from observed citizen science data32. These herbarium-based results provide evidence that phenological sensitivity differs across the temperate forest biome (but see ref. 33 for evidence of differences in response to warming and chilling accumulation). To our knowledge, our study is the first to contrast overstory and understory phenology across multiple continents and, therefore, to find differences in phenological sensitivity between trees and forest wildflowers across continents. We recommend future studies explore these differences using alternative approaches and methodologies that focus on the physiological basis for and mechanisms that underlie these patterns.Fig. 2: Posterior estimated means and 95% credible intervals for spring temperature sensitivity.Shapes represent parameter estimates for wildflower First Flower Date (FFD, blue circles; n = 1418, 618, and 1060 for Asia, Europe, and North America, respectively) and canopy tree Leaf Out Date (LOD, yellow triangles; n = 899, 532, and 995, for Asia, Europe, and North America, respectively). Estimates are considered different from 0 if credible intervals do not overlap the dashed 0 line and are considered different from each other if credible intervals do not overlap.Full size imageIn contrast to trees, wildflower sensitivity to spring temperature was similar across all three continents and exhibited no strong differences (i.e., overlap in 95% Bayesian credible intervals) among continents (means and 95% credible intervals in brackets: North America = −3.14, [−3.28, −3.00]; Europe = −3.02, [−3.48, −2.56]; Asia = −3.12, [−3.36, −2.86]; Fig. 2). These values are also generally consistent with those reported elsewhere in the literature (i.e., 95% credible intervals overlap with those reported in other studies; −2.2, [−3.7, −0.76] days °C−1 in North America7 and −3.6, [−4.04, −3.18] days °C−1 in Europe9), although we are unaware of any studies that have estimated phenological sensitivity for Asian forest wildflowers in days °C−1. Ge et al.3 report herbaceous plant sensitivity of −5.71 days per decade in Asia (±7.90 standard deviation; based primarily on long-term observational data), which appears to be roughly consistent with our model results, but the difference in units makes this more speculative than the other comparisons. Discrepancies in mean responses between this study and others may be due in part to different types of data (herbarium specimens versus field observations) and to choice in focal taxa, as temperature sensitivity has been shown to vary widely across taxa28.Particularly noticeable in our results was that r2 coefficients of predicted versus observed phenology were much higher in North America (0.70 and 0.76 for wildflower and tree models, respectively) compared to Asian (0.40 and 0.44, respectively) and European models (0.41 and 0.25, respectively). This difference in model performance could be due to the higher interannual variability of spring temperatures in North America33, leading to greater selective pressure for strong sensitivity to spring temperatures in North American plants. This difference could explain why North American species exhibit higher correlation of phenology with average spring temperatures (Table S4). Alternatively, European and Asian species may have stronger phenological responses to alternative spring forcing windows, winter chilling temperatures, or photoperiod, relative to the March–April temperature period used in this study (see Methods). We think the latter explanation is unlikely, given the strong correlations of phenology with spring temperature across all continents (see Supplementary Information – Justification for March–April Temperature Window).Herbarium-based phenological models may be improved by accounting for spatial autocorrelation within the dataset. For example, Willems et al.9 found that including spatial autocorrelation significantly improved predictability of European herbaceous flowering phenology, even when accounting for multiple drivers of spring phenology. We followed a similar approach as their study and found similar improvements in model performance with the addition of spatial autocorrelation (Tables S3–S4) that had substantial positive effects on r2 values of Asian and European models. However, spatial distributions of specimens differed substantially among continents (see Figs. S2–S4), and these differences could lead to artifacts that make results unreliable to interpret (see Supplementary Information). Therefore, we focus here on results for models without spatial autocorrelation while acknowledging that spatial aggregation of herbarium specimens in Europe and Asia may be partially responsible for the relatively lower r2 values. We encourage other researchers to explore this question further both with our data set and other datasets.Climate change and spring light windowsThe relative difference between wildflower and tree sensitivity varied substantially among continents, with wildflowers being approximately equally as sensitive to spring temperature as trees in Asia and Europe but substantially less sensitive (i.e., 95% BCI do not overlap) than trees in North America (Fig. 2). Importantly, these differences were driven by changes in tree phenological sensitivities among continents and resulted in different expectations for spring light window duration (i.e., the difference in time between estimated wildflower flowering date and canopy tree leaf out date) on different continents under current climate conditions (Fig. 3), based on modeled leaf out and flowering under a climate scenario derived from average climate conditions from 2009–2018 (Fig. S5).Fig. 3: Current estimated phenological escape duration in northern temperate deciduous forests.Estimated mean difference between wildflower First Flower Date (FFD) and canopy tree Leaf Out Date (LOD) (in days) under current climate conditions (averaged from 2009–2018, see methods) in a Asia, b Europe, and c North America. Negative values indicate tree LOD is estimated to occur before wildflower FFD. Estimations were cropped by the estimated area of broadleaf and mixed-broadleaf forest (see methods). Dark gray regions indicate areas where the consensus land classification is More

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    Natural selection under conventional and organic cropping systems affect root architecture in spring barley

    Root morphological traitsThe wild-type parent ISR42-8 produced longer root length (RL) than the modern cultivar parent Golf and tested lines (Table S2, Fig. 1A.h,A.f [h = hydroponic; f = field]). The tested lines of the two evolving barley populations displayed significant variations under hydroponic conditions. Barley lines evolved under OCS had on average 3484 mm longer roots compared to CCS under hydroponic treatment (Fig. 1A.h, Table 2). Complementary results under field conditions show as well higher RL for the OCS lines, even though the variance was significantly less pronounced (Fig. 1A.f). In addition, a less evident variance was observed in the field within both groups compared to the hydroponic (Fig. 1A.d). Across both experimental setups, the observed range of RL was higher in the OCS lines [Standard deviation (SD)OCS = 883, SDCCS = 597] (Table 2).Figure 1Significantly variant root morphological phenotypes. Boxplots illustrate the overall distribution of observed data points for the parents Golf and ISR 42-8 as well as for the conventional (CCS) and organic (OCS) lines. Density plots highlight the overall distribution of organic and conventional adapted lines. (A)—Root length (RL)—the sum of all roots harvested in millimeters (mm), illustrated for all four groups. (A.h)—root length measured in the hydroponic experiment; (A.f)—field experiment; A.d—distribution histogram for root length in both field and hydroponic experiment for CCS and OCS adapted lines. (B)—the ratio of root length to volume (L/V). Data available for hydroponics (B.h), field (B.f), and distribution of the ratio of root length to volume illustrated in B.d. (C.f)—Root mass density (RMD) from the field; (D.f)—Root angle (RA) from the field, distribution of the root angle illustrated in (D.d); (E.f)—root tip per plant count from the field, corresponding histogram visualized in (E.d). (F.f)—root fork per plant count from the field.Full size imageTable 2 Comparison of organic and conventional population root phenotypes under field and hydroponic evaluation.Full size tableThe root length to volume (L/V) is an important indicator of the soil volume that can be explored by the roots. Under hydroponics conditions, variations were found for L/V between the parental genotypes as well as between the OCS and CCS populations (Tables 2 & S3). The organic lines were characterized by a significantly higher L/V, indicating a much more distinct exploration of the soil by these lines (Fig. 1B.h,B.f). In comparison to field, highest diversity in L/V was found under hydroponic experiments within both OCS and CCS populations (Fig. 1B.d).The root mass density (RMD) is the ratio of root volume for a given root mass and is a key indicator of root thickness. Although significant variations existed between ISR42-8 and Golf under hydroponics conditions, such significant variations were not found between the OCS and CCS groups (P = 0.09) (Fig. 1C.f and Tables 2 and S3).The root angle (RA) measurements were only performed under field conditions since plants grown under hydroponics conditions were placed in uniform growing vessels and the direction of root growth is restricted by tubes. Significant variation was observed for the RA between the two parental lines, which was also reflected in the CCS and OCS lines (Fig. 1D.f). ISR42-8 was characterized by an 11.5° average narrower RA than Golf (Table S3). The RA was 4.1° bigger in the OCS compared to the CCS population (P = 0.005) (Table 2). However, a higher diversity in RA was observed in the OCS compared to the CCS lines (Fig. 1D.d, Table 2).In addition to the RA, the number of root tips and forks was measured under field conditions only. Both tips and forks indicate a similar pattern, where the OCS lines produced on average more for both PForks = 0.014, PTips = 0.0041 (Fig. 1E.f,F.f). After applying a P-adjustment, the number of forks count was no longer significantly different between OCS and CCS (PForks = 0.07, Table 2). Complementary, ISR 42-8 was observed to produce more tips and forks than Golf, which remained highly significant even after probability adjustment (Fig. 1E.f,F.f, Table S3). The distribution and the standard deviation of observed phenotypes highlight once more the fact that the OCS lines tend to have a higher variation (Fig. 1E.d, Table 2). Similarly, a significant increasing trend was recorded in root surface area (RSA) and root average diameter (RAD) by ISR42-8 as compared to Golf under hydroponics (Table S3). Contrasting to the parental genotypes, no variation was observed between OCS and CCS lines for RSA (Table 2).Root anatomical traitsWithin the observed anatomical traits, four were considered due to their relevance and variation between the systems. In both hydroponic and field experiments, significant variations were observed for the late metaxylem number (LMXN) between the parental lines as well as OCS and CCS lines (Tables 2 & S3, Fig. 2A.h,A.f). An increased LMXN for ISR 42-8 compared to Golf was observed (Fig. 2A.h). Regarding the CCS and OCS lines, a heterogenic scenario was presented over both experimental setups. While the median LMXN under CCS was identical with ISR 42-8 in the seedling stages of plant development (Fig. 2A.h), it was much lower in flowering stages under field conditions (Fig. 1A.f). Additionally, the LMXN was significantly higher in the CCS lines in the seedling stage compared to OCS lines, vice-versa LMXN was observed at the flowering time point (Fig. 2A.d).Figure 2Significantly variant root anatomical traits. Boxplots illustrate the overall distribution of observed data points for the parents Golf and ISR 42-8 as well as for the conventional (CCS) and organic (OCS) lines. (A) –Late metaxylem number (LMN)—the sum of all roots harvested and expressed by plant−1, illustrated for all four groups. A.h—Late metaxylem number measured in the hydroponic experiment; A.f—field experiment; A.d—distribution histogram for late metaxylem number in both field and hydroponic experiment for CCS and OCS adapted lines. (B)—Aerenchyma area (AA). Data available for hydroponics (B.h), field (B.f), and distribution of the aerenchyma area illustrated in (B.d). (C.f)—Total cortical area (TCA) from the field; (D.f)—Root cross-section area (RA) from the field, distribution of the total cortical area and root cross-section area illustrated in C.d and D.d, respectively.Full size imageThe intercellular space, represented by the aerenchyma area (AA), was observed to be significantly more pronounced in the tested CCS compared to OCS lines in both environments (Fig. 2B.h,B.f). Furthermore, the OCS population did not show significant differences to both parents under hydroponics conditions, however, when grown under field conditions, it was noted that Golf had a significantly higher AA mean value as compared to the OCS population (Table S3). As illustrated by the values, the AA expended from early to late stages by a magnitude of 10-folds (Fig. 2B.d). In general, although the two parents did not indicate phenotypic variations, OCS and CCS lines showed significant variations (Table S4).A 0.12 mm2 decreased average total cortical area (TCA) was recorded in the OCS compared to the CCS population under field conditions (P = 0.003, Fig. 2C.f), although substantial variations for TCA was observed within OCS and CCS populations (Fig. 2C.d). The root cross-section area (RXA) is a two-dimensional axis of the root which is an important indicator of root thickness. In the hydroponic examination of the seedling stage, significant variations existed between the CCS and ISR42-8 as well as OCS population (Tables 2 and S3). The complementary study under field conditions observed a noticeable variation for OCS from both parental genotypes and the CCS (Table S3). About 0.13 mm2 increased average value for RXA was identified for CCS (Fig. 2D.f), while consistent significant variations were also observed between the populations in the under field experiment, where 0.13 mm2 increased average value for RXA was identified for CCS (Fig. 2D.f). Analog (Fig. 2D.d). Analogue to the AA, the RXA indicates a lower root extension in the OCS compared to the CCS population. For the stele area (SA), significant variations were only observed at the flowering stage, where ISR42-8 generally had the highest SA and varies significantly between Golf and its progeny lines (Tables 2 and S3).Shoot-related traitsBeyond the root-related phenotypic observations, above-ground characteristics were also recorded to assess the root-borne shoot dynamics (Figs. 3 and S2). Among the OCS and CCS populations and the parents, ISR42-8 had the longest duration of emergence. While CCS-adapted lines took on average 5.8 days of emergence (DE), OCS-adapted lines emerged 1.8 days later (7.6 days) (Fig. S2). No variation was observed for the tiller number (TN) throughout all tested groups, while ISR 42-8 tends to produce much more leaf number (LN), accompanied by a lower plant height (PH) and higher shoot dry weight (SDW) (Table S4, Fig. S2). The OCS and CCS plants significantly differed in PH as well as SDW (Fig. 3B,C). The LN was marginally above the probability threshold of 0.05 (p = 0.058, Fig. 3A), with a clear tendency of increased variability in phenotypic variation (Fig. 3D). Similar trend was recorded for the SDW (Fig. 3F).Figure 3Above-ground plant characteristics. Boxplots illustrate the overall distribution of observed data points for the parents Golf and ISR 42-8 as well as for the conventional (CCS) and organic (OCS) lines under the hydroponic experiment. (A)—Leaf number (LN) expressed by; (B)—Plant height (PH) and C-Shoot dry weight (SDW). The data distribution of the leaf number, plant height and shoot dry weight is illustrated in (D,E,F), respectively.Full size imageInterconnection of root-shoot traitsWe performed inter-trait correlation analysis to unravel association among root traits and in between root and shoot phenotypes (Fig. 4). Pearson correlation coefficient revealed significant correlations among root-shoot traits. LN, PH and SDW had strong positive associations with all root architectural traits under hydroponic conditions (P  0.30) in both CCS and OCS, while DE has negative association with all shoot traits (r = −0.17 to −0.48) (Fig. 4A.h,B.h). A consistent negative relationship was observed for L/V with shoot traits such as LN, PH and SDW and root morphological traits such as RL, RSA and RAD in both CCS and OSC populations (Fig. 4A.h,B.h). A strong negative association existed between RL and all shoot morphological, root architectural and anatomical traits in both populations, except for L/V where a weak negative (r = −0.09) association was displayed only in the OCS. Likewise, all above-ground traits and all root architectural traits exhibited significant positive associations with all root anatomical features in both groups with an exception for the AA (Fig. 4A.h,B.h). Moreover, correlation analysis revealed strong positive relationships in both groups of SDW and root dry weight (RDW) to all above-ground traits, below-ground traits including, RL, SA, and RAD, as well as in all root anatomical traits (Fig. 4A.h,B.h). This means that the growth of tissue and organ is proportional to the increase in total dry biomass. More importantly, we observed a significant positive correlation among most of the root morphological, architectural, and anatomical traits in both OCS and CCS adapted populations, with few exceptions such as L/V (Fig. 4A.h,B.h).Figure 4Correlation matrix for shoot morphological (only in hydroponic conditions; A.h and B.h), root architectural and anatomical traits in two groups of barley populations and their parental lines grown across two growing conditions. (A)—conventional and (B)—organic cropping systems. (A.h)—conventional under hydroponic, (B.h)—organic under hydroponic, (A.f)—conventional under field, and B.f—organic under field conditions. The color scale represents Spearman’s ranked correlation coefficient. A larger circle size indicates a smaller p-value; blank cells represent that correlation was non-significant at P  −0.90) and RDW (r =   > −0.80) (Fig. 4A.f,B.f) for CCS and OCS populations respectively, which means that narrower the angle of the nodal roots, the longer was the root system. The two root branching traits, the number of tips and number of roots forks which were known to be associated and dependent on the RL have a strong positive correlation reflected by r = 0.81 and 0.90 in CCS and r = 0.74 and 0.84 in OCS developed lines, respectively, while they have a significant negative correlation with RA (r = −0.72 to −0.77) in both barley groups. In addition, RA had also strong negative relationship to RDW contributing architectural traits including, RMD (r = −0.81 to −0.84) and L/V (r = −0.34 to −0.44). However, no positive associations were observed for RA and all root anatomical traits in both OCS and CCS populations (Fig. 4A.f,B.f).Allometry analysisThe correlation analysis identified interconnection among root and shoot-related traits. Therefore, we checked if these correlations can be explained by allometric relations (Tables 3 and 4).Table 3 Summary of allometric analysis of root-shoot system traits under hydroponic condition.Full size tableTable 4 Summary of allometric analysis of root-shoot system traits under field condition.Full size tableIn the hydroponic environment, we observed a total of ten allometric relations, from which six were annotated to the PH. The PH was allometrically related to the SDW, the RSA, the RV, the RDW, the SRL, and the RMD (Table 3). Besides, the SDW was allometrically associated with the RSD. Furthermore, the TCA was related to the RXA. Finally, an allometry relationship was detected between the LMXN and AA (Table 3).In the field experimental setup, we detected in total ten allometric relations (P  More