Parsimonious estimation of hourly surface ozone concentration across China during 2015-2020.

Surface ozone is an important air pollutant detrimental to human health and vegetation productivity, particularly in China. However, high resolution surface ozone concentration data is still lacking, largely hindering accurate assessment of associated environmental impacts. Here, we collected hourly ground ozone observations (over 6 million records), remote sensing products, meteorological data, and social-economic information, and applied recurrent neural networks to map hourly surface ozone data (HrSOD) at a 0.1° × 0.1° resolution across China during 2015-2020. The coefficient of determination (R2) values in sample-based, site-based, and by-year cross-validations were 0.72, 0.65 and 0.71, respectively, with the root mean square error (RMSE) values being 11.71 ppb (mean = 30.89 ppb), 12.81 ppb (mean = 30.96 ppb) and 11.14 ppb (mean = 31.26 ppb). Moreover, it exhibits high spatiotemporal consistency with ground-level observations at different time scales (diurnal, seasonal, annual), and at various spatial levels (individual sites and regional scales). Meanwhile, the HrSOD provides critical information for fine-resolution assessment of surface ozone impacts on environmental and human benefits.

In-vitro colonic fermentation of Kakadu plum (Terminalia ferdinandiana) fruit powder: Microbial biotransformation of phenolic compounds and cytotoxicity.

Kakadu plum (Terminalia ferdinandiana) (KP) is an indigenous fruit used as a functional ingredient in powdered form. Three KP doses (1, 2.5 and 5 g) were digested in a dynamic in vitro gut digestion model over 48 h. Faecal water digests from the colonic reactors were assessed for total soluble polyphenols (TSP), ferric reducing antioxidant power (FRAP), phenolic metabolites and short-chain fatty acids (SCFAs). Effects of digests on cell viability were tested against Caco-2 intestinal and HepG2 hepatic cells. All doses of KP fermentation produced castalagin, corilagin, chebulagic acid, chebulinic acid, and gallic acid. TSP and FRAP significantly increased in 5 g KP digests at 0 and 48 h of fermentation. SCFA concentrations significantly increased after 48 h. Cytotoxic effects of 2.5 and 5 g KP digests diminished significantly after 12 h. Overall, colonic fermentation increased antioxidant activity and polyphenolic metabolites of 5 g KP powder for 48 h.

Genome-Wide Identification and Phylogenetic Analysis of TRP Gene Family Members in Saurian.

The transient receptor potential plays a critical role in the sensory nervous systems of vertebrates in response to various mechanisms and stimuli, such as environmental temperature. We studied the physiological adaptive evolution of the TRP gene in the saurian family and performed a comprehensive analysis to identify the evolution of the thermo-TRPs channels. All 251 putative TRPs were divided into 6 subfamilies, except TRPN, from the 8 saurian genomes. Multiple characteristics of these genes were analyzed. The results showed that the most conserved proteins of TRP box 1 were located in motif 1, and those of TRP box 2 were located in motif 10. The TRPA and TRPV in saurian tend to be one cluster, as a sister cluster with TRPC, and the TRPM is the root of group I. The TRPM, TRPV, and TRPP were clustered into two clades, and TRPP were organized into TRP PKD1-like and PKD2-like. Segmental duplications mainly occurred in the TRPM subfamily, and tandem duplications only occurred in the TRPV subfamily. There were 15 sites to be under positive selection for TRPA1 and TRPV2 genes. In summary, gene structure, chromosomal location, gene duplication, synteny analysis, and selective pressure at the molecular level provided some new evidence for genetic adaptation to the environment. This result provides a basis for identifying and classifying TRP genes and contributes to further elucidating their potential function in thermal sensors.

Effect of light intensities on the photosynthesis, growth and physiological performances of two maple species.

Photoinhibition decreases photosynthetic capacity and can therefore affect the plant survival, growth, and distribution, but little is known about how it affects on kindred tree species. We conducted field experiments to measure the photosynthetic, growth and physiological performances of two maple species (Acer mono and A. pseudosieboldianum) seedlings at four light intensities (100%, 75%, 55%, and 20% of full light) and evaluated the adaptability of seedlings. We found that: (1) A. mono seedlings have larger light saturated photosynthetic rates (A max), the light saturation point (LSP), and lower light compensation point (LCP) than A. pseudosieboldianum seedlings, thus indicating that the former has a stronger light utilization ability. (2) A. mono seedlings under 75% light intensity and had higher seedling height (SH), basal stem diameter (BSD), leaf number (LN), leaf area per plant (LAPP) and total dry weight (TDW), while A. pseudosieboldianum seedling at 55% light intensity displayed greater growth advantages, which agreed with their response of light saturated photosynthetic rate. Morphological plasticity adjustments such as decreased root shoot ratio (RSR) and increased specific leaf area (SLA) showed how seedlings adapt to weak light environments. (3) 100% and 20% light intensities increased the malondialdehyde (MDA) content of two maple seedlings, indicating that very strong or very weak light could lead to the imbalance of reactive oxygen species (ROS) metabolism. The regulation of antioxidant enzyme activities such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), as well as the content of osmoregulation substances such as free proline and soluble protein, are the main mechanisms of plant adaptation to light stress. Although both A. mono and A. pseudosieboldianum are highly shade tolerant, subtle differences in the photosynthetic, morphological and physiological traits underpinning their shade tolerance suggest A. pseudosieboldianum has the advantage to deal with the light threat. Future studies should focus on the expression level of photosynthesis-related genes and cell, to better understand the adaptation mechanism of plants to light variation which facilitates forest development, either natural or via silvicultural practices. This information expands our understanding of the light-regulating mechanism of trees, which contributes to develop management practices to support natural forest regeneration.

The mitochondrial genome and phylogenetic analysis of Rhacophorus rhodopus.

Classification of the genus Rhacophorus has been problematic. In particular there has been considerable controversy surrounding the phylogenetic relationships among Rhacophorus rhodopus, R. bipunctatus, and R. reinwardtii. To examine the relationship among these Rhacophorus species, we assembled the complete mitochondrial genome sequence of R. rhodopus. The R. rhodopus genome is 15,789 bp in length with 12 protein-coding genes (PCGs) (losing ND5), two ribosomal genes, 22 transfer RNA genes, and a control region (D-loop). Base composition of the overall sequence was 60.86% for A + T content and 39.14% for C + G content. Most of the PCGs used ATG as a start codon, except for the COX I gene, which used the ATA start codon. COX I and ND6 used AGG and ATP8 stop codons respectively, while ND3 and ND4L used the TAA stop codon. For the remaining seven genes, the stop codons was incomplete. In addition, both 5' and 3' of the control areas had distinct repeating regions. Based on three datasets and two methods (Bayesian inference (BI) and maximum likelihood (ML)), we reconstructed three phylogenetic trees to explore the taxonomic status of the species and the phylogenetic relationship among R. rhodopus, R. bipunctatus and R. reinwardtii. Our results indicated that these three species are non-monophyletic; thus, the phylogenetic relationship among them is complex and difficult to determine. Further, R. rhodopus is divided into three lineages from different parts of China. The two Rhacophorus samples showed very close phylogenetic relationship with R. rhodopus. Our results add to the mitochondrial genome database of amphibians and will help to disentangle the phylogenetic relationships within the Rhacophoridae.

Unscrambling the Provenance of Eggs by Combining Chemometrics and Near-Infrared Reflectance Spectroscopy.

Issues related to food authenticity, traceability, and fraud have increased in recent decades as a consequence of the deliberate and intentional substitution, addition, tampering, or misrepresentation of food ingredients, where false or misleading statements are made about a product for economic gains. This study aimed to evaluate the ability of a portable NIR instrument to classify egg samples sourced from different provenances or production systems (e.g., cage and free-range) in Australia. Whole egg samples (n: 100) were purchased from local supermarkets where the label in each of the packages was used as identification of the layers' feeding system as per the Australian legislation and standards. The spectra of the albumin and yolk were collected using a portable NIR spectrophotometer (950-1600 nm). Principal component analysis (PCA) and linear discriminant analysis (LDA) were used to analyze the NIR data. The results obtained in this study showed how the combination of chemometrics and NIR spectroscopy allowed for the classification of egg albumin and yolk samples according to the system of production (cage and free range). The proposed method is simple, fast, environmentally friendly and avoids laborious sample pre-treatment, and is expected to become an alternative to commonly used techniques for egg quality assessment.

Lessons from the diet: Captivity and sex shape the gut microbiota in an oviparous lizard (Calotes versicolor).

Studies have indicated that the abundance and community structure of gut microbiota are altered by diet. In this study, next-generation sequencing of the 16S rRNA gene amplicon was performed to evaluate variations in the gut microbiota of wild and captive individuals of both sexes of Calotes versicolor. The results showed that there was a significant sex difference in microbial community structure for wild C. versicolor, Bacteroide was the dominant genus in wild females (WF), whereas Ochrobactrum was the dominant genus in wild males (WM). Acinetobacter and Hymenobacter were the dominant genera in WF, while Clostridium was the dominant genus in captive females (CF). The results indicated that differences in diet between wild and captive C. versicolor also resulted in variations in gut microbiota. Thus, it was not surprising that captivity and sex shape the gut microbiota in C. versicolor. In summary, the fundamental information presented about the gut microbiota of both sexes of wild (and captive females) C. versicolor, indicates that the artificial environments are not suitable for the wild C. versicolor.

Environmental drivers of the leaf nitrogen and phosphorus stoichiometry characteristics of critically endangered Acer catalpifolium.

Acer catalpifolium is a perennial deciduous broad-leaved woody plant, listed in the second-class protection program in China mainly distributed on the northwest edge of Chengdu plain. However, extensive anthropogenic disturbances and pollutants emissions (such as SO2, NH3 and NOX) in this area have created a heterogeneous habitat for this species and its impacts have not been systematically studied. In this study, we investigated the leaf nitrogen (N) and phosphorus (P) content of A. catalpifolium in the natural distribution areas, and a series of simulation experiments (e.g., various water and light supply regimes, different acid and N deposition levels, reintroduction management) were conducted to analyze responses of N and P stoichiometric characteristics to environmental changes. The results showed that leaf nitrogen content (LNC) was 14.49 ~ 25.44 mg g-1, leaf phosphorus content (LPC) was 1.29~3.81 mg g-1 and the N/P ratio of the leaf (L-N/P) was 4.87~13.93. As per the simulation experiments, LNC of A. catalpifolium is found to be relatively high at strong light conditions (80% of full light), high N deposition (100 and 150 kg N ha-1), low acidity rainwater, reintroduction to understory area or N fertilizer applications. A high level of LPC was found when applied with 80% of full light and moderate N deposition (100 kg N ha-1). L-N/P was high under severe shade (8% of full light), severe N deposition (200 kg N ha-1), and reintroduction to gap and undergrowth habitat; however, low L-N/P was observed at low acidity rainwater or P fertilizer application. The nutrient supply facilitates corresponding elements uptake, shade tends to induce P limitation and soil acidification shows N limitation. Our results provide theoretical guidance for field management and nutrient supply regimes for future protection, population rejuvenation of this species and provide guidelines for conservation and nutrient management strategies for the endangered species.

Whole-Genome Sequencing of Acer catalpifolium Reveals Evolutionary History of Endangered Species.

Acer catalpifolium is an endangered species restricted to remote localities of West China. Understanding the genomic content and evolution of A. catalpifolium is essential to conservation efforts of this rare and ecologically valuable plant. Here, we report a high-quality genome of A. catalpifolium consisting of ∼654 Mbp and ∼35,132 protein-coding genes. We detected 969 positively selected genes in two Acer genomes compared with four other eudicots, 65 of which were transcription factors. We hypothesize that these positively selected mutations in transcription factors might affect their function and thus contribute to A. catalpifolium's decline-type population. We also identified 179 significantly expanded gene families compared with 12 other eudicots, some of which are involved in stress responses, such as the FRS-FRF family. We inferred that A. catalpifolium has experienced gene family expansions to cope with environmental stress in its evolutionary history. Finally, 109 candidate genes encoding key enzymes in the lignin biosynthesis pathway were identified in A. catalpifolium; of particular note were the large range and high copy number of cinnamyl alcohol dehydrogenase genes. The chromosome-level genome of A. catalpifolium presented here may serve as a fundamental genomic resource for better understanding endangered Acer species, informing future conservation efforts.

Morphological, Physiological and Photophysiological Responses of Critically Endangered Acer catalpifolium to Acid Stress.

Acid rain deposition (AR) has long-lasting implications for the community stability and biodiversity conservation in southwest China. Acer catalpifolium is a critically endangered species in the rain zone of Western China where AR occurs frequently. To understand the effects of AR on the morphology and physiology of A. catalpifolium, we conducted an acid stress simulation experiment for 1.5 years. The morphological, physiological, and photosynthetic responses of A. catalpifolium to the acidity, composition, and deposition pattern of acid stress was observed. The results showed that simulated acid stress can promote the growth of A. catalpifolium via the soil application mode. The growth improvement of A. catalpifolium under nitric-balanced acid rain via the soil application mode was greater than that of sulfuric-dominated acid rain via the soil application mode. On the contrary, the growth of A. catalpifolium was significantly inhibited by acid stress and the inhibition increased with the acidity of acid stress applied via leaf spraying. The inhibitory impacts of nitric-balanced acid rain via the leaf spraying of A. catalpifolium were greater than that of sulfur-dominant acid rain via leaf spraying. The observations presented in this work can be utilized for considering potential population restoration plans for A. catalpifolium, as well as the forests in southwest China.

Environment-Dependent Variation in Gut Microbiota of an Oviparous Lizard (Calotes versicolor).

Vertebrates maintain complex symbiotic relationships with microbiota living within their gastrointestinal tracts which reflects the ecological and evolutionary relationship between hosts and their gut microbiota. However, this understanding is limited in lizards and the spatial heterogeneity and co-occurrence patterns of gut microbiota inside the gastrointestinal tracts of a host and variations of microbial community among samples remain poorly understood. To address this issue and provide a guide for gut microbiota sampling from lizards, we investigated the bacteria in three gut locations of the oriental garden lizard (Calotes versicolor) and the data were analyzed for bacterial composition by 16S ribosomal RNA (16S rRNA) gene amplicon sequencing. We found the relative abundance of the dominant phyla exhibited an increasing trend from the small intestine to the large intestine, and phyla Firmicutes, Bacteroidetes and Proteobacteria were the three primary phyla in the gut microbiota of C. versicolor. There were a higher abundance of genus Bacteroides (Class: Bacteroidia), Coprobacillus and Eubacterium (Class: Erysipelotrichia), Parabacteroides (Family: Porphyromonadaceae) and Ruminococcus (Family: Lachnospiraceae), and Family Odoribacteraceae and Rikenellaceae in the sample from the hindgut. The secondary bile acid biosynthesis, glycosaminoglycan degradation, sphingolipid metabolism and lysosome were significantly higher in the hindgut than that in the small intestine. Taken together our results indicate variations of gut microbiota composition and metabolic pathway in different parts of the oriental garden lizard.

Body temperature and time of day both affect nocturnal lizard performance: An experimental investigation.

The locomotor performance of reptiles is profoundly influenced by temperature, but little is known about how the time of day when the animal is usually active may influence performance. Time of day may be particularly relevant for studies on nocturnal reptiles that thermoregulate by day, but are active at night when ambient temperatures are cooler. If selection favours individuals that match their performance to activity times, then nocturnal species should perform better during the night, when they are normally active, than during the day. To test this hypothesis, we investigated how the time of day and body temperature affected the locomotor performance of adult females of the velvet gecko (Amalosia lesueurii). We measured the sprint speeds, running speeds and number of stops of 43 adult females at four different body temperatures (20, 25, 30 and 35 °C) during the day and at night. At night, sprint speeds were higher at 20 and 35 °C but sprint speeds were similar at 25 and 30 °C. By day, sprint speed increased with body temperature, peaking at 30 °C, before declining at 35 °C. However, gecko speeds over 1 m was higher at night at all four test temperatures than by day. Number of stops showed broadly similar patterns and females stopped almost twice as often on the racetrack during the day than they did at night. Furthermore, the thermal breadth of performance differed depending on when geckos were tested. Our results demonstrate that both body temperature and the time of day affects the behaviour and locomotor performance of female velvet geckos, with geckos running faster at night, the time of day when they are usually active. This study adds to evidence that both body temperature and the time of day are crucial for estimating the performance of ectotherms and evaluations and predictions of their vulnerability to climate warming should consider the context of laboratory experimental design.

Predicting the effect of climate change on a range-restricted lizard in southeastern Australia.

Climate change is ranked as one of the most severe threats to global biodiversity. This global phenomenon is particularly true for reptiles whose biology and ecology are closely linked to climate. In this study, we used over 1,300 independent occurrence points and different climate change emission scenarios to evaluate the potential risk of changing climatic conditions on the current and future potential distribution of a rock-dwelling lizard; the velvet gecko. Furthermore, we investigated if the current extent of protected area networks in Australia captures the full range distribution of this species currently and in the future. Our results show that climate change projections for the year 2075 have the potential to alter the distribution of the velvet gecko in southeastern Australia. Specifically, climate change may favor the range expansion of this species to encompass more suitable habitats. The trend of range expansion was qualitatively similar across the different climate change scenarios used. Additionally, we observed that the current network of protected areas in southeast Australia does not fully account for the full range distribution of this species currently and in the future. Ongoing climate change may profoundly affect the potential range distribution of the velvet gecko population. Therefore, the restricted habitat of the velvet geckos should be the focus of intensive pre-emptive management efforts. This management prioritization should be extended to encompass the increases in suitable habitats observed in this study in order to maximize the microhabitats available for the survival of this species.

Hotter nests produce hatchling lizards with lower thermal tolerance.

In many regions, the frequency and duration of summer heatwaves is predicted to increase in future. Hotter summers could result in higher temperatures inside lizard nests, potentially exposing embryos to thermally stressful conditions during development. Potentially, developmentally plastic shifts in thermal tolerance could allow lizards to adapt to climate warming. To determine how higher nest temperatures affect the thermal tolerance of hatchling geckos, we incubated eggs of the rock-dwelling velvet gecko, Amalosia lesueurii, at two fluctuating temperature regimes to mimic current nest temperatures (mean 23.2°C, range 10-33°C, 'cold') and future nest temperatures (mean 27.0°C, range 14-37°C, 'hot'). Hatchlings from the hot incubation group hatched 27 days earlier and had a lower critical thermal maximum (CTmax 38.7°C) and a higher critical thermal minimum (CTmin 6.2°C) than hatchlings from cold incubation group (40.2 and 5.7°C, respectively). In the field, hatchlings typically settle under rocks near communal nests. During the hatching period, rock temperatures ranged from 13 to 59°C, and regularly exceeded the CTmax of both hot- and cold-incubated hatchlings. Because rock temperatures were so high, the heat tolerance of lizards had little effect on their ability to exploit rocks as retreat sites. Instead, the timing of hatching dictated whether lizards could exploit rocks as retreat sites; that is, cold-incubated lizards that hatched later encountered less thermally stressful environments than earlier hatching hot-incubated lizards. In conclusion, we found no evidence that CTmax can shift upwards in response to higher incubation temperatures, suggesting that hotter summers may increase the vulnerability of lizards to climate warming.

Incubation under climate warming affects learning ability and survival in hatchling lizards.

Despite compelling evidence for substantial individual differences in cognitive performance, it is unclear whether cognitive ability influences fitness of wild animals. In many animals, environmental stressors experienced in utero can produce substantial variation in the cognitive abilities of offspring. In reptiles, incubation temperatures experienced by embryos can influence hatchling brain function and learning ability. Under climate warming, the eggs of some lizard species may experience higher temperatures, which could affect the cognitive abilities of hatchlings. Whether such changes in cognitive abilities influence the survival of hatchlings is unknown. To determine whether incubation-induced changes in spatial learning ability affect hatchling survival, we incubated velvet gecko, Amalosia lesueurii, eggs using two fluctuating temperature regimes to mimic current (cold) versus future (hot) nest temperatures. We measured the spatial learning ability of hatchlings from each treatment, and released individually marked animals at two field sites in southeastern Australia. Hatchlings from hot-incubated eggs were slower learners than hatchlings from cold-incubated eggs. Survival analyses revealed that hatchlings with higher learning scores had higher survival than hatchlings with poor learning scores. Our results show that incubation temperature affects spatial learning ability in hatchling lizards, and that such changes can influence the survival of hatchlings in the wild.

Effects of pregnancy on body temperature and locomotor performance of velvet geckos.

Pregnancy is a challenging period for egg laying squamates. Carrying eggs can encumber females and decrease their locomotor performance, potentially increasing their risk of predation. Pregnant females can potentially reduce this handicap by selecting higher temperatures to increase their sprint speed and ability to escape from predators, or to speed up embryonic development and reduce the period during which they are burdened with eggs ('selfish mother' hypothesis). Alternatively, females might select more stable body temperatures during pregnancy to enhance offspring fitness ('maternal manipulation hypothesis'), even if the maintenance of such temperatures compromises a female's locomotor performance. We investigated whether pregnancy affects the preferred body temperatures and locomotor performance of female velvet geckos Amalosia lesueurii. We measured running speed of females during late pregnancy, and one week after they laid eggs at four temperatures (20°, 25°, 30° and 35°C). Preferred body temperatures of females were measured in a cost-free thermal gradient during late pregnancy and one week after egg-laying. Females selected higher and more stable set-point temperatures when they were pregnant (mean =29.0°C, Tset =27.8-30.5°C) than when they were non-pregnant (mean =26.2°C, Tset =23.7-28.7°C). Pregnancy was also associated with impaired performance; females sprinted more slowly at all four test temperatures when burdened with eggs. Although females selected higher body temperatures during late pregnancy, this increase in temperature did not compensate for their impaired running performance. Hence, our results suggest that females select higher temperatures during pregnancy to speed up embryogenesis and reduce the period during which they have reduced performance. This strategy may decrease a female's probability of encountering predatory snakes that use the same microhabitats for thermoregulation. Selection of stable temperatures by pregnant females may also benefit embryos, but manipulative experiments are necessary to test this hypothesis.

Communal nesting under climate change: fitness consequences of higher incubation temperatures for a nocturnal lizard.

Communal nesting lizards may be vulnerable to climate warming, particularly if air temperatures regulate nest temperatures. In southeastern Australia, velvet geckos Oedura lesueurii lay eggs communally inside rock crevices. We investigated whether increases in air temperatures could elevate nest temperatures, and if so, how this could influence hatching phenotypes, survival, and population dynamics. In natural nests, maximum daily air temperature influenced mean and maximum daily nest temperatures, implying that nest temperatures will increase under climate warming. To determine whether hotter nests influence hatchling phenotypes, we incubated eggs under two fluctuating temperature regimes to mimic current 'cold' nests (mean = 23.2 °C, range 10-33 °C) and future 'hot' nests (27.0 °C, 14-37 °C). 'Hot' incubation temperatures produced smaller hatchlings than did cold temperature incubation. We released individually marked hatchlings into the wild in 2014 and 2015, and monitored their survival over 10 months. In 2014 and 2015, hot-incubated hatchlings had higher annual mortality (99%, 97%) than cold-incubated (11%, 58%) or wild-born hatchlings (78%, 22%). To determine future trajectories of velvet gecko populations under climate warming, we ran population viability analyses in Vortex and varied annual rates of hatchling mortality within the range 78- 96%. Hatchling mortality strongly influenced the probability of extinction and the mean time to extinction. When hatchling mortality was >86%, populations had a higher probability of extinction (PE: range 0.52- 1.0) with mean times to extinction of 18-44 years. Whether future changes in hatchling survival translate into reduced population viability will depend on the ability of females to modify their nest-site choices. Over the period 1992-2015, females used the same communal nests annually, suggesting that there may be little plasticity in maternal nest-site selection. The impacts of climate change may therefore be especially severe on communal nesting species, particularly if such species occupy thermally challenging environments.