Haplotype-resolved genome assembly of Phanera championii reveals molecular mechanisms of flavonoid synthesis and adaptive evolution.

Phanera championii is a medicinal liana plant that has successfully adapted to hostile karst habitats. Despite extensive research on its medicinal components and pharmacological effects, the molecular mechanisms underlying the biosynthesis of critical flavonoids and its adaptation to karst habitats remain elusive. In this study, we performed high-coverage PacBio and Hi-C sequencing of P. championii, which revealed its high heterozygosity and phased the genome into two haplotypes: Hap1 (384.60 Mb) and Hap2 (383.70 Mb), encompassing a total of 58 612 annotated genes. Comparative genomes analysis revealed that P. championii experienced two whole-genome duplications (WGDs), with approximately 59.59% of genes originating from WGD events, thereby providing a valuable genetic resource for P. championii. Moreover, we identified a total of 112 genes that were strongly positively selected. Additionally, about 81.60 Mb of structural variations between the two haplotypes. The allele-specific expression patterns suggested that the dominant effect of P. championii was the elimination of deleterious mutations and the promotion of beneficial mutations to enhance fitness. Moreover, our transcriptome and metabolome analysis revealed alleles in different tissues or different haplotypes collectively regulate the synthesis of flavonoid metabolites. In summary, our comprehensive study highlights the significance of genomic and morphological adaptation in the successful adaptation of P. championii to karst habitats. The high-quality phased genomes obtained in this study serve as invaluable genomic resources for various applications, including germplasm conservation, breeding, evolutionary studies, and elucidation of pathways governing key biological traits of P. championii.

Cosmogenic nuclide dating of Australopithecus at Sterkfontein, South Africa.

Sterkfontein is the most prolific single source of Australopithecus fossils, the vast majority of which were recovered from Member 4, a cave breccia now exposed by erosion and weathering at the landscape surface. A few other Australopithecus fossils, including the StW 573 skeleton, come from subterranean deposits [T. C. Partridge et al., Science 300, 607-612 (2003); R. J. Clarke, K. Kuman, J. Hum. Evol. 134, 102634 (2019)]. Here, we report a cosmogenic nuclide isochron burial date of 3.41 ± 0.11 million years (My) within the lower middle part of Member 4, and simple burial dates of 3.49 ± 0.19 My in the upper middle part of Member 4 and 3.61 ± 0.09 My in Jacovec Cavern. Together with a previously published isochron burial date of 3.67 ± 0.16 My for StW 573 [D. E. Granger et al., Nature 522, 85-88 (2015)], these results place nearly the entire Australopithecus assemblage at Sterkfontein in the mid-Pliocene, contemporaneous with Australopithecus afarensis in East Africa. Our ages for the fossil-bearing breccia in Member 4 are considerably older than the previous ages of ca. 2.1 to 2.6 My interpreted from flowstones associated with the same deposit. We show that these previously dated flowstones are stratigraphically intrusive within Member 4 and that they therefore underestimate the true age of the fossils.

Effects of different karst fissures and rainfall distribution on the biomass, mineral nutrient elements, antioxidant substances, and photosynthesis of two coniferous seedlings.

BACKGROUND: Studying the physiological growth status of Pinus yunnanensis Franch and Pinus elliottii Engelm. seedlings under different karst fissure thicknesses and rainfall distributions is of great significance for the management, vegetation restoration, and tree species selection in karst rocky desertification areas. In this study, we used a two-factor block experiment and set different rainfall durations, namely reduced rainfall duration (I3d), natural rainfall duration (I6d), and extended rainfall duration (I9d); Different karst small habitats, i.e., stone-free soil (S0), less stone and more soil (S1/4), and half stone and half soil (S1/2), are simulated at these three levels. Analyze the changes in physiological growth and photosynthetic characteristics in two coniferous seedlings under different treatments with different karst thicknesses. RESULTS: The results showed that with the increase of karst thickness, the growth volumes of height and diameter of P. yunnanensis seedlings, the biomass of various organs, and the accumulation of K+, Ca2+, Na+, and Mg2+ showed a significant change pattern of first increasing and then decreasing (P < 0.05); P. elliottii seedlings show a gradually decreasing trend (except for Ca2+). The biomass accumulation of each organ in two coniferous seedlings showed that leaves > stems > roots. The K+, Ca2+, and Mg2+ content in various organs of P. yunnanensis seedlings showed that leaves > roots > stems, while Na+ shows the order of roots > leaves > stems. The accumulation of mineral elements in various organs of P. elliottii seedlings is manifested as roots > stems > leaves and the accumulation of mineral elements in both coniferous seedlings is manifested as Ca2+ > Mg2+ > K+ > Na+. Root length, root volume, root surface area, root diameter, SOD, POD, SP, photosynthetic pigment content, fluorescence parameters, and gas exchange parameters of P. yunnanensis seedlings gradually increase with the increase of karst thickness (except for the 9-day rainfall duration), while those of P. elliottii seedlings gradually decrease. The light saturation point of P. yunnanensis seedlings is highest under the I6dS1/2 treatment, while that of P. elliottii is highest under the I3dS0 treatment. CONCLUSIONS: In summary, prolonging rainfall duration has an inhibitory effect on the growth of two types of coniferous seedlings. Increasing karst thickness inhibits the growth of P. elliottii seedlings, and to some extent, promotes the growth and development of P. yunnanensis seedlings. I6dS1/4 and I3dS0 treatments have the best growth effects on P. yunnanensis and P. elliottii seedlings. Therefore, we give priority to P. yunnanensis as the tree species for vegetation restoration or rocky desertification management in karst areas. Our study reveals the role of limestone-filled different karst fissures in mitigating the effects of drought as "containers" for plant growth. These findings help us understand the response of plants to drought stress and provide valuable insights for vegetation restoration in karst environments affected by global climate change. Therefore, further experiments with various karst fissure sizes are necessary to test the universality of the reactions of various plants under different karst fissures.

The application of varying amount of green manure combined with nitrogen fertilizer altered the soil bacterial community and rice yield in karst paddy areas.

Long-term application of green manure (GM) and nitrogen (N) fertilizers markedly improved soil fertility and boosted rice yield in ecologically fragile karst paddy fields. However, the precise response mechanisms of the soil bacterial community to varying amounts of green manure alone and in combination with N fertilizer in such environments remain poorly elucidated. In this study, we investigated the soil bacterial communities, keystone taxa, and their relationship with soil environmental variables across eight fertilization treatments. These treatments included group without N addition (N0M0, no N fertilizer and no GM; N0M22.5, 22.5 t/ha GM; N0M45, 45 t/ha GM, N0M67.5, 67.5 t/ha GM) and group with N addition (NM0, N fertilizer and no GM; NM22.5, N fertilizer and 22.5 t/ha GM; NM45, N fertilizer and 45 t/ha GM; NM67.5, N fertilizer and 67.5 t/ha GM). The results revealed that increasing green manure input significantly boosted rice yield by 15.51-22.08% and 21.84-35% in both the group without and with N addition, respectively, compared to N0M0 treatment. Moreover, with escalating green manure input, soil TN, AN, AK, and AP showed an increasing trend in the group without N addition. However, following the addition of N fertilizer, TN and AN content initially rose, followed by a decline due to the enhanced nutrient availability for rice. Furthermore, the application of a large amount of N fertilizer decreased the C: N ratio in the soil, resulting in significant changes in both the soil microbial community and its function. Particularly noteworthy was the transition of keystone taxa from their original roles as N-fixing and carbon-degrading groups (oligotrophs) to roles in carbon degradation (copiotrophs), nitrification, and denitrification. This shift in soil community and function might serve as a primary factor contributing to enhanced nutrient utilization efficiency in rice, thus significantly promoting rice yield.

Variations of soil metal content, soil enzyme activity and soil bacterial community in Rhododendron delavayi natural shrub forest at different elevations.

BACKGROUND: Rhododendron delavayi is a natural shrub that is distributed at different elevations in the karst region of Bijie, China, and that has an important role in preventing land degradation in this region. In this study, we determined the soil mineral element contents and soil enzyme activities. The composition of the soil bacterial community of R. delavayi at three elevations (1448 m, 1643 m, and 1821 m) was analyzed by high-throughput sequencing, and the interrelationships among the soil bacterial communities, mineral elements, and enzyme activities were determined. RESULTS: The Shannon index of the soil bacterial community increased and then decreased with increasing elevation and was highest at 1643 m. Elevations increased the number of total nodes and edges of the soil bacterial community network, and more positive correlations at 1821 m suggested stronger intraspecific cooperation. Acidobacteria, Actinobacteria and Proteobacteria were the dominant phyla at all three elevations. The Mantel test and correlation analysis showed that Fe and soil urease significantly affected bacterial communities at 1448 m; interestingly, Chloroflexi was positively related to soil urease at 1448 m, and Actinobacteria was positively correlated with Ni and Zn at 1821 m. Fe and soil urease significantly influenced the bacterial communities at lower elevations, and high elevation (1821 m) enhanced the positive interactions of the soil bacteria, which might be a strategy for R. delavayi to adapt to high elevation environments. CONCLUSION: Elevation significantly influenced the composition of soil bacterial communities by affecting the content of soil mineral elements and soil enzyme activity.

Species-Specific Root Distribution and Leaf Iso/Anisohydric Tendencies Shape Transpiration Patterns Across Heterogeneous Karst Habitats.

The driving forces of transpiration are not only atmospheric evaporation but also root zone water supply and stomatal regulation among species. However, the biophysiological drivers of transpiration remain incompletely understood in heterogeneous karst habitats. This study investigated the commonly coexisting tree species Mallotus philippensis and Celtis biondii in two typical karst habitats: rock-dominated (RD) habitat and control soil-dominated (SD) habitat. Over 2 years, soil moisture, transpiration, root distribution, and leaf water potential were measured. The results showed that soil moisture in the RD habitat was significantly lower than in the SD habitat. Transpiration patterns also differed between habitats, with species-specific distinctions driven by biophysiological traits. M. philippensis showed small hydroscape areas and its root system mainly distributed in the soil zone in both habitats. The isohydric behaviour and lower root density in the RD habitat drove M. philippensis to reduce transpiration in response to soil water deficiency. Conversely, C. biondii had large hydroscape areas and roots capable of penetrating bedrock. It transpired higher relying on ample accessible water through anisohydric behaviour and having a more robust root system both in soil and bedrock zones in the RD habitat. Our study highlights the critical role of root water accessibility and leaf iso/anisohydric tendencies in driving transpiration.

Bacterial isolation and genome analysis of a novel Klebsiella quasipneumoniae phage in southwest China's karst area.

BACKGROUND: Southwest China is one of the largest karst regions in the world. Karst environment is relatively fragile and vulnerable to human activities. Due to the discharge of sewage and domestic garbage, the karst system may be polluted by pathogenic bacteria. The detection of bacterial distribution and identification of phage capable of infecting them is an important approach for environmental assessment and resource acquisition. METHODS: Bacteria and phages were isolated from karst water in southwest China using the plate scribing and double plate method, respectively. Isolated phage was defined by transmission electron microscopy, one-step growth curve and optimal multiplicity of infection (MOI). Genomic sequencing, phylogenetic analysis, comparative genomic and proteomic analysis were performed. RESULTS: A Klebsiella quasipneumoniae phage was isolated from 32 isolates and named KL01. KL01 is morphologically identified as Caudoviricetes with an optimal MOI of 0.1, an incubation period of 10 min, and a lysis period of 60 min. The genome length of KL01 is about 45 kb, the GC content is 42.5%, and it contains 59 open reading frames. The highest average nucleotide similarity between KL01 and a known Klebsiella phage 6939 was 83.04%. CONCLUSIONS: KL01 is a novel phage, belonging to the Autophagoviridae, which has strong lytic ability. This study indicates that there were not only some potential potentially pathogenic bacteria in the karst environment, but also phage resources for exploration and application.

Yeast Biodiversity of Karst Waters: Interest of Four Culture Media and an Improved MALDI-TOF MS Database.

Karst aquifers are a significant source of drinking water and highly vulnerable to pollution and microbial contamination. Microbiological regulations for the quality of drinking water mostly focus on bacterial levels and lack guidance concerning fungal contamination. Moreover, there is no standardised microbial analysis methodology for identifying fungi in water. Our main objective was to establish the most effective culture and identification methodology to examine yeast diversity in karst waters. We assessed the comparative efficacy of four culture media (CHROMagar Candida, dichloran glycerol 18% [DG18], dichloran rose Bengal chloramphenicol [DRBC], and SYMPHONY agar) for yeast isolation from karst water samples. Furthermore, we investigated the comprehensiveness of databases used in MALDI-TOF mass spectrometry (MALDI-TOF MS) for identifying environmental yeast species. In total, we analysed 162 water samples, allowing the identification of 2479 yeast isolates. We demonstrate that a combination of four culture media, each with distinct specifications, more efficiently covers a wide range of yeast species in karst water than a combination of only two or three. Supplementation of a MALDI-TOF MS database is also critical for analysing environmental microbial samples and improved the identification of yeast biodiversity. This study is an initial step towards standardising the analysis of fungal biodiversity in karst waters, enabling a better understanding of the significance of this environmental reservoir in relation to public health.

Dynamic Changes of Soil Microbial Communities During the Afforestation of Pinus Armandii in a Karst Region of Southwest China.

Clarifying the response of soil microbial communities to vegetation restoration is essential to comprehend biogeochemical processes and ensure the long-term viability of forest development. To assess the variations in soil microbial communities throughout the growth of Pinus armandii plantations in the karst region, we utilized the "space instead of time" approach and selected four P. armandii stands with ages ranging from 10 to 47 years, along with a grassland control. The microbial community structure was determined by conducting Illumina sequencing of the 16 S rRNA gene and the ITS gene, respectively. The results demonstrated that afforestation with P. armandii significantly influenced soil microbial communities, as indicated by notable differences in bacterial and fungal composition and diversity between the plantations and the control. However, soil microbe diversity did not display significant variation across stand ages. Moreover, the bacterial community exhibited higher responsiveness to age gradients compared to the fungal community. Soil physicochemical factors play a critical role in elucidating microbial diversity and community composition variations during restoration processes. TN, AN, TP, AP, SOC, AK, and pH were the most significant influencing factors for the composition of bacterial community, while TC, SOC, pH, and TCa were the most significant influencing factors for the composition of fungal community. Our findings indicate substantial changes in soil bacterial and fungal communities across successive stages of development. Additionally, the changes in dominant bacteria and fungi characteristics across the age gradient were primarily attributed to variations in the prevailing soil conditions and chemical factors.

Characteristics and whole-genome analysis of a novel Pseudomonas syringae pv. tomato bacteriophage D6 isolated from a karst cave.

Pseudomonas syringae is a gram-negative plant pathogen that infects plants such as tomato and poses a threat to global crop production. In this study, a novel lytic phage infecting P. syringae pv. tomato DC3000, named phage D6, was isolated and characterized from sediments in a karst cave. The latent period of phage D6 was found to be 60 min, with a burst size of 16 plaque-forming units per cell. Phage D6 was stable at temperatures between 4 and 40 °C but lost infectivity when heated to 70 °C. Its infectivity was unaffected at pH 6-10 but became inactivated at pH ≤ 5 or ≥ 12. The genome of phage D6 is a linear double-stranded DNA of 307,402 bp with a G + C content of 48.43%. There is a codon preference between phage D6 and its host, and the translation of phage D6 gene may not be entirely dependent on the tRNA library provided by the host. A total of 410 open reading frames (ORFs) and 14 tRNAs were predicted in its genome, with 92 ORFs encoding proteins with predicted functions. Phage D6 showed low genomic similarity to known phage genomes in the GenBank and Viral sequence databases. Genomic and phylogenetic analyses revealed that phage D6 is a novel phage. The tomato plants were first injected with phage D6, and subsequently with Pst DC3000, using the foliar spraying and root drenching inoculum approach. Results obtained after 14 days indicated that phage D6 inoculation decreased P. syringae-induced symptoms in tomato leaves and inhibited the pathogen's growth in the leaves. The amount of Pst DC3000 was reduced by 150- and 263-fold, respectively. In conclusion, the lytic phage D6 identified in this study belongs to a novel phage within the Caudoviricetes class and has potential for use in biological control of plant diseases.

Karst fissures mitigate the negative effects of drought on plant growth and photosynthetic physiology.

Hard limestone substrates, which are extensively distributed, are believed to exacerbate drought and increase the difficulty of restoration in vulnerable karst regions. Fissures in such substrates may alleviate the negative effect of drought on plants, but the underlying mechanisms remain poorly understood. In a two-way factorial block design, the growth and photosynthesis of 2-year-old Phoebe zhennan seedlings were investigated in two water availabilities (high versus low) and three stimulated fissure habitat groups (soil, soil-filled fissure and non-soil-filled fissure). Moreover, the fissure treatments included both small and big fissures. Compared to the soil group, the non-soil-filled fissure group had decreased the total biomass, root biomass, total root length, and the root length of fine roots in the soil layer at both water availabilities, but increased net photosynthetic rate (Pn) and retained stable water use efficiency (WUE) at low water availability. However, there were no significant differences between the soil-filled fissure group and soil group in the biomass accumulation and allocation as well as Pn. Nevertheless, the SF group decreased the root distribution in total and in the soil layer, and also increased WUE at low water availability. Across all treatments, fissure size had no effect on plant growth or photosynthesis. Karst fissures filled with soil can alleviate drought impacts on plant root growth, which involves adjusting root distribution strategies and increasing water use efficiency. These results suggest that rock fissures can be involved in long-term plant responses to drought stress and vegetation restoration in rocky mountain environments under global climate change.

Dissolved carbon in effluent of wastewater treatment plants and its potential impacts in the receiving karst river.

The dissolved carbon cycling in river system fueled by wastewater treatment plant effluent have been a research hotspot. However, the composition of dissolved carbon (DC) in wastewater effluents from karst regions remains poorly understood, resulting in a lack of clarity regarding its impact on the dynamics of dissolved carbon in karst rivers. To address this knowledge gap, this study investigated variations of dissolved inorganic (DIC) and organic C (DOC) components in effluent in karst regions and preliminarily discussed their influence on the DC cycling in karst rivers. The results showed that bicarbonate (HCO3-) in WWTP effluents makes more than 90% of the total dissolved inorganic carbon (DIC). The partial pressure of aqueous CO2 (pCO2) of the effluent reached 14450 ± 10084µtam, and pCO2 level declined with increasing river distance from the effluent discharge, effluent acted as a strong CO2 emitter to the atmosphere. Stable carbon isotope and water chemistry evidence revealed that organic matter degradation made important contributions to the high CO2 concentrations in effluent. PHREEQC mixing simulation together with filed samples data indicated that the DIC species can be changed, and pCO2 increased in receiving karst river water after mixed with effluent. The dissolved organic carbon (DOC) of effluent contained humic-like and protein-tryptophan-like, both of them appeared important and recent autochthonous, which could interfere the distinguish the sources of DOC in receiving karst river water. Thus, these findings highlight that the effluent can be an essential factor for the changes of the karst riverine DC pool, which advance our understanding on karst riverine DC evolution under anthropogenic activities. As more than 30% of the earth surface in China, northern America, and Europe are covered by carbonate rocks, this study has relevant implications for other karst regions as it underscores the influence of WWTP effluents on the carbon cycle in karst rivers. Such information and knowledge are valuable for monitoring and managing effluent-receiving river in other karst regions in the world.

Dual carbon isotopes (δ13C and Δ14C) were used to reveal the main sources and input fluxes of dissolved inorganic carbon in a karst reservoir in winter.

In karst areas, the dissolved inorganic carbon (DIC) concentrations in aquatic systems are typically higher than that in non-karst areas due to intensive carbonate rock weathering. Understanding the sources and input fluxes of DIC in karst reservoirs is crucial for regional carbon cycle studies. This study utilized dual carbon isotopes (δ13CDIC and Δ14CDIC) to estimate the contribution rates and input fluxes of DIC from various sources in Aha Reservoir (AHR), located in southwestern China. Our results indicated that the DIC concentrations (22.33-32.79 mg L-1) and δ13CDIC values (-10.02‰ to -8.55‰) were nearly homogeneous both vertically and laterally in the reservoir (p > 0.05). The Δ14CDIC values (-246.31‰ to -137.86‰) were homogeneous along the vertical profile (p > 0.05), but showed significant horizontal variation (p < 0.05), with values decreasing from -149.57 ± 10.27‰ to -232.85 ± 2.37‰ at the mouths of the inflowing rivers. We found that the inflowing rivers were the primary DIC sources to AHR, contributing 70% of the total input, while groundwater and atmospheric CO2 contributions were relatively minor, at 18% and 12%, respectively. The Jinzhong River (JZR), influenced by industrial and domestic wastewater discharge, contributed the largest DIC input flux at 2.01 t/(km2·mon). In contrast, the Youyu River (YYR), influenced by acidic mine drainage, and the Baiyan River (BYR), influenced by agricultural activities, contributed relatively smaller DIC input fluxes of 1.29 t/(km2·mon) and 1.03 t/(km2·mon), respectively. This study highlights the significant impact of anthropogenic activities on DIC input in AHR, with industrial and domestic wastewater discharges having a greater influence than agricultural activities and acidic mine wastewater inputs. These findings underscore the critical need to manage and mitigate the impacts of human activities on karst reservoir ecosystems.

High-frequency dynamics of CO2 emission flux and its influencing factors in a subtropical karst groundwater-fed reservoir, south China.

Revealing the magnitude, dynamics, and influencing factors of CO2 emissions across the water-air interface in karst water with high frequency is crucial for accurately assessing the carbon budget in a karst environment. Due to the limitations of observation methods, the current research is still very insufficient. To solve the above problems and clarify the main influencing factors of CO2 emission in karst water, this study selected Dalongdong (DLD) Reservoir, located in the typical karst peak and valley area in southwest China, to carry out a multi-parameter high-frequency monitoring study from January to December 2021, and used the thin boundary model method to estimate the CO2 flux across the water-air interface (CF). The average annual flux of DLD reservoir is 84.48 mmol·(m2·h)-1, which represents a CO2 source overall. However, during the stratification period in August, there is a transient carbon sink due to negative CO2 emission. The alteration of thermal stratification in water is crucial in regulating the seasonal variation of CF. Meanwhile, the diurnal variation is significantly influenced by changes in hydrochemical parameters during the thermal stratification stage. Compared to low wind speeds (<3 m/s), high wind speeds (≥3 m/s) have a greater impact on the CO2 flux. Furthermore, high-frequency continuous data revealed that the reservoir triggered a CO2 pulse emission during the turnover process, primarily at night, leading to unusually high CO2 flux values. It is of great significance to monitor and reveal the process, flux, and control factors of CO2 flux in land water at a high-frequency strategy. They will help improve the accuracy of regional or watershed carbon budgets and clarify the role of global land water in the global carbon budget.

Methane sink of subterranean space in an integrated atmosphere-soil-cave system.

CH4 serves as an important greenhouse gas, yet limited knowledge is available in global and regional CH4 cycling, particularly in widely distributed karst terrain. In this study, we investigated an upland in Puding Karst Ecosystem Research Station, and explored CH4 concentration and/or flux in atmosphere, soil and cave using a closed static chamber method and an eddy covariance system. Meanwhile, we monitored atmospheric temperature, precipitation, temperature and wind velocity in the cave entrance. The results demonstrated that atmospheric CH4 and actual soil CH4 fluxes in the source area of eddy covariance system were -0.19 ± 8.64 nmols-1m-2 and -0.16 nmols-1m-2 respectively. The CH4 concentrations in Shawan Cave exhibited 10 âˆ¼ 100-fold lower than that of the external atmosphere. CH4 oxidation rate dominated by methane-oxidizing bacteria was 1.98 nmols-1m-2 in Shawan Cave when it combined with temperature difference between cave and external atmosphere. Therefore, CH4 sink in global karst subterranean spaces was estimated at 106.2 Tg CH4 yr-1. We supplemented an understanding of CH4 cycling paths and fluxes in karst terrain, as well as CH4 sinks in karst subterranean space. Further works require to establish a karst ecosystem observation network to conduct long-term integrated studies on CH4 fluxes regarding atmosphere, soils, plants and caves.

Impact of permanganate with polyaluminium chloride on algae-laden karst water: Behaviors and disinfection by-products control.

The removal of algal organic matter (AOM) through water treatment processes is a major approach of reducing the formation of disinfection by-products (DBP). Here, the formation of DBP from AOM in karst water under different combination of potassium permanganate (KMnO4) and polyaluminium chloride (PACl) was investigated. The effect of divalent ions (Ca2+ and Mg2+) on DBP formation was traced by AOM chemistry variations. For DBP formation after KMnO4 preoxidation, total carbonaceous DBPs (C-DBPs) decreased by 12.9% but nitrogen-containing DBPs (N-DBPs) increased by 18.8%. Conversely, the C-DBPs further increased by 3.3% but N-DBPs reduced by 10.7% after the addition of PACl besides KMnO4 preoxidation. The variations of aromatic protein-like, soluble microbial products-like compounds and ultraviolet absorbance at 254 nm (UV254) were highly correlated with the formation of DBPs, which suggest aromatic substances strongly affect DBP behaviors at different treatment conditions. In the presence of divalent ions (Ca2+ = 135.86 mg/L, Mg2+ = 18.51 mg/L), the combination of KMnO4 and PACl was more effective in controlling DBP formation compared to the situation without Ca2+ and Mg2+. Specifically, trichloromethane formation was largely inhibited compared to the other tested DBPs, which may refer to complexation of electron-donating groups via divalent ions. While Ca2+ and Mg2+ may not affect the nature of α-carbon and amine groups, so the variation of haloacetonitriles (HANs) was not obvious. The study enhances the understanding of the DBP formation patterns, transformation of carbon and nitrogen by preoxidation-coagulation (KMnO4-PACl) treatment in algae-laden karst water.

Aquatic photosynthetic carbon pump driven by periodic temperature variations affects dissolved inorganic carbon and hydrochemical characteristics in a groundwater-fed reservoir.

Under the influence of periodic temperature variations, biogeochemical cycling in water bodies is markedly affected by the periodic thermal stratification processes in subtropical reservoirs or lakes. In current studies, there is insufficient research on the influence and mechanism of dissolved inorganic carbon (DIC) distribution in karst carbon-rich groundwater-fed reservoirs under the coupled effects of thermal structure stratification and the biological carbon pump (BCP) effect. To address this issue, the Dalongdong (DLD) reservoir in the subtropical region of southern China was chosen as the site for long-term monitoring and research on relevant physicochemical parameters of water, DIC, and its stable carbon isotope (δ13CDIC), CO2 emission flux, as well as the reservoir's thermal stratification index. The results show that: (1) the DLD reservoir is a typical warm monomictic reservoir, which exhibits regular variations of mixing period-stratification period-mixing period on a yearly scale due to thermal structure changes; (2) DIC was consumed by aquatic photosynthetic organisms in the epilimnion during the stratification period, leading to a decrease in DIC concentration, partial pressure of CO2 (pCO2) and CO2 emission flux, and an increase in stable carbon isotope (δ13CDIC). During the mixing period, the trend was reversed; (3) During the thermal stratification, aquatic photosynthesis and water temperature were the primary factors controlling DIC variations in both the epilimnion and thermocline. Regarding the hypolimnion, calcite dissolution, organic matter decomposition, and water temperature were the dominant controlling factors. These results indicate that although carbon-rich karst groundwater provides a plentiful supply of DIC in the DLD reservoir, its availability is still influenced by variations in the reservoir's thermal structure and the metabolic processes of aquatic photosynthetic organisms. Therefore, to better estimate the regional carbon budget in a reservoir or lake, future studies should especially consider the combined effects of BCP and thermal structure variations on carbon variations.

A New Species of the Blind Cave Loach Genus Protocobitis (Cypriniformes: Cobitidae), Protocobitis longicostatus sp. nov., from Guangxi, China.

Protocobitis species are typical cave-dwelling fish, exhibiting distinctive morphological adaptations such as colorless body, lack of eyes, and reduced scales and ribs in response to their extreme cave habitats. Distinct from the recorded species, P. anteroventris, P. polylepis, and P. typhlops, a new species, Protocobitis longicostatus sp. nov., is described from Guangxi Zhuang Autonomous Region, China. Protocobitis longicostatus sp. nov. can easily be distinguished from all known congeners by the following characteristics: whole body covered by scales except head, 12 branched caudal fin rays, and long ribs. These species face threats from habitat degradation, hydrological changes, and environmental pollution. Thus, the conservation of cavefish in China has become an urgent issue.

Removal of potentially toxic elements from water with the moss-tufa micro-filtration system.

Mosses are an integral component in the tufa sedimentary landscape. In this study, we investigated the use of the porous moss-tufa structure as a filtration system for removing potentially toxic elements (PTEs) from water samples. Three species of mosses that commonly grow on tufa were selected, and the PTEs filtered by the moss-tufa system were identified by inductively coupled plasma mass spectrometry. The bioconcentration factor (BCF) of mosses was calculated to compare the enrichment effects of different mosses on PTEs. Likewise, the level of PTEs flowing through the moss-tufa system was measured, and the water quality removal rate (C) was calculated accordingly. The results revealed that the moss-tufa system was mainly composed of Fissidens grandifrons Brid., Hydrogonium dixonianum P. C. Chen, and Cratoneuron filicinum (Hedw.) Spruce var. filicinum. Among these, Fissidens grandifrons Brid. reported the highest retention capacity for PTEs. Collectively, the moss-tufa filtration system displayed a strong retention capacity and removal rate of Mn, Pb, and Ni from the water sample. The removal of PTEs by the moss-tufa system was mainly based on the enrichment of mosses and the adsorption-retention ability of tufa. In conclusion, the moss-tufa micro-filtration system displayed the effective removal of PTEs from water samples and could be applied to control the levels of toxic elements in karst water bodies.

A Novel 3D Reconstruction Sensor Using a Diving Lamp and a Camera for Underwater Cave Exploration.

Aquifer karstic structures, due to their complex nature, present significant challenges in accurately mapping their intricate features. Traditional methods often rely on invasive techniques or sophisticated equipment, limiting accessibility and feasibility. In this paper, a new approach is proposed for a non-invasive, low-cost 3D reconstruction using a camera that observes the light projection of a simple diving lamp. The method capitalizes on the principles of structured light, leveraging the projection of light contours onto the karstic surfaces. By capturing the resultant light patterns with a camera, three-dimensional representations of the structures are reconstructed. The simplicity and portability of the equipment required make this method highly versatile, enabling deployment in diverse underwater environments. This approach is validated through extensive field experiments conducted in various aquifer karstic settings. The results demonstrate the efficacy of this method in accurately delineating intricate karstic features with remarkable detail and resolution. Furthermore, the non-destructive nature of this technique minimizes disturbance to delicate aquatic ecosystems while providing valuable insights into the subterranean landscape. This innovative methodology not only offers a cost-effective and non-invasive means of mapping aquifer karstic structures but also opens avenues for comprehensive environmental monitoring and resource management. Its potential applications span hydrogeological studies, environmental conservation efforts, and sustainable water resource management practices in karstic terrains worldwide.