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    Effect of land use, habitat suitability, and hurricanes on the population connectivity of an endemic insular bat

    1.Ceballos, G. Mammal population losses and the extinction crisis. Science 296, 904–907 (2002).ADS 
    CAS 
    PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    2.Meyer, C. F. J., Struebig, M. J. & Willig, M. R. Responses of tropical bats to habitat fragmentation, logging, and deforestation. In Bats in the Anthropocene: Conservation of Bats in a Changing World (eds Voigt, C. C. & Kingston, T.) 63–103 (Springer, 2016). https://doi.org/10.1007/978-3-319-25220-9_4.
    Google Scholar 
    3.Torres-Romero, E. J., Giordano, A. J., Ceballos, G. & López-Bao, J. V. Reducing the sixth mass extinction: understanding the value of human-altered landscapes to the conservation of the world’s largest terrestrial mammals. Biol. Conserv. 249, 108706 (2020).Article 

    Google Scholar 
    4.Mittermeier, R. A., Turner, W. R., Larsen, F. W., Brooks, T. M. & Gascon, C. Global biodiversity conservation: the critical role of hotspots BT—biodiversity hotspots: distribution and protection of conservation priority areas. In (eds Zachos, F. E. & Habel, J. C.) 3–22 (Springer, Berlin, 2011). https://doi.org/10.1007/978-3-642-20992-5_1.5.Bosso, L., Mucedda, M., Fichera, G., Kiefer, A. & Russo, D. A gap analysis for threatened bat populations on Sardinia. Hystrix Ital. J. Mammal. 27, 212–214 (2016).
    Google Scholar 
    6.Upham, N. S. Past and present of insular Caribbean mammals: understanding Holocene extinctions to inform modern biodiversity conservation. J. Mammal. 98, 913–917 (2017).Article 

    Google Scholar 
    7.Gould, W. A., Castro-Prieto, J. & Álvarez-Berríos, N. L. Climate change and biodiversity conservation in the Caribbean islands. In Encyclopedia of the World’s Biomes (eds Goldstein, M. & DellaSala, D.) 114–125 (Elsevier, 2020). https://doi.org/10.1016/B978-0-12-409548-9.12091-3.
    Google Scholar 
    8.Schoener, T. W., Spiller, D. A. & Losos, J. B. Variable ecological effects of hurricanes: the importance of seasonal timing for survival of lizards on Bahamian islands. Proc. Natl. Acad. Sci. 101, 177 LP – 181 (2004).ADS 
    Article 
    CAS 

    Google Scholar 
    9.Barnosky, A. D. et al. Has the Earth’s sixth mass extinction already arrived?. Nature 471, 51–57 (2011).ADS 
    CAS 
    PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    10.Pimm, S. L. et al. The biodiversity of species and their rates of extinction, distribution, and protection. Science 344, 1246752–1246752 (2014).CAS 
    PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    11.Turvey, S. T., Kennerley, R. J., Nuñez-Miño, J. M. & Young, R. P. The Last Survivors: current status and conservation of the non-volant land mammals of the insular Caribbean. J. Mammal. 98, 918–936 (2017).Article 

    Google Scholar 
    12.Andermann, T., Faurby, S., Turvey, S. T., Antonelli, A. & Silvestro, D. The past and future human impact on mammalian diversity. Sci. Adv. 6, eabb313 (2020).Article 

    Google Scholar 
    13.Turvey, S. T. & Crees, J. J. Extinction in the anthropocene. Curr. Biol. 29, R982–R986 (2019).CAS 
    PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    14.Donihue, C. M. et al. Hurricane effects on neotropical lizards span geographic and phylogenetic scales. Proc. Natl. Acad. Sci. 117, 10429 LP – 10434 (2020).Article 
    CAS 

    Google Scholar 
    15.Gannon, M. R., Kurta, A., Rodríguez-Durán, A. & Willig, M. R. Bats of Puerto Rico: An Island Focus and a Caribbean Perspective (Texas Tech University Press, 2005).
    Google Scholar 
    16.Miller, G. L. & Lugo, A. E. Guide to the ecological systems of Puerto Rico. IITF-GTR-35. (2009).17.Guzmán-Colón, D. K., Pidgeon, A. M., Martinuzzi, S. & Radeloff, V. C. Conservation planning for island nations: using a network analysis model to find novel opportunities for landscape connectivity in Puerto Rico. Glob. Ecol. Conserv. 23, e01075 (2020).Article 

    Google Scholar 
    18.Gould, W. A. et al. The Puerto Rico Gap Analysis Project Volume 1: Land Cover, Vertebrate Species Distributions, and Land Stewardship. General technical reports IITF-39 vol. 1 https://www.fs.usda.gov/treesearch/pubs/38430 (2008).19.Gould, W. A. Puerto Rico gap analysis project. GAP Anal. Bull. 16, 71–79 (2009).
    Google Scholar 
    20.Gould, W. A., Quiñones, M., Solorzano, M., Alcobas, W. & Alarcon, C. Protected Natural Areas of Puerto Rico. Res. Map IITF-RMAP-02. Rio Piedras, PR US Dep. Agric. For. Serv. Int. Inst. Trop. For. (2011).21.Junta de Planificación. Plan de Uso de Terrenos, Guías de Ordenación del Territorio. 220 (2015).22.Gould, W. A., Wadsworth, F. H., Quiñones, M., Fain, S. J. & Álvarez-Berríos, N. L. Land use, conservation, forestry, and agriculture in Puerto Rico. Forests 8, 242–263 (2017).Article 

    Google Scholar 
    23.QGIS.org. QGIS Geographic Information System (2016).24.Martinuzzi, S., Gould, W. A., González, O. M. R., Quiñones, M. & Jiménez, M. E. Urban and rural land use in Puerto Rico. Res. Map IITF-RMAP-01. Rio Piedras, PR US Dep. Agric. For. Serv. Int. Inst. Trop. For. (2008).25.Gould, W. A., Martinuzzi, S. & González, O. M. R. High and low density development in Puerto Rico. Res. Map IITF-RMAP-11. Rio Piedras, PR US Dep. Agric. For. Serv. Int. Inst. Trop. For. (2008).26.Gannon, M. R. & Willig, M. R. The effects of Hurricane Hugo on bats of the Luquillo experimental forest of Puerto Rico. Biotropica 26, 320 (1994).Article 

    Google Scholar 
    27.Gannon, M. R. & Willig, M. R. Long-term monitoring protocol for bats: lessons from the Luquillo Experimental Forest of Puerto Rico. For. Biodivers. North Cent. South Am. Caribbean. Res. Monit. Man Biosph. Ser. 21, 271–291 (1998).
    Google Scholar 
    28.Gannon, M. R. & Willig, M. R. Island in the storm: disturbance ecology of plant-visiting bats on the hurricane-prone island of Puerto Rico. In Island Bats: Evolution, Ecology, and Conservation (eds Fleming, T. H. & Racey, P.) 281–301 (University of Chicago Press, 2009).
    Google Scholar 
    29.Jones, K. E., Barlow, K. E., Vaughan, N., Rodríguez-Durán, A. & Gannon, M. R. Short-term impacts of extreme environmental disturbance on the bats of Puerto Rico. Anim. Conserv. 4, 59–66 (2001).Article 

    Google Scholar 
    30.Rodríguez-Durán, A. & Vázquez, R. The bat Artibeus jamaicensis in Puerto Rico (West Indies): seasonality of diet, activity, and effect of a hurricane. Acta Chiropterologica 3, 53–61 (2001).
    Google Scholar 
    31.Rodríguez-Durán, A., Nieves, N. A. & Avilés-Ruiz, Y. Hurricane-mediated extirpation of a bat from an Antillean Island. Caribb. Nat. 78, 1–7 (2020).
    Google Scholar 
    32.Genoways, H. H. & Baker, R. J. Stenoderma rufum. Mamm. Species https://doi.org/10.2307/3503991 (1972).Article 

    Google Scholar 
    33.Kwiecinski, G. G. & Coles, W. C. Presence of Stenoderma rufum beyond the Puerto Rican bank. Occas. Pap. Museum Texas Tech Univ. https://doi.org/10.5962/bhl.title.156896 (2007).Article 

    Google Scholar 
    34.Liu, X. et al. Litterfall production prior to and during Hurricanes Irma and Maria in four Puerto Rican forests. Forests 9, 367 (2018).Article 

    Google Scholar 
    35.Rodríguez-Durán, A. Stenoderma rufum. IUCN Red List Threat. Species e.T20743A22065638 https://doi.org/10.2305/IUCN.UK.2016-1.RLTS.T20743A22065638.en (2016).Article 

    Google Scholar 
    36.Gannon, M. R. Foraging Ecology, Reproductive Biology, and Systematics of the Red Fig-Eating Bat (Stenoderma rufum) in the Tabonuco Rain Forest of Puerto Rico (Texas Tech University, 1991).
    Google Scholar 
    37.Meyer, C. F. J. & Kalko, E. K. V. Assemblage-level responses of phyllostomid bats to tropical forest fragmentation: land-bridge islands as a model system. J. Biogeogr. 35, 1711–1726 (2008).Article 

    Google Scholar 
    38.Estrada-Villegas, S., Meyer, C. F. J. & Kalko, E. K. V. Effects of tropical forest fragmentation on aerial insectivorous bats in a land-bridge island system. Biol. Conserv. 143, 597–608 (2010).Article 

    Google Scholar 
    39.Feng, Y., Negrón-Juárez, R. I. & Chambers, J. Q. Remote sensing and statistical analysis of the effects of hurricane María on the forests of Puerto Rico. Remote Sens. Environ. 247, 111940 (2020).ADS 
    Article 

    Google Scholar 
    40.Soto-Centeno, J. A. & Steadman, D. W. Fossils reject climate change as the cause of extinction of Caribbean bats. Sci. Rep. 5, 7971 (2015).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    41.Razgour, O. Beyond species distribution modeling: a landscape genetics approach to investigating range shifts under future climate change. Ecol. Inform. 30, 250–256 (2015).Article 

    Google Scholar 
    42.Rodríguez-Durán, A. Bat assemblages in the West Indies: the role of caves. In Island Bats: Evolution, Ecology and Conservation (eds Fleming, T. H. & Racey, P.) 265–280 (University of Chicago Press, 2009).
    Google Scholar 
    43.Nassar, J. M., Aguirre, L. F., Rodríguez-Herrera, B. & Medellín, R. A. Threats, status, and conservation perspectives for leaf-nosed bats. In Phyllostomid Bats: A Unique Mammalian Radiation (eds Fleming, T. H. et al.) 470 (University of Chicago Press, 2020).
    Google Scholar 
    44.Rodríguez-Durán, A. Nonrandom aggregations and distribution of cave-dwelling bats in Puerto Rico. J. Mammal. 79, 141–146 (1998).Article 

    Google Scholar 
    45.Rodríguez-Durán, A. & Padilla-Rodríguez, E. New records for the bat fauna of Mona Island, Puerto Rico, with notes on their natural history. Caribb. J. Sci. 46, 102–105 (2010).Article 

    Google Scholar 
    46.Rodríguez-Durán, A. & Feliciano-Robles, W. Conservation value of remnant habitat for neotropical bats on islands. Caribb. Nat. 35, 1–10 (2016).
    Google Scholar 
    47.Gómez-Ruiz, E. P. & Lacher, T. E. Modelling the potential geographic distribution of an endangered pollination corridor in Mexico and the United States. Divers. Distrib. 23, 67–78 (2017).Article 

    Google Scholar 
    48.Shah, V. B. & McRae, B. H. Circuitscape: a tool for landscape ecology. In Proceedings of the 7th Python in Science Conference, vol. 7, 62–66 (SciPy Conference California, 2008).49.McRae, B. H., Dickson, B. G., Keitt, T. H. & Shah, V. B. Using circuit theory to model connectivity in ecology, evolution, and conservation. Ecology 89, 2712–2724 (2008).PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    50.Carroll, C., McRae, B. H. & Brookes, A. Use of linkage mapping and centrality analysis across habitat gradients to conserve connectivity of Gray wolf populations in Western North America. Conserv. Biol. 26, 78–87 (2012).PubMed 
    Article 
    PubMed Central 

    Google Scholar 
    51.Theobald, D. M., Reed, S. E., Fields, K. & Soulé, M. Connecting natural landscapes using a landscape permeability model to prioritize conservation activities in the United States. Conserv. Lett. 5, 123–133 (2012).Article 

    Google Scholar 
    52.Dutta, T., Sharma, S., McRae, B. H., Roy, P. S. & DeFries, R. Connecting the dots: mapping habitat connectivity for tigers in central India. Reg. Environ. Change 16, 53–67 (2016).Article 

    Google Scholar 
    53.Mallory, C. D. & Boyce, M. S. Prioritization of landscape connectivity for the conservation of Peary caribou. Ecol. Evol. 9, 2189–2205 (2019).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    54.Osipova, L. et al. Using step-selection functions to model landscape connectivity for African elephants: accounting for variability across individuals and seasons. Anim. Conserv. 22, 35–48 (2019).Article 

    Google Scholar 
    55.GBIF.org. GBIF Occurrence Download (2019). https://doi.org/10.15468/dl.atjvik56.Fick, S. E. & Hijmans, R. J. WorldClim 2: new 1-km spatial resolution climate surfaces for global land areas. Int. J. Climatol. 37, 4302–4315 (2017).Article 

    Google Scholar 
    57.Vermote, E. & NOAA CDR Program. NOAA Climate Data Record (CDR) of AVHRR Normalized Difference Vegetation Index (NDVI), Version 5 (2019). https://doi.org/10.7289/V5ZG6QH9.58.de Moraes, W. M. & Viveiros Grelle, C. E. Does environmental suitability explain the relative abundance of the tailed tailless bat, Anoura caudifer. Nat. Conserv. 10, 221–227 (2012).Article 

    Google Scholar 
    59.Gutiérrez, E. E., Boria, R. A. & Anderson, R. P. Can biotic interactions cause allopatry? Niche models, competition, and distributions of South American mouse opossums. Ecography 37, 741–753 (2014).Article 

    Google Scholar 
    60.Gutiérrez, E. E. et al. The taxonomic status of Mazama bricenii and the significance of the Táchira depression for mammalian endemism in the Cordillera de Mérida, Venezuela. PLoS ONE 10, 1–24 (2015).
    Google Scholar 
    61.Ancillotto, L., Mori, E., Bosso, L., Agnelli, P. & Russo, D. The Balkan long-eared bat (Plecotus kolombatovici) occurs in Italy—first confirmed record and potential distribution. Mamm. Biol. 96, 61–67 (2019).Article 

    Google Scholar 
    62.Alberdi, A., Aizpurua, O., Aihartza, J. & Garin, I. Unveiling the factors shaping the distribution of widely distributed alpine vertebrates, using multi-scale ecological niche modelling of the bat Plecotus macrobullaris. Front. Zool. 11, 77 (2014).PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    63.Phillips, S. J., Anderson, R. P. & Schapire, R. E. Maximum entropy modeling of species geographic distributions. Ecol. Model. 190, 231–259 (2006).Article 

    Google Scholar 
    64.Phillips, S. J. & Dudík, M. Modeling of species distributions with Maxent: new extensions and a comprehensive evaluation. Ecography (Cop.) 31, 161–175 (2008).Article 

    Google Scholar 
    65.R Core Team. R: A Language and Environment for Statistical Computing (2018).66.Muscarella, R. et al. ENMeval: an R package for conducting spatially independent evaluations and estimating optimal model complexity for Maxent ecological niche models. Methods Ecol. Evol. 5, 1198–1205 (2014).Article 

    Google Scholar 
    67.Hirzel, A. H., Le Lay, G., Helfer, V., Randin, C. & Guisan, A. Evaluating the ability of habitat suitability models to predict species presences. Ecol. Model. 199, 142–152 (2006).Article 

    Google Scholar  More

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    Isolation and screening of multifunctional phosphate solubilizing bacteria and its growth-promoting effect on Chinese fir seedlings

    Plant materialsIn April 2019, 2-year-old Chinese fir seedlings were collected from the Yalin Center of the Chinese Academy of Forestry in good condition and free from pests and diseases. (The use of Chinese fir seedlings in the experiment complies with national regulations).MediumPikovskava (PVK) solid medium: glucose 10 g, Ca3(PO4) 25 g, CaCO35g, (NH4)2SO40.5 g, NaCl 0.2 g, MgSO4 7H2O 0.1 g, KCl 0.1 g, MnSO4 0.002 g, FeSO4 7H2O 0.002 g, agar 18 g, Distilled water 1000 mL, pH 7.0.PVK liquid medium: PVK solid medium without agar.Luria-Bertan (LB) medium: tryptone 10 g, yeast extract powder 5 g, NaCl 10 g, agar 18 g, distilled water 1000 mL, pH7.0.LB liquid medium:LB solid medium without agar.Isolation and purification of endophytes from Chinese fir seedlingsThe roots, stems, and leaves of the selected Chinese fir seedlings were washed away with running water to remove the surface soil, and then washed with running water for 24 h to 36 h, and the surface moisture was absorbed with sterile filter paper. Weigh 1 g of roots, stems and leaves in a petri dish, and then carry out surface disinfection in a sterile operating table with 75% alcohol (C2H5OH) for 30 s, 5% sodium hypochlorite (NaClO) 10 min, wash the sterile water 7 times, and use sterile filter paper to absorb the water. The material after surface sterilization is cut into 2 mm × 2 mm with sterile surgical scissors, placed in a sterilized mortar and grated with a small amount of sterile quartz sand and ground into a homogenate, then diluted with sterile water to 10–1, 10–2 and 10–3, pipette to draw 150 µL of sample grinding fluid, spread on the medium, set the sterile water of the last rinse of Chinese fir tissue as a blank control, incubate with other plates under the same conditions, and verify whether the surface disinfection. Cultivate in a 28 ℃ incubator according to the characteristics of the colony phenotype, and use the streak separation method to further purify and isolate the strains until the isolation of the colony morphology is uniform for each isolate.Note: 75% C2H5OH: 75 mL absolute ethanol + 25 mL sterile deionized water; 5% NaClO: sodium hypochlorite solution with 10% available chlorine: sterile deionized water = 1:1.Characterization of PGP traitsDetermination of phosphorus solubilizing abilityThe strain was inoculated into PVK liquid medium and cultured at 28 ℃ for 180 days/min on a reciprocating shaker for 7 days, and then the pH value of the medium was measured. The culture solution was centrifuged at 8000 rpm for 15 min to remove bacterial cells. Take the supernatant and use the molybdenum antimony scandium colorimetric method23 to determine the soluble phosphorus content in the culture broth.Determination of nitrogenase activityAn aliquot of 200 µL fresh culture was inoculated to 20 mL of nutrient broth and incubated overnight at 30℃. Bacterial growth was collected by centrifugation and was washed twice using sterile water, and resuspended by liquid limited nitrogen culture medium (OD600 = 0.2). The 3 mL suspension was transferred to a 25 mL sterilized serum vial and 2.4 mL acetylene gas (99.9999%) was driven into the serum bottle, and then incubated at 30 °C for 12 h. The ethylene content and the protein of bacterial suspension were determined as You et al.24.1-Aminocyclopropane-1-carboxylate (ACC) deaminase activity determinationACC deaminase activity was determined by the method of Glick et al.25 using N-free medium (Nfb)26 for bacteria and minimal medium (MM)27 for actinomycetes containing 0.3 m mol L−1 ACC (Sigma, USA) as a sole nitrogen source. MM with 0.1% (w/v) NH4(SO4)2 was used as a positive control and cultivation without ACC was used as a negative control. After incubation at 28 ℃ for 7 days for non-actinomycete bacteria and 14 days for actinomycetes, colony growth on Nfb or MM with addition of ACC indicated ACC deaminase activity.Indole-3-acetic acid (IAA) productionIAA production was measured by colorimetric assay27. Bacterial isolates were cultured for 3 days in TY broth (without L-tryptophan or supplemented with 500 μg/mL of l-tryptophan) in the dark at 28 °C. Cells were removed from the culture medium by centrifugation at 13,000×g for 10 min; then, 1 mL of the supernatant was mixed vigorously with 2 mL of Salkowski’s reagent (4.5 g of FeCl3 per L in 10.8 M H2SO4). Samples were incubated at room temperature for 30 min and the IAA production was estimated from the optical density at 600 nm (OD600) by comparison with a standard curve prepared from known concentrations of IAA.Siderophore productionSiderophore production was examined by using chrome azurol S (CAS) agar28. Isolate was inoculated onto CAS agar, cultured at 28 °C for 2 days, and the positive strain was indicated by an orange halo around the bacterial colony. Determine the ratio (D/d) of orange aperture diameter (D) to colony diameter (d) to determine the iron-producing carrier capacity of the strain.Physiological and biochemical tests of phosphate-solubilizing bacteriaThe conventional physiological and biochemical identification of PSB is carried out according to the methods in the “Common Bacterial System Identification Manual”, which mainly includes Gram stain, glucose hydrolysis test, lactose hydrolysis test, methyl red test, Voges-Proskauer (VP) test, hydrogen sulfide production test, gelatin liquefaction Test, citrate utilization test, malonate utilization test, denitrification test.16 SrRNA gene sequencingTaking the screened multifunctional PSB as the object, the bacterial genomic DNA extraction kit of Beijing Bomed Biotechnology Co., Ltd. was used to extract the DNA of the strain, using the DNA as a template, and using the bacterial universal primer 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′) PCR amplification, the amplification system is as follows: DNA template 1 uL, primer 27F 0.5 µL, 1492R 0.5 µL, 2 × TaqMix 12.5 µL, ddH2O10.5 µL. The PCR procedure is as follows: 93℃ for 3 min, 93℃ for 30 s, 56 ℃ for 30 s, 72℃ for 2 min, 32 cycles; 72 ℃ for 7 min. The amplified products were sequenced bidirectionally by BGI. After splicing the measured 16SrDNA sequences in ContigExpress, search in GenBank, EzTaxon, BIGSdb databases respectively, select the model strains with high homology. The phylogenetic tree was constructed by the neighbor-joining method using MEGA version 7.0 with the Kimura 2-parameter model29, the robustness of the tree was evaluated by performing bootstrap analyses based on 1000 replications30.Evaluation of plant growth promotion by individual inoculationPreparation of inoculumThe single colonies were picked out and incubated in LB broth at 180 rpm at 28 ℃ for 12 h. The above solution was inserted into 200 mL of LB broth at 1% inoculation, incubated at 180 rpm at 28℃ for 48 h and the cell pellet was resuspended in sterile distilled water and made up to a final concentration of 3 × 108 CFU/mL.Experimental seedlings and soilChinese fir seedlings were provided by the Experimental Center of Subtropical Forestry, Chinese Academy of Forestry (117° 67′ E, 27° 82′ N), Jiangxi Province, China. The use of Chinese fir seedlings in the experiment complies with national regulations. Five months old seedlings with vigorous and apparently disease and pest free were used. The height and root collar diameters of seedlings were 8.7 cm and 1.36 mm, respectively. The seedling container was made of non-woven fabric, the specification was 4.5 cm × 8.0 cm (Diameter × Height). The soil was consisted of nursery medium and loess at a ratio of 9:1, was thoroughly mixed and homogenized with 3 kg slow-release fertilizer per cubic. The slow-release fertilizer is produced by American Abbes (180 g kg−1 total N, 80 g kg−1 available P, and 80 g kg−1 total K, the fertilizer effect period is 9 months). The soil exhibited the following properties: 6.34 g kg−1 total N, 0.80 g kg−1 total P, 2.50 g kg−1 total K, and a pH-value of 6.00.Test designThe SSP2, JRP22 and HRP2, which were confirmed to have the characteristics of promoting plant growth, so pot experiments were conducted. To determine the effectiveness of phosphorus-solubilizing bacteria in plant growth of Chinese fir, a pot culture experiment was conducted between August and November 2019 in an open-sided greenhouse in Experimental Center of Subtropical Forestry, Chinese Academy of Forestry, Jiangxi Province, China. The experiment was carried out in three-factor orthogonal design with five replications for each treatment. The orthogonal experimental design of experiment is provided in Table 1, and strain, dilution ratio, inoculation method contained 3 levels. The pots with water was used as control (CK). For the irrigation of the rhizosphere (IR) treatments, 30 mL of diluted inoculum was added to the soil in the vicinity of the roots of Chinese fir. For the foliar spray (FS) treatments, 30 mL diluted bacterial cell suspension was inoculated in the leaves of Chinese fir by using a syringe. For the rhizosphere + foliar spray (IS) treatments, 15 mL of diluted inoculum was inoculated in the rhizosphere of seedlings, 15 mL was added to the leaves of seedlings by foliar spray. Each treatment contained sixteen seedlings for a total of nine treatments. A total of 3 inoculations were given in the middle of each month. The plant height and stem diameter were recorded before the first inoculation. The plants were harvested after 90 days (16 plantlets/replicate/treatment, i.e., a total of 80 plantlets per treatment) and the root biomass, stem biomass, leaf biomass, plant height and stem diameter were measured.Table 1 L9(34) Orthogonal design of experiment.Full size tableDetermination of growth indicatorsDuring the test, before each inoculation, the height of the seedlings was measured with a ruler and the ground diameter of the seedlings was measured with a vernier caliper. After the experiment, 10 plants of Chinese fir seedlings in average growth were randomly selected from each treatment, a total of 30 plants were washed with clean water to remove surface impurities, the filter paper was dried and the roots, stems, and leaves were put into paper bags respectively at 105 °C. After being degraded for 0.5 h, dried at 70 °C to a constant weight, weighed and recorded the biomass of each part.Determination of leaf and soil nutrient contentTotal N content of leaf and soil was measured by a 2300 Kjeltec Analyzer Unit (FOSS, Höganäs, Sweden). Total P and total K, total Mg and total Fe of leaf, soil TP, TK, AP and AK were extracted according to literature31 and were determined by ICP (Kleve, Germany).Determination of soil enzyme activitiesActivities of the soil urease, cellulase, sucrase, dehydrogenase and acid phosphatase were determined by spectrophotometry. Firstly, 0.05 g of soil was added to 450 mL of phosphate buffer solution (PBS, 0.1 mol L−1, pH 7.4). Then the solution was mixed by shaking, and centrifuged at 2000 rpm at 4 ℃ for 10 min and supernatant was collected with a new centrifugal tube. The supernatant and reagents were added according to the kit instructions (Shanghai Enzyme-linked Biotechnology Co., Ltd., Shanghai, China). Absorbance at 450 nm was measured on a SpectraMax Paradigm Multi-Mode detection platform (Molecular devices, San Jose, CA, USA).Data processing and analysisAll statistical analyses were performed using SPSS24. Data are presented in terms of means (± SE; standard error). Statistical differences were tested by one-factor ANOVA to evaluate the differences in the nutrient content of soil and plant growth status. In MEGA7.0, the Neighbor-Joining method was used to construct the phylogenetic tree, and the Bootstrap value was 1000. More

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    Evidence for magnesium–phosphorus synergism and co-limitation of grain yield in wheat agriculture

    1.Elser, J. J. et al. Global analysis of nitrogen and phosphorus limitation of primary producers in freshwater, marine and terrestrial ecosystems. Ecol. Lett. 10, 1135–1142 (2007).Article 

    Google Scholar 
    2.Mengel, K. & Kirkby, E. A. Principles of Plant Nutrition (Kluwer Academic Publishers, 2001).Book 

    Google Scholar 
    3.Reich, M., Aghajanzadeh, T. & De Kok, L. J. Physiological basis of plant nutrient use efficiency—Concepts, opportunities and challenges for its improvement. In Nutrient Use Efficiency in Plants: Concepts and Approaches (eds Hawkesford, M. J. et al.) (Springer, 2014).
    Google Scholar 
    4.Agren, G. I. Ideal nutrient productivities and nutrient proportions in plant growth. Plant Cell Environ. 11, 613–620 (1988).Article 

    Google Scholar 
    5.Weih, M., Hamner, K. & Pourazari, F. Analyzing plant nutrient uptake and utilization efficiencies: Comparison between crops and approaches. Plant Soil 430, 7–21 (2018).CAS 
    Article 

    Google Scholar 
    6.Sterner, R. W. & Elser, J. J. Ecological stoichiometry: The biology of elements from molecules to the biosphere (2002).7.Reich, P. B. et al. Evidence of a general 2/3-power law of scaling leaf nitrogen to phosphorus among major plant groups and biomes. Proc. R. Soc. B Biol. Sci. 277, 877–883 (2010).CAS 
    Article 

    Google Scholar 
    8.Hutchinson, G. E. Population studies—Animal ecology and demography—Concluding remarks. Cold Spring Harbor. Symp. Quant. Biol. 22, 415–427 (1957).Article 

    Google Scholar 
    9.Agren, G. I. & Weih, M. Multi-dimensional plant element stoichiometry-looking beyond carbon, nitrogen, and phosphorus. Front. Plant Sci. 11, 23 (2020).Article 

    Google Scholar 
    10.Niklas, K. J. Plant allometry, leaf nitrogen and phosphorus stoichiometry, and interspecific trends in annual growth rates. Ann. Bot. 97, 155–163 (2006).CAS 
    Article 

    Google Scholar 
    11.Hou, E. et al. Global meta-analysis shows pervasive phosphorus limitation of aboveground plant production in natural terrestrial ecosystems. Nat. Commun. 11, 637 (2020).ADS 
    CAS 
    Article 

    Google Scholar 
    12.Ryan, P. R. et al. Early vigour improves phosphate uptake in wheat. J. Exp. Bot. 66, 7089–7100 (2015).CAS 
    Article 

    Google Scholar 
    13.Wiel, CCMvd., Linden, CGvd & Scholten, O. E. Improving phosphorus use efficiency in agriculture: Opportunities for breeding. Euphytica 207, 1–22 (2016).Article 

    Google Scholar 
    14.Bilal, H. M., Aziz, T., Maqsood, M. A., Farooq, M. & Yan, G. Categorization of wheat genotypes for phosphorus efficiency. PLoS ONE 13, e0205471 (2018).Article 

    Google Scholar 
    15.Wang, Z. et al. Magnesium fertilization improves crop yield in most production systems: A meta-analysis. Front. Plant Sci. 10, 1727 (2020).Article 

    Google Scholar 
    16.Hauer-Jakli, M. & Traenkner, M. Critical leaf magnesium thresholds and the impact of magnesium on plant growth and photo-oxidative defense: a systematic review and meta-analysis from 70 years of research. Front. Plant Sci. 10, 766 (2019).Article 

    Google Scholar 
    17.Chawade, A. et al. A transnational and holistic breeding approach is needed for sustainable wheat production in the Baltic Sea region. Physiol. Plant. 164, 442–451 (2018).CAS 
    Article 

    Google Scholar 
    18.Weih, M., Pourazari, F. & Vico, G. Nutrient stoichiometry in winter wheat: Element concentration pattern reflects developmental stage and weather. Sci. Rep. 6, 35958–35958 (2016).ADS 
    CAS 
    Article 

    Google Scholar 
    19.Hamner, K., Weih, M., Eriksson, J. & Kirchmann, H. Influence of nitrogen supply on macro- and micronutrient accumulation during growth of winter wheat. Field Crop Res. 213, 118–129 (2017).Article 

    Google Scholar 
    20.Jia, X., Liu, P. & Lynch, J. P. Greater lateral root branching density in maize improves phosphorus acquisition from low phosphorus soil. J. Exp. Bot. 69, 4961–4970 (2018).CAS 
    Article 

    Google Scholar 
    21.Kumar, A. et al. Root trait plasticity and plant nutrient acquisition in phosphorus limited soil. J. Plant Nutr. Soil Sci. 182, 945–952 (2019).CAS 
    Article 

    Google Scholar 
    22.Lynch, J. P. Root phenes for enhanced soil exploration and phosphorus acquisition: Tools for future crops. Plant Physiol. 156, 1041–1049 (2011).CAS 
    Article 

    Google Scholar 
    23.Lynch, J. P. Steep, cheap and deep: An ideotype to optimize water and N acquisition by maize root systems. Ann. Bot. 112, 347–357 (2013).CAS 
    Article 

    Google Scholar 
    24.Lambers, H., Shane, M., Cramer, M., Pearse, S. & Veneklaas, E. Root structure and functioning for efficient acquisition of phosphorus: Matching morphological and physiological traits. Ann. Bot. 98, 693–713 (2006).Article 

    Google Scholar 
    25.Trachsel, S., Kaeppler, S. M., Brown, K. M. & Lynch, J. P. Maize root growth angles become steeper under low N conditions. Field Crop Res 140, 18–31 (2013).Article 

    Google Scholar 
    26.Jobbagy, E. G. & Jackson, R. B. The distribution of soil nutrients with depth: Global patterns and the imprint of plants. Biogeochemistry 53, 51–77 (2001).CAS 
    Article 

    Google Scholar 
    27.Sun, B. R., Gao, Y. Z. & Lynch, J. P. Large crown root number improves topsoil foraging and phosphorus acquisition. Plant Physiol. 177, 90–104 (2018).CAS 
    Article 

    Google Scholar 
    28.Weih, M., Asplund, L. & Bergkvist, G. Assessment of nutrient use in annual and perennial crops: A functional concept for analyzing nitrogen use efficiency. Plant Soil 339, 513–520 (2011).CAS 
    Article 

    Google Scholar 
    29.Malhi, S. S., Johnston, A. M., Schoenau, J. J., Wang, Z. H. & Vera, C. L. Seasonal biomass accumulation and nutrient uptake of wheat, barley and oat on a Black Chernozern soil in Saskatchewan. Can. J. Plant Sci. 86, 1005–1014 (2006).Article 

    Google Scholar 
    30.Maeoka, R. E. et al. Changes in the phenotype of winter wheat varieties released between 1920 and 2016 in response to in-furrow fertilizer: Biomass allocation, yield, and grain protein concentration. Front. Plant Sci. 10, 1786 (2020).Article 

    Google Scholar 
    31.Pourazari, F., Vico, G., Ehsanzadeh, P. & Weih, M. Contrasting growth pattern and nitrogen economy in ancient and modern wheat varieties. Can. J. Plant Sci. 95, 851–860 (2015).Article 

    Google Scholar 
    32.Rietra, R. P. J. J., Heinen, M., Dimkpa, C. O. & Bindraban, P. S. Effects of nutrient antagonism and synergism on yield and fertilizer use efficiency. Commun. Soil Sci. Plant Anal. 48, 1895–1920 (2017).CAS 
    Article 

    Google Scholar 
    33.Pedro, A., Savin, R. & Slafer, G. A. Crop productivity as related to single-plant traits at key phenological stages in durum wheat. Field Crop Res. 138, 42–51 (2012).Article 

    Google Scholar 
    34.Cakmak, I. & Yazici, A. M. Magnesium: A forgotten element in crop production. Better Crops Plant Food 94, 23–25 (2010).
    Google Scholar 
    35.Lancashire, P. D. et al. A uniform decimal code for growth-stages of crops and weeds. Ann. Appl. Biol. 119, 561–601 (1991).Article 

    Google Scholar 
    36.Trachsel, S., Kaeppler, S. M., Brown, K. M. & Lynch, J. P. Shovelomics: High throughput phenotyping of maize (Zea mays L.) root architecture in the field. Plant Soil 341, 75–87 (2011).CAS 
    Article 

    Google Scholar 
    37.Colombi, T. & Walter, A. Root responses of triticale and soybean to soil compaction in the field are reproducible under controlled conditions. Funct. Plant Biol. 43, 114–128 (2016).Article 

    Google Scholar  More

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    Sight of parasitoid wasps accelerates sexual behavior and upregulates a micropeptide gene in Drosophila

    We asked whether the mating of male and female fruit flies would be affected by the presence of parasitoid wasps. We placed a pair of D. melanogaster flies in a small Petri dish, either with or without parasitoid wasps (Fig. 1a). In an initial experiment we used the wasp Leptopilina boulardi, which specializes on D. melanogaster and on closely related fly species14.Fig. 1: Exposure of Drosophila to wasps accelerates sexual behavior.a Courtship arena containing a male and virgin female fly with (left) and without (right) two wasps, one male and one female. b Copulation latency of D. melanogaster. p  More

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    China’s wildlife protection: add annual reviews and oversight

    Now that China has finally updated its List of Wildlife under Special State Protection, a more nimble and responsive approach is needed to aid conservation. The list should be reviewed every year, as well as subjected to the planned five-yearly updates. Species can quickly become endangered in times of rapid development.The latest additions are the first in more than 30 years (see go.nature.com/2q7sfga). During that time, China has changed profoundly, but the list of protected species has not kept pace. This lag has been disastrous for some animals that were not given the protection they needed.At least 33 species became extinct in China and many more are critically endangered (Y. Xie & W. Sung Integr. Zool. 2, 26–35; 2007; Z. Jiang et al. Biodivers. Sci. 24, 500–551; 2016).An independent government committee should be created to oversee amendments. When making decisions, it could refer to appendices of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) and the ‘red lists’ of threatened species curated by the Chinese Academy of Sciences and the International Union for Conservation of Nature (IUCN). These steps would build on the more forceful approach to managing wildlife that China has taken since the start of the COVID-19 pandemic. More

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    Continuous presence of proto-cereals in Anatolia since 2.3 Ma, and their possible co-evolution with large herbivores and hominins

    Vegetation history of the Acıgöl areaOur palynological analyses of 72 regularly spaced samples show a diversified vegetal landscape alternately wooded and open, in response to orbitally driven climatic cyclicity. However, arboreal pollen values remain almost constantly below 50% of the Pollen Sum (PS) (average 27.5%, median 22.8%), which corresponds to an overall open landscape (Fig. 3). Among herbaceous plants, the dominant taxa are steppics such as Artemisia, heliophilous and halophilous taxa including Calystegia, several Compositae, Convolvulus, Linum, Plantago ssp., Poaceae and Chenopodiaceae that could develop on the saline shores of Acıgöl lake during evaporitic periods. Forests are composed of a mixture of conifers, Mediterranean Pinus, Abies, Cedrus, Cupressaceae and Picea, associated with broadleaved trees dominated by Mediterranean oaks, i.e. deciduous and evergreen Quercus, with some Olea. Riverine trees such as Alnus, Salix, Populus, Tamarix, Juglans and Platanus have also been identified. Few Tertiary or megathermic relictual taxa (Carya, Liquidambar, Parrotia, Pterocarya fraxinifolia, Taxodiaceae, Tsuga, Zelkova) were identified so far in the pollen assemblages, mostly before 2.2 Ma, due to climatic cooling17,18 since the end of Tertiary which led to a decline in global biodiversity19,20.Figure 3Simplified pollen and NPP diagram in percentages of Acıgöl, core 3, based on the age model of Demory et al. [1]. Equidistant scale. Values are in percentages calculated on a pollen sum without Non-Pollen Palynomorphs (NPP), Ferns, Bryophytes and Algae. The beige rectangle corresponds to the date of the presence of Homo erectus at Kocabaş (Lebatard et al. [4]).Full size imageThe vast freshwater stretch of Acıgöl, located in a predominantly arid limestone hills environment, seems to have been a crucial resource for the mammalian fauna, which probably concentrated around the site in search of water and pastures. Indeed, low percentages of arboreal pollen imply that the landscape remained open throughout the sequence and suggest a marked grazing pressure by herbivores in addition to climatic factors21,22,23.Coprophilous fungi spores, cereals and other ancestors of cultivated plantsCoprophilous fungi spores are excellent indicators of herbivorous mega-mammal herds since they grow exclusively on dung deposited by these animals24. At Acıgöl, a wide variety of coprophilous fungi spores has been identified throughout the pollen record including: Sporormiella sp., Podospora sp., Delitschia sp., Sordaria sp. and Valsaria variospora (Figs. 3, 4). They provide evidence for a continuous presence of large herbivorous mammals around the lake throughout Quaternary.Figure 4Coprophilous fungi spores of Acıgöl, core 3. Equidistant scale. Age model is from Demory et al. [1]. In red: coprophilous fungi taxa..Full size imagePollens of Poaceae, such as Secale (rye) and Cerealia-type, have been identified throughout the sequence (Figs. 3, 5). Unexpectedly, they present the same morphological characteristics as that of modern cereal grains25,26, namely an average size of ≥ 40 µm and a large pore + annulus (≥ 8 µm). As by definition cereals are cultivated plants, we will call the corresponding plants “proto-cereals” to highlight that their pollen are identical to those of cereals. This resemblance can be seen clearly in Fig. 5, where we have brought together fossil cereals from Acιgöl (Fig. 5, photos 1–7), from Roman time (Fig. 5, photo 8), not modified by modern agricultural practices, and from the current wheat field of the Lauragais agricultural plain, Gardouch, France (Fig. 5, photo 9). Cerealia-type frequencies reach a maximum of 9% of the PS around 2.2 Ma and can be as abundant as wild Poaceae pollen (Fig. 3). The Cerealia/Poaceae ratio shows that 24.66% of all Poaceae are proto-cereals from 2.0 to 2.3 Ma (Supplementary Table 1). Such high proto-cereal rates are almost never reached in pollen records, even in recent periods and in the presence of agriculture, because of the very low pollen dispersal capacity of cereals27. A lowering of frequencies down to 2–4% range is recorded in younger periods (Fig. 3), as well as a step like decrease of the Cerealia/Poaceae ratio (Fig. 6). This change may be related to the Middle-Pleistocene Transition (MPT) cooling and to the mega-mammal fauna change from a Villafranchian to a Galerian type28. MPT and faunal changes occurred around 0.9–1.0 Ma, while a decrease in our proto-cereal starts around 1.5 Ma, however signs of cooling and amplified climatic cycles predate the MPT28.Figure 5Pollen grain of Cerealia and Triticum sp. from Acıgöl (ACI), core 3 (photos 1–7), the Roman site of La Verrerie, Arles, France (photo 8) and Gardouch, France, current wheat field (photo 9). Photographies with a photonic (photo 1 – 4 and 8) and a confocal microscope (photos 5-7 and 9). 1) sample ACI 239 m, age: 0.871 Ma. 2) sample ACI 435.50 m, age: 1.709 Ma. 3) sample ACI 532.44 m, age: 2.122 Ma. 4) sample ACI 509.50 m, age: 2.026 Ma. 5) sample ACI 552.57 m, age 2.206. 6) sample ACI 552.57 m, age: 2.206 Ma. 7) sample ACI 429.50 m, age: 1.681 Ma. 8) sample La Verrerie 1455, age: 50-70 BC (Roman). 9) current pollen of Triticum sp., age: 2000 AD. L: maximal length (µm).Full size imageFigure 6Cerealia/Poaceae ratio in %, % cultivated tree ancestors and % Olea of Acıgöl, core 3.Full size imageThe histogram of wild Poaceae and proto-cereal pollen size (Fig. 7a) shows that there are a number of pollen populations modes around 30, 37.5, 45–50, supporting the idea that the larger grain sizes cannot be interpreted as a tail of ‘anomalous’ wild Poaceae pollen. Moreover, comparison with the present-day pollen rain recorded in moss pollsters, sampled around the lake of Acıgöl (Fig. 7b and Supplementary Table 2), show that the large pollen size mode (≥ 40 µm) is nowadays nearly absent (0–0.97% of the PS, Cerealia/Poaceae ratio of 4.52%, Supplementary Tables 3 and 4), even in biotopes with wild Poaceae considered to be ancestors of cereals (Aegilops, sample 2a, cereal rate: 0.97% of the PS) or with cereals such as Hordeum (sample 3a, b and 4, cereal rate 0.31, 0.00, 0.33 of the PS respectively, Supplementary Tables 2 and 3).Figure 7a) Pollen size of wild Poaceae and proto-cereal of Acıgöl, core 3. The measurements were made on the 10 samples with the highest cereal pollen content. A total of 991 grains of pollen were measured. b) Current pollen rain at the Acıgöl lake and surroundings. 8 moss samples were collected and 354 measurements of the longest axis of the wild Poaceae and cereal pollen grain were made.Full size imageOur interpretation is that proto-cereals recorded throughout the Acıgöl sequence derive from wild Poaceae. Their emergence and predominance may have been favoured by the impact of large herbivore herds attracted to Acıgöl lake shores, and through genetic drift. Through the process of trampling, nitrogen enrichment of soils and browsing, large mammal herds could have altered the genotype of proto-cereals naturally present in Acıgöl and thus, favoured the emergence of modern cereals. For genetic reasons, the descendants of these proto-cereals are not represented today among cultivated Poaceae because domestication bottlenecks eliminate genetic variation29.Is there a relationship between the size of proto-cereal pollen and climate? To our knowledge, the genetic literature does not show any relationship between the increase in pollen size and temperature. However, there does seem to be a relationship with atmospheric drought30,31 which is said to have favoured the appearance of polyploidy in certain species of Poaceae. It cannot be excluded that climate has had an influence on the proto-cereal genome, but only the interaction between herds of large herbivores and proto-cereal steppes can explain why proto-cereal pollen has never been found in such abundance elsewhere in Pleistocene pollen records.The ancestors of cultivated trees (Olea sp., Juglans sp., Castanea sp., Corylus sp., Prunus t.), typical of the modern Mediterranean agriculture, are also present in the Acıgöl sequence (Fig. 3 and Supplementary Table 5). Their amount increases after 1.5 Ma, mainly due to Olea (Fig. 6). Other potentially edible plants such as Ephedra, Hippophae, all the Compositae and the Fagaceae have been identified in the pollen assemblages. They correspond to 54.4% of plants identified in the pollen assemblages. Among these plants, there are 72% grasses and 28% trees and, among edible organs, 51% are vegetables and 20% are seeds (Supplementary Fig. 1a,b). These results testify to the potential wealth of accessible food resources that human and animal populations could feed on. Interestingly, studies carried out in Spain on the present-day consumption of wild plants lead to results close to those obtained at Acıgöl, with 87% grasses and 13% trees32.In recent years, new biological and archaeological data obtained from sites with human occupation have improved our knowledge of the beginnings of agriculture and the modalities of its implementation. In the Levant, the Ohalo II site highlights the presence of proto-cereal seeds, and flint tools to harvest, as early as 23,000 years before the present33. Further north, on the archaeological site of Gesher Benot Ya’aqov, proto-cereal seeds (oats, Avena) as well as pollen from cereals and trees currently cultivated, were identified over a period ranging from 750,000 to 820,000 years34,35. Moreover, recent genetic data indicate that the emergence of agriculture did not occur at a single location at the onset of the Neolithic (e.g. the “Fertile Crescent” hypothesis) but is, on the contrary, an evolutionary and multi-regional long-term phenomenon36,37,38. Alternatively, or simultaneously, are the hominins also partly responsible by having developed episodes of a form of transitory “proto-agriculture”? We already know that this domestication process was discontinuous with shutdown and restart phases37,39. Acheulean lithic tools, characterised by symmetrically shaped bifaces, testify to the rather advanced cognitive capacities of early Pleistocene populations that may have visited the lakeshore of Acıgöl5. Hominin populations may also have benefited from this opportunity to diversify their food regime with easily harvested and nutrient-rich wild plants (Supplementary Table 5), as it is the case today for hunter-gatherer populations in Africa and elsewhere in the world. More

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    Reef foraminifera as bioindicators of coral reef health in southern South China Sea

    1.Hoegh-Guldberg, O. Climate change, coral bleaching and the future of the world’s coral reefs. Mar. Freshwater Res. 50(8), 839–866. https://doi.org/10.1071/MF99078 (1999).Article 

    Google Scholar 
    2.Moberg, F. & Folke, C. Ecological goods and services of coral reef ecosystems. Ecol. Econ. 29(2), 215–233. https://doi.org/10.1016/S0921-8009(99)00009-9 (1999).Article 

    Google Scholar 
    3.Shahbudin, S., Fikri Akmal, K. F., Faris, S., Normawaty, M. N. & Mukai, Y. Current status of coral reefs in Tioman Island Peninsular Malaysia. Turk. J. Zool. 41(2), 294–305. https://doi.org/10.3906/zoo-1511-42 (2017).Article 

    Google Scholar 
    4.Anthony, K. R. N. et al. Operationalizing resilience for adaptive coral reef management under global environmental change. Glob. Change Biol. 21(1), 48–61. https://doi.org/10.1111/gcb.12700,Pubmed:25196132 (2015).ADS 
    Article 

    Google Scholar 
    5.Bruno, J. F. & Selig, E. R. Regional decline of coral cover in the Indo-Pacific: timing, extent, and subregional comparisons. PLoS ONE 2(8), e711. https://doi.org/10.1371/journal.pone.0000711,Pubmed:17684557 (2007).ADS 
    Article 
    PubMed Central 
    PubMed 

    Google Scholar 
    6.Cowburn, B., Samoilys, M. A. & Obura, D. The current status of coral reefs and their vulnerability to climate change and multiple human stresses in the Comoros Archipelago Western Indian Ocean. Mar. Pollut. Bull. 133, 956–969. https://doi.org/10.1016/j.marpolbul.2018.04.065,Pubmed:29778407 (2018).CAS 
    Article 

    Google Scholar 
    7.Schueth, J. D. & Frank, T. D. Reef foraminifera as bioindicators of coral reef health: Low Isles Reef, northern Great Barrier Reef Australia. J. Foram. Res. 38(1), 11–22. https://doi.org/10.2113/gsjfr.38.1.11 (2008).Article 

    Google Scholar 
    8.Uthicke, S., Thompson, A. & Schaffelke, B. Effectiveness of benthic foraminiferal and coral assemblages as water quality indicators on inshore reefs of the Great Barrier Reef Australia. Coral Reefs 29(1), 209–225. https://doi.org/10.1007/s00338-009-0574-9 (2010).ADS 
    Article 

    Google Scholar 
    9.Natsir, S. M. & Subkhan, M. The distribution of benthic foraminifera in coral reefs community and seagrass bad of Belitung Islands based on FORAM Index. J. Coast. Dev. 15(1), 51–58 (2012).
    Google Scholar 
    10.Alve, E. Benthic foraminiferal responses to estuarine pollution: a review. J. Foram. Res. 25(3), 190–203. https://doi.org/10.2113/gsjfr.25.3.190 (1995).Article 

    Google Scholar 
    11.Hallock, P., Lidz, B. H., Cockey-Burkhard, E. M. & Donnelly, K. B. Foraminifera as bioindicators in coral reef assessment and monitoring: the FORAM index. Foraminifera in reef assessment and monitoring. Environ. Monit. Assess. 81(1–3), 221–238 (2003).Article 

    Google Scholar 
    12.Sen Gupta, B. K. Systematics of modern Foraminifera. In Sen Gupta, B.K. (ed.) Modern Foraminifera (Springer, 2003) 7–36. https://doi.org/10.1007/0-306-48104-9.13.Carnahan, E. A. Foraminiferal Assemblages as Bioindicators of Potentially Toxic Elements in Biscayne Bay, Florida. M.Sc. thesis (U.S.A.: University of South Florida, 2005)14.Barbosa, C. F., Prazeres, M. D. F., Ferreira, B. P. & Seoane, J. C. S. Foraminiferal assemblage and Reef Check census in coral reef health monitoring of East Brazilian margin. Mar. Micropaleontol. 73(1–2), 62–69. https://doi.org/10.1016/j.marmicro.2009.07.002 (2009).ADS 
    Article 

    Google Scholar 
    15.Dimiza, M. D., Koukousioura, O., Triantaphyllou, M. V. & Dermitzakis, M. D. Live and dead benthic foraminiferal assemblages from coastal environments of the Aegean Sea (Greece): distribution and diversity. Rev. Micropaleontol. 59(1), 19–32. https://doi.org/10.1016/j.revmic.2015.10.002 (2016).Article 

    Google Scholar 
    16.Uthicke, S. & Nobes, K. Benthic foraminifera as ecological indicators for water quality on the Great Barrier Reef. Estuarine Coast. Shelf Sci. 78(4), 763–773. https://doi.org/10.1016/j.ecss.2008.02.014 (2008).ADS 
    Article 

    Google Scholar 
    17.Renema, W. Terrestrial influence as a key driver of spatial variability in large benthic foraminiferal assemblage composition in the Central Indo-Pacific. Earth Sci. Rev. 177, 514–544. https://doi.org/10.1016/j.earscirev.2017.12.013 (2018).ADS 
    Article 

    Google Scholar 
    18.Förderer, M. & Langer, M. R. Exceptionally species-rich assemblages of modern larger benthic foraminifera from nearshore reefs in northern Palawan (Philippines). Rev. Micropaleontol. 100, 65. https://doi.org/10.1016/j.revmic.2019.100387 (2019).Article 

    Google Scholar 
    19.Eichler, P. P. B. & de Moura, D. S. Symbiont-bearing foraminifera as health proxy in coral reefs in the equatorial margin of Brazil. Environ. Sci. Pollut. Res. 27(12), 13637–13661. https://doi.org/10.1007/s11356-019-07483-y,Pubmed:32034594 (2020).CAS 
    Article 

    Google Scholar 
    20.Renema, W. Is increased calcarinid (foraminifera) abundance indicating a larger role for macro-algae in Indonesian Plio-Pleistocene coral reefs?. Coral Reefs 29(1), 165–173. https://doi.org/10.1007/s00338-009-0568-7 (2010).ADS 
    Article 

    Google Scholar 
    21.Chen, C. & Lin, H. L. Applying benthic Foraminiferal assemblage to evaluate the coral reef condition in Dongsha Atoll lagoon. Zool. Stud. 56, e20. https://doi.org/10.6620/ZS.2017.56-20,Pubmed:31966219 (2017).Article 
    PubMed Central 
    PubMed 

    Google Scholar 
    22.Hallock, P. Interoceanic differences in foraminifera with symbiotic algae: a result of nutrient supplies. Mar. Sci. Faculty Publication 1228 (1988). https://scholarcommons.usf.edu/msc_facpub/122823.Langer, M. R., Weinmann, A. E., Lötters, S., Bernhard, J. M. & Rödder, D. Climate-driven range extension of Amphistegina (protista, foraminiferida): Models of current and predicted future ranges [Protista, Foraminiferida]. PLoS ONE 8(2), e54443. https://doi.org/10.1371/journal.pone.0054443,Pubmed:23405081 (2013).ADS 
    CAS 
    Article 
    PubMed Central 
    PubMed 

    Google Scholar 
    24.Culver, S. J. et al. Distribution of foraminifera of the Poverty continental margin, New Zealand: implications for sediment transport. J. Foram. Res. 42(4), 305–326. https://doi.org/10.2113/gsjfr.42.4.305 (2012).Article 

    Google Scholar 
    25.Szarek, R. Biodiversity and Biogeography of Recent Benthic Foraminiferal Assemblages in the South Western South China Sea (Sunda Shelf). Doctoral dissertation (Kiel, Kiel, Germany: Christian-Albrechts Universität, 2001).26.Prazeres, M., Martínez-Colón, M. & Hallock, P. Foraminifera as bioindicators of water quality: the FoRAM Index revisited. Environ. Pollut. 257, 113612. https://doi.org/10.1016/j.envpol.2019.113612,Pubmed:31784269 (2020).CAS 
    Article 

    Google Scholar 
    27.Toda, T. et al. Community structures of coral reefs around Peninsular Malaysia. J. Oceanogr. 63(1), 113–123. https://doi.org/10.1007/s10872-007-0009-6 (2007).Article 

    Google Scholar 
    28.Zakai, D. & Chadwick-Furman, N. E. Impacts of intensive recreational diving on reef corals at Eilat, northern Red Sea. Biol. Conserv. 105(2), 179–187. https://doi.org/10.1016/S0006-3207(01)00181-1 (2002).Article 

    Google Scholar 
    29.Carnahan, E. A., Hoare, A. M., Hallock, P., Lidz, B. H. & Reich, C. D. Foraminiferal assemblages in Biscayne Bay, Florida, USA: responses to urban and agricultural influence in a subtropical estuary. Mar. Pollut. Bull. 59(8–12), 221–233. https://doi.org/10.1016/j.marpolbul.2009.08.008 (2009).CAS 
    Article 

    Google Scholar 
    30.Oliver, L. M. et al. Contrasting responses of coral reef fauna and foraminiferal assemblages to human influence in la Parguera Puerto Rico. Mar. Environ. Res. 99, 95–105. https://doi.org/10.1016/j.marenvres.2014.04.005 (2014).CAS 
    Article 

    Google Scholar 
    31.Unsworth, R. K., Clifton, J. & Smith, D. J. Marine Research and Conservation in the Coral Triangle: The Wakatobi National Park (Nova Science Publishers, 2010).
    Google Scholar 
    32.Praveena, S. M., Siraj, S. S. & Aris, A. Z. Coral reefs studies and threats in Malaysia: a mini review. Rev. Environ. Sci. Bio Technol. 11(1), 27–39. https://doi.org/10.1007/s11157-011-9261-8 (2012).Article 

    Google Scholar 
    33.Akmal, K. F., Shahbudin, S., Faiz, M. H. M. & Hamizan, Y. M. Diversity and abundance of scleractinian corals in the East Coast of peninsular Malaysia: a case study of Redang and Tioman Islands. Ocean Sci. J. 54(3), 435–456. https://doi.org/10.1007/s12601-019-0018-6 (2019).ADS 
    CAS 
    Article 

    Google Scholar 
    34.Oron, S., Abramovich, S., Almogi-Labin, A., Woeger, J. & Erez, J. Depth related adaptations in symbiont bearing benthic foraminifera: new insights from a field experiment on Operculina ammonoides. Sci. Rep. 8(1), 9560. https://doi.org/10.1038/s41598-018-27838-8,Pubmed:29934603 (2018).ADS 
    Article 
    PubMed Central 
    PubMed 

    Google Scholar 
    35.Oliver, J. K., Berkelmans, R. & Eakin, C. M. Coral bleaching in space and time in (eds van Oppen, M. J. H. & Lough, J. M.) Coral Bleaching. Ecological Studies. 205 (Springer, 2009) 21–39.36.Harborne, A., Fenner, D., Barnes, A., Beger, M., Harding, S. & Roxburgh, T. Status Report on the Coral Reefs of the East Coast of Peninsular Malaysia. Report Prepared to Department of Fisheries Malaysia, Kuala Lumpur, Malaysia, 361–369 (2000)37.Akhir, M., Fadzil, M., Zakaria, N. Z. & Tangang, F. Intermonsoon variation of physical characteristics and current circulation along the east coast of Peninsular Malaysia. Int. J. Oceanogr., 1–9 (2014)38.Chu, P. C., Qi, Y., Chen, Y., Shi, P. & Mao, Q. South China sea wind-wave characteristics. Part I: validation of WAVEWATCH-III using TOPEX/Poseidon data. J. Atmos. Ocean. Technol. 21(11), 1718–1733. https://doi.org/10.1175/JTECH1661.1 (2004).ADS 
    Article 

    Google Scholar 
    39.Marghany, M. Velocity bunching model for modelling wave spectra along east coast of Malaysia. J. Indian Soc. Remote Sens. 32(2), 185–198. https://doi.org/10.1007/BF03030875 (2004).Article 

    Google Scholar 
    40.Department of Marine Park, Malaysia. Laporan Tahunan Jabatan Taman Laut Malaysia. Annual report (2012)41.Chia, K. W., Ramachandran, S., Ho, J. A. & Ng, S. S. I. Conflicts to consensus: Stakeholder perspectives of Tioman Island tourism sustainability. Int. J. Bus. Soc. 19, 159 (2018).
    Google Scholar 
    42.Game, E. T., Meijaard, E., Sheil, D. & McDonald-Madden, E. Conservation in a wicked complex world; challenges and solutions. Conserv. Lett. 7(3), 271–277. https://doi.org/10.1111/conl.12050 (2014).Article 

    Google Scholar 
    43.Murray, J. W. Ecology and applications of benthic foraminifera. Cambridge University Press (2006)44.Loeblich, A. R. & Tappan, H. Foraminiferal Genera and Their Classification (Van Nostrand Reinhold, 1987).
    Google Scholar 
    45.Szarek, R., Kuhnt, W., Kawamura, H. & Kitazato, H. Distribution of recent benthic foraminifera on the Sunda Shelf (South China Sea). Mar. Micropaleontol. 61(4), 171–195. https://doi.org/10.1016/j.marmicro.2006.06.005 (2006).ADS 
    Article 

    Google Scholar 
    46.Martin, S. Q. et al. Announcements. J. Foram. Res. 48(4), 388–389. https://doi.org/10.2113/gsjfr.48.4.388 (2018).Article 

    Google Scholar 
    47.Folk, R. L. Petrology of Sedimentary Rocks (Hemphill Publishing Company, 1980)48.Dean, W. E. Determination of carbonate and organic matter in calcareous sediments and sedimentary rocks by loss on ignition; comparison with other methods. J. Sediment. Res. 44(1), 242–248 (1974).CAS 

    Google Scholar 
    49.Heiri, O., Lotter, A. F. & Lemcke, G. Loss on ignition as a method for estimating organic and carbonate content in sediments: reproducibility and comparability of results. J. Paleolimnol. 25(1), 101–110. https://doi.org/10.1023/A:1008119611481 (2001).ADS 
    Article 

    Google Scholar 
    50.Romesburg, C. Cluster Analysis for Researchers (Lulu Press, 2004).
    Google Scholar 
    51.Milker, Y. et al. Distribution of recent benthic foraminifera in shelf carbonate environments of the western Mediterranean Sea. Mar. Micropaleontol. 73(3–4), 207–225. https://doi.org/10.1016/j.marmicro.2009.10.003 (2009).ADS 
    Article 

    Google Scholar  More

  • in

    Pupal cannibalism by worker honey bees contributes to the spread of deformed wing virus

    1.Grassly, N. C. & Fraser, C. Mathematical models of infectious disease transmission. Nat. Rev. Microbiol. 6, 477–487 (2008).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    2.Cressler, C. E., McLeod, D. V., Rozins, C., Van Den Hoogen, J. & Day, T. The adaptive evolution of virulence: A review of theoretical predictions and empirical tests. Parasitology 143, 915–930 (2016).PubMed 
    Article 

    Google Scholar 
    3.Lanzi, G. et al. Molecular and biological characterization of deformed wing virus of honeybees (Apismellifera L.). J. Virol. 80, 4998–5009 (2006).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    4.Dainat, B., Evans, J. D., Chen, Y. P., Gauthier, L. & Neumann, P. Dead or alive: Deformed wing virus and Varroa destructor reduce the life span of winter honeybees. Appl. Environ. Microbiol. 78, 981–987 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    5.Highfield, A. C. et al. Deformed wing virus implicated in overwintering honeybee colony losses. Appl. Environ. Microbiol. 75, 7212–7220 (2009).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    6.Le Conte, Y., Ellis, M. & Ritter, W. Varroa mites and honey bee health: Can Varroa explain part of the colony losses?. Apidologie 41, 353–363 (2010).Article 

    Google Scholar 
    7.De Miranda, J. R. & Genersch, E. Deformed wing virus. J. Invertebr. Pathol. 103, S48–S61 (2010).PubMed 
    Article 
    CAS 

    Google Scholar 
    8.Martin, S. J. & Brettell, L. E. Deformed wing virus in honeybees and other insects. Annu. Rev. Virol. 6, 49–69 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    9.Sumpter, D. J. & Martin, S. J. The dynamics of virus epidemics in Varroa-infested honey bee colonies. J. Anim. Ecol. 73, 51–63 (2004).Article 

    Google Scholar 
    10.Ramsey, S. D. et al. Varroa destructor feeds primarily on honey bee fat body tissue and not hemolymph. Proc. Natl. Acad. Sci. 116, 1792–1801 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    11.Yang, X. & Cox-Foster, D. L. Impact of an ectoparasite on the immunity and pathology of an invertebrate: Evidence for host immunosuppression and viral amplification. Proc. Natl. Acad. Sci. 102, 7470–7475 (2005).ADS 
    CAS 
    PubMed 
    Article 

    Google Scholar 
    12.Rosenkranz, P., Aumeier, P. & Ziegelmann, B. Biology and control of Varroa destructor. J. Invertebr. Pathol. 103, S96–S119 (2010).PubMed 
    Article 

    Google Scholar 
    13.Wilfert, L. et al. Deformed wing virus is a recent global epidemic in honeybees driven by Varroa mites. Science 351, 594–597 (2016).ADS 
    CAS 
    PubMed 
    Article 

    Google Scholar 
    14.Dalmon, A. et al. Evidence for positive selection and recombination hotspots in deformed wing virus (DWV). Sci. Rep. 7, 1–12 (2017).Article 
    CAS 

    Google Scholar 
    15.Martin, S. J. et al. Global honey bee viral landscape altered by a parasitic mite. Science 336, 1304–1306 (2012).ADS 
    CAS 
    PubMed 
    Article 

    Google Scholar 
    16.Moore, J. et al. Recombinants between deformed wing virus and Varroa destructor virus-1 may prevail in Varroa destructor-infested honeybee colonies. J. Gen. Virol. 92, 156–161 (2011).CAS 
    PubMed 
    Article 

    Google Scholar 
    17.Ryabov, E. V. et al. A virulent strain of deformed wing virus (DWV) of honeybees (Apis mellifera) prevails after Varroa destructor-mediated, or in vitro, transmission. PLoS Pathog. 10, e1004230 (2014).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    18.Ryabov, E. V. et al. Dynamic evolution in the key honey bee pathogen deformed wing virus: Novel insights into virulence and competition using reverse genetics. PLoS Biol. 17, e3000502 (2019).MathSciNet 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    19.Mondet, F. et al. Specific cues associated with honey bee social defence against Varroa destructor infested brood. Sci. Rep. 6, 25444 (2016).ADS 
    CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    20.Spivak, M. & Danka, R. G. Perspectives on hygienic behavior in Apismellifera and other social insects. Apidologie https://doi.org/10.1007/s13592-020-00784-z (2020).Article 

    Google Scholar 
    21.Spivak, M. & Gilliam, M. Facultative expression of hygienic behaviour of honey bees in relation to disease resistance. J. Apic. Res. 32, 147–157 (1993).Article 

    Google Scholar 
    22.Baracchi, D., Fadda, A. & Turillazzi, S. Evidence for antiseptic behaviour towards sick adult bees in honey bee colonies. J. Insect Physiol. 58, 1589–1596 (2012).CAS 
    PubMed 
    Article 

    Google Scholar 
    23.Traynor, K. S. et al. Varroa destructor: A complex parasite, crippling honey bees worldwide. Trends Parasitol. 36, 592–606 (2020).CAS 
    PubMed 
    Article 

    Google Scholar 
    24.Sun, Q. & Zhou, X. Corpse management in social insects. Int. J:. Biol. Sci. 9, 313 (2013).
    Google Scholar 
    25.Van Allen, B. G. et al. Cannibalism and infectious disease: Friends or foes?. Am. Nat. 190, 299–312 (2017).PubMed 
    Article 

    Google Scholar 
    26.Bourke, A. F. Queen behaviour, reproduction and egg cannibalism in multiple-queen colonies of the ant Leptothorax acervorum. Anim. Behav. 42, 295–310 (1991).Article 

    Google Scholar 
    27.Pulliainen, U., Helanterä, H., Sundström, L. & Schultner, E. The possible role of ant larvae in the defence against social parasites. Proc. R. Soc. B 286, 20182867 (2019).CAS 
    PubMed 
    Article 

    Google Scholar 
    28.Evans, H. & West-Eberhard, M. The Wasps (Univ. Michigan, 1970).
    Google Scholar 
    29.Schmickl, T. & Crailsheim, K. Cannibalism and early capping: Strategy of honeybee colonies in times of experimental pollen shortages. J. Comp. Physiol. A 187, 541–547 (2001).CAS 
    PubMed 
    Article 

    Google Scholar 
    30.Webster, T. C., Peng, Y. S. & Duffey, S. S. Conservation of nutrients in larval tissue by cannibalizing honey bees. Physiol. Entomol. 12, 225–231 (1987).CAS 
    Article 

    Google Scholar 
    31.Woyke, J. Cannibalism and brood-rearing efficiency in the honeybee. J. Apic. Res. 16, 84–94 (1977).Article 

    Google Scholar 
    32.Chouvenc, T. Limited survival strategy in starving subterranean termite colonies. Insectes Soc. 67, 71–82 (2020).Article 

    Google Scholar 
    33.Raina, A. K., Park, Y. I. & Lax, A. Defaunation leads to cannibalism in primary reproductives of the Formosan subterranean termite, Coptotermes formosanus (Isoptera: Rhinotermitidae). Ann. Entomol. Soc. Am. 97, 753–756 (2004).Article 

    Google Scholar 
    34.Schmickl, T. & Crailsheim, K. Inner nest homeostasis in a changing environment with special emphasis on honey bee brood nursing and pollen supply. Apidologie 35, 249–263 (2004).Article 

    Google Scholar 
    35.Meunier, J. Social immunity and the evolution of group living in insects. Philos. Trans. R. Soc. B Biol. Sci. 370, 20140102 (2015).Article 

    Google Scholar 
    36.Rueppell, O., Hayworth, M. K. & Ross, N. Altruistic self-removal of health-compromised honey bee workers from their hive. J. Evol. Biol. 23, 1538–1546 (2010).CAS 
    PubMed 
    Article 

    Google Scholar 
    37.Halling, L. & Oldroyd, B. P. Do policing honeybee (Apis mellifera) workers target eggs in drone comb?. Insectes Soc. 50, 59–61 (2003).Article 

    Google Scholar 
    38.Santomauro, G., Oldham, N. J., Boland, W. & Engels, W. Cannibalism of diploid drone larvae in the honey bee (Apis mellifera) is released by odd pattern of cuticular substances. J. Apic. Res. 43, 69–74 (2004).Article 

    Google Scholar 
    39.Imdorf, A., Rickli, M., Kilchenmann, V., Bogdanov, S. & Wille, H. Nitrogen and mineral constituents of honey bee worker brood during pollen shortage. Apidologie 29, 315–325 (1998).Article 

    Google Scholar 
    40.Rudolf, V. H. & Antonovics, J. Disease transmission by cannibalism: Rare event or common occurrence?. Proc. R. Soc. B Biol. Sci. 274, 1205–1210 (2007).Article 

    Google Scholar 
    41.Chapman, J. W. et al. Age-related cannibalism and horizontal transmission of a nuclear polyhedrosis virus in larval Spodoptera frugiperda. Ecol. Entomol. 24, 268–275 (1999).Article 

    Google Scholar 
    42.Hamano, K. et al. Waterborne and cannibalism-mediated transmission of the Yellow head virus in Penaeus monodon. Aquaculture 437, 161–166 (2015).Article 

    Google Scholar 
    43.Möckel, N., Gisder, S. & Genersch, E. Horizontal transmission of deformed wing virus: Pathological consequences in adult bees (Apis mellifera) depend on the transmission route. J. Gen. Virol. 92, 370–377 (2011).PubMed 
    Article 
    CAS 

    Google Scholar 
    44.Ryabov, E. V. et al. Development of a honey bee RNA virus vector based on the genome of a deformed wing virus. Viruses 12, 374 (2020).CAS 
    PubMed Central 
    Article 
    PubMed 

    Google Scholar 
    45.Posada-Florez, F. et al. Deformed wing virus type A, a major honey bee pathogen, is vectored by the mite Varroa destructor in a non-propagative manner. Sci. Rep. 9, 1–10 (2019).CAS 
    Article 

    Google Scholar 
    46.Bull, J. C. et al. A strong immune response in young adult honeybees masks their increased susceptibility to infection compared to older bees. PLoS Pathog. 8, e1003083 (2012).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    47.Shi, M. et al. Redefining the invertebrate RNA virosphere. Nature 540, 539–543 (2016).ADS 
    CAS 
    PubMed 
    Article 

    Google Scholar 
    48.Masterman, R., Ross, R., Mesce, K. & Spivak, M. Olfactory and behavioral response thresholds to odors of diseased brood differ between hygienic and non-hygienic honey bees (Apis mellifera L.). J. Comp. Physiol. A 187, 441–452 (2001).CAS 
    PubMed 
    Article 

    Google Scholar 
    49.Crailsheim, K. Trophallactic interactions in the adult honeybee (Apis mellifera L.). Apidologie 29, 97–112 (1998).Article 

    Google Scholar 
    50.Nixon, H. & Ribbands, C. R. Food transmission within the honeybee community. Proc. R. Soc. Lond. Ser. B Biol. Sci. 140, 43–50 (1952).ADS 
    CAS 
    Article 

    Google Scholar 
    51.Arathi, H. & Spivak, M. Influence of colony genotypic composition on the performance of hygienic behaviour in the honeybee, Apis mellifera L. Anim. Behav. 62, 57–66 (2001).Article 

    Google Scholar 
    52.Knecht, D. & Kaatz, H. Patterns of larval food production by hypopharyngeal glands in adult worker honey bees. Apidologie 21, 457–468 (1990).Article 

    Google Scholar 
    53.Li, Z. et al. Transcriptional and physiological responses of hypopharyngeal glands in honeybees (Apis mellifera L.) infected by Nosema ceranae. Apidologie 50, 51–62 (2019).CAS 
    Article 

    Google Scholar 
    54.Lass, A. & Crailsheim, K. Influence of age and caging upon protein metabolism, hypopharyngeal glands and trophallactic behavior in the honey bee (Apis mellifera L.). Insectes Soc. 43, 347–358 (1996).Article 

    Google Scholar 
    55.Chiou, S.-S. & Chen, W.-J. Mutations in the NS3 gene and 3′-NCR of Japanese encephalitis virus isolated from an unconventional ecosystem and implications for natural attenuation of the virus. Virology 289, 129–136 (2001).CAS 
    PubMed 
    Article 

    Google Scholar 
    56.Steel, A., Gubler, D. J. & Bennett, S. N. Natural attenuation of dengue virus type-2 after a series of island outbreaks: A retrospective phylogenetic study of events in the South Pacific three decades ago. Virology 405, 505–512 (2010).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    57.de Souza, F. S., Allsopp, M. H. & Martin, S. J. Deformed wing virus prevalence and load in honeybees in South Africa. Arch. Virol. 166, 237–241 (2020).PubMed 
    PubMed Central 
    Article 
    CAS 

    Google Scholar 
    58.Martin, S. J. et al. Varroa destructor reproduction and cell re-capping in mite-resistant Apis mellifera populations. Apidologie 51, 369–381 (2020).CAS 
    Article 

    Google Scholar 
    59.Kulhanek, K. et al. Survey-derived best management practices for backyard beekeepers improve colony health and reduce mortality. PLoS ONE 16, e0245490 (2021).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    60.Peck, D. T. & Seeley, T. D. Mite bombs or robber lures? The roles of drifting and robbing in Varroa destructor transmission from collapsing honey bee colonies to their neighbors. PLoS ONE 14, e0218392 (2019).CAS 
    PubMed 
    PubMed Central 
    Article 

    Google Scholar 
    61.Ryabov, E. V. et al. Recent spread of Varroa destructor virus-1, a honey bee pathogen, in the United States. Sci. Rep. 7, 1–10 (2017).CAS 
    Article 

    Google Scholar 
    62.Abràmoff, M. D., Magalhães, P. J. & Ram, S. J. Image processing with ImageJ. Biophoton. Int. 11, 36–42 (2004).
    Google Scholar  More