Linking coral fluorescence phenotypes to thermal bleaching in the reef-building Galaxea fascicularis from the northern South China Sea

Abstract

Coral fluorescence phenotypes have been suggested as an adaptation to a broad range of environmental conditions, yet the mechanisms linking thermal bleaching tolerance in reef-building coral populations, associated with fluorescence phenotypes due to GFP-like proteins, remains unclear. In this study, the relationship between the thermal sensitivity and phenotypic plasticity of corals was investigated using two phenotypes of Galaxea fascicularis, green and brown. The results reveal that brown G. fascicularis was more susceptible to bleaching than green G. fascicularis when exposed to a higher growth temperature of 32 °C. Both phenotypes of G. fascicularis were associated with the thermotolerant Symbiodiniaceae symbiont, Durusdinium trenchii. However, the brown G. fascicularis showed a significant decrease in Symbiodiniaceae cell density and a significant increase in pathogenic bacteria abundance when the growth temperature was raised from 29 to 32 °C. The physiological traits and transcriptomic profiles of Symbiodiniaceae were not notably affected, but there were differences in the transcriptional levels of certain genes between the two phenotype hosts of G. fascicularis. Under heat stress of 32 °C, the gene encoding green fluorescent protein (GFP)-like and chromosome-associated proteins, as well as genes related to oxidative phosphorylation, cell growth and death showed lower transcriptional levels in the brown G. fascicularis compared to the green G. fascicularis. Overall, the results demonstrate that the green form of G. fascicularis is better able to tolerate ocean warming and defend against pathogenic bacteria, likely due to higher gene transcription levels and defense ability.

Keywords

Fluorescence phenotype / Thermal bleaching / Microbiome / Galaxea fascicularis / Ocean warming

References

[1] Abe M, Watanabe T, Suzuki Y, Hidaka M. Genetic and morphological differentiation in the hermatypic coral Galaxea fascicularis in Okinawa, Japan. Plankton Benthos Res, 2008, 3: 174-179,

[2] Aihara Y, Maruyama S, Baird AH, Iguchi A, Takahashi S, Minagawa J. Green fluorescence from cnidarian hosts attracts symbiotic algae. Proc Natl Acad Sci, 2019, 116: 2118-2123,

[3] Alieva NO, Konzen KA, Field SF, Meleshkevitch EA, Hunt ME, Beltran-Ramirez V, Miller DJ, Wiedenmann J, Salih A, Matz MV. Diversity and evolution of coral fluorescent proteins. PLoS ONE, 2008, 3,

[4] Baird AH, Birrel CL, Hughes TP, Mcdonald A, Nojima S, Page CA, Prachett MS, Yamasaki H. Latitudinal variation in reproductive synchrony in Acropora assemblages: Japan vs. Australia. Galaxea J Coral Reef Stud, 2009, 11: 101-108,

[5] Barott KL, Huffmyer AS, Davidson JM, Lenz EA, Matsuda SB, Hancock JR, Innis T, Drury C, Putnam HM, Gates RD. Bleaching resistant corals retain heat tolerance following acclimatization to environmentally distinct reefs. Cold Spring Harb Lab, 2020,

[6] Benoit M, Desnues B, Mege J-L. Macrophage polarization in bacterial infections. J Immunol, 2008, 181: 3733-3739,

[7] Ben-Zvi O, Eyal G, Loya Y. Light-dependent fluorescence in the coral Galaxea fascicularis. Hydrobiologia, 2015, 759: 15-26,

[8] Blackall LL, Wilson B, van Oppen MJH. Coral-the world’s most diverse symbiotic ecosystem. Mol Ecol, 2015, 24: 5330-5347,

[9] Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 2014, 30: 2114-2120,

[10] Bollati E, D’Angelo C, Alderdice R, Pratchett M, Ziegler M, Wiedenmann J. Optical feedback loop involving dinoflagellate symbiont and scleractinian host drives colorful coral bleaching. Curr Biol, 2020, 30: 2433-2445.e3,

[11] Botana MT (2019) The role of Symbiodinium membrane lipids in response to heat shock: implications for coral bleaching. https://doi.org/10.13140/RG.2.2.18789.35041

[12] Buchfink B, Xie C, Huson DH. Fast and sensitive protein alignment using DIAMOND. Nat Methods, 2015, 12: 59-60,

[13] Buerger P, Alvarez-Roa C, Coppin CW, Pearce SL, Chakravarti LJ, Oakeshott JG, Edwards OR, van Oppen MJH. Heat-evolved microalgal symbionts increase coral bleaching tolerance. Sci Adv, 2020, 6: eaba2498,

[14] Buitrago-López C, Mariappan KG, Cárdenas A, Gegner HM, Voolstra CR. The Genome of the cauliflower coral Pocillopora verrucosa. Genome Biol Evol, 2020, 12: 1911-1917,

[15] Claar DC, Starko S, Tietjen KL, Epstein HE, Cunning R, Cobb KM, Baker AC, Gates RD, Baum JK. Dynamic symbioses reveal pathways to coral survival through prolonged heatwaves. Nat Commun, 2020, 11: 6097,

[16] D’Angelo C, Denzel A, Vogt A, Matz M, Oswald F, Salih A, Nienhaus G, Wiedenmann J. Blue light regulation of host pigment in reef-building corals. Mar Ecol Prog Ser, 2008, 364: 97-106,

[17] D’Angelo C, Smith EG, Oswald F, Burt J, Tchernov D, Wiedenmann J. Locally accelerated growth is part of the innate immune response and repair mechanisms in reef-building corals as detected by green fluorescent protein (GFP)-like pigments. Coral Reefs, 2012, 31: 1045-1056,

[18] Douglas AE. Coral bleaching––how and why?. Mar Pollut Bull, 2003, 46: 385-392,

[20] Dove SG, Hoegh-Guldberg O, Ranganathan S. Major colour patterns of reef-building corals are due to a family of GFP-like proteins. Coral Reefs, 2001, 19: 197-204,

[21] Eakin CM, Sweatman HPA, Brainard RE. The 2014–2017 global-scale coral bleaching event: insights and impacts. Coral Reefs, 2019, 38: 539-545,

[22] Finn RD, Bateman A, Clements J, Coggill P, Eberhardt RY, Eddy SR, Heger A, Hetherington K, Holm L, Mistry J, Sonnhammer ELL, Tate J, Punta M. Pfam: the protein families database. Nucleic Acids Res, 2014, 42: D222-D230,

[23] Fukami H, Budd AF, Paulay G, Solé-Cava A, Allen Chen C, Iwao K, Knowlton N. Conventional taxonomy obscures deep divergence between Pacific and Atlantic corals. Nature, 2004, 427: 832-835,

[24] Gittins JR, D’Angelo C, Oswald F, Edwards RJ, Wiedenmann J. Fluorescent protein-mediated colour polymorphism in reef corals: multicopy genes extend the adaptation/acclimatization potential to variable light environments. Mol Ecol, 2015, 24: 453-465,

[25] Gong S, Chai G, Xiao Y, Xu L, Yu K, Li J, Liu F, Cheng H, Zhang F, Liao B, Li Z. Flexible symbiotic associations of symbiodinium with five typical coral species in tropical and subtropical reef regions of the northern South China Sea. Front Microbiol, 2018, 9: 2485,

[26] Gong S, Jin X, Ren L, Tan Y, Xia X. Unraveling heterogeneity of coral microbiome assemblages in tropical and subtropical corals in the South China Sea. Microorganisms, 2020, 8: 604,

[27] Gong S, Jin X, Xiao Y, Li Z. Ocean acidification and warming lead to increased growth and altered chloroplast morphology in the Thermo-Tolerant Alga Symbiochlorum hainanensis. Front Plant Sci, 2020, 11,

[28] González-Pech RA, Stephens TG, Chen Y, Mohamed AR, Cheng Y, Shah S, Dougan KE, Fortuin MDA, Lagorce R, Burt DW, Bhattacharya D, Ragan MA, Chan CX. Comparison of 15 dinoflagellate genomes reveals extensive sequence and structural divergence in family Symbiodiniaceae and genus Symbiodinium. BMC Biol, 2021, 19: 73,

[29] Grottoli AG, Warner ME, Levas SJ, Aschaffenburg MD, Schoepf V, McGinley M, Baumann J, Matsui Y. The cumulative impact of annual coral bleaching can turn some coral species winners into losers. Glob Change Biol, 2014, 20: 3823-3833,

[30] Gurskaya NG, Savitsky AP, Yanushevich YG, Lukyanov SA, Lukyanov KA. Color transitions in coral’s fluorescent proteins by site-directed mutagenesis. BMC Biochem, 2001,

[31] Hoogenboom MO, Campbell DA, Beraud E, DeZeeuw K, Ferrier-Pagès C. Effects of light, food availability and temperature stress on the function of photosystem II and photosystem I of coral Symbionts. PLoS ONE, 2012, 7,

[32] Huerta-Cepas J, Szklarczyk D, Forslund K, Cook H, Heller D, Walter MC, Rattei T, Mende DR, Sunagawa S, Kuhn M, Jensen LJ, von Mering C, Bork P. eggNOG 4.5: a hierarchical orthology framework with improved functional annotations for eukaryotic, prokaryotic and viral sequences. Nucleic Acids Res, 2016, 44: D286-D293,

[33] Hughes TP, Kerry JT, Baird AH, Connolly SR, Chase TJ, Dietzel A, Hill T, Hoey AS, Hoogenboom MO, Jacobson M, Kerswell A, Madin JS, Mieog A, Paley AS, Pratchett MS, Torda G, Woods RM. Global warming impairs stock-recruitment dynamics of corals. Nature, 2019, 568: 387-390,

[34] Hume BCC, Smith EG, Ziegler M, Warrington HJM, Burt JA, LaJeunesse TC, Wiedenmann J, Voolstra CR. SymPortal: a novel analytical framework and platform for coral algal symbiont next-generation sequencing ITS2 profiling. Mol Ecol Resour, 2019, 19: 1063-1080,

[35] Iglesias-Prieto R, Matta JL, Robins WA, Trench RK. Photosynthetic response to elevated temperature in the symbiotic dinoflagellate Symbiodinium microadriaticum in culture. Proc Natl Acad Sci, 1992, 89: 10302-10305,

[36] Jarett JK, MacManes MD, Morrow KM, Pankey MS, Lesser MP. Comparative genomics of color morphs in the coral Montastraea cavernosa. Sci Rep, 2017, 7: 16039,

[37] Jones AM, Berkelmans R, van Oppen MJH, Mieog JC, Sinclair W. A community change in the algal endosymbionts of a scleractinian coral following a natural bleaching event: field evidence of acclimatization. Proc R Soc B Biol Sci, 2008, 275: 1359-1365,

[38] Kahng SE, Salih A. Localization of fluorescent pigments in a nonbioluminescent, azooxanthellate octocoral suggests a photoprotective function. Coral Reefs, 2005, 24: 435-435,

[39] Karim W, Seidi A, Hill R, Chow WS, Minagawa J, Hidaka M, Takahashi S. Novel characteristics of photodamage to PSII in a high-light-sensitive Symbiodinium Phylotype. Plant Cell Physiol, 2015, 56: 1162-1171,

[40] Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol, 2018, 35: 1547-1549,

[41] LaJeunesse TC, Parkinson JE, Gabrielson PW, Jeong HJ, Reimer JD, Voolstra CR, Santos SR. Systematic revision of Symbiodiniaceae highlights the antiquity and diversity of coral endosymbionts. Curr Biol, 2018, 28: 2570-2580.e6,

[42] Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods, 2012, 9: 357-359,

[43] Leutenegger A, D’Angelo C, Matz MV, Denzel A, Oswald F, Salih A, Nienhaus GU, Wiedenmann J. It’s cheap to be colorful: Anthozoans show a slow turnover of GFP-like proteins. FEBS J, 2007, 274: 2496-2505,

[44] Leveque S, Afiq-Rosli L, Ip YCA, Jain SS, Huang D. Searching for phylogenetic patterns of Symbiodiniaceae community structure among Indo-Pacific Merulinidae corals. PeerJ, 2019, 7,

[45] Li B, Dewey CN. RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform, 2011, 12: 323,

[46] Li X, Liu S, Huang H, Huang L, Jing Z, Zhang C. Coral bleaching caused by an abnormal water temperature rise at Luhuitou fringing reef, Sanya Bay, China. Aquat Ecosyst Health Manag, 2012, 15: 227-233,

[47] Li D, Liu C-M, Luo R, Sadakane K, Lam T-W. MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph. Bioinformatics, 2015, 31: 1674-1676,

[48] Liu W, Hao Z, Huang L, Chen L, Wei Q, Cai L, Liang S. Comparative expression profile of microRNAs in Anopheles anthropophagus midgut after blood-feeding and Plasmodium infection. Parasit Vectors, 2017, 10: 86,

[49] Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol, 2014, 15: 550,

[50] Luna GM, Bongiorni L, Gili C, Biavasco F, Danovaro R. Vibrio harveyi as a causative agent of the White Syndrome in tropical stony corals: Vibrio pathogens of stony corals. Environ Microbiol Rep, 2009, 2: 120-127,

[51] Morikawa MK, Palumbi SR. Using naturally occurring climate resilient corals to construct bleaching-resistant nurseries. Proc Natl Acad Sci, 2019, 116: 10586-10591,

[52] Niu W, Huang H, Lin R, Chen C-H, Shen K-N, Hsiao C-D. The complete mitogenome of the Galaxy Coral, Galaxea fascicularis (Cnidaria: Oculinidae). Mitochondrial DNA Part B, 2016, 1: 10-11,

[53] Ogata H, Goto S, Sato K, Fujibuchi W, Bono H, Kanehisa M. KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res, 1999, 27: 29-34,

[54] Osman EO, Smith DJ, Ziegler M, Kürten B, Conrad C, El-Haddad KM, Voolstra CR, Suggett DJ. Thermal refugia against coral bleaching throughout the northern Red Sea. Glob Change Biol, 2018, 24: e474-e484,

[55] Oswald F, Schmitt F, Leutenegger A, Ivanchenko S, D’Angelo C, Salih A, Maslakova S, Bulina M, Schirmbeck R, Nienhaus GU, Matz MV, Wiedenmann J. Contributions of host and symbiont pigments to the coloration of reef corals: coloration of reef corals. FEBS J, 2007, 274: 1102-1122,

[56] Paley AS (2012) Bleaching condition varies among Acropora millepora color morphs. https://www.icrs2012.com/proceedings/manuscripts/ICRS2012_9A_9.pdf

[57] Palmer CV, Roth MS, Gates RD. Red fluorescent protein responsible for pigmentation in trematode-infected Porites compressa tissues. Biol Bull, 2009, 216: 68-74,

[58] Parks DH, Tyson GW, Hugenholtz P, Beiko RG. STAMP: statistical analysis of taxonomic and functional profiles. Bioinformatics, 2014, 30: 3123-3124,

[59] Qin Z, Yu K, Chen B, Wang Y, Liang J, Luo W, Xu L, Huang X. Diversity of symbiodiniaceae in 15 coral species from the southern South China Sea: potential relationship with coral thermal adaptability. Front Microbiol, 2019, 10: 2343,

[60] Raina J-B, Tapiolas DM, Forêt S, Lutz A, Abrego D, Ceh J, Seneca FO, Clode PL, Bourne DG, Willis BL, Motti CA. DMSP biosynthesis by an animal and its role in coral thermal stress response. Nature, 2013, 502: 677-680,

[61] Ritson-Williams R, Gates RD. Coral community resilience to successive years of bleaching in Kāne‘ohe Bay, Hawai‘i. Coral Reefs, 2020, 39: 757-769,

[62] Roberty S, Furla P, Plumier J-C. Differential antioxidant response between two Symbiodinium species from contrasting environments: antioxidant responses of Symbiodinium sp. Plant Cell Environ, 2016, 39: 2713-2724,

[63] Satoh N, Kinjo K, Shintaku K, Kezuka D, Ishimori H, Yokokura A, Hagiwara K, Hisata K, Kawamitsu M, Koizumi K, Shinzato C, Zayasu Y (2020) Color morphs of the coral, Acropora tenuis, show different responses to environmental stress and different expression profiles of fluorescent-protein genes. G3 (Bethesda) 11:jkab018

[64] Shinzato C, Khalturin K, Inoue J, Zayasu Y, Kanda M, Kawamitsu M, Yoshioka Y, Yamashita H, Suzuki G, Satoh N. Eighteen coral genomes reveal the evolutionary origin of Acropora strategies to accommodate environmental changes. Mol Biol Evol, 2021, 38: 16-30,

[65] Skirving WJ, Heron SF, Marsh BL, Liu G, De La Cour JL, Geiger EF, Eakin CM. The relentless march of mass coral bleaching: a global perspective of changing heat stress. Coral Reefs, 2019, 38: 547-557,

[66] Smith-Keune C, Dove S. Gene expression of a green fluorescent protein homolog as a host-specific biomarker of heat stress within a reef-building coral. Mar Biotechnol, 2008, 10: 166-180,

[67] Tamames J, Puente-Sánchez F. SqueezeMeta, A highly portable, fully automatic metagenomic analysis pipeline. Front Microbiol, 2019, 9: 3349,

[68] Thompson FL, Hoste B, Thompson CC, Huys G, Swings J. The coral bleaching Vibrio shiloi Kushmaro et al. 2001 is a Later Synonym of Vibrio mediterranei Pujalte and Garay. Syst Appl Microbiol, 1986, 24: 516-519,

[69] Voolstra CR. Coral bleaching: a colorful struggle for survival. Curr Biol, 2020, 30: R768-R770,

[70] Xu L, Yu K, Li S, Liu G, Tao S, Shi Q, Chen T, Zhang H. Interseasonal and interspecies diversities of Symbiodinium density and effective photochemical efficiency in five dominant reef coral species from Luhuitou fringing reef, northern South China Sea. Coral Reefs, 2017, 36: 477-487,

[71] Zhong H, Sun H, Liu R, Zhan Y, Huang X, Ju F, Zhang X-H. Comparative genomic analysis of Labrenzia aggregata (Alphaproteobacteria) strains isolated from the mariana trench: insights into the metabolic potentials and biogeochemical functions. Front Microbiol, 2021, 12,