Co-cultures of Oophila amblystomatis between Ambystoma maculatum and Ambystoma gracile hosts show host-symbiont fidelity
Tóm tắt
A unique symbiosis occurs between embryos of the spotted salamander (Ambystoma maculatum) and a green alga (Oophila amblystomatis). Unlike most vertebrate host-symbiont relationships, which are ectosymbiotic, A. maculatum exhibits both an ecto- and an endo-symbiosis, where some of the green algal cells living inside egg capsules enter embryonic tissues as well as individual salamander cells. Past research has consistently categorized this symbiosis as a mutualism, making this the first example of a “beneficial” microbe entering vertebrate cells. Another closely related species of salamander, Ambystoma gracile, also harbors beneficial Oophila algae in its egg capsules. However, our sampling within the A. gracile range consistently shows this to be a strict ectosymbiotic interaction—with no sign of tissue or presumably cellular entry. In this study we swapped cultured algae derived from intracapsular fluid of different salamander hosts to test the fidelity of tissue entry in these symbioses. Both A. maculatum and A. gracile embryos were raised in cultures with their own algae or algae cultured from the other host. Under these in vitro culture conditions A. maculatum algae will enter embryonic A. maculatum tissues. Additionally, although at a much lower frequency, A. gracile derived algae will also enter A. maculatum host tissues. However, neither Oophila strain enters A. gracile hosts in these co-culture conditions. These data reveal a potential host-symbiont fidelity that allows the unique endosymbiosis to occur in A. maculatum, but not in A. gracile. However, preliminary trials in our study found that persistent endogenous A. maculatum algae, as opposed to the cultured algae used in subsequent trials, enters host tissues at a higher frequency. An analysis of previously published Oophila transcriptomes revealed dramatic differences in gene expression between cultured and intracapsular Oophila. These include a suite of genes in protein and cell wall synthesis, photosynthesis, central carbon metabolism suggesting the intracapsular algae are assimilating ammonia for nitrogen metabolism and may be undergoing a life-cycle transition. Further refinements of these co-culture conditions could help determine physiological differences between cultured and endogenous algae, as well as rate-limiting cues provided for the alga by the salamander.
Tài liệu tham khảo
Akhtar TA, Orsomando G, Mehrshahi P, Lara-Núñez A, Bennett MJ, Gregory JF, Hanson AD (2010) A central role for gamma-glutamyl hydrolases in plant folate homeostasis. Plant J 64:256–266
Alexa A, Rahnenfuhrer J (2016) topGO: enrichment analysis for gene ontology. R package version 232.0
Bachmann M, Carlton RG, Burkholder J, Wetzel RG (1985) Symbiosis between salamander eggs and green algae: microelectrode measurements inside eggs demonstrate effect of photosynthesis on oxygen concentration. Can J Zool 64:1586–1588
Baldan B, Girard-Bascou J, Wollman FA, Olive J (1991) Evidence for thylakoid membrane fusion during zygote formation in Chlamydomonas reinhardtii. J Cell Biol 114:905–915
Bianchini K, Tattersall GJ, sashaw J, Porteus CS, Wright PA (2012) Acid water interferes with salamander-green algae symbiosis during early embryonic development. Physiol Biochem Zool 85:470–480
Bishop CD, Miller AG (2014) Dynamics of the growth, life history transformation and photosynthetic capacity of Oophila amblystomatis (Chlorophyceae), a green algal symbiont associated with embryos of the northeastern yellow spotted salamander Ambystoma maculatum (Amphibia). Symbiosis 63:47–57
Black CK, Mihai DM, Washington I (2014) The photosynthetic eukaryote Nannochloris eukaryotum as an intracellular machine to control and expand functionality of human cells. Nano Lett 14:2720–2725
Burns J, Zhang H, Hill E, Kim E, Kerney RR (2017) Transcriptome analysis illuminates the nature of the intracellular interaction in a vertebrate-algal symbiosis. eLife 6:e22054
Burr HS (1916) The effects of the removal of the nasal pits in Amblystoma. J Exp Zool 20:27–51
Carrier G, Garnier M, Le Cunff L, Bougaran G, Probert I, De Vargas C, Corre E, Cadoret JP, Saint-Jean B (2014) Comparative transcriptome of wild type and selected strains of the microalgae Tisochrysis lutea provides insights into the genetic basis, lipid metabolism and the life cycle. PLoS One 9:e86889
Cliburn JW, Ward BQ (1963) Occurrence of Oophila amblystomatis (a symbiotic alga) in Ambystoma maculatum of the lower gulf coastal plain. Am Midl Nat 69:508
Dodge JD (1973) The fine structure of algal cells. Academic Press, New York
Fang W, Si Y, Douglass S, Casero D, Merchant SS, Pellegrini M, Ladunga I, Liu P, Spalding MH (2012) Transcriptome-wide changes in Chlamydomonas reinhardtii gene expression regulated by carbon dioxide and the CO2-concentrating mechanism regulator CIA5/CCM1. Plant Cell 24:1876–1893
Gibbs M (2003) Axial patterning: using retinoic acid to disrupt homeobox gene expression in axolotls. In: A Practical Guide to Developmental Biology Oxford, England. p. 28–31
Gilbert P (1942) Observations on the eggs of Ambystoma maculatum with especial reference to the green algae found within the egg envelopes. Ecology 23:215–227
Gilbert PW (1944) The alga-egg relationship in Ambystoma maculatum: a case of symbiosis. Ecology 25:366–369
Goff LJ, Stein JR (1976) Preliminary studies on the green alga Oophila in salamander egg masses. J Phycol 12(suppl):23
Guy RD, Vanlerberghe GC, Turpin DH (1989) Significance of phosphoenolpyruvate carboxylase during ammonium assimilation: carbon isotope discrimination in photosynthesis and respiration by the N-limited green alga Selenastrum minutum. Plant Physiol 89:1150–1157
Hale RE, Miller N, Francis RA, Kennedy C (2016) Does breeding ecology alter selection on developmental and life history traits? A case study in two Ambystomatid salamanders. Evol Ecol 30(3):503–517
Hale RE, Kennedy C, Winkelman D, Brown C (2017) An advantage of clear over white egg mass morphs in metabolically demanding microhabitats suggests a role of symbiotic algae in the maintenance of a polymorphism in the spotted salamander (Ambystoma maculatum). Evol Ecol Res 18:637–650
Harrison RG (1969) Harrison stages and description of the normal development of the spotted salamander, Amblystoma punctatum. In: Wilens S (ed) Organization and development of the embryo. Yale University Press, New Haven, p 44–66
Hutchison V (1971) On the Ambystoma egg-alga relationship. Herp Rev 3:82
IUCN, Conservation International, and NatureServe. 2004. Global Amphibian Assessment. IUCN, Conservation International and NatureServe, Washington, DC and Arlington
Jeong SW, Nam SW, HwangBo K, Jeong WJ, Jeong B-R, Chang YK, Park Y-I (2017) Transcriptional regulation of cellulose biosynthesis during the early phase of nitrogen deprivation in Nannochloropsis salina. Sci Rep 7:5264
Kerney R (2011) Symbioses between salamander embryos and green algae. Symbiosis 54:107–119
Kerney R, Kim E, Hangarter RP, Heiss AA, Bishop CD, Hall BK (2011) Intracellular invasion of green algae in a salamander host. Proc Natl Acad Sci U S A 108:6497–6502
Kerney R, Burns JB, Kim E (2017) Chapter 7: investigating mechanisms of algal entry into salamander cells. In: Algal and Cyanobacteria Symbioses. Grube M, Seckbach J, Muggia L (eds) World Scientific Press, London, p 209–239
Kim E, Lin Y, Kerney R, Blumenberg L, Bishop C (2014) Phylogenetic analysis of algal symbionts associated with four North American amphibian egg masses. PLoS One 9:e108915
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lewis LA, Landberg T (2014) Evolutionary diversity of the symbiotic salamander algae, Oophila (Chlorophyta). Unpublished Genbank Submission
Lewis L, Lo C, Urban M, Schwenk K, Xue C, Landberg T (2013) Natural history of the green algae-salamander symbiosis. Phycologia 52(4):62
Lin Y, Bishop CD (2015) Identification of free-living Oophila amblystomatis (Chlorophyceae) from yellow spotted salamander and wood frog breeding habitat. Phycologia 54:183–191
Mandel MJ, Wollenberg MS, Stabb EV, Visick KL, Ruby EG (2009) A single regulatory gene is sufficient to alter bacterial host range. Nature 458:215–218
Marco A, Blaustein AR (2000) Symbiosis with green algae affects survival and growth of northwestern salamander embryos. J Herp 34:617–621
Mills N, Barnhart M (1999) Effects of hypoxia on embryonic development in two Ambystoma and two Rana species. Physiol Biochem Zool 72:179–188
Moran NA, Yun Y (2015) Experimental replacement of an obligate insect symbiont. Proc Natl Acad Sci U S A 112:2093–2096
Muto K, Nishikawa K, Kamikawa R, Miyashita H (2017) Symbiotic green algae in eggs of Hynobius nigrescens, an amphibian endemic to Japan. Phycol Res 65:171–174
Nema M, Hanson ML, Müller KM (2018) Phylogeny of the egg-loving green alga Oophila amblystomatis (Chlamydomonadales) and its response to the herbicides atrazine and 2,4-D. Symbiosis Online ahead of print. https://doi.org/10.1007/s13199-018-0564-1
Ohkawa H, Hashimoto N, Furukawa S, Kadono T, Kawano T (2011) Forced symbiosis between Synechocystis spp. PCC 6803 and apo-symbiotic Paramecium bursaria as an experimental model for evolutionary emergence of primitive photosynthetic eukaryotes. Plant Signal Behav 6:773–776
Oliver KM, Degnan PH, Burke GR, Moran NA (2010) Facultative symbionts in aphids and the horizontal transfer of ecologically important traits. Annu Rev Entomol 55:247–266
Orsomando G, de la Garza RD, Green BJ, Peng M, Rea PA, Ryan TJ, Gregory JF, Hanson AD (2005) Plant gamma-glutamyl hydrolases and folate polyglutamates: characterization, compartmentation, and co-occurrence in vacuoles. J Biol Chem 280:28877–28884
Owen OE, Kalhan SC, Hanson RW (2002) The key role of anaplerosis and cataplerosis for citric acid cycle function. J Biol Chem 277:30409–30412
Pinder A, Friet S (1994) Oxygen transport in egg masses of the amphibians Rana sylvatica and Ambystoma maculatum: convection, diffusion and oxygen production by algae. J Exp Biol 197:1–14
Ramírez-Romero R, Rodríguez-Tovar LE, Nevárez-Garza AM, Lopez A (2015) Chlorella infection in a sheep in Mexico and minireview of published reports from humans and domestic animals. Mycopathologia 169:461–466
Ravanel S, Douce R, Rébeillé F (2011) Chapter 3 - Metabolism of folates in plants, In: Rébeillé F, Douce R (eds) Advances in botanical research. Elsevier, Amsterdam, p 67–106
Robinson MD, McCarthy DJ, Smyth GK (2010) edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26:139–140
Rodriguez-Gil JL, Brain R, Baxter L, Ruffell S, McConkey B, Solomon K, Hanson M (2014) Optimization of culturing conditions for toxicity testing with the alga Oophila sp. (Chlorophyceae), an amphibian endosymbiont. Envir Tox Chem 33:2566–2575
Schultz N (2016) The symbiotic green algae, Oophila (Chlamydomonadales, Chlorophyceae): a heterotrophic growth study and taxonomic history. Master’s Thesis. University of Connecticut. Mansfeld, CT
Sive HL, Grainger RM, Harland RM (2000) Early development of Xenopus laevis. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press
Small DP, Bennett RS, Bishop CD (2014) The roles of oxygen and ammonia in the symbiotic relationship between the spotted salamander Ambystoma maculatum and the green alga Oophila amblystomatis during embryonic development. Symbiosis 64:1–10
Twitty VC (1932) Influence of the eye on the growth of its associated structures, studied by means of heteroplastic transplantation. J Exp Zool 61:333–374
Voolstra CR, Schwarz JA, Schnetzer J, Sunagawa S, Desalvo MK, Szmant AM, Coffroth MA, Medina M (2009) The host transcriptome remains unaltered during the establishment of coral-algal symbioses. Mol Ecol 18:1823–1833
Walker T, Johnson PH, Moreira LA, Iturbe-Ormaetxe I, Frentiu FD, McMeniman CJ, Leong YS, Dong Y, Axford J, Kriesner P, Lloyd AL, Ritchie SA, O'Neill SL, Hoffmann AA (2011) The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations. Nature 476:450–453
Walter W, Sanchez-Cabo F, Ricote M (2015) GOplot: an R package for visually combining expression data with functional analysis. Bioinformatics 17:2912–2914
Wantanabe MM, Kawachi M, Hiroki M, Kasai F (2000) NIES collection list of strains. 6 ed. Japan. 1 p
Wilhelm C, Wild A (1982) Growth and photosynthesis of Nanochlorum eucaryotum, a new and extremely small eucaryotic green alga. Z Naturforsch 37:115–119