DOCSLIB.ORG

This Paper Must Be Cited As

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
This Paper Must Be Cited As Document downloaded from: http://hdl.handle.net/10251/121824 This paper must be cited as: Molins, A.; Moya, P.; García-Breijo, F.; Reig-Armiñana, J.; Barreno, E. (2018). A multi-tool approach to assess microalgal diversity in lichens. isolation, Sanger sequencing, HTS and ultrastructural correlations. The Lichenologist. 50(1):123-138. https://doi.org/10.1017/S0024282917000664 The final publication is available at http://doi.org/10.1017/S0024282917000664 Copyright Cambridge University Press Additional Information The Lichenologist A multi -tool approach to assess microalgal diversity in lichens: isolation, Sanger sequencing, HTS and ultrastructural correlations Journal: The Lichenologist ManuscriptManuscript ID LICH-Mar-17-SP-0768.R1 For Review Manuscript Type: Standard Paper Date Submitted by the Author: n/a Complete List of Authors: Molins , Arántzazu; Botanica Moya, Patricia; Botanica García-Breijo, Francisco; Universitat Politecnica de Valencia Reig-Armiñana, Jose Barreno, Eva Keywords: coexistence, propagation, 454-pyrosequencing, Trebouxia Cambridge University Press Page 1 of 40 The Lichenologist 1 ORIGINAL ARTICLES 2 3 Title: A multi-tool approach to assess microalgal diversity in lichens: isolation, Sanger 4 sequencing, HTS and ultrastructural correlations. 5 6 Authors:Manuscript Arántzazu Molins 1, ┼┼┼, Patricia Moya For 1, ┼┼┼, Francisco Review J. García-Breijo 2, José Reig- 7 Armiñana 1 and Eva Barreno 1. 8 9 1 Dpto. Botánica, ICBIBE & Jardí Botànic, Fac. CC. Biológicas, Universitat de València, C/ 10 Dr. Moliner 50. 46100-Burjassot, Valencia, Spain. 11 12 2 Dpto. Ecosistemas Agroforestales, Universidad Politécnica de Valencia, Camino de Vera s/n, 13 46022-Valencia, Spain. 14 15 Running Head: Algal diversity by a multi-tool approach 16 17 For correspondence: A. Molins, Dpto. Botánica, ICBIBE, Fac. CC. Biológicas, Universitat de 18 València, C/ Dr. Moliner 50. 46100-Burjassot, Valencia, Spain 19 Phone number: +34 963544376; Fax number +34 963544082; email: [email protected] . 20 21 ┼ These two authors contributed equally to this work. 22 23 1 Cambridge University Press The Lichenologist Page 2 of 40 24 25 ABSTRACT 26 Lichen thalli represent the most relevant examples of fungal–algal interactions. Studies 27 that describe phycobiont diversity within entire thalli are based mainly on Sanger sequencing. 28 In some Manuscriptlichen species, this technique could undereForstimate Review the intrathalline coexistence of 29 multiple microalgae. In this study, different multi-tool approaches were applied to two lichen 30 taxa, Circinaria hispida and Flavoparmelia soredians , to detect algal coexistence. Here, we 31 combined Sanger sequencing, a specific polymerase chain reaction (PCR) primer, 454- 32 pyrosequencing, phycobiont isolation and ultrastructural characterisation. Furthermore, we 33 compared pyrenoid ultrastructural features of lichenised phycobionts with microalgae isolated 34 in culture. An improved methodology was used to isolate and propagate phycobionts which, 35 in combination with fast genetic identification, resulted in a considerable reduction in time 36 and cost to complete the process. This isolation method, coupled with a specific PCR primer, 37 allowed for the detection of coexisting algae in C. hispida (four Trebouxia lineages). 454- 38 pyrosequencing detected only a fraction of such diversity, while Sanger sequencing identified 39 only the primary phycobiont. Ultrastructural features of the isolated algae were observed by 40 transmission electron microscopy; the maintenance of the pyrenoid characteristics suggested 41 the existence of different Trebouxia lineages. In F. soredians , a single Trebouxia lineage was 42 identified using all these approaches. 43 In cases of lichens with algal coexistence, a combination of different molecular and 44 ultrastructural approaches may be required to reveal the underlying algal diversity within a 45 single thallus. The approaches proposed in this study provide information about the 46 relationship between molecular and ultrastructural data, and they represent an improvement in 2 Cambridge University Press Page 3 of 40 The Lichenologist 47 the delimitation of taxonomical features which is needed to recognise intrathalline Trebouxia 48 diversity. 49 50 KEY WORDS 51 coexistence;Manuscript propagation; 454-pyrosequencing; For Trebouxia. Review 52 53 INTRODUCTION 54 Lichens are classical symbiotic associations (holobiont) that result in a complex 55 organism with a specific biological organisation (Chapman & Margulis 1998). Such 56 associations involve at least two very different organisms, a heterotrophic fungus (mycobiont) 57 and a photosynthetic (photobiont). Photobionts can be cyanobacteria (cyanobionts) or green 58 microalgae (phycobionts). The presence of microalgal communities, lichen-associated fungi, 59 bacteria and yeast highlight the intrathalline complexity of lichen thalli (Arnold et al . 2009; 60 U´Ren et al . 2010, 2012; Park et al . 2015; Aschenbrenner et al . 2016; Muggia et al. 2016a; 61 Spribille et al . 2016; Moya et al . 2017). 62 Lichen literature has reported the occurrence of different phycobiont lineages inside 63 the thalli in several lichen species (Blaha et al . 2006; Ohmura et al . 2006; Piercey-Normore 64 2006; Muggia et al . 2010, 2014; Schmull et al . 2011; Molins et al . 2013; Leavitt et al . 2015). 65 The lichen Ramalina farinacea has proven to be a suitable model to study the multiplicity of 66 microalgae in lichen thalli due to the recurrent coexistence of Trebouxia sp. TR9 and T. 67 jamesii in long-distance populations (del Campo et al. 2010; Casano et al. 2011). Moreover, a 68 recent study using a deep sequencing technique revealed a much higher microalgal diversity 3 Cambridge University Press The Lichenologist Page 4 of 40 69 associated with a single lichen thallus of Ramalina farinacea (22 operational taxonomic units 70 [OTUs]) (Moya et al . 2017). 71 In order to understand the functions of microalgal communities in lichen thalli, 72 phycobiont isolation is a key step in the experimental design. The isolation and culture of 73 symbioticManuscript organisms offers an opportunity to undersFortand theReview mechanisms underlying the 74 selection of partners in symbiotic interactions (Yoshimura et al . 2002). Cultures of lichen 75 microalgae involve time-consuming techniques necessary for the successful propagation of 76 these symbionts (Rafat et al . 2015). However, phycobiont isolation is an essential procedure 77 to study lichen physiology, perform morphologic characterisation and obtain pure material for 78 molecular identification. 79 Studies dealing with phycobiont diversity in lichen thalli are primarily based on 80 Sanger sequencing. However, this technique could underestimate phycobiont diversity in 81 some lichen species (del Campo et al . 2010; Molins et al . 2013). There has been an increase 82 in the use of deep sequencing techniques to analyse the diversity of microalgal communities 83 (Bates et al . 2012; Park et al . 2015) because these techniques allow for the detection of a vast 84 number of genotypes that would otherwise remain underestimated in conventional PCR 85 amplifications (Stewart & Cavanaugh 2009). 86 To understand the hidden intrathalline phycobiont diversity and the role of each 87 phycobiont type in a symbiotic relationship, multi-tool and multidisciplinary approaches are 88 needed to obtain comparative results (Catalá et al. 2015). In particular, we focused on three 89 specific aims: i) to upgrade a methodology to propagate phycobionts that, coupled to fast 90 molecular identification, will significantly reduce the time needed to complete the process; ii) 91 to evaluate the phycobiont diversity using Sanger sequencing, 454-pyrosequencing and 4 Cambridge University Press Page 5 of 40 The Lichenologist 92 isolation; and iii) to compare the ultrastructural characterisation between lichenised 93 phycobionts and microalgae isolated in culture, and to correlate these ultrastructural traits 94 using molecular analyses. To achieve these objectives, we selected Circinaria hispida 95 (Mereschk.) A. Nordin, S. Savi ć and Tibell and Flavoparmelia soredians (Nyl.) Hale, two 96 lichen speciesManuscript with different biotypes that occur For in different Review ecosystems. 97 98 MATERIALS AND METHODS 99 Lichen material 100 The Circinaria hispida specimen was collected from Zaorejas (Guadalajara, Spain) 101 (40°46’02”N, 2°11’40”W, 1105 m ), and the Flavoparmelia soredians specimen was collected 102 on Quercus suber cork from Chóvar (Castellón, Spain) (39°51’13,68”N, 0°19’09.60”W). 103 Samples were dried out for one day and then stored at -20ºC until their processing. 104 105 Experimental design 106 Two lichen thalli (one per species) were examined under a stereo microscope to 107 exclude surface contamination. Samples were rehydrated with Milli-Q sterile water one day 108 before being processed, and they were stored in a growth chamber at 21ºC under a 12 h/12 h 109 light/dark cycle (lighting conditions: 15 µmol/m 2s). Lichen thalli were vortexed three times 110 for 5 min at 2,000 rpm with Milli-Q sterile water. Fragments from different parts of each 111 thallus were randomly excised and homogenised together, and each pool was labelled as 112 sample A, B, C or D. The mycobiont and primary phycobiont were identified by Sanger 113 sequencing from sample A. Isolation protocol and Sanger identification was performed for 5 Cambridge University Press The Lichenologist Page 6 of 40 114 sample B. Sample C was analysed
Load more

Recommended publications
  • Species Relationships in the Lichen Alga Trebouxia (Chlorophyta, Trebouxiophyceae): Molecular Phylogenetic Analyses of Nuclear-E
    Symbiosis, 23 (1997) 125-148 125 Balaban, Philadelphia/Rehovot Species Relationships in the Lichen Alga Trebouxia (Chlorophyta, Trebouxiophyceae): Molecular Phylogenetic Analyses of Nuclear-Encoded Large Subunit rRNA Gene Sequences THOMAS FRIEDL* and CLAUDIA ROKITTA Fachbereich Biologie, Allgemeine Botanik, Universitiit Kaiserslautern, POB 3049, 67653 Kaiserslautern, Germany, Tel. +49-631-2052360, Fax. +49-631-2052998, E-mail. [email protected] Received December 11, 1996; Accepted May 22, 1997 Abstract Sequences of the 5' region of the nuclear-encoded large subunit (26S) rRNA genes were determined for seven species of Trebouxia to investigate the evolutionary relationships among these coccoid green algae that form the most frequently occurring photobiont in lichen symbiosis. Phylogenies inferred from these data substantiate the importance of certain chloroplast characters for tracing species relationships within Trebouxia. The monophyletic origin of the "Trebouxia cluster" which comprises only those species that have centrally located chloroplasts and distinct pyrenoid matrices interdispersed by a thylakoid tubule network is clearly resolved. However, those species of Trebouxia with a chloroplast closely appressed to the cell wall at certain stages and an indistinct pyrenoid containing regular thylakoids are distantly related to the Trebouxia cluster; these species may represent an independent genus. These findings are corroborated by analyses of available complete 18S rDNA sequences from Trebouxia spp. There are about 1.5 times more variable positions in the partial 26S rDNA sequences than in the full 18S sequences, and most of these positions are .. The author to whom correspondence should be sent. Presented at the Third International Lichenological Symposium (IAL3), September 1-7, 1996, Salzburg, Austria 0334-5114/97 /$05.50 ©1997 Balaban 126 T.
    [Show full text]
  • The Puzzle of Lichen Symbiosis
    Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1503 The puzzle of lichen symbiosis Pieces from Thamnolia IOANA ONUT, -BRÄNNSTRÖM ACTA UNIVERSITATIS UPSALIENSIS ISSN 1651-6214 ISBN 978-91-554-9887-0 UPPSALA urn:nbn:se:uu:diva-319639 2017 Dissertation presented at Uppsala University to be publicly examined in Lindhalsalen, EBC, Norbyvägen 14, Uppsala, Thursday, 1 June 2017 at 09:15 for the degree of Doctor of Philosophy. The examination will be conducted in English. Faculty examiner: Associate Professor Anne Pringle (University of Wisconsin-Madison, Department of Botany). Abstract Onuț-Brännström, I. 2017. The puzzle of lichen symbiosis. Pieces from Thamnolia. Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology 1503. 62 pp. Uppsala: Acta Universitatis Upsaliensis. ISBN 978-91-554-9887-0. Symbiosis brought important evolutionary novelties to life on Earth. Lichens, the symbiotic entities formed by fungi, photosynthetic organisms and bacteria, represent an example of a successful adaptation in surviving hostile environments. Yet many aspects of the lichen symbiosis remain unexplored. This thesis aims at bringing insights into lichen biology and the importance of symbiosis in adaptation. I am using as model system a successful colonizer of tundra and alpine environments, the worm lichens Thamnolia, which seem to only reproduce vegetatively through symbiotic propagules. When the genetic architecture of the mating locus of the symbiotic fungal partner was analyzed with genomic and transcriptomic data, a sexual self-incompatible life style was revealed. However, a screen of the mating types ratios across natural populations detected only one of the mating types, suggesting that Thamnolia has no potential for sexual reproduction because of lack of mating partners.
    [Show full text]
  • A Metagenomic Approach to Reconstruct the Holo-Genome Of
    bioRxiv preprint doi: https://doi.org/10.1101/810986; this version posted October 21, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. What is in a lichen? A metagenomic approach to reconstruct the holo-genome of Umbilicaria pustulata Bastian Greshake Tzovaras1,2,3, Francisca H.I.D. Segers1,4, Anne Bicker5, Francesco Dal Grande4,6, Jürgen Otte6, Seyed Yahya Anvar7, Thomas Hankeln5, Imke Schmitt4,6,8, and Ingo Ebersberger1,4,6,# 1Applied Bioinformatics Group, Institute of Cell Biology and Neuroscience, Goethe University Frankfurt, Max-von-Laue Str. 13, Frankfurt am Main, 60438, Germany 2Lawrence Berkeley National Laboratory, Berkeley, United States 3Center for Research & Interdisciplinarity, Université de Paris, France 4LOEWE Center for Translational Biodiversity Genomics, Frankfurt, Germany 5 Institute for Organismic and Molecular Evolution, Molecular Genetics and Genome Analysis, Johannes Gutenberg University Mainz, J. J. Becher-Weg 30A, 55128 Mainz, Germany 6Senckenberg Biodiversity and Climate Research Centre (SBiK-F), Senckenberg Anlage 25, 60325 Frankfurt, Germany 7Department of Human Genetics, Leiden University Medical Center, Leiden 2300 RC, Netherlands 8Molecular Evolutionary Biology Group, Institute of Ecology, Diversity, and Evolution, Goethe University Frankfurt, Max-von-Laue Str. 13, Frankfurt am Main, 60438, Germany 1 bioRxiv preprint doi: https://doi.org/10.1101/810986; this version posted October 21, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. # Correspondence to: [email protected] Keywords: Metagenome-assembled genome, Sequencing error, Symbiosis, Chlorophyta, Mitochondria, Gene loss, organellar ploidy levels 2 bioRxiv preprint doi: https://doi.org/10.1101/810986; this version posted October 21, 2019.
    [Show full text]
  • Chromatic Adaptation in Lichen Phyco- and Photobionts
    Biologia 65/4: 587—594, 2010 Section Botany DOI: 10.2478/s11756-010-0058-y Chromatic adaptation in lichen phyco- and photobionts Bazyli Czeczuga,EwaCzeczuga-Semeniuk & Adrianna Semeniuk Department of General Biology, Medical University, Kili´nskiego 1,PL-15-089 Bialystok, Poland; e-mail: [email protected] Abstract: The effect of light quality on the photosynthetic pigments as chromatic adaptation in 8 species of lichens were examined. The chlorophylls, carotenoids in 5 species with green algae as phycobionts (Cladonia mitis, Hypogymnia physodes, H. tubulosa var. tubulosa and subtilis, Flavoparmelia caperata, Xanthoria parietina) and the chlorophyll a, carotenoids and phycobiliprotein pigments in 3 species with cyanobacteria as photobionts (Peltigera canina, P. polydactyla, P. rufescens) were determined. The total content of photosynthetic pigments was calculated according to the formule and particular pigments were determined by means CC, TLC, HPLC and IEC chromatography. The total content of the photosynthetic pigments (chlorophylls, carotenoids) in the thalli was highest in red light (genus Peltigera), yellow light (Xanthoria parietina), green light (Cladonia mitis) and at blue light (Flavoparmelia caperata and both species of Hypogymnia). The biggest content of the biliprotein pigments at red and blue lights was observed. The concentration of C-phycocyanin increased at red light, whereas C-phycoerythrin at green light. In Trebouxia phycobiont of Hypogymnia and Nostoc photobiont of Peltigera species the presence of the phytochromes was observed. Key words: carotenoids; chlorophylls; chromatic adaptation; lichens; photobionts; phycobiliproteins; phycobionts; phy- tochromes. Introduction Taking the above into account, we decided to in- vestigate the chromatic adaptation of common lichen Lichens can be found all over our planet, inhabiting species in the Knyszy´nska Forest to various light con- diverse biotopes and tolerating even extreme environ- ditions during the vegetative period.
    [Show full text]
  • International Journal of Scientific Research and Reviews
    Zachariah Sonia Anna et al., IJSRR 2018, 7(3), 1160-1169 Review article Available online www.ijsrr.org ISSN: 2279–0543 International Journal of Scientific Research and Reviews The Lichen Symbiosis: A Review Zachariah Sonia Anna* and Varghese K. Scaria Postgraduate and Research Department of Botany, SB College, Changanacherry, Kerala, INDIA ABSTRACT Organisms belonging to two different kingdoms are mutually associated to form a structure with particular morphology and anatomy. Lichens are not much eye- catching in its external morphology, but spectacular in its internal structure. Morphological studies don’t reveal the mutualistic associations of individual partners while anatomical study reveals the beauty of its symbiotic association. The corroboration of recent studies on lichen and their symbiotic association suggests that, other micro communities such as algae, fungi and bacterial bionts are associated with many lichens in addition to the main photobiont and mycobiont. Microbial association of lichen helps them to develop a stable and successful symbiotic life, which can adapt in its natural habitat. KEYWORDS: Lichen bionts, bacterial bionts, microbial community, symbiotic association *Corresponding author Sonia Anna Zachariah Department of Botany SB College, Changanacherry, Kerala- 686101 Email: [email protected], Mob. 9947841655 IJSRR, 7(3) July – Sep., 2018 Page 1160 Zachariah Sonia Anna et al., IJSRR 2018, 7(3), 1160-1169 INTRODUCTION 1. Lichen: A successful holobiont Lichens are holobionts with more than one participant in the association. They are usually comprised of a filamentous fungal partner called mycobiont and an algal partner called photobiont which can be either a eukaryotic chlorobiont (green algae) or a prokaryotic cyanobiont (cyanobacteria) and a very few are from other classes of algae.
    [Show full text]
  • Does the Lichen Alga Trebouseia Occur Free-Living in Nature: Further Immunological Evidence
    Symbiosis, 17 (1994) 247-253 247 Balaban, Philadelphia/Rehovot Does the Lichen Alga Trebouseia Occur Free-Living in Nature: Further Immunological Evidence A. MUKHTAR, J. GARTY* and M. GALUN Department of Botany tuul " Institute for Nature Conservation Research, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv 69978, Israel Tel. 972 3 6409163, Fax 972 3 6409380 Received November 29, 1994; Accepted December 19, 1994 Abstract Morphological and immunological methods have revealed free-living Trebouxia cells among the first settlers in an area that has been completely sterilized by a forest fire. These cells could be detected three years after the fire, and before any lichen colony had been established. We provide evidence that these free• living Trebouzia cells are identical by morphological and immunological criteria with the photobiont of Xanthoria parietina and of Buellia sp. that developed in another area of the same region which was exposed to a forest fire four years earlier. It appeared that the X. parietina colony examined contained two ( or more) different Trebouxios. Keywords: lichen, Trebouxia, photobiont, Buellia sp., immunoassay 1. Introduction De Bary (1887) claimed "that lichen algae have become adapted to sym• biosis with fungi and cannot exist in the free-living state." A consequence of this statement would be, that lichens reproduce only vegetatively, by dias• pores which contain both components - the fungus and the alga. There are indeed many lichen species with vegetative reproduction units, such as soredia and isidia (Jahns, 1988). However, a large number of foliose species and the ·0334-5114/94 /$05.50 @1994 Balaban 248 A.
    [Show full text]
  • Unravelling the Symbiotic Microalgal Diversity in Buellia Zoharyi (Lichenized Ascomycota) from the Iberian Peninsula and Balearic Islands Using DNA Metabarcoding
    diversity Article Unravelling the Symbiotic Microalgal Diversity in Buellia zoharyi (Lichenized Ascomycota) from the Iberian Peninsula and Balearic Islands Using DNA Metabarcoding Patricia Moya * , Salvador Chiva , Arantzazu Molins , Isaac Garrido-Benavent and Eva Barreno Botánica, ICBIBE, Fac. CC. Biológicas, Universitat de València, C/Dr. Moliner, 50, 46100 Valencia, Spain; [email protected] (S.C.); [email protected] (A.M.); [email protected] (I.G.-B.); [email protected] (E.B.) * Correspondence: [email protected]; Tel.: +34-963-544-376 Abstract: Buellia zoharyi is a crustose placodioid lichen, usually occurring on biocrusts of semiarid ecosystems in circum-Mediterranean/Macaronesian areas. In previous work, we found that this lichenized fungus was flexible in its phycobiont choice in the Canary Islands. Here we test whether geography and habitat influence phycobiont diversity in populations of this lichen from the Iberian Peninsula and Balearic Islands using Sanger and high throughput sequencing (HTS). Additionally, three thallus section categories (central, middle and periphery) were analyzed to explore diversity of microalgal communities in each part. We found that B. zoharyi populations hosted at least three different Trebouxia spp., and this lichen can associate with distinct phycobiont strains in different Citation: Moya, P.; Chiva, S.; Molins, habitats and geographic regions. This study also revealed that the Trebouxia composition of this A.; Garrido-Benavent, I.; Barreno, E. lichen showed significant differences when comparing the Iberian Peninsula with the Balearics thalli. Unravelling the Symbiotic Microalgal No support for differences in microalgal communities was found among thallus sections; however, Diversity in Buellia zoharyi several thalli showed different predominant Trebouxia spp.
    [Show full text]
  • Photosynthetic Carbon Acquisition in the Lichen Photobionts Coccomyxa and Trebouxia (Chlorophyta)
    PHYSIOLOGIA PLANTARUM 101: 67-76. 1997 Copyright © Physiologia Plantarum 1997 Printed in Dettmark - all rights reserved ISSN 0031-9317 Photosynthetic carbon acquisition in the lichen photobionts Coccomyxa and Trebouxia (Chlorophyta) Kristin Palmqvist, Asuncion de los Rios, Carmen Ascaso and Goran Samuelsson Palmqvist, K., de los Rios, A., Ascaso, C. and Samuelsson, G. 1997. Photosynthetic carbon acquisition in the lichen photobionts Coccomyxa and Trebouxia (Chlorophyta). - Physiol. Plant. 101: 67-76. Processes involved in photosynthetic CO2 acquisition were characterised for the iso- lated lichen photobiont Trebouxia erici (Chlorophyta, Trebouxiophyceae) and com- pared with Coccomyxa (Chlorophyta), a lichen photobiont without a photosynthetic COi-concentrating mechanism. Comparisons of ultrastructure and immuno-gold la- belling of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco; EC 4.1.1.39) showed that the chloroplast was larger in T. erici and that the majority of Rubisco was located in its centrally located pyrenoid. Coccomyxa had no pyrenoid and Rubisco was evenly distributed in its chloroplast. Both species preferred CO2 rather than HCO3 as an external substrate for photosynthesis, but T. erici was able to use CO2 concentra- tions below 10-12 \xM more efficiently than Coccomyxa. In T. erici, the lipid-insolu- ble carbonic anhydrase (CA; EC 4.2.1.1) inhibitor acetazolamide (AZA) inhibited photosynthesis at CO2 concentrations below 1 \xM, while the lipid-soluble CA inhibi- tor ethoxyzolamide (EZA) inhibited C02-dependent O2 evolution over the whole CO2 range. EZA inhibited photosynthesis also in Coccomyxa, but to a much lesser extent below 10-12 |iM CO2. The intemal CA activity of Trebouxia, per unit chlorophyll (Chi), was ca 10% of that of Coccomyxa.
    [Show full text]
  • Vulcanochloris (Trebouxiales, Trebouxiophyceae), a New Genus of Lichen Photobiont from La Palma, Canary Islands, Spain
    Phytotaxa 219 (2): 118–132 ISSN 1179-3155 (print edition) www.mapress.com/phytotaxa/ PHYTOTAXA Copyright © 2015 Magnolia Press Article ISSN 1179-3163 (online edition) http://dx.doi.org/10.11646/phytotaxa.219.2.2 Vulcanochloris (Trebouxiales, Trebouxiophyceae), a new genus of lichen photobiont from La Palma, Canary Islands, Spain LUCIE VANČUROVÁ1*, ONDŘEJ PEKSA2, YVONNE NĚMCOVÁ1 & PAVEL ŠKALOUD1 1Charles University in Prague, Faculty of Science, Department of Botany, Benátská 2, 128 01 Prague 2, Czech Republic 2The West Bohemian Museum in Pilsen, Kopeckého sady 2, 301 00 Plzeň, Czech Republic * Corresponding author (E-mail: [email protected]) Abstract This paper describes a new genus of lichen photobionts, Vulcanochloris, with three newly proposed species, V. canariensis, V. guanchorum and V. symbiotica. These algae have been discovered as photobionts of lichen Stereocaulon vesuvianum growing on slopes of volcanos and lava fields on La Palma, Canary Islands, Spain. Particular species, as well as the newly proposed genus, are delimited based on ITS rDNA, 18S rDNA and rbcL sequences, chloroplast morphology, and ultrastruc- tural features. Phylogenetic analyses infer the genus Vulcanochloris as a member of Trebouxiophycean order Trebouxiales, in a sister relationship with the genus Asterochloris. Our data point to the similar lifestyle and morphology of these two genera; however, Vulcanochloris can be well distinguished by a unique formation of spherical incisions within the pyrenoid. Mycobiont specificity and geographical distribution of the newly proposed genus is further discussed. Introduction The class Trebouxiophyceae, originally circumscribed by ultrastructural features as Pleurastrophyceae, is currently defined phylogenetically, predominantly by a similarity in 18S rDNA sequence data.
    [Show full text]
  • Green Microalga Trebouxia Sp. Produces Strigolactone
    bioRxiv preprint doi: https://doi.org/10.1101/195883; this version posted September 29, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 1 Green microalga Trebouxia sp. produces strigolactone- 2 related compounds 3 4 Smýkalová I.1, Ludvíková M.1, Ondráčková E.1, Klejdus B.2, Bonhomme S.3, Kronusová O.4, 5 Soukup A.5, Rozmoš M.6, Guzow-Krzemińska B.7, Matúšová R.8 6 7 1 Agritec Plant Research Ltd., Zemědělská 16, Šumperk, 787 01, Czech Republic; 8 2 Department of Chemistry and Biochemistry, Mendel University in Brno, Zemědělská 1, 9 Brno, 613 00 Brno, Czech Republic; 10 3 Institut Jean-Pierre Bourgin, INRA, AgroParisTech, CNRS, Université Paris-Saclay, Route 11 de St-Cyr 10, 780 26 Versailles Cedex, France. 12 4 Ecofuel Laboratories Ltd., Ocelářská 9, 190 02 Prague, Czech Republic; 13 5 Department of Experimental Plant Biology, Faculty of Science Charles University in 14 Prague, Viničná 5, 128 44 Prague 2, Czech Republic; 15 6 Institute of Experimental Biology, Faculty of Plant Science, Masaryk University, Kotlářská 16 267/2 , Brno, 611 37, Czech Republic; 17 7 Department of Plant Taxonomy and Narture Conservation, Faculty of Biology, University 18 of Gdańsk, Wita Stwosza 59, 80-308 Gdańsk, Poland; 19 8 Plant Science and Biodiversity Center SAS, Institute of Plant Genetics and Biotechnology, 20 Akademicka 2, PO Box 39A, 95007 Nitra, Slovak Republic; 21 22 Corresponding authors: [email protected]; [email protected] 23 Running title 24 Production of SLs in microalga 25 Highlight 26 In lichenized alga Trebouxia arboricola there are produced SLs-related compounds inducing 27 germination of parasitic weed Phelipanche aegyptiaca; T.
    [Show full text]
  • The Lichen Symbiosis Re-Viewed Through the Genomes of Cladonia
    Armaleo et al. BMC Genomics (2019) 20:605 https://doi.org/10.1186/s12864-019-5629-x RESEARCH ARTICLE Open Access The lichen symbiosis re-viewed through the genomes of Cladonia grayi and its algal partner Asterochloris glomerata Daniele Armaleo1* , Olaf Müller1,2, François Lutzoni1, Ólafur S. Andrésson3, Guillaume Blanc4, Helge B. Bode5, Frank R. Collart6, Francesco Dal Grande7, Fred Dietrich2, Igor V. Grigoriev8,9, Suzanne Joneson1,10, Alan Kuo8, Peter E. Larsen6, John M. Logsdon Jr11, David Lopez12, Francis Martin13, Susan P. May1,14, Tami R. McDonald1,15, Sabeeha S. Merchant9,16, Vivian Miao17, Emmanuelle Morin13, Ryoko Oono18, Matteo Pellegrini19, Nimrod Rubinstein20,21, Maria Virginia Sanchez-Puerta22, Elizabeth Savelkoul11, Imke Schmitt7,23, Jason C. Slot24, Darren Soanes25, Péter Szövényi26, Nicholas J. Talbot27, Claire Veneault-Fourrey13,28 and Basil B. Xavier3,29 Abstract Background: Lichens, encompassing 20,000 known species, are symbioses between specialized fungi (mycobionts), mostly ascomycetes, and unicellular green algae or cyanobacteria (photobionts). Here we describe the first parallel genomic analysis of the mycobiont Cladonia grayi and of its green algal photobiont Asterochloris glomerata.We focus on genes/predicted proteins of potential symbiotic significance, sought by surveying proteins differentially activated during early stages of mycobiont and photobiont interaction in coculture, expanded or contracted protein families, and proteins with differential rates of evolution. Results: A) In coculture, the fungus upregulated small secreted proteins, membrane transport proteins, signal transduction components, extracellular hydrolases and, notably, a ribitol transporter and an ammonium transporter, and the alga activated DNA metabolism, signal transduction, and expression of flagellar components. B) Expanded fungal protein families include heterokaryon incompatibility proteins, polyketide synthases, and a unique set of G- protein α subunit paralogs.
    [Show full text]
  • A Glimpse Into Genetic Diversity and Symbiont Interaction Patterns in Lichen Communities from Areas with Different Disturbance Histories in Białowie˙Zaforest, Poland
    microorganisms Article A Glimpse into Genetic Diversity and Symbiont Interaction Patterns in Lichen Communities from Areas with Different Disturbance Histories in Białowie˙zaForest, Poland Garima Singh 1,*, Martin Kukwa 2 , Francesco Dal Grande 1 , Anna Łubek 3, Jürgen Otte 1 and Imke Schmitt 1,4,* 1 Senckenberg Biodiversity and Climate Research Centre (SBiK-F), 60325 Frankfurt am Main, Germany 2 Department of Plant Taxonomy and Nature Conservation, Faculty of Biology, University of Gda´nsk, 80-308 Gda´nsk,Poland 3 Jan Kochanowski University in Kielce, Institute of Biology, 25-406 Kielce, Poland 4 Department of Biological Sciences, Institute of Ecology, Evolution and Diversity, Goethe Universität, 60325 Frankfurt am Main, Germany * Correspondence: [email protected] (G.S.); [email protected] (I.S.) Received: 12 April 2019; Accepted: 22 August 2019; Published: 9 September 2019 Abstract: Anthropogenic disturbances can have strong impacts on lichen communities, as well as on individual species of lichenized fungi. Traditionally, lichen monitoring studies are based on the presence and abundance of fungal morphospecies. However, the photobionts, as well photobiont mycobiont interactions also contribute to the structure, composition, and resilience of lichen communities. Here we assess the genetic diversity and interaction patterns of algal and fungal partners in lichen communities along an anthropogenic disturbance gradient in Białowie˙zaForest (Poland). We sampled a total of 224 lichen thalli in a protected, a managed, and a disturbed area of the forest, and sequenced internal transcribed spacer (ITS) ribosomal DNA (rDNA) of both, fungal and algal partners. Sequence clustering using a 97% similarity threshold resulted in 46 fungal and 23 green algal operational taxonomic units (OTUs).
    [Show full text]