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Transcriptome profile in naturally hydrated and desiccated fronds of Porphyra columbina(Plantae, Rhodophyta)

January 2012
BMC Genomics

January 2012

Authors:

Loretto Contreras-Porcia

Universidad Andrés Bello

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Desiccation tolerance in Pyropia columbina 933

Lat. Am. J. Aquat. Res., 41(5): 933-958, 2013

DOI: 103856/vol41-issue5-fulltext-13

Research Article



Differential gene expression in Pyropia columbina (Bangiales, Rhodophyta)

under natural hydration and desiccation conditions

Loretto Contreras-Porcia1, Camilo López-Cristoffanini1,2, Carlos Lovazzano1

María Rosa Flores-Molina3, Daniela Thomas1, Alejandra Núñez1, Camila Fierro1, Eduardo Guajardo1

Juan A. Correa2, Michael Kube4 & Richard Reinhardt5

1Departamento de Ecología y Biodiversidad, Facultad de Ecología y Recursos Naturales

Universidad Andres Bello, República 470, Santiago, Chile

2Departamento de Ecología, Center for Advanced Studies in Ecology and Biodiversity (CASEB)

Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile

Postal code 6513677, Santiago, Chile

3Instituto de Ciencias Marinas y Limnológicas, Facultad de Ciencias, Universidad Austral de Chile

P.O. Box 567, Valdivia, Chile

4Department of Crop and Animal Sciences, Faculty of Agriculture and Horticulture Humboldt

Universität zu Berlin, Lentzeallee 55/57, 14195 Berlin, Germany

5Max-Planck Institute for Molecular Genetics, Ihnestr. 63-73, Berlin, Germany

ABSTRACT. In rocky shores, desiccation is triggered by daily tide changes, and experimental evidence

suggests that local distribution of algal species across the intertidal rocky zone is related to their capacity to

tolerate desiccation. In this context, the permanence of Pyropia columbina in the high intertidal rocky zone is

explained by its exceptional physiological tolerance to desiccation. This study explored the metabolic

pathways involved in tolerance to desiccation in the Chilean P. columbina, by characterizing its transcriptome

under contrasting conditions of hydration. We obtained 1,410 ESTs from two subtracted cDNA libraries in

naturally hydrated and desiccated fronds. Results indicate that transcriptome from both libraries contain

transcripts from diverse metabolic pathways related to tolerance. Among the transcripts differentially

expressed, 15% appears involved in protein synthesis, processing and degradation, 14.4% are related to

photosynthesis and chloroplast, 13.1% to respiration and mitochondrial function (NADH dehydrogenase and

cytochrome c oxidase proteins), 10.6% to cell wall metabolism, and 7.5% are involved in antioxidant activity,

chaperone and defense factors (catalase, thioredoxin, heat shock proteins, cytochrome P450). Both libraries

highlight the presence of genes/proteins never described before in algae. This information provides the first

molecular work regarding desiccation tolerance in P. columbina, and helps, to some extent, explaining the

classical patterns of ecological distribution described for algae across the intertidal zone.

Keywords: Pyropia, desiccation stress, ESTs, seaweeds, transcriptomics, proteins.

Expresión diferencial de genes en Pyropia columbina (Bangiales, Rhodophyta)

bajo hidratación y desecación natural

RESUMEN. En zonas rocosas costeras, la desecación es gatillada por cambios diarios en los niveles de

marea, y la evidencia experimental indica que la distribución de las algas en la zona intermareal está

relacionada con su capacidad para tolerar la desecación. En este contexto, la presencia de Pyropia columbina

en la zona alta del intermareal se explica por su excepcional tolerancia fisiológica a la desecación. Este estudio

explora las vías metabólicas involucradas en la tolerancia a la desecación en P. columbina, a través de la

caracterización de su transcriptoma bajo condiciones de hidratación contrastantes. Se obtuvo 1,410 ESTs

provenientes de dos librerías de substracción de cDNA de frondas naturalmente hidratadas y desecadas. Los

transcriptomas de ambas librerías contienen transcritos de diversas rutas metabólicas relacionadas a la

tolerancia. Entre los transcritos expresados 15% están involucrados en la síntesis de proteínas, su

procesamiento y degradación, 14,4% asociados a fotosíntesis y cloroplasto, 13,1% a respiración y función

mitocondrial, 10,6% al metabolismo de la pared celular y 7,5% a la actividad antioxidante, proteínas

chaperonas y factores de defensa (catalasa, tiorredoxina, proteínas de shock térmico, citocromo P450). En

ambas librerías se destaca la presencia de genes/proteínas no descritos en algas. Esta información proporciona

934 Latin American Journal of Aquatic Research

el primer trabajo molecular que estudia la tolerancia a desecación en P. columbina y sus resultados ayudan a

explicar los patrones clásicos de distribución descritos para algas en la zona intermareal.

Palabras clave: Pyropia, estrés por desecación, ESTs, macroalgas, transcriptómica, proteínas.

___________________

Corresponding author: Loretto Contreras-Porcia (lorettocontreras@unab.cl)

INTRODUCTION

Red algae (Rhodophyta), the most ancient lineage of

photosynthetic eukaryotes (Baldauf et al., 2000; Yoon

et al., 2004), are distributed worldwide and include

several commercially important species. Porphyra and

Pyropia spp. are rhodophytes that represent an annual

value of over US$1.3 billion (Blouin et al., 2011). In

Chile, Pyropia columbina (Montagne) W.A. Nelson

(formerly Porphyra columbina (Sutherland et al.,

2011) is one of the economically important species,

together with members of the rhodophycean genera

Mazzaella, Gracilaria and Gelidium (Santelices, 1989;

Hoffmann & Santelices, 1997; Buschmann et al.,

2008), and it is found along the Chilean coast from

20° to 54°S (Hoffmann & Santelices, 1997; Guiry &

Guiry, 2013). This species has a biphasic life history

that includes a microscopic sporophyte generation (2n,

conchocelis stage) alternating with a macroscopic

generation of male and female gametophytes (n). The

foliose gametophytes constitute the edible “Nori”.

Water, and its intracellular balance, is a critical

factor for all living organisms in both terrestrial and

marine ecosystems. Mobile animals actively avoid

desiccation, induced by water deficiency, while other

organisms, such as resurrection plants ―a small group

of angiosperms that live in the most arid habitats of

the world― are adapted to tolerate water losses of up

to 90% (Gaff, 1987). This adaptation, in general

terms, is based on the ability of an organism to

equilibrate its internal water potential with the dry

environment, and re-start normal functions when re-

hydrated (Alpert, 2000). Several studies using

resurrection plants as model have been conducted to

fully understand their impressive adaptation to

desiccation (Scott, 2000). Recent advances in our

understanding of the mechanisms of tolerance in these

organisms have revealed changes at the morphological

level, osmolites and protein synthesis, and a decline in

ROS (reactive oxygen species) production and

photosynthesis rate (Ingram & Bartels, 1996; Hoekstra

et al., 2001; Bernacchia & Furini, 2004; Vicré et al.,

2004; Dinakar et al., 2012). More specifically, it has

been observed that the plant hormone ABA is

accumulated in desiccated resurrection plants, which

induces the expression of several proteins related to

desiccation (Bartels et al., 1990; Dinakar et al., 2012).

Also, leaves of resurrection plants tend to curl to

reduce water loss and minimize oxidative damage due

to desiccation (Vicré et al., 2004; Farrant et al., 2007;

Toldi et al., 2009). Oxidative damage is attenuated or

avoided by increasing antioxidant activity of some

enzymes (e.g., ascorbate peroxidase, glutathione

reductase, superoxide dismutase, among others) and

levels of antioxidant compounds (e.g., anthocyanins)

(Farrant et al., 2007; Toldi et al., 2009; Dinakar et al.,

2012). Additionally, these plants reduce photo-

synthetic activity to minimize photo-oxidative damage

that could lead to increased ROS levels. Thus, it seems

clear that diverse metabolic pathways are involved in

attenuating the oxidative stress condition caused by

desiccation.

In the Chilean coastal ecosystems, P. columbina

grows abundantly along the upper intertidal zone

(Alveal, 1970; Santelices, 1989; Hoffmann &

Santelices, 1997), where it is exposed to a wide range

of environmentally stressful conditions, mainly

desiccation-driven stress induced by low tides and air

exposure (Contreras-Porcia et al., 2011a). P.

columbina is well adapted to daily extremes, which

range from exposure to water (full hydration) during

high tides to long exposure to air (maximum

desiccation) during low tides. Natural exposure to

these extreme regimes have been described in other

organisms exposed to desiccation. It is already known

that Porphyra and Pyropia species have high tolerance

to desiccation, and quickly recover photosynthetic

activity once rehydrated after a period of desiccation

(e.g., Smith & Berry, 1986; Kim et al., 2008;

Contreras-Porcia et al., 2011a; Gao & Wang, 2012).

Basic physiology of these organisms, including the

mechanisms to tolerate environmentally stressful

conditions, remains poorly studied. However, it is

known that, in P. columbina, desiccation induces

losses ca. 96% of the water content and enhanced

significantly the production of ROS (Contreras-Porcia

et al., 2011a). The quick return of ROS to their basal

levels during high tide is explained by an efficient

activation of the antioxidant system. In comparison,

species inhabiting the lower intertidal zone [e.g.,

Desiccation tolerance in Pyropia columbina 935

Mazzaella laminarioides (Bory de Saint-Vincent)

Fredericq, Ulva compressa Linnaeus and Lessonia

spicata (Suhr) Santelices] are more sensitive to

desiccation, and this seems related to the absence of

efficient mechanisms to attenuate the over-production

of ROS during rehydration (Contreras-Porcia et al.,

2012; López-Cristoffanini et al., 2013). Flores-Molina

et al. (unpublished data) recently provided some

physiological and biochemical bases that help explain

the role of desiccation on species distribution across

the intertidal zone. They reported that sensitive

species displayed i) inactivation of antioxidant

enzymes, ii) over-oxidation of biomolecules, and iii)

inactivation of the photosystem II. For example, Ulva

compressa (Chlorophyta, Plantae) and Scytosiphon

lomentaria (Lyngbye) Link (Ochrophyta, Chromista)

inhabiting the mid-intertidal zone have lower

tolerance to desiccation than P. columbina. However,

they tolerate desiccation better than Lessonia spicata

(Ochrophyta, Chromista) and Gelidium rex Santelices

& I.A. Abbott (Rhodophyta, Plantae), both lower

intertidal species.

Given the ecological and economic relevance of

Pyropia and that some basic biochemical and

physiological information on the mechanisms

involved in tolerance to desiccation is known, we

focused this work in determining the genes/proteins

that are differentially expressed in P. columbina

during the hydration-desiccation cycle, using

Suppression Subtractive Hybridization (SSH) and

ESTs determined by pyrosequencing (454 Life

Sciences, Roche). The results of this study will help to

elucidate the genetic basis underlying the high

tolerance to desiccation displayed by this species, and

will broaden our knowledge of the molecular

biology/ecology of this organism and other macroalgal

species.

MATERIALS AND METHODS

Ethics statement

No specific permits were required for the described

field studies. The study area is unrestricted to public

access and use, and is not privately owned or

designated as a protected area (reserves or parks). No

protected or endangered species were involved in this

study. Fronds of P. columbina were collected along

200-300 m of coastline during high (naturally

hydrated plants, 100% relative water content (RWC))

and low tide (naturally desiccated plants, ca. 4%

RWC) in Maitencillo beach, Valparaiso (32º39.5´S,

71º26.6´W). After collection, fronds were quickly

rinsed (15-20 s) in 0.22 µm-filtered seawater,

manually cleaned and frozen on site with liquid

nitrogen.

RNA extraction

Total RNA was isolated from 20-30 g of fresh tissue

of naturally hydrated and desiccated, pooled fronds of

P. columbina. Tissue, frozen in liquid nitrogen, was

homogenized in 25 mL of lysis buffer containing 4 M

guanidinium thiocyanate, 25 mM EDTA, 200 mM

sodium acetate, 2% polyvinylpyrrolidone (PVP-40)

and 1% 2-mercaptoethanol. The homogenate was

incubated for 10 min at 70°C with constant agitation

in the presence of 20% sarcosin, and then centrifuged

for 5 min at 16000 g. The RNA present in the

supernatant solution was purified and re-extracted

using an RNeasy mini Kit (Qiagen, Hilden, Germany),

according to the manufacturer protocol. RNA quality

and yield was assessed by spectrophotometry

(NanoDropTM 1000 Spectrophotometer, Thermo

Scientific, DE, USA) and denaturing agarose gel

electrophoresis. Finally, the mRNA was obtained from

the total RNA extracted (ca. 290-300 µg) using

DynaBeads (Invitrogen, Oregon, USA). Prior to RNA

extraction, all material was treated in 0.1% DEPC

water.

Preparation of the cDNA libraries by SSH and

next-generation sequencing

The synthesis of cDNAs, from both natural conditions

(i.e., natural hydration and desiccation stress), were

obtained using a SMARTTM cDNA Library

Construction Kit (Clontech, Mountain View, CA,

USA), as described in Wellenreuther et al. (2004).

Then, two subtracted cDNA libraries were

constructed: one with genes expressed exclusively

under hydration (UH) and the other by those

expressed under desiccation stress (UD), as described

in Diatchenko et al. (1996). cDNAs were then

sequenced by 454-pyrosequencing (Margulies et al.,

2005). cDNAs were ligated to 454 self-made adaptors

with Multiplex Identifier Adaptors (MIDs) for the GS

FLX Titanium Chemistry following Roche's technical

bulletin TCB 09004 introducing SfiI-sites. The 454

libraries were immobilized on beads and clonally

amplified using a "GS FLX Titanium LV emPCR

Kit". The libraries were then sequenced using a "GS

FLX Titanium Sequencing Kit XLR70" and a "GS

FLX Titanium PicoTiterPlate Kit". All kits were

purchased from Roche and used according to the

manufacturer protocols.

Data assembly and bioinformatics analysis

Readings from both libraries were processed by self-

written Perl scripts and assembled into coatings, i.e.,

936 Latin American Journal of Aquatic Research

representing putative transcripts using MIRA 3

assembly (Contreras-Porcia et al., 2011b). The

expressed sequences tags (ESTs) determined from

both libraries were subjected to separated bioinfor-

matics analyses. ESTs were analyzed for sequence

similarities using the BLASTX program (NCBI, MD,

USA). Reading frames with the highest sequence

similarity scores were used to analyze protein identity

using the BLASTP program. Threshold values were

set above 50 for high-scoring segment pairs, with a

minimum significance at least of e ≤ 10-4 and an

identity higher than 30%. ESTs coding for known

proteins were classified into functional categories by

the KO (KEGG Orthology) database for ortholog

grouping and hierarchical classification of genes,

according their functionality (Kanehisa et al., 2004)

by using the BLAST2GO software (Götz et al., 2008).

Finally, the putative subcellular localization of the

proteins was determined using the TARGETP

(http://www.cbs.dtu.dk/services/TargetP Emanuelsson

et al., 2000), WOLFPSORT (http://psort.ims.u-

tokyo.ac.jp), ChloroP (http://www.cbs.dtu.dk/services/

ChloroP Emanuelsson et al., 1999) and PSORTb

(http://www.psort.org/psortb Yu et al., 2010) servers.

RESULTS

EST sequencing and assembly

A total of 8,054 sequence reads were obtained from

the library enriched with hydration-responsive

exclusive transcripts (UH library, Table 1). Moreover,

8,432 sequence reads were obtained from the library

enriched with desiccation-responsive exclusive

transcripts (UD library). In the UH library, 49.2% of

ESTs ranged from 200 to 500 bp, with an average size

of 423 bp. In the UD library, 46% of the ESTs ranged

from 200 to 500 bp, with an average size of 385 bp.

The rest of the transcripts in both libraries ranged

between 100 to 190 bp. The sequences of both

libraries are available on the EMBL Nucleotide

Sequence Database (http://www.ebi.ac.uk/ embl) with

accession numbers HE858615 to HE859412 for UH

and HE859413 to HE859937 for UD libraries.

Gene ontology from naturally hydrated and

desiccated P. columbina transcriptome

Almost 59% of the total ESTs (i.e., 491) from UH

displayed no similarity with sequences available in

databases; whereas the remaining 41% (i.e., 347) were

similar to registered proteins (Table 1). Similarities

(68.8% of them) concentrated in sequences reported

for Arthropoda (e.g., Culex quinquefasciatus),

Chordata (e.g. Danio rerio) and Mollusca (e.g.,

Littorina saxatilis). Another 18% of the total ESTs

were similar to proteins from the kingdom Plantae,

mainly Rhodophyta (e.g., Porphyra purpurea) and

Tracheophyta (e.g., Vigna unguiculata), and 10.7% to

the kingdom Bacteria, mainly Proteobacteria (e.g.,

Haemophilus influenzae) and Cyanobacteria (e.g.,

Microcystis aeruginosa). Only 2.5% of the sequences

were similar to proteins from Protozoa (i.e.,

Dictyostelium discoideum), Fungi (i.e.,

Saccharomyces cerevisiae) and Chromista (i.e.,

Pylaiella littoralis). Finally, analysis of the amino

terminal sequences showed that 51.8% of the

identified proteins were potentially assignable to the

cytosol, 28.6% to mitochondria, 13% to the

chloroplast, and 4.4% to the nucleus (Table 1).

In the UD library, 72% of the total ESTs (i.e., 412)

displayed no similarity with previously reported

sequences (Table 2). Of the remaining 28% (i.e., 160

ESTs) showing similarity with proteins registered in

databases: 37% were similar to those reported for

Plantae, mainly Rhodophyta (e.g., Porphyra purpu-

rea) and Tracheophyta (e.g., Arabidopsis thaliana),

29.4% with proteins from Bacteria, mainly

Proteobacteria (e.g., Burkholderia multivorans) and

Cyanobacteria (e.g., Thermosynechococcus elonga-

tus), 21% with proteins from Animalia, mainly

Arthropoda (e.g., Aedes aegypti), Chordata (e.g.,

Mauremys mutica) and Mollusca (e.g., Crassostrea

virginica), and 7.5% with proteins from Chromista,

mainly Ochrophyta (e.g., Ectocarpus siliculosus).

Only 5% of the sequences were similar to proteins

from Protozoa (i.e., Dictyostelium discoideum) and

Fungi (i.e., Leptosphaeria maculans). Finally, analysis

of the amino terminal sequences showed that 47% of

the identified proteins were potentially assignable to

cytosol, 21-23% to both mitochondria and chloroplast,

and 7% to the nucleus (Table 2).

Functional categorization

The identified genes/proteins were classified into

thirteen functional categories according their

functionality (Fig. 1). For example, most ESTs from

hydrated fronds (Table 1) matched with proteins

involved in protein synthesis, processing and

degradation (ca. 24%, peptidylprolyl cis-trans

isomerases, ubiquitin and proteasome proteins),

respiration and mitochondria (ca. 14%), antioxidant

function, chaperone and defense factors (ca. 9%, e.g.,

peroxiredoxin (PRX), arachidonate 5-lipoxygenase, a

glutathione S-transferase, and several cytochrome

P450 and HSPs), cell motility (ca. 9%) and basal

metabolism (ca. 8%; e.g., glyceraldehyde 3-phosphate

dehydrogenase and L-lactate dehydrogenase). However,

the transcriptome induced by desiccation (Fig. 1)

Desiccation tolerance in Pyropia columbina 937

Table 1. Functional category, identity and potential cellular destination of proteins, and accession number of identified

ESTs in P. columbina under natural hydration (UH library). E value: the best (lowest) Expect value (E value) of all

alignments from that database sequence, D*: Putative destination.

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

1. Signal transduction Phosphatidylinositol 3-kinase 1 Culex quinquefasciatus /

XP_001847784.1 4.00E-07 nuc HE859298

MAP kinase activated protein-kinase-2 Glossina morsitans morsitans /

ABC25082.1 2.00E-47 cyt HE859158

Low-density lipoprotein receptor-

related protein 1B-like

Xenopus (Silurana) tropicalis /

XP_002937402.1 5.00E-14 cyt HE858680

Low-density lipoprotein receptor-

related protein 2 precursor

Danio rerio / NP_001181916.1 2.00E-19 cyt HE858962

Mitogen-activated protein kinase

kinase kinase 15

Apis mellifera / XP_003250315.1 4.00E-19 cyt HE859035

Neurochondrin Drosophila melanogaster /

NP_649658.1 4.00E-20 n.d. HE859071

Neurochondrin-like protein Harpegnathos saltator /

EFN88263.1 2.00E-16 n.d. HE859267

Serine/threonine kinase Culex quinquefasciatus /

XP_001844678.1 6.00E-33 nuc HE859315

Protein serine/threonine kinase,

putative

Aedes aegypti / XP_001660772.1 2.00E-21 nuc HE858859

Ras-related protein, isoform B Drosophila melanogaster /

NP_726881.1 4.00E-07 nuc HE859339

Serine/threonine kinase receptor-

associated protein

Glossina morsitans morsitans /

ADD19360.1 3.00E-70 nuc HE858825

Serine/threonine protein kinase Aedes aegypti / XP_001655486.1 5.00E-44 cyt HE859285

Serine/threonine-protein phosphatase 5 Culex quinquefasciatus /

XP_001850926.1 4.00E-45 cyt HE859130

SNF4/AMP-activated protein kinase

gamma subunit, isoform M

Drosophila melanogaster /

NP_001097854.1 4.00E-26 cyt HE858743

Troponin C type IIIa Apis mellifera / NP_001011651.1 9.00E-21 mit HE859041

Calmodulin Pyropia yezoensis / ABN41559.1 2.00E-52 cyt HE858656

Cyclic AMP phosphoprotein Scophthalmus maximus /

ABJ98640.1 1.00E-10 cyt HE858976

GAF sensor signal transduction

histidine kinase

Microcoleus vaginatus FGP-2

/ ZP_08492202.1 5.00E-18 cyt HE858774

Guanine nucleotide binding protein

beta polypeptide 2- like 1

Lethenteron camtschaticum/

BAE93065.1 9.00E-109 cyt HE858703

Putative guanine nucleotide binding

protein beta polypeptide 2-like 1

protein

Ovis aries / ABY75292.1 2.00E-43 cyt HE858908

Troponin T Pediculus humanus corporis /

XP_002424248.1 3.00E-31 cyt HE859343

Calreticulin Eisenia andrei / ABI74618.1 6.00E-42 mit HE859085

Adenylyl cyclase associated protein Gossypium herbaceum subsp.

africanum / ADZ16114.1 2.00E-22 cyt HE858826

Transcription factor BTF3 homolog 4 Salmo salar / ACI69109.1 5.00E-45 nuc HE858889

2. Transcription, splicing,

and replication

Transcription factor Brugia malayi / XP_001891758.1 4.00E-26 nuc HE858778

TFIIH basal transcription factor

complex P44 subunit

Culex quinquefasciatus /

XP_001845822.1 2.00E-07 nuc HE858942

RNA polymerase beta subunit Pyropia yezoensis / YP_536931.1 1.00E-21 chl HE858844

Homeotic protein spalt-major Camponotus floridanus /

EFN74164.1 3.00E-12 n.d. HE859115

Nascent polypeptide associated

complex protein alpha subunit

Glossina morsitans morsitans /

ADD19571.1 8.00E-17 n.d. HE858969

Similar to nuclear histone binding

protein

Strongylocentrotus purpuratus /

XP_791433.2 1.00E-11 nuc HE858932

orf544 reverse transcriptase Porphyra purpurea / NP_049292.1 6.00E-47 mit HE858715

Reverse transcriptase al1 Saccharomyces cerevisiae S288c /

NP_009310.1 1.00E-12 mit HE858909

Reverse transcriptase homolog Pylaiella littoralis / BAG06161.1 2.00E-05 mit HE858919

RNA-directed DNA polymerase Moorea producens 3L /

ZP_08427361.1 6.00E-05 mit HE859330

938 Latin American Journal of Aquatic Research

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

RNA-directed DNA polymerase Microcystis aeruginosa NIES-843

/ YP_001659893.1 3.00E-19 mit HE859084

RNA-directed DNA polymerase

(reverse transcriptase)

Microcoleus vaginatus FGP-2 /

ZP_08493544.1 7.00E-05 mit HE859136

RNA-directed DNA polymerase

(reverse transcriptase)

Arthrospira maxima CS-328 /

ZP_03271771.1 2.00E-11 mit HE858655

Retrotransposon protein Oryza sativa Indica Group /

ABR26094.1 5.00E-14 nuc HE858617

Retrotransposon protein Oryza sativa Indica Group /

ABR26094.1 3.00E-09 mit HE859408

RNA-binding protein 8A Apis mellifera / XP_395245.2 2.00E-35 cyt HE859164

rRNA intron-encoded homing

endonuclease

Thalassiosira pseudonana

CCMP1335 / XP_002294430.1 8.00E-12 mit HE858662

Ribonuclease R Weeksella virosa DSM 16922 /

YP_004238302.1 3.00E-23 cyt HE858827

rRNA promoter binding protein Brugia malayi / XP_001891797.1 7.00E-05 mit HE859073

ATP-dependent helicase Zobellia galactanivorans

/ CAZ98388.1 5.00E-14 n.d. HE859151

rRNA intron-encoded homing

endonuclease

Medicago truncatula /

XP_003614389.1 2.00E-05 cyt HE858668

Metal-dependent RNase Streptococcus pyogenes M49 591 /

ZP_00366321.1 2.00E-20 cyt HE858664

3. Basal metabolism Mitochondrial Malate dehy

drogenase 2

Culex quinquefasciatus /

XP_001849862.1 2.00E-26 mit HE858747

Glyceraldehyde 3-phosphate

dehydrogenase

Simulium nigrimanum /

ACZ28396.1 4.00E-05 cyt HE858837

Glyceraldehyde 3-phosphate

dehydrogenase

Tribolium castaneum /

XP_974181.1 5.00E-32 cyt HE858850

Pyruvate dehydrogenase Aedes aegypti / XP_001648922.1 3.00E-94 mit HE859341

Medium chain acyl-coenzyme A

dehydrogenase

Pachycara brachycephalum /

AEC32921.1 3.00E-34 mit HE859282

L-lactate dehydrogenase-like Apis mellifera / XP_394662.4 3.00E-37 cyt HE858775

Acetyl-CoA carboxylase beta subunit Porphyra purpurea / NP_053808.1 7.00E-33 chl HE859176

NAD-dependent

epimerase/dehydratase

Zea mays / NP_001148959.1 1.00E-13 cyt HE859024

Phosphogluconate dehydrogenase Tipula abdominalis / ACH95392.1 5.00E-35 cyt HE859302

Phosphoglycerate mutase Porphyra purpurea / NP_053989.1 7.00E-34 chl HE858842

Mitochondrial Succinyl-CoA ligase

[ADP-forming] subunit beta

Camponotus floridanus /

EFN67099.1 5.00E-43 mit HE859064

Succinyl-CoA ligase [GDP-forming]

subunit alpha

Anolis carolinensis /

XP_003221906.1 2.00E-10 mit HE858963

Transcript antisense to ribosomal RNA

protein 1(Tar1p)

Saccharomyces cerevisiae S288c /

NP_690845.1 7.00E-06 mit HE858643

Trehalose-6-phosphate synthase Aedes aegypti / XP_001657813.1 1.00E-22 cyt HE858687

Trehalose-6-phosphate synthase 1 Culex quinquefasciatus /

XP_001850996.1 2.00E-06 cyt HE858920

2,5-diketo-D-gluconic acid reductase

Rhodococcus erythropolis SK121 /

ZP_04383522.1 2.00E-05 cyt HE859383

3,4-dihydroxy-2-butanone-4-

phosphate synthase

Haemophilus influenzae HK1212 /

ZP_06222660.1 5.00E-16 cyt HE858684

3,4-dihydroxy-2-butanone-4-

phosphate synthase

Haemophilus influenzae HK1212 /

ZP_06222660.1 1.00E-13 cyt HE858659

3,4-dihydroxy-2-butanone-4-

phosphate synthase

Haemophilus influenzae HK1212 /

ZP_06222660.1 3.00E-20 cyt HE858708

Amylo-1, 6-glucosidase, 4-alpha-

glucanotransferase

Danio rerio / NP_001166124.1 2.00E-39 cyt HE858838

Arginine kinase Philaethria wernickei /

ACZ26817.1 1.00E-42 cyt HE858779

Arginine kinase Glossina morsitans morsitans /

ADD19663.1 6.00E-67 cyt HE859110

Putative enolase Lutzomyia longipalpis /

ABV60328.1 3.00E-39 mit HE858686

Putative enolase Myrmecocystus depilis /

ABY55706.1 5.00E-08 mit HE859204



Desiccation tolerance in Pyropia columbina 939

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

Glutamate dehydrogenase Bombyx mori / NP_001040245.1 3.00E-43 mit HE859167

Glycerol kinase Candidatus Liberibacter

americanus str. Sao Paulo /

ACD87749.1

4.00E-22 mit HE858676

Short-chain dehydrogenase Aedes aegypti / XP_001663676.1 1.00E-17 cyt HE859135

4. Antioxidant,

chaperone and defense

factors

Peroxiredoxin (PRX) family, typical

2-Cys

Pyropia yezoensis / YP_536954.1 3.00E-20 chl HE858809

Arachidonate 5-lipoxygenase Strongylocentrotus purpuratus /

XP_001198535.1 1.00E-13 cyt HE858870

Glutathione S-transferase sigma 5 Locusta migratoria / AEB91977.1 7.00E-41 n.d. HE858928

28 kDa heat- and acid-stable

phosphoprotein

Harpegnathos saltator /

EFN84845.1 2.00E-13 cyt HE858971

Apolipoprotein D Culex quinquefasciatus /

XP_001848365.1 2.00E-43 n.d. HE858692

Probable Bax Inhibitor 1 Harpegnathos saltator /

EFN76935.1 3.00E-14 n.d. HE859251

Ferritin heavy chain-like Ailuropoda melanoleuca /

XP_002929594.1 1.00E-06 cyt HE859190

Ferritin heavy chain-like protein Phlebotomus papatasi /

ABV44737.1 1.00E-33 cyt HE858813

Small heat shock protein IbpA Trichinella pseudospiralis /

ABJ55915.1 2.00E-28 mit HE858802

Heat shock protein 23 beta Ceratitis capitata / ACG58884.1 1.00E-05 cyt HE858848

Heat shock protein 27 Drosophila melanogaster /

NP_524000.1 7.00E-05 cyt HE859216

Heat shock protein 27 Drosophila bipectinata /

AEB40325.1 5.00E-15 cyt HE858649

Heat shock protein 27 Drosophila parabipectinata /

AEB40323.1 5.00E-08 cyt HE858750

Heat shock protein 27 Ceratitis capitata / ACD76913.1 2.00E-29 cyt HE859304

Heat shock protein 40 Bactrocera dorsalis / ADO30472.1 6.00E-17 n.d. HE858885

Heat shock protein 70 Porphyra purpurea / NP_053925.1 5.00E-71 chl HE858621

Heat shock protein 70 Pyropia haitanensis / Q06W39.1 5.00E-26 chl HE858874

Heat shock protein 70 Pyropia haitanensis / ACF71814.1 5.00E-36 cyt HE858890

Heat shock protein 70 Ostreococcus lucimarinus /

XP_001417572.1 6.00E-65 n.d. HE858770

Heat shock protein 70 Raphidiopsis brookii D9 /

ZP_06305357.1 9.00E-10 cyt HE859296

Heat shock protein 70B Dunaliella salina / ACJ24805.1 3.00E-12 cyt HE859318

Chloroplast Heat shock protein 70 Pyropia yezoensis / ABF54971.1 2.00E-47 chl HE858982

Heat shock protein 70 Gracilaria tenuistipitata var. liui /

YP_063608.1 4.00E-64 chl HE858640

Heat shock protein 70- type chaperone

(chloroplast)

Guillardia theta / AAC35702.1 7.00E-54 chl HE859033

Heat shock protein 70C Ascaris suum / ADY47984.1 3.00E-36 cyt HE858821

Heat shock protein 82 Philodina roseola / ACC43981.1 8.00E-20 n.d. HE859067

Heat shock protein 90 Spodoptera exigua / ACQ78181.1 9.00E-84 cyt HE859242

Heat shock protein 90 Thitarodes pui/ ADA61011.1 2.00E-43 n.d. HE858745

Stress-induced-phosphoprotein 1 Ciona intestinalis /

XP_002128875.1 3.00E-28 cyt HE858818

Cytochrome P450 4C1-like Acyrthosiphon pisum /

XP_001945361.2 8.00E-19 mit HE859003

Cytochrome P450 like_TBP Citrullus lanatus / BAD26579.1 1.00E-12 mit HE858931

Putative cytochrome P450 like protein

precursor

Phillyrea latifolia / CAK18871.1 8.00E-19 mit HE858629

5. Protein synthesis,

processing and

degradation

20S proteasome alpha subunit Scylla paramamosain /

ACY66486.1 4.00E-34 cyt HE859122

26S protease regulatory subunit 8 Acromyrmex echinatior /

EGI70168.1 7.00E-57 cyt HE858682

26S proteasome non-ATPase

regulatory subunit

Aedes aegypti / XP_001662445.1 2.00E-28 cyt HE859333

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940 Latin American Journal of Aquatic Research

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

26S proteasome non-ATPase

regulatory subunit 14

Aedes albopictus / ADB43603.1 2.00E-47 cyt HE858896

26S proteasome regulatory complex

subunit RPN5/PSMD12

Glossina morsitans morsitans /

ADD19088.1 5.00E-28 cyt HE859277

Proteasome subunit beta type 3 Culex quinquefasciatus /

XP_001843139.1 1.00E-33 cyt HE859180

Proteasome subunit alpha type 3 Oncorhynchus mykiss /

ACO07671.1 2.00E-19 cyt HE859121

Eukaryotic translation initiation

factor 3

Culex quinquefasciatus /

XP_001851152.1 3.00E-37 cyt HE859058

Eukaryotic translation initiation factor

3 subunit 10

Culex quinquefasciatus /

XP_001844367.1 6.00E-23 cyt HE859324

Putative 23S ribosomal RNA Vigna unguiculata / CAO02532.1 4,93E-09 cyt HE858807

Putative ribosomal protein S3 Vigna unguiculata / CAO02550.1 8.00E-09 cyt HE858624

Ribosomal protein S8 Polaribacter irgensii 23-P /

ZP_01119260.1 2.00E-28 cyt HE858921

30S ribosomal protein S10 Polaribacter sp.MED152 /

ZP_05108264.1 5.00E-06 cyt HE859208

Ribosomal protein RPS3a Eurythoe complanata /

ABW23199.1 2.00E-68 mit HE858801

40S ribosomal protein S4 Mytilus edulis / ABA55738.1 9.00E-47 cyt HE859005

Ribosomal protein S9 Haliotis discus discus /

ABX26127.1 5.00E-62 mit HE858760

Ribosomal protein S13 Xenopus (Silurana) tropicalis /

NP_001016602.1 1.00E-62 cyt HE859014

Ribosomal protein S15 isoform A Lysiphlebus testaceipes /

AAX62477.1 7.00E-56 cyt HE858833

40S ribosomal protein S16 Glossina morsitans morsitans /

ADD20564.1 6.00E-34 cyt HE859173

Putative 40S ribosomal protein RPS16 Novocrania anomala /

ACD65104.1 2.00E-67 cyt HE858899

40S ribosomal protein S30 Aedes aegypti / XP_001653913.1 2.00E-14 cyt HE858808

40S ribosomal protein SA-like Nasonia vitripennis /

XP_001608082.1 7.00E-56 nuc HE859192

Ribosomal protein RPL14 Arenicola marina / ABW23170.1 6.00E-55 mit HE859118

Ribosomal protein L32 Lepidochitona cinerea /

ACR24953.1 8.00E-48 cyt HE859166

60S ribosomal protein L6 Harpegnathos saltator /

EFN79475.1 7.00E-26 cyt HE858776

60S ribosomal protein L10-3 Zea mays / NP_001149336.1 1.00E-11 cyt HE858638

60S ribosomal protein L11 Glossina morsitans morsitans /

ADD20563.1 4.00E-35 mit HE858806

60S ribosomal protein L12 Zea mays / ACG35173.1 3.00E-38 nuc HE859020

Ribosomal protein RPL17 Eurythoe complanata /

ABW23221.1 3.00E-16 nuc HE858940

Large subunit ribosomal protein 23 Pristionchus pacificus /

ABR87202.1 2.00E-51 cyt HE859259

Putative 60S ribosomal protein RPL27 Novocrania anomala /

ACD65128.1 2.00E-39 mit HE858729

Ribosomal protein L10a, component

of cytosolic 80S ribosome and 60S

large subunit

Chlamydomonas reinhardtii /

XP_001699807.1 2.00E-46 cyt HE858749

Putative 23S ribosomal RNA Vigna unguiculata / CAO02532.1 3.00E-13 mit HE858674

Putative ribosomal protein S3 Vigna unguiculata / CAO02550.1 4.00E-13 mit HE859345

Putative ribosomal protein S3 Vigna unguiculata / CAO02550.1 8.00E-15 mit HE859397

Ubiquitin C Schistosoma japonicum /

CAX79699.1 2.00E-53 cyt HE858679

Ubiquitin C Equus caballus / NP_001075331.1 5.00E-24 cyt HE858977

Putative ubiquitin C variant 1 Taeniopygia guttata / ACH45550.1 7.00E-38 cyt HE858934

Predicted protein (ubiquitin isoform 1) Physcomitrella patens subsp.

patens / XP_001764949.1 4.00E-14 chl HE858820

Ubiquitin-conjugating enzyme E2 Zea mays / NP_001140410.1 3.00E-33 chl HE858941

E3 Ubiquitin-protein ligase UHRF1-

Meleagris gallopavo /

XP_003213379.1 2.00E-08 cyt HE858646

Desiccation tolerance in Pyropia columbina 941

Continuation

Functional category Putative Identity Species and accession number E value D*

Pyropia accession

number

Hypothetical protein 382 Pyropia yezoensis / YP_537017.1 1.00E-14 cyt HE858985

Molybdopterin biosynthesis protein Porphyra purpurea / NP_053945.1 9.00E-10 cyt HE859052

Peptidase membrane zinc

metallopeptidase

Pelodictyon phaeoclathratiforme

BU-1 / YP_002018463.1 3.00E-12 cyt HE858858

Protein disulfide isomerase Dictyostelium discoideum AX4 /

XP_635206.1 2.00E-16 cyt HE858804

Serine peptidase 2 Radix peregra / ABL67951.1 1.00E-22 ext HE858888

Serine protease Aedes aegypti / XP_001655711.1 1.00E-07 cyt HE859141

Serine-type endopeptidase Culex quinquefasciatus /

XP_001846627.1 8.00E-12 cyt HE858654

Putative serine-type endopeptidase Aedes aegypti / XP_001663439.1 4.00E-18 cyt HE858699

Signal peptidase complex catalytic

subunit SEC11A

Danio rerio / NP_001002521.1 7.00E-08 cyt HE859034

Signal peptidase complex subunit 3 Bombyx mori / NP_001091763.1 3.00E-32 cyt HE859075

Signal recognition particle subunit

Srp68

Glossina morsitans morsitans /

ADD18348.1 4.00E-10 cyt HE858914

Signal sequence receptor, beta-like Saccoglossus kowalevskii /

XP_002740523.1 6.00E-14 cyt HE858810

T-complex protein 1 subunit epsilon Harpegnathos saltator /

EFN84860.1 2.00E-54 cyt HE859168

Translation initiation factor Anopheles gambiae str. PEST /

XP_316499.2 2.00E-30 cyt HE859078

Protein translation factor Griffithsia japonica / AAM93956.1 8.00E-40 cyt HE858711

Translation initiation factor 2 gamma

subunit

Allacma fusca / CAG29667.1 1.00E-24 cyt HE859111

Histidine-tRNA synthetase Porphyra purpurea / NP_053958.1 1.00E-47 chl HE858867

Histidyl-tRNA synthetase Moorea producens 3L /

ZP_08428961.1 2.00E-10 chl HE859222

60S acidic ribosomal protein P0 Haliotis diversicolor / ABY87386.1 1.00E-58 cyt HE859006

60S acidic ribosomal protein P2 Spodoptera frugiperda /

AAL62467.1 5.00E-18 cyt HE859081

Transcript antisense to ribosomal RNA

protein 1 (Tar1p)

Saccharomyces cerevisiae S288c /

NP_690845.1 2.00E-07 mit HE859051

Venom serine carboxypeptidase

precursor

Apis mellifera / NP_001152775.1 1.00E-25 cyt HE858876

Cathepsin L1-like isoform 1 (cysteine

protease)

Danio rerio / XP_001341714.2 5.00E-07 mit HE858698

Cathepsin L2 cysteine protease Pinctada fucata / ADC52431.1 8.00E-32 mit HE859305

Cysteine proteinase cathepsin F Glossina morsitans morsitans /

ADD19167.1 3.00E-22 cyt HE859086

Elongation factor 1 gamma Caenorhabditis brenneri /

ACE00305.1 1.00E-13 cyt HE859382

Putative elongation factor 1 gamma Ixodes scapularis /

XP_002410199.1 5.00E-39 cyt HE859183

Elongation factor 1-beta Culex tarsalis / ACJ64291.1 5.00E-26 cyt HE858793

Elongation factor 1 beta Simulium nigrimanum /

ACZ28393.1 2.00E-30 cyt HE858987

Elongation factor 1-beta Artemia salina / P12262.3 4.00E-16 cyt HE858954

Elongation factor-1 gamma Trichinella spiralis /

XP_003375107.1 2.00E-09 cyt HE859353

Elongation factor-1 gamma Bothriocyrtum californicum /

ABG88914.1 6.00E-30 cyt HE859385

Elongation factor-1 gamma Aliatypus plutonis / ABG88916.1 5.00E-44 cyt HE859214

Elongation factor-1 gamma Deinopis spinosa / ABG88956.1 3.00E-12 cyt HE859387

Elongation factor-1 gamma Aptostichus sp. 4 NAA-2006 /

ABG88917.1 2.00E-64 cyt HE859398

Elongation factor-2 Harbansus paucichelatus /

AAR01294.1 5.00E-12 cyt HE859262

Peptidyl prolyl cis-trans isomerase B Conus novaehollandiae /

ADC80506.1 2.00E-61 cyt HE858895

Peptidyl-prolyl cis-trans isomerase Culex quinquefasciatus /

XP_001846632.1 2.00E-23 cyt HE859125

Peptidyl-prolyl isomerase-1 Gryllus firmus / ACD69575.1 6.00E-12 cyt HE859254

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942 Latin American Journal of Aquatic Research

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Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

Phenylalanyl-tRNA synthetase beta

chain

Porphyra purpurea / NP_053956.1 9.00E-42 chl HE859109

Ribosome biogenesis protein Nsa2

homolog

Ictalurus punctatus /

NP_001187984.1 3.00E-84 nuc HE858869

Metallocarboxypeptidase inhibitor Medicago truncatula /

XP_003616487.1 3.00E-52 cyt HE858630

6. Cell motility Intermediate filament protein Biomphalaria glabrata /

AAZ39528.1 2.00E-25 cyt HE859009

Myomesin Aedes aegypti / XP_001655591.1 1.00E-13 cyt HE859273

Myosin light chain Gryllotalpa orientalis /

AAW22542.1 2.00E-43 cyt HE858829

Myosin 1 light chain Palaemonetes varians /

ACR54116.1 3.00E-16 cyt HE859189

Myosin heavy chain isoform 3 Daphnia pulex / EFX87106.1 8.00E-35 n.d. HE859089

Paramyosin, short form Harpegnathos saltator /

EFN75172.1 9.00E-68 n.d. HE858678

Paramyosin, short form Camponotus floridanus /

EFN63022.1 6.00E-35 n.d. HE859063

Tropomodulin Culex quinquefasciatus /

XP_001867000.1 1.00E-28 cyt HE859137

Tropomodulin, isoform K Drosophila melanogaster /

NP_001189319.1 2.00E-33 cyt HE858787

Tropomyosin-2 isoform 1 Bombyx mori / NP_001103782.1 1.00E-60 cyt HE858718

Actin Pyropia yezoensis / BAB64309.1 1.00E-14 cyt HE859049

Actin Apis mellifera / XP_003251465.1 6.00E-43 cyt HE858761

Actin-2, partial Ascaris suum / ADY47269.1 9.00E-73 cyt HE858791

Actin 4 Tetranychus urticae / ACN53544.1 8.00E-35 cyt HE859406

Actin 57B Glossina morsitans morsitans /

ADD19431.1 7.00E-13 cyt HE858814

Actin 88F Drosophila melanogaster /

NP_524367.1 5.00E-78 cyt HE858619

Actin Timema tahoe / ADX66580.1 1.00E-74 cyt HE858642

Actin E2 Drosophila virilis / AAK25829.1 2.00E-41 cyt HE859379

Alpha actinin, isoform A Drosophila melanogaster /

NP_477484.2 1.00E-29 cyt HE859142

Beta-actin Diabolocatantops pinguis /

ACV32627.1 5.00E-78 cyt HE858722

Beta-actin Diabolocatantops pinguis /

ACV32627.1 9.00E-28 cyt HE858763

Beta-actin Channa punctata / AEA50896.1 9.00E-40 cyt HE859079

Actin 87E Glossina morsitans morsitans /

ADD19714.1 4.00E-76 cyt HE858840

Actin type 1 Ostrea edulis / CAL69229.1 1.00E-72 cyt HE859022

Actin, cytoplasmic 2-like isoform 1 Callithrix jacchus /

XP_002758560.1 2.00E-54 cyt HE858700

Alpha-tubulin Boltenia villosa / AAM76122.1 2.00E-45 cyt HE858975

Ankyrin-1-like Apis mellifera / XP_397331.2 3.00E-60 cyt HE859178

Dynein heavy chain Culex quinquefasciatus /

XP_001843519.1 2.00E-04 cyt HE859030

Stretchin-Mlck, isoform L Drosophila melanogaster /

NP_001188951.1 3.00E-06 cyt HE858663

Up-regulated during skeletal muscle

growth protein 5 – like

Acyrthosiphon pisum /

XP_003247952.1 6.00E-04 cyt HE858948

Inhibitor of apoptosis protein Bombyx mori / AAN46650.1 6.00E-20 cyt HE858854

7. Cell growth and death Translationally-controlled tumor

protein

Cyprinus carpio / ABC59222.1 1.00E-11 cyt HE858712

Cathepsin D Pteria penguin / AEI58895.1 3.00E-17 cyt HE858983

Death-associated small cytoplasmic

leucine-rich protein

Bombyx mori / NP_001138799.1 3.00E-27 cyt HE858839

CHK1 checkpoint-like protein Perca flavescens / ADX97237.1 1.00E-04 cyt HE859132

8. Membrane transporters Transmembrane channel-like 2 Taeniopygia guttata /

XP_002196817.1 2.00E-04 n.d. HE859194

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Desiccation tolerance in Pyropia columbina 943

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

Protein-export membrane protein

Secd/Secf

Psychroflexus torquis ATCC

700755 / ZP_01252230.1 4.00E-19 n.d. HE859043

RND superfamily resistance-

nodulation-cell division: proton (H+)

antiporter

Lactobacillus rhamnosus LMS2-1

/ ZP_04439763.1 6.00E-06 n.d. HE859400

Sugar transporter Haliscomenobacter hydrossis

DSM 1100 / YP_004445152.1 2.00E-08 n.d. HE858852

Transport protein Paenibacillus sp. HGF5 /

ZP_08281960.1 1.00E-03 n.d. HE859386

9. Vesicular transport and

metabolism

Putative Ras-related protein Rab-2A Aedes aegypti / XP_001650005.1 4.00E-15 gol HE859113

Rtnl1, isoform C Drosophila melanogaster /

NP_787988.1 1.00E-36 n.d. HE859153

Transport protein Sec61 subunit alpha

Culex quinquefasciatus /

XP_001850184.1 3.00E-113 n.d. HE859332

10. Cell wall metabolism Heparanase precursor Rattus norvegicus / NP_072127.1 4.00E-05 cyt HE858732

Cell wall-associated hydrolase Escherichia sp. 3253FAA /

ZP_04532936.1 3.00E-21 cyt HE859007

Cell wall-associated hydrolase Burkholderia multivorans ATCC

17616 / YP_001949468.1 1.00E-29 cyt HE858683

Cell wall-associated hydrolase Microscilla marina ATCC 23134/

ZP_01689674.1 7.00E-20 cyt HE858990

Cell wall-associated hydrolase Roseobacter sp. AzwK-3b /

ZP_01900972.1 5.00E-16 cyt HE859054

Cell wall-associated hydrolase Escherichia sp.3253FAA/

ZP_04532936.1 3.00E-12 cyt HE859275

Cell wall-associated hydrolase Helicobacter canadensis /

ZP_07804970.1 2.00E-05 cyt HE859290

Cell wall-associated hydrolase Streptomyces sp. e14 /

ZP_06708251.1 4.00E-09 cyt HE859401

Glycoside hydrolases Aedes aegypti / XP_001650341.1 1.00E-58 cyt HE858995

Glycosyl transferase family protein Acidiphilium cryptum JF-5 /

YP_001233531.1 8.00E-06 cyt HE858973

30S ribosomal protein S13 Pyropia yezoensis / ABJ91310.1 2.00E-26 chl HE859292

11. Photosynthesis and

chloroplast proteins

Ribosomal protein L3 Porphyra purpurea / NP_053924.1 2.00E-25 chl HE858764

Ribosomal protein L13 Porphyra purpurea / NP_053902.1 3.00E-43 chl HE858795

Ribosomal protein L35 Pyropia yezoensis / YP_536952.1 7.00E-07 chl HE859002

Photosystem I P700 chlorophyll A

apoprotein A1

Arthrospira platensis str. Paraca /

ZP_06383599.1 4.00E-40 chl HE859186

Photosystem I P700 chlorophyll A

apoprotein A1

Bangia fuscopurpurea /

AAM62007.1 7.00E-70 chl HE858916

Photosystem I P700 chlorophyll A

apoprotein A1

Porphyra purpurea / NP_053894.1 1.00E-52 chl HE859114

Photosystem II cytochrome C550 Porphyra purpurea / NP_053809.1 1.00E-37 chl HE858727

Photosystem II D2 protein Pyropia yezoensis / YP_537038.1 1.00E-06 chl HE859365

Photosystem II protein D1 Pyropia yezoensis / YP_536893.1 9.00E-103 chl HE858641

Photosystem II protein D1 Porphyra purpurea / NP_053822.1 1.00E-93 chl HE858733

Photochlorophyllide reductase subunit

Porphyra purpurea / NP_053888.1 9.00E-35 chl HE858824

Photochlorophyllide reductase subunit

Porphyra purpurea / NP_053797.1 2.00E-32 chl HE859157

Photochlorophyllide reductase subunit

Porphyra purpurea / NP_053798.1 2.00E-27 chl HE859287

Phycobilisome linker polypeptide Cyanothece sp. PCC 7822 /

YP_003889235.1 1.00E-03 chl HE859143

Phycoerythrin beta subunit Porphyra purpurea / NP_053977.1 3.00E-08 chl HE858627

Phycoerythrin beta subunit Ceramium boydenii / AAM88398.1 1.00E-50 chl HE858639

Putative rubisco expression protein Rhodomonas salina /

YP_001293520.1 3.00E-75 chl HE859066

Allophycocyanin subunit alpha Microcystis aeruginosa NIES-843

/ YP_001656041.1 1.00E-12 chl HE858886

ATP synthase CF1 delta subunit Porphyra purpurea / NP_053853.1 5.00E-12 chl HE858967

ATP synthase CF0 A subunit Pyropia yezoensis / YP_536928.1 9.00E-11 chl HE859243

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944 Latin American Journal of Aquatic Research

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Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

ATP synthase CF0 subunit I Vigna unguiculata / CAO02552.1 7.00E-09 chl HE858944

Beta-Ig-H3/fasciclin Acidiphilium cryptum JF-5 /

YP_001235357.1 3.00E-07 chl HE858860

Clp protease ATP binding subunit Pyropia yezoensis / YP_537014.1 2.00E-56 chl HE858999

12. Respiration and

mitochondrial proteins

Cytochrome P450 like_TBP Nicotiana tabacum / BAA10929.1 5.00E-30 chl HE858796

Glycine cleavage system protein H Hydrogenivirga sp. 128-5-R1-1 /

ZP_02176878.1 3.00E-05 chl HE859272

Mitochondrial processing peptidase

beta subunit

Glossina morsitans morsitans /

ADD20239.1 4.00E-62 mit HE858961

Cytochrome B-C1 complex subunit 9-

Anolis carolinensis /

XP_003230444.1 1.00E-05 mit HE858900

NADH dehydrogenase [ubiquinone] 1

beta subcomplex subunit 8

mitochondrial

Nasonia vitripennis /

NP_001161450.1 4.00E-14 mit HE858968

NADH dehydrogenase subunit 1 Littorina saxatilis / CAM57998.1 6.00E-52 mit HE858930

NADH dehydrogenase subunit 2 Fusiturris similis /

YP_003204746.1 8.00E-12 mit HE858904

NADH dehydrogenase subunit 2 Porphyra purpurea / NP_049304.1 5.00E-04 mit HE859276

NADH-ubiquinone oxidoreductase 39

kDa subunit

Culex quinquefasciatus /

XP_001865472.1 2.00E-37 mit HE858694

NADH-ubiquinone oxidoreductase Fe-

S protein 2 (Ndufs2)

Aedes aegypti / XP_001655316.1 2.00E-69 mit HE859008

Cytochrome oxidase subunit I Lucilia cuprina / CBX78584.1 2.00E-52 mit HE858780

Cytochrome oxidase subunit I Dicranomyia variabilis /

ABV71159.1 1.00E-35 mit HE858720

Cytochrome oxidase subunit I Littorina saxatilis / CAA10593.1 1.00E-65 mit HE858856

Cytochrome oxidase subunit I Echinolittorina novaezelandiae /

CAF22173.1 1.00E-61 mit HE859031

Cytochrome oxidase subunit II Geranomyia advena / ABV71230.1 5.00E-42 mit HE858740

Cytochrome oxidase subunit II Ceratitis ditissima / ADB55739.1 3.00E-10 mit HE859328

Cytochrome oxidase subunit II Dicranomyia stygipennis /

ABV71214.1 8.00E-06 mit HE858917

Cytochrome oxidase subunit II Dicranomyia swezeyi /

ABV71215.1 2.00E-10 mit HE859117

Cytochrome oxidase subunit II Littorina saxatilis / CAA10594.1 2.00E-22 mit HE858884

Cytochrome C oxidase subunit 3 Potamopyrgus antipodarum /

ADB93443.1 1.00E-65 mit HE858669

Cytochrome C oxidase subunit III Cymatium parthenopeum /

YP_003204809.1 7.00E-25 mit HE859331

Cytochrome C oxidase subunit III Aedes albopictus / YP_194916.1 2.00E-38 mit HE858725

Cytochrome C oxidase subunit IV Drosophila simulans /

ABM88269.1 5.00E-29 mit HE858714

Cytochrome C oxidase subunit

Va/COX6

Simulium nigrimanum /

ACZ28403.1 1.00E-32 mit HE858693

Cytochrome C oxidase polypeptide

VIa

Oncorhynchus mykiss /

ACO08369.1 4.00E-08 mit HE858746

Cytochrome C oxidase subunit 6a

polypeptide 1

Saccoglossus kowalevskii /

XP_002738468.1 1.00E-22 mit HE859201

F1F0-ATP synthase subunit

C/ATP9/proteolipid

Simulium nigrimanum /

ACZ28249.1 1.00E-14 mit HE858739

F1F0-ATP synthase subunit

epsilon/ATP15

Glossina morsitans morsitans /

ADD20505.1 4.00E-16 mit HE858689

F1F0-ATP synthase subunit

OSCP/ATP5

Glossina morsitans morsitans /

ADD18988.1 3.00E-54 mit HE858984

Mitochondrial matrix protein P32 Bombyx mori / ABD36320.1 2.00E-11 mit HE858994

Mitochondrial succinate

dehydrogenase cytochrome B subunit

Culex quinquefasciatus /

XP_001859594.1 4.00E-10 mit HE859236

ADP / ATP carrier protein Ricinus communis /

XP_002517886.1 5.00E-41 mit HE858960

ADP / ATP translocase Apis mellifera / NP_001010975.1 1.00E-58 mit HE858665

ATP synthase alpha subunit precursor Drosophila silvestris / ABY55748.1 4.00E-36 mit HE859342

ATP synthase alpha subunit precursor Strongylocentrotus purpuratus /

NP_999743.1 4.00E-30 mit HE858957



Desiccation tolerance in Pyropia columbina 945

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

ATP synthase b Spodoptera exigua / ACL77780.1 1.00E-10 mit HE859072

ATPSyn-beta Drosophila simulans /

XP_002105781.1 2.00E-36 mit HE858786

Mitochondrial ATP synthase delta

subunit

Aedes aegypti / XP_001655448.1 1.00E-20 mit HE859029

ATP synthase delta subunit, isoform A Drosophila melanogaster /

NP_524402.1 2.00E-25 mit HE858979

ATP synthase, H+ transporting,

mitochondrial F1 complex, O subunit

Xenopus (Silurana) tropicalis /

NP_001037877.1 4.00E-17 mit HE859056

Cytochrome B Bolinus brandaris /

YP_003204844.1 2.00E-38 mit HE858728

Cytochrome B Drosophila mojavensis /

DAA06245.1 5.00E-44 mit HE858782

Cytochrome B Liriomyza trifolii /

YP_003734909.1 3.00E-06 mit HE859193

Cytochrome C Glossina morsitans morsitans /

ADD19108.1 4.00E-34 mit HE858933

Cytochrome C1 Tribolium castaneum /

EFA01361.1 3.00E-74 mit HE858766

Cytochrome C1 Glossina morsitans morsitans /

ADD19528.1 1.00E-18 mit HE859010

ATP synthase F0 subunit 6 Drosophila littoralis /

YP_002327406.1 3.00E-07 mit HE858843

H+ transporting ATP synthase

subunit e

Bombyx mori / NP_001091812.1 2.00E-08 mit HE859061

Predicted ubiquinol-cytochrome C

reductase core protein I

Oryctolagus cuniculus /

XP_002713430.1 4.00E-20 mit HE859147

Voltage-dependent anion-selective

channel

Culex quinquefasciatus /

XP_001842637.1 3.00E-47 mit HE859045

13. Others Putative senescence-associated protein Lilium longiflorum / ABO20851.1 2.00E-15 n.d. HE858777

Putative senescence-associated protein Cupressus sempervirens /

ACA30301.1 1.00E-23 mit HE859069

Putative senescence-associated protein Perkinsus marinus ATCC 50983/

XP_002771156.1 3.00E-07 n.d. HE859266

Senescence-associated protein Chlorella variabilis / EFN58441.1 1.00E-05 mit HE859270

Kazal domain-containing peptide Anopheles darlingi / ACI30205.1 1.00E-06 n.d. HE859175

Putative secreted protein 10 kDa Argas monolakensis / ABI52743.1 7.00E-15 cyt HE859146

AGAP009754-PA (angiotensin-

converting enzyme 4)

Anopheles gambiae /

XP_001238056.2 9.00E-71 cyt HE859209

Another B-box affiliate, isoform A Drosophila melanogaster /

NP_611390.2 8.00E-26 ext HE859269

AGAP008459-PA (cuticular protein 9,

low complexity family)

Anopheles gambiae /

XP_001688982.1 1.00E-05 ext HE858653

AGAP009872-PA (cuticular protein

133, RR-1 family)

Anopheles gambiae str. PEST/

XP_001689165.1 1.00E-21 ext HE858951

Putative pupal cuticle protein Aedes aegypti / XP_001661677.1 2.00E-17 ext HE858625

Fibroleukin-like Meleagris gallopavo /

XP_003201937.1 2.00E-26 cyt HE858705

Cuticular protein analogous to

peritrophins 3-A1

Acyrthosiphon pisum /

NP_001156724.1 2.00E-29 ext HE859090

Vitellogenin Lethocerus deyrollei / BAG12118.1 3.06E-31 mit HE858626

Vitellogenin 1b Culex tarsalis / ADH04225.1 2.00E-05 n.d. HE858633

Vitellogenin 1a Culex tarsalis / ADH04224.1 2.00E-13 cyt HE859037

Vitellogenin 2a Culex tarsalis / ADH04226.1 4.00E-22 chl HE858675

Vitellogenin A1 Culex quinquefasciatus /

XP_001843136.1 2.00E-07 cyt HE858849

Vitellogenin 2b Culex tarsalis / ADH04227.1 5.00E-36 cyt HE858632

Vitellogenin B Ochlerotatus atropalpus /

AAV31927.1 1.00E-06 ext HE858657

Vitellogenin C Toxorhynchites amboinensis /

AAV31932.1 4.00E-09 cyt HE859170

Vitellogenin C Anopheles albimanus /

AAV31933.1 5.00E-47 mit HE858622

Vitellogenin precursor Tenebrio molitor / AAU20328.2 1.00E-06 n.d. HE859404

946 Latin American Journal of Aquatic Research

Continuation

Functional category Putative Identity Species and accession number E value D* Pyropia accession

number

Vitellogenin C Aedes polynesiensis / AAV31926.1 7.00E-32 cyt HE858631

CDGSH iron sulfur domain-containing

protein 2-like protein

Camponotus floridanus /

EFN65337.1 2.00E-33 mit HE858716

Opsin-1 Culex quinquefasciatus /

XP_001845697.1 8.00E-06 n.d. HE859197

Rhodopsin Aedes aegypti / XP_001657619.1 4.00E-46 n.d. HE858950

Lethal(2)essential for life protein, l2efl Aedes aegypti / XP_001663499.1 2.00E-36 n.d. HE858887

Similar to fibropellin Ia Strongylocentrotus purpuratus /

XP_001199485.1 3.00E-06 cyt HE858672

Cob(I)yrinic acid a,c-diamide

adenosyltransferase

Selenomonas noxia ATCC43541 /

ZP_06604478.1 2.00E-05 n.d. HE858754

revealed that ESTs with higher representation are

involved in protein synthesis, processing and

degradation (ca. 15%, e.g., Clp-protease),

photosynthesis and chloroplast structure (ca. 14.4%;

e.g., ferredoxin NADP+ reductase), respiration and

mitochondria (ca. 13.1%), proteins involved in cell

wall metabolism (ca. 10.6%, cell-wall hydrolases,

glycosyl transferases and chitin deacetylases) and

antioxidant activity, chaperone and defense factors

(7.5%; thioredoxin (TRX), catalase and HSPs) (Table

1). Finally, with an important number of sequences it

was not possible to find clear functional similarity

with known proteins (ca. 11%).

DISCUSSION

The hydration-desiccation cycle in P. columbina

generates an unbalance in the intracellular redox

potential (Contreras-Porcia et al., 2011a), a situation

that must be controlled by a coordinated cascade of

responses that are induced differentially in each

condition. Our results show that, during hydration,

predominantly expressed genes were those involved in

protein metabolism. Among them, we highlight

several ribosomal proteins, translation initiation

factors and elongation factors, proteases, the

proteasome system and several ubiquitins (Table 1).

Of particular relevance is the ubiquitin-proteasome

system, present in all eukaryotes, which is activated in

response to several abiotic stress factors and

participates in tolerance mechanisms by removing

unfolded proteins and proteins damaged during

oxidative stress (Dreher & Callis, 2007; Pena et al.,

2007; Kurepa et al., 2008). In this functional group

three types of peptidylprolyl cistrans-isomerases

(PPIases) were identified and found to be similar to

those from Arthropoda and Mollusca. These proteins

catalyze the cis-trans isomerisation of prolines

(C5H9NO2), and have been described in the processes

of cellular signalling (with a calmodulin-binding

domain), regulation of gene transcription, and acting

as chaperones and folding catalysts. Specifically, in

the gastropod Conus novaehollandiae these proteins

facilitate the oxidative folding of several neurotoxic

peptides (Safavi-Hemami et al., 2010). In plants, these

proteins are involved in flowering (Wang et al., 2010)

and in controlling cell proliferation, since PPIase

expression increases in the presence of cytokinin

(Vittorioso et al., 1998). PPIases are induced by

wounding, heat and salt stress (Vucich & Gasser,

1996; Kurek et al., 1999), and have been directly

involved in membrane protein folding (i.e., chloroplast

and mitochondria) (Breiman et al., 1992). Thus, the

quick re-establishment of the normal condition after

desiccation in P. columbina could be in part explained

by both: i) the re-folding of structural proteins with

important functions such as those involved in

transcriptional regulation, and ii) by the removal of

oxidized proteins. In fact, levels of oxidised proteins

measured during the hydration-desiccation cycle were

consistently lower in P. columbina compared to those

in sensitive species (Contreras-Porcia et al., 2011a,

Flores-Molina et al., unpublished data).

Proteins that form part of the energy metabolism

were more highly represented during hydration [i.e.,

Cytochrome C Oxidase subunits, NADH Dehydro-

genase subunits, NADH-Ubiquinone Oxidoreductase

subunits, F1F0-ATP Synthase subunits and ADP/ATP

carrier proteins (Table 1)] than during desiccation

stress (Table 2). Theoretically, maintaining the

tolerance mechanisms required to buffer the effects of

a stress imposes energy costs (e.g., Zagdańska, 1995).

Acclimation to oxidative stress depends on a high

availability of NADPH and ATP, since most of the

intracellular metabolic reactions require energy. For

example, the ubiquitin- mediated system for

intracellular protein degradation is ATP-dependent in

all organisms (Ciechanover et al., 1984). Additionally,

the structure of the chromosomes plays a critical role

in transcriptional regulation where the chromatin

Desiccation tolerance in Pyropia columbina 947

Table 2. Functional category, identity and potential cellular destination of proteins and accession number of identified

ESTs in Pyropia columbina under natural desiccation (UD library). Data from the hydrated library (UH) is presented in

Table 1. E value: the best (lowest) Expect value (E value) of all alignments from that database sequence, D*: Putative

destination.

Functional category Putative identity Species and accession number E value D*

Pyropia

accession

number

1. Signal transduction Calmodulin Pyropia yezoensis / ABN41559 8.00E-11 cyt HE859451

Enkurin TRPC channel interacting

protein

Saccoglossus kowalevskii /

NP_001158494 2.00E-11 cyt HE859515

Guanine nucleotide binding protein 3 Dictyostelium discoideum AX4 /

XP_637356.1 2.00E-38 cyt HE859861

Hexamerin 2 beta Aedes aegypti / XP_001659531.1 4.00E-27 cyt HE859661

Serine/threonine protein phosphatase 1 Malus domestica / AAD56010.1 4.00E-45 cyt HE859472

2. Transcription, splicing

and replication

RNA-directed DNA polymerase

(reverse transcriptase)

Ktedonobacter racemifer DSM 44963

/ ZP_06971690.1 8.00E-13 nuc HE859542

Reverse transcriptase Cyanothece sp. ATCC 51142 /

YP_001801525.1 9.00E-11 nuc HE859708

RNA-directed DNA polymerase

(reverse transcriptase)

Arthrospira maxima CS-328 /

ZP_03273642 1.00E-24 nuc HE859482

DNA-directed RNA polymerase, beta

subunit

Crocosphaera watsonii WH 8501 /

ZP_00516049.1 8.00E-48 nuc HE859717

RNA polymerase alpha subunit Porphyra purpurea / NP_053903.1 3.00E-09 nuc HE859795

Argonaute2 (AGO2) Albugo laibachii Nc14 / CCA27516.1 4.00E-21 cyt HE859580

Predicted metal-dependent RNase Streptococcus pyogenes M49 591 /

ZP_00366321.1 8.00E-17 nuc HE859892

Group II intron reverse

transcriptase/maturase

Streptococcus dysgalactiae subsp.

dysgalactiae ATCC 27957 /

EFY02523.1

8.00E-11 nuc HE859734

Histone H3.3 Harpegnathos saltator / EFN83153.1 4.00E-38 nuc HE859642

Retrotransposon protein Oryza sativa Indica Group /

ABR26094.1 2.00E-22 nuc HE859423

3. Basal metabolism Aconitate hydratase Camponotus floridanus / EFN71522.1 2.00E-45 mit HE859793

Lipase Aedes aegypti / XP_001652527.1 3.00E-25 ext HE859755

Fructose-1,6-biphosphatase F-II Griffithsia japonica / AAP80707.1 2.00E-07 cyt HE859780

Fructose-bisphosphate aldolase Sphingobacterium sp. 21 /

YP_004319198.1 6.00E-59 n.d. HE859506

Alpha-amylase B Culex quinquefasciatus /

XP_001846541.1 6.00E-26 ext HE859517

3-oxoacyl-ACP reductase Streptomyces avermitilis MA-4680 /

NP_824830.1 3.00E-06 ext HE859599

C-4 sterol methyl oxidase 2 Nicotiana benthamiana / AAQ83692.1 4.00E-11 mit HE859431

Amine oxidase Plasmodium yoelii yoelii 17XNL /

XP_730508.1 8.00E-13 cyt HE859420

Scavenger receptor cysteine-rich

protein

Culex quinquefasciatus /

XP_001866937.1 2.00E-05 n.d. HE859437

Vacuolar H+-ATPase C chain Pyropia tenera / JC7151 4.00E-50 n.d. HE859667

4. Antioxidant, chaperone

and defense factors

Thioredoxin Hyperthermus butylicus DSM 5456 /

YP_001013307.1 2.00E-05 mit HE859628

Catalase Pyropia yezoensis / ADO23652.1 4.00E-06 mit HE859586

Heme oxygenase Porphyra purpurea / NP_053881.1 2.00E-05 chl HE859574

Cytochrome P450-Like TBP protein Lilium longiflorum / ABO20848.1 4.00E-19 cyt HE859591

Heat shock protein 70 Pyropia yezoensis / YP_536996.1 2.00E-63 chl HE859563

dnaK gene product (Heat shock

protein 70)

Thermosynechococcus elongatus BP-

1 / NP_682523.1 1.00E-39 chl HE859424

Heat shock protein Hsp70 Raphidiopsis brookii D9 /

ZP_06305357.1 2.00E-24 chl HE859837

Molecular chaperones HSP70/HSC70,

HSP70

Ectocarpus siliculosus / CBN79394.1 2.00E-82 mit HE859570

Heat shock protein Hsp70 Cylindrospermopsis raciborskii CS-

505 / ZP_06308882.1 3.00E-47 chl HE859573

Heat shock protein 90, partial Thraustotheca clavata / AAX10950.1 6.00E-79 nuc HE859869

Heat shock protein 90 Cellulophaga algicola DSM 14237 /

YP_004165199.1 1.00E-61 cyt HE859588



948 Latin American Journal of Aquatic Research

Continuation

Functional category Putative identity Species and accession number E value D*

Pyropia

accession

number

Molecular chaperones

GRP78/Bip/KAR2, HSP70

superfamily

Ectocarpus siliculosus / CBJ48460.1 1.00E-34 cyt HE859467

5. Protein synthesis,

processing and

degradation

Ubiquitin Oncorhynchus mykiss / ACO07546.1 4.00E-04 mit HE859794

Ubiquitin protein Triticum aestivum / AAQ08322 3.00E-36 cyt HE859488

Ubiqutin Phanerochaete chrysosporium /

CAA83244 1.00E-06 cyt HE859630

Ubiquitin Drosophila persimilis /

XP_002023314.1 8.00E-04 cyt HE859858

Ubiquitin-conjugating enzyme E2-17

kDa

Zea mays / ACG37110.1 7.00E-39 cyt HE859634

Proteasome subunit beta type-6 Phytophthora infestans T30-4 /

XP_002901169.1 6.00E-11 cyt HE859840

Molybdopterin biosynthesis protein Porphyra purpurea / NP_053945.1 2.00E-35 chl HE859454

30S ribosomal protein S18 Gemella sanguinis M325 /

ZP_08260903 2.00E-10 n.d. HE859511

Ribosomal protein L35 Pyropia yezoensis / YP_536952.1 1.00E-06 chl HE859735

Protein translation factor Griffithsia japonica/ AAM93956 2.00E-47 cyt HE859466

Eukaryotic translation elongation

factor 1 alpha 1

Chlamydomonas reinhardtii /

XP_001696568.1 2.00E-58 cyt HE859552

EF2, translation elongation factor 2 Ectocarpus siliculosus / CBJ32863.1 1.00E-36 cyt HE859801

Elongation factor-like protein Pseudoperkinsus tapetis /

ADE62443.1 3.00E-35 cyt HE859728

Clp protease ATP binding subunit Pyropia yezoensis / YP_537014.1 2.00E-58 cyt HE859649

Cysteine synthase Porphyra purpurea / AAP97124.1 1.00E-74 cyt HE859626

Der F 3 allergen Dermatophagoides farinae /

AAP35076.1 2.00E-05 n.d. HE859604

trpB gene product Acaryochloris marina MBIC11017 /

YP_001519094.1 5.00E-10 n.d. HE859936

Isoleucyl-tRNA synthetase Lacinutrix sp. 5H-3-7-4 /

YP_004579101.1 2.00E-36 cyt HE859750

Prefolding-like protein Opisthacanthus cayaporum /

CAX51417.1 3.00E-22 cyt HE859714

Protease inhibitor G11A6 Mayetiola destructor / ABB70517.1 1.00E-05 cyt HE859462

Signal peptidase, catalytic subunit Chlamydomonas reinhardtii /

XP_001697614.1 1.00E-22 cyt HE859469

6. Cell motility Actin Helicoverpa armigera / ADN84930.1 3.00E-24 cyt HE859471

Actin Pyropia yezoensis / BAB64309.1 7.00E-13 cyt HE859826

Actin Undaria pinnatifida / ADW66613.1 5.00E-97 cyt HE859564

Actin Pyropia yezoensis / BAG71158.1 7.00E-24 cyt HE859659

Actin 1 Paxillus involutus / ABQ85639.1 9.00E-17 cyt HE859928

Beta-actin Mauremys mutica / ADX86815.1 1.00E-21 cyt HE859680

Tropomyosin Crassostrea virginica / AAC61869.1 3.00E-24 mit HE859676

Troponin T-1 Drosophila melanogaster /

AAR24583.1 1.00E-05 cyt HE859762

Beta-tubulin Phytophthora palmivora / AAW58084 4.00E-64 cyt HE859549

Calponin Haliotis diversicolor / ABU53030 9.00E-40 cyt HE859480

7. Cell growth and death Cell division protein Pyropia yezoensis / YP_537009.1 2.00E-17 chl HE859522

Cell division protein FtsA Lacinutrix sp. 5H-3-7-4 /

YP_004578825.1 6.00E-67 cyt HE859665

CHK1 checkpoint homolog Xenopus (Silurana) tropicalis /

CAJ83813.1 8.00E-04 cyt HE859790

CHK1 checkpoint-like protein Helicoverpa armigera / ABK29471.1 1.00E-11 cyt HE859593

CHK1 checkpoint-like Protein Perca flavescens / ADX97237.1 2.00E-10 cyt HE859702

8. Membrane transporters ABC Transporter B family member 13 Arabidopsis thaliana / NP_174115 9.00E-16 n.d. HE859505

ABC Transporter Thalassiosira pseudonana

CCMP1335 / XP_002288593 1.00E-06 n.d. HE859535

ATP-binding cassette (ABC)

superfamily

Ectocarpus siliculosus / CBJ32723.1 1.00E-05 cyt HE859694

9. Vesicular transport and

metabolism

Endoplasmic reticulum vesicle

transporter protein

Arabidopsis thaliana / NP_564162 6.00E-06 cyt HE859464

Desiccation tolerance in Pyropia columbina 949

Continuation

Functional category Putative identity Species and accession number E value D*

Pyropia

accession

number

GTPase SAR1 Triticum aestivum / ACD03831.1 2.00E-04 mit HE859693

Small GTP-binding protein Solanum lycopersicum / AAA80679 8.00E-06 cyt HE859512

10. Cell wall metabolism Cell wall-associated hydrolase Microscilla marina ATCC 23134 /

ZP_01689674.1 1.00E-17 cyt HE859576

Cell wall-associated hydrolase Escherichia sp 3_2_53FAA /

ZP_04532936.1 3.00E-39 cyt HE859695

Cell wall-associated hydrolase Escherichia sp. 3_2_53FAA /

ZP_04532936.1 3.00E-09 cyt HE859906

Cell wall-associated hydrolase Escherichia sp. 3_2_53FAA /

ZP_04532936.1 2.00E-11 cyt HE859921

Cell wall-associated hydrolase Escherichia sp. 3_2_53FAA /

ZP_04532936.1 3.00E-13 cyt HE859926

Cell wall-associated hydrolase Escherichia sp. 3_2_53FAA /

ZP_04532936.1 2.00E-12 cyt HE859929

Cell wall-associated hydrolase Escherichia sp. 3_2_53FAA /

ZP_04532936.1 2.00E-15 cyt HE859930

Cell wall-associated hydrolase Microscilla marina ATCC 23134 /

ZP_01689674.1 5.00E-19 cyt HE859413

Putative cell wall-Associated hydrolase Campylobacter jejuni subsp. jejuni

BH-01-0142 / ZP_03217789.1 9.00E-11 cyt HE859421

Cell wall-associated hydrolase Prevotella bryantii B14 /

ZP_07059369.1 9.00E-25 cyt HE859434

Cell wall-associated hydrolase Burkholderia multivorans

ATCC17616 / YP_001949468.1 1.00E-25 cyt HE859495

Cell wall-associated hydrolase Erysipelothrix rhusiopathiae ATCC

19414 / ZP_08082416.1 2.00E-18 cyt HE859498

Cell wall-associated Hydrolase Brucella abortus NCTC 8038 /

ZP_05822976.1 2.00E-14 mit HE859566

Cell wall-associated hydrolase Brucella suis bv. 4 str. 40 /

ZP_05839398.1 7.00E-19 cyt HE859629

Chitin deacetylase 7 precursor Tribolium castaneum /

NP_001104012.1 7.00E-10 cyt HE859430

Glycine cleavage system protein H Hydrogenivirga sp. 128-5-R1-1 /

ZP_02176878.1 7.00E-15 cyt HE859890

Glycosyl transferase Rhodospirillum photometricum DSM

122 / CCG06616.1 1.00E-11 n.d. HE859904

11. Photosynthesis and

chloroplast proteins

Photosystem I P700 chlorophyll A

apoprotein A1

Porphyra purpurea / NP_053894.1 8.00E-72 chl HE859902

Photosystem II protein D1 Pyropia yezoensis / YP_536893.1 1.00E-41 chl HE859757

Photosystem II protein D2 Pyropia yezoensis / YP_537038.1 4.00E-62 chl HE859806

Photosystem II protein D2 Porphyra purpurea / NP_053967 5.00E-93 chl HE859500

Ferredoxin NADP+ reductase Pisum sativum / AAB59349.1 6.00E-05 chl HE859729

23S Ribosomal RNA Vigna unguiculata / CAO02530.1 2.00E-09 chl HE859509

Ribosomal protein S3 Vigna unguiculata / CAO02550.1 5.00E-12 chl HE859706

Ribosomal protein L4 Pyropia yezoensis / YP_536993.1 5.00E-24 chl HE859613

30S ribosomal protein S13 Trichodesmium erythraeum IMS101 /

YP_722618.1 4.00E-23 chl HE859559

Ribosomal protein L2 Chlamydomonas reinhardtii /

ACS16390.1 1.00E-35 chl HE859868

30S ribosomal protein S12 Porphyra purpurea / NP_049316.1 6.00E-16 cyt HE859584

40S ribosomal protein S16 Phytophthora infestans T30-4 /

XP_002899087.1 2.00E-13 cyt HE859787

50S ribosomal protein L3P Cyanobacterium sp.UCYN-A /

YP_003422083.1 3.00E-05 chl HE859842

50S ribosomal protein L3 Prochlorococcus marinus str. MIT

9515 / YP_001012054.1 5.00E-15 chl HE859881

50S ribosomal protein L4 Lyngbya sp. PCC 8106 /

ZP_01619314.1 3.00E-12 chl HE859818

50S ribosomal protein L4 Cyanothece sp. PCC 7424 /

YP_002378953.1 1.00E-06 chl HE859828

50S ribosomal protein L13 Arthrospira platensis NIES-39 /

BAI90083.1 4.00E-37 chl HE859578

60S ribosomal protein L29-1 Glycine max / XP_003546330 2.00E-20 cyt HE859481

RNA polymerase beta subunit Porphyra purpurea / NP_053860.1 3.00E-32 chl HE859802

950 Latin American Journal of Aquatic Research

Continuation

Functional category Putative identity Species and accession number E value D*

Pyropia

accession

number

ATP synthase beta subunit Trebouxia photobiont / ABP98704.1 9.00E-11 chl HE859691

ATP synthase CF1 delta subunit Porphyra purpurea / NP_053853.1 1.00E-14 chl HE859731

Cell division protein Porphyra purpurea / NP_053937.1 5.00E-16 chl HE859460

Ferredoxin, chloroplast precursor Micromonas sp. RCC299 /

XP_002508604.1 2.00E-22 chl HE859587

Photochlorophyllide reductase subunit

Porphyra purpurea / NP_053888.1 2.00E-12 chl HE859810

Photochlorophyllide reductase subunit

Pyropia yezoensis / YP_536960.1 4.00E-52 chl HE859555

HESB-like domain-containing protein

Zea mays / NP_001149194 8.00E-38 chl HE859536

12. Respiration and

mitochondrial proteins

NADH dehydrogenase subunit 4L Porphyra purpurea / NP_049322.1 7.00E-06 mit HE859443

NADH dehydrogenase subunit 7 Coccomyxa subellipsoidea C-169 /

YP_004339024.1 3.00E-14 mit HE859913

Cytochrome C oxidase Ephedra viridis / AAD01658.1 4.00E-09 mit HE859770

Cytochrome C oxidoreductase subunit

Porphyra purpurea / NP_049294.1 7.00E-36 mit HE859672

Cytochrome C oxidase subunit III Tricula hortensis / YP_003434166.1 2.00E-57 mit HE859617

Cytochrome C oxidase subunit IV Aedes aegypti / XP_001663204.1 7.00E-16 mit HE859683

Ribosomal protein S3 Porphyra purpurea / NP_049318.1 4.00E-12 mit HE859602

Ribosomal protein S5 Griffithsia japonica / AAP80699.1 7.00E-64 mit HE859456

40S ribosomal protein S13-2 Arabidopsis thaliana / NP_567151.1 8.00E-43 mit HE859742

40S ribosomal protein S15 Oryza sativa Japonica Group /

BAD30388 3.00E-24 mit HE859501

50S ribosomal protein L16 Psychrobacter arcticus 273-4 /

YP_263793.1 5.00E-06 mit HE859479

50S ribosomal protein L16 Porphyra purpurea / NP_049319.1 8.00E-15 mit HE859664

conserved hypothetical protein Clostridium botulinum NCTC 2916/

ZP_02955128.1 3.00E-08 mit HE859899

HNH endonuclease Arthrospira maxima CS-328 /

ZP_03274502.1 3.00E-15 mit HE859427

Reverse transcriptase Synechococcus sp. PCC 7335 /

ZP_05039776.1 4.00E-12 mit HE859447

Reverse transcriptase Arthrospira platensis NIES-39 /

BAI90930.1 3.00E-10 mit HE859590

Reverse transcriptase Porphyra purpurea / NP_049296.1 7.00E-40 mit HE859800

F-Type H-ATPase beta subunit Ectocarpus siliculosus / CBJ32298.1 1.00E-45 mit HE859457

Dehydration responsive protein Corchorus olitorius / BAJ11784.1 1.00E-13 mit HE859843

Putative microvillar-like protein 5 Phlebotomus perniciosus /

ADJ57676.1 6.00E-06 mit HE859901

Tumor differentially expressed protein Haplopelma schmidti / ACH48223.1 2.00E-06 mit HE859663

13. Others Senescence-associated protein Cupressus sempervirens /

ACA30301.1 5.00E-33 cyt HE859606

Lipoprotein Lactobacillus rhamnosus GG /

CAR86203.1 5.00E-31 n.d HE859514

Lipoprotein Lactobacillus rhamnosus GG /

CAR86203.1 9.00E-11 n.d HE859908

Low-density lipoprotein receptor-

related protein

Ascaris suum / ADY39765.1 2.00E-13 n.d. HE859807

Apolipoprotein D isoform 2 Apis mellifera / XP_623787.2 3.00E-08 n.d. HE859474

Putative apolipoprotein D Aedes aegypti / XP_001660232.1 5.00E-10 n.d. HE859648

Dermatopontin 2 Biomphalaria glabrata / AAZ80785.1 1.00E-05 n.d. HE859882

Cysteine-rich venom protein Aedes aegypti / XP_001655503.1 5.00E-07 n.d. HE859698

Venom allergen Aedes aegypti / XP_001655382 1.00E-36 n.d. HE859494

Leucine rich protein Arachis hypogaea / ABH09321.1 2.00E-12 n.d HE859468

Pg1 protein Staphylococcus aureus subsp. aureus

MN8 / ZP_06947416 4.00E-24 n.d. HE859493

Short-chain collagen C4-like Amphimedon queenslandica /

XP_003391538.1 4.00E-22 cyt HE859623

Similar To DUF221 domain protein Leptosphaeria maculans / CBX98851 1.00E-19 n.d. HE859532

Microvillar-like protein 5 Phlebotomus perniciosus /

ADJ57676.1 4.00E-05 cyt HE859418

Desiccation tolerance in Pyropia columbina 951

Continuation

Functional category Putative identity Species and accession number E value D*

Pyropia

accession

number

Tep1 Anopheles arabiensis / ACG68562.1 5.00E-11 cyt HE859562

Viral A-type inclusion protein Trichomonas vaginalis G3 /

XP_001319569.1 1.00E-06 cyt HE859685

Yip1 domain-containing protein Dictyostelium discoideum /

XP_639001 3.00E-31 n.d. HE859496

Figure 1. Functional categorization of ESTs obtained from hydration and desiccation P. columbina libraries. Percentage

values indicate the number of ESTs grouped in each functional group in relation to the total number of sequences

obtained in each library.

remodeling is also ATP-dependent (Luo & Dean,

1999). In the case of P. columbina it seems reasonable

to hypothesize the occurrence of a higher energy

production by the mitochondrial system during

hydration, as many metabolic reactions are necessary

to maintain a homeostatic redox state and, as a result,

a healthy physiological condition during the

hydration-desiccation cycle. Moreover, the decline in

ATP production resulting from a general metabolic

slow down during desiccation could be beneficial in

preventing ROS production triggered by the electronic

alterations induced by low water potential. In vascular

plants under water stress, ROS production has been

detected in apoplast, xylem vessels, chloroplasts and

mitochondria (Mittler et al., 2004; Toldi et al., 2009).

Therefore, it is possible that desiccation in P.

columbina induces ROS production in the organelles,

a hypothesis that needs to be experimentally

demonstrated. Additionally, the mitochondrial meta-

bolism should be measured in order to demonstrate a

decay-activation sequence of this system during the

desiccation-hydration cycle.

In the context of ROS attenuation by the

antioxidant system during the hydration-desiccation

cycle, several antioxidant enzymes should be expre-

ssed. In fact, a peroxiredoxin (PRX) typical 2-Cys, an

arachidonate 5-lipoxygenase, a glutathione S-transfe-

rase, and several cytochrome P450 and HSPs (heat

shock proteins, 27, 40, 70, 80, 90 types) were detected

during hydration. Moreover, during desiccation,

enzymes such as thioredoxin (TRX), catalase and low

variants of HSPs (70 and 90, gene sequences different

952 Latin American Journal of Aquatic Research

from hydrated fronds) were expressed. PRXs are

involved in detoxification of hydrogen peroxide,

alkylhydroperoxides and peroxinitrites (Hall et al.,

2009). These enzymes react at low peroxide

concentrations and may become inactive at higher

concentrations. In plants, PRX transcripts increase in

response to different abiotic stresses such as salinity,

drought, and heavy metals (Wood et al., 2003; Dietz,

2011). Their expression in algae has been poorly

studied, although some studies indicate they are

regulated by light, oxygen, copper, desiccation and

redox state (Goyer et al., 2002; Contreras-Porcia et

al., 2011a, 2001b; Lovazzano et al., 2013; see section

3 in Contreras-Porcia & López-Cristoffanini, 2012). In

vascular plants, several PRXs have been described

based on their catalytic mechanisms and subcellular

localization (Baier & Dietz, 1997). In this study, PRX

expresses mainly when fronds are hydrated, is

localized exclusively in the chloroplast, and its

reduction is TRX-dependent. The chloroplast TRX

activity was also recorded during desiccation (Table

2). Therefore, it seems likely that P. columbina PRX

play an important role in buffering oxidative stress

and in post-desiccation detoxification of lipoperoxides

in the chloroplast. However, other attenuation systems

might also be operating in both environmental

conditions in order to normalize the redox state

imbalance. For example, a chlorophycean ferredoxin

was identified during desiccation (Table 2). This is a

small protein that plays a key role in electron

distribution in the chloroplast (Schürmann &

Buchanan, 2008) by regulating the chloroplast

metabolic network through the TRX system, and

contributes directly to ascorbate antioxidant protection

(i.e., antioxidant compound and ascorbate peroxidase

substrate) and PRX regeneration (Ceccoli et al., 2011;

Dietz, 2011).

Another enzyme detected in hydrated fronds was a

lipoxygenase (LOX, Table 1), a dioxygenase which

peroxidates polyunsaturated fatty acids (Gigon et al.,

2004). Lipid molecules produced by lipid degrading

enzymes, such as oxylipins, can act as secondary

messengers of stress-response signal transduction

pathways (Blée, 2002; Vellosillo et al., 2007).

However, a hyper-stimulation of lipoxygenase activity

could induce an accumulation of lipope-roxides,

which leads to cell damage and organelle dysfunction.

For example, Contreras et al. (2009) demonstrated that

hyper-activity of LOX led to an over-production of

lipoperoxides, and at the end, to cell death in sensitive

species under copper-induced oxidative stress. The

particular expression of this enzyme during hydration

could explain the exceptional control of the lipid

peroxidation in P. columbina (Contreras-Porcia et al.,

2011a) in comparison with several other algae (Flores-

Molina et al., unpublished data). Moreover, the

involvement of PRX during the hydration-desiccation

cycle may additionally explain the effective ROS and

lipoperoxide attenuation in this species.

Several cytochrome P450 variants were found in P.

columbina under natural conditions of hydration and

desiccation. These enzyme variants are present in all

living species and catalyze the oxygenation of a high

variety of substrates (Anzenbacher & Anzenbacherová,

2003). It has been demonstrated that P450s are

induced by abiotic and biotic stress (Narusaka et al.,

2004; Stolf-Moreira et al., 2011). Some P450s have

been identified as ABA 8´hydroxylase that degrades

ABA (abscidic acid) during the hydration-desiccation

cycle (Kushiro et al., 2004; Shinozaki & Yamaguchi-

Shinozaki, 2007). In vascular plants, ABA is over-

produced during desiccation (as in P. columbina,

Guajardo et al. pers. comm.), causes stomatal closure,

and induces stress related genes. However, via the

ABA 8´hydroxylase-P450, ABA concentration is

reduced to basal levels during the transition from

desiccation to hydration (Kushiro et al., 2004). ABA

has been recently identified in several algal species

(i.e., Tarakhovskaya et al., 2007; Yokoya et al., 2010),

although its role in regulating the expression of genes

associated with tolerance to abiotic/biotic stress has

not being explored. In P. columbina the functional

role of P450 could open new avenues to learn on

tolerance pathways involved in managing

environmental stressors, such as the ABA involvement

in transcription of regulatory networks of desiccation

stress signals and gene expression. Moreover, ABA in

P. columbina could be involved in the up-regulation of

several compounds like sugar, prolines and

polyamines (e.g., putrescine, spermidine and

spermine), which are known to increase their

expression under water stress in vascular plants and

algae (Guill & Tuteja, 2010; Alcázar et al., 2011;

Kumar et al., 2011). Thus, it is also possible to

hypothesize regarding the participation of the ABA-

independent pathways in the regulatory response to

dehydration stress. Indeed, a Clp-protease was

identified during desiccation (Table 2), and the clp

gene not only was induced by dehydration, but was

also up-regulated during natural senescence

(Nakashima et al., 1997). Analysis of the clp gene in

transgenic plants indicates that the clp promoter

contains cis-acting element(s) involved not only in

ABA independent stress-responsive gene expression

but also in senescence-activated gene expression

(Simpson et al., 2003). Thus, several tolerance

pathways, previously unknown in algae and other

organisms, could be synergistically activated under

particular environmental stress conditions.

Desiccation tolerance in Pyropia columbina 953

Proteases are indispensable for the normal

functioning of cells and tissues in all living organisms.

However, their activities need to be correctly

regulated (Habib & Fazili, 2007). During the

hydration-desiccation cycle in P. columbina, several

peptidases were identified (Tables 1 and 2). Even

though the expression of these proteins is required for

protein metabolism, an over-expression can be

potentially harmful. During hydration, a

metallocarboxypeptidase inhibitor (CPI) was

identified, that forms a less active or fully inactive

enzyme. In vascular plants, CPIs are activated by

metals, mechanical wounds or insect injury

(Villanueva et al., 1998; Habib & Fazili, 2007; Harada

et al., 2010), and its expression is ABA-regulated

(Villanueva et al., 1998). Thus, this type of protease in

P. columbina could be part of the ABA genes that are

regulated during the hydration-desiccation cycle

needed to maintain cellular integrity under water

deficiency.

ABC (ATP binding cassette) transporter sequences

were identified in desiccated fronds. These

transporters are involved in translocation of a wide

variety of compounds across cell membranes,

including ions, carbohydrates, lipids, xenobiotics,

drugs, and heavy metals (Ehrmann et al., 1998; Sipos

& Kuchler, 2006; Contreras et al., 2010). These

transporters have been reported during metal tolerance

in vascular plants, carrying metal complexes from

cytosol to the vacuole (Clemens, 2001). Also, in

humans they appear associated with the protection of

placental tissue by preventing cellular accumulation of

cytotoxic compounds (Aye & Keelan, 2013). The

participation of these transporters in desiccated P.

columbina was unexpected, despite information which

recently demonstrated that tolerance to desiccation in

the free-living soil bacterium Rhizobium legume-

nosarum was associated with ABC-transporter activity

(Vanderlinde et al., 2010). In the bacterium, a

mutation in the ATP-binding component of a

previously uncharacterized ATP transporter (Young et

al., 2006) decreased the tolerance to desiccation due to

low exopolysaccharides levels in the cell wall

envelope of the mutant. That study demonstrated the

crucial role of polysaccharides and their transport in

organisms tolerant to desiccation. Therefore, our

results suggest, for the first time in algae, the

involvement of ABC-transporters in desiccation

tolerance, possibly through cell wall stabilization

during desiccation stress.

Several HSP were identified during hydration and

desiccation stress. These proteins are recognized in

prokaryotes and eukaryotes during responses to

different physiological and environmental stress

conditions (Feder & Hofmann, 1999). HSPs induction

leads to a state of resistance for subsequent stress in

the cell by preventing protein aggregation (Feder &

Hofmann, 1999) and suppressing apoptosis (e.g.,

HSP90 see Beere et al., 2000 and Ravagnan et al.,

2001). HSP70 blocks apoptosis by binding apoptosis

activating factor-1 (Apaf-1), thereby preventing the

formation of the apoptosome complex (Ravagnan et

al., 2001). In fish hepatocytes, HSPs induction is

involved in stress tolerance by modulating the action

of key proteins and kinases in the signal transduction

pathways (Padmini & Usha-Rani, 2011). In the

seaweeds Fucus serratus and F. vesiculosus, a small

HSP has also been recorded in response to abiotic

stress, but its specific role remains undetermined

(Pearson et al., 2010).

During desiccation sequences of cell-wall

hydrolases, glycosyl transferases, and chitin deacety-

lases (CDA) ―all involved in cell wall metabolism―

were identified. CDA, a type of carbohydrate esterase,

hydrolyzes the acetamide group in the N-

acetylglucosamine polymers derived from glucose

(e.g., chitin), and promotes the formation of

glucosamine units (e.g., chitosan). The substrates for

this enzyme come from the carbon (fructose 6-

phosphate) and nitrogen (glutamine) metabolism

(Ghormade et al., 2010). CDA was first discovered in

extracts of the fungus Mucor rouxii (Araki & Ito,

1975). It was further reported associated with cell wall

synthesis. CDA has also been reported in association

with spore formation in yeast and attack-defense

systems in plant-pathogen interactions. In P.

columbina an over-activation of this enzyme is likely

to occur, due to the induction, under desiccation stress,

of a fructose-1,6-phosphatase (Table 2) that can over-

produce fructose 6-phosphate, an important precursor

of the CDA substrate. Thus, the potential induction of

CDA in desiccated P. columbina may help in

remodeling and maintaining cell wall integrity during

growth, survival, and pathogenesis.

CONCLUSIONS

Despite their crucial ecological role as primary

producers, molecular information on stress responses

in intertidal macroalgae remains limited. Pyropia

columbina, such as others Porphyra and Pyropia

species, is a good model for unravelling some of the

biological and molecular responses associated with

desiccation and other environmental conditions that

may cause oxidative stress. In this context, two

subtracted EST libraries were constructed in order to

understand the metabolic pathways active during the

hydration-desiccation cycle in this species. Results

954 Latin American Journal of Aquatic Research

showed that a significant portion of the transcripts had

no known homologues in algae or other organisms as

far as sequence data were available. These sequences

are interesting since they could represent genes unique

to this species. On the other hand, several

genes/proteins not previously described in algae were

differentially expressed in both environmental

conditions. This information contributes to a better

understanding the molecular mechanisms involved in

tolerance to desiccation. However, a confirmation of

the expression profiles of the reported genes by qPCR

is needed to characterize, for example, temporality of

the gene expression profile during the hydration-

desiccation cycle. Additionally, our study provides for

the first time, a set of candidate genes for further

examination of the physiological responses to other

environmental stressors. This genetic background will

broaden our understanding on physiological

differences may contribute, or even determine, the

ecological features of macroalgae inhabiting the

intertidal rocky zone.

ACKNOWLEDGEMENTS

Funding provided by FONDECYT 11085019 and

partially by FONDECYT 1120117, to LCP, and

Marine Genomics Europe Technology Platforms Bid

no. 43, to LCP, JC, MK and RR. Additional funding

comes from FONDAP 1501-0001 (CONICYT), to the

Center for Advanced Studies in Ecology &

Biodiversity (CASEB) Program 7, and to JC.

Authors' Contributions: LCP, JC, and RR

conceived and designed the project. MK and RR were

responsible for library construction and sequencing.

CLC, CL, MFM, EG, DT, and AN performed

functional annotation analyses. LCP wrote the

manuscript. All authors edited the manuscript. All

authors read and approved the manuscript.

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Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome

Article

Full-text available

Jan 2000

Corrigendum: Genome sequencing in microfabricated high-density picolitre reactors

Article

Full-text available

Jan 2006

Estudios ficoecológicos en la región costera de Valparaíso

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Krisler Alveal

Impact of phyto-oxylipins in plant defense

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Jan 2004
TRENDS PLANT SCI

E. Blée

Algas Marinas de Chile, Distribución, Ecología, Utilización y Diversidad. Ediciones Universidad Católica de Chile, Santiago

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Bernabe Santelices

The ubiquitin-mediated proteolytic pathway and mechanisms of energy-dependent intracellular protein degradation

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In this review we briefly describe the lysosomal system, consider the evidence for multiplicity of protein degradation pathways in vivo, discuss in detail the ubiquitin‐mediated pathway of intracellular ATP‐dependent protein degradation, and also the possible significance of ubiquitin‐histone conjugates in chromatin. For detailed discussions of the various characteristics and physiological roles of intracellular protein breakdown, the reader is referred to earlier reviews [1–7] and reports of recent symposia [8–10]. Information on the ubiquitin system prior to 1981 was described in an earlier review [11]. Hershko has briefly reviewed more recent information [12].

Reactive oxygen gene network of plants [J]

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Jan 2004
TRENDS PLANT SCI

Investigaciones. Algas Marinas de Chile: Distribucion, Ecologica, Utilizacion, Diversidad

Article

Aug 1990

Molecular Cloning of Abscisic Acid-Modulated Genes which are Induced during Desiccation of the Resurrection Plant

Article

Apr 1990

Leaves of the resurrection plant Craterostigma plantagineum Hochst, can be desiccated up to 1% relative water content and are still viable after rehydration. To clone genes related to this extreme desiccation tolerance, an in-vitro system was first developed which allows the induction of the same resurrection response in callus tissue upon treatment with abscisic acid (ABA). Several proteins and in-vitro-synthesized polypeptides were then identified which can be induced both in desiccation-tolerant, naturally dried leaves and in ABA-treated calli surviving after rehydration. Complementary-DNA clones corresponding to mRNAs expressed only in desiccation-tolerant tissues were obtained and classified into several gene families. In hybrid-selected translation experiments, representative cDNA clones were associated with water stress and ABA-inducible polypeptides abundantly expressed in dried leaves and ABA-treated calli. The expression pattern of several of these abundant transcripts was analyzed in RNA-hybridization experiments. Upon stress or ABA treatment the transcription levels increased rapidly, but they declined after relief from the stress state. This, together with data on genomic copy numbers indicated that a set of abundantly expressed genes are involved in the desiccation process of resurrection plants. Data on endogenous ABA contents before and after stress applications and on the physiological effects of exogenous ABA treatments indicate that in Craterostigma plantagineum the induction of an extreme desiccation tolerance is mediated by this plant hormone.

Physiological activity of Porphyra in relation to eulittoral zonation

Article

Oct 2008
J EXP MAR BIOL ECOL

Eulittoral seaweeds at different tidal elevations are exposed to various frequencies and durations of emergence. Their physiological activities (e.g. nutrient uptake) may be affected by water loss during emersion. We used three Porphyra species from different tidal elevations to test whether species at different vertical elevations on the shore respond differently to the increasingly non-marine environment, in terms of their physiological activities including nutrient uptake, tissue carbon, nitrogen and phycoerythrin contents. Simulated tidal cycles produced water losses of 0%, 40±10% and 90±5% tissue water. Emersion was stressful for all species regardless of their habitat. It was more stressful to nitrate and phosphate uptake for the sublittoral species P. yezoensis than eulittoral species, P. umbilicalis and P. leucosticta. Interestingly, tissue N for thalli that had been emerged and then re-submerged was significantly higher than those of continuously submerged individuals. During exposure, tissue N contents of all species declined but recovered quickly (e.g. within 30 min) after re-submergence. This result suggests that emersion-induced N release may constitute an undescribed biogeochemical pathway linking marine, terrestrial, and atmospheric N reservoirs.

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