Experimental fossilisation of viruses from extremophilic Archaea

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n

Discussion Paper | Discussion Paper | Discussion Paper | Discussion Paper |

Bio

geoscie

nces

Dis

cuss.,

8,2235–2257,2011

ww

w.b

iogeoscie

nces-d

iscuss.n

et/8/2

235/2

011/

doi:10.5

194/b

gd-8

-2235-2

011

©A

uth

or(

s)

2011.

CC

Attri

bution

3.0

Lic

ense.

Bio

ge

os

cie

nc

es

Dis

cu

ss

ion

s

This

dis

cussio

npa

per

is/h

as

been

under

revie

wfo

rth

ejo

urn

alB

iogeoscie

nces

(BG

).

Ple

ase

refe

rto

the

corr

espondin

gfinalpa

per

inB

Gif

availa

ble

.

Exp

eri

men

talfo

ssilis

ati

on

of

vir

uses

fro

m

extr

em

op

hilic

Arc

haea

F.O

ran

ge

1,2

,A

.C

hab

in1,A

.G

orl

as

3,S

.L

ucas-S

taat4

,C

.G

eslin

3,

M.L

eR

om

an

cer3

,D

.P

ran

gis

hvili4

,P.F

ort

err

e4,an

dF.W

esta

ll1,2

1C

en

tre

de

Bio

physiq

ue

Mo

lecu

laire,

CN

RS

,U

PR

43

01

,R

ue

Ch

arl

es

Sa

dro

n,

45

07

1O

rle

an

s

Ce

dex

2,

Fra

nce

2O

bse

rva

toire

de

sS

cie

nce

sd

el’U

niv

ers

en

reg

ion

Ce

ntr

e,

UM

S3

11

6,

1A

Ru

ed

ela

Fe

rolle

rie,

45

07

1O

rle

an

sC

ed

ex

2,

Fra

nce

3U

niv

ers

ite

de

Bre

tag

ne

Occid

enta

le,

CN

RS

–U

MR

65

39

,In

stitu

tU

niv

ers

ita

ire

Eu

rop

ee

nd

e

laM

er,

Te

ch

no

po

leB

rest-

Iro

ise,

Ru

eD

um

on

td

’Urv

ille,

29

28

0P

lou

za

ne

,F

ran

ce

4M

ole

cu

lar

Bio

log

yo

fth

eG

en

ein

Extr

em

op

hile

sU

nit,

Institu

tP

aste

ur,

rue

du

Do

cte

ur

Ro

ux

25

,7

57

24

Pa

ris

Ce

dex

15

,F

ran

ce

Re

ce

ive

d:

16

Fe

bru

ary

20

11

–A

cce

pte

d:

17

Fe

bru

ary

20

11

–P

ublis

he

d:

4M

arc

h2

01

1

Co

rre

sp

on

de

nce

to:

F.O

ran

ge

(fra

nco

is.o

ran

ge

@cn

rs-o

rle

an

s.f

r)

Pu

blis

he

dby

Co

pe

rnic

us

Pu

blic

atio

ns

on

be

ha

lfo

fth

eE

uro

pe

an

Ge

oscie

nce

sU

nio

n.

2235

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Ab

str

act

The

role

of

vir

uses

at

diff

ere

nt

sta

ges

of

the

ori

gin

of

life

has

recently

been

reconsid

-

ere

d.

Itappears

thatvir

uses

may

have

accom

panie

dth

eearl

iestfo

rms

oflif

e,allo

win

g

the

transitio

nfr

om

an

RN

Ato

aD

NA

worl

dand

possib

lybein

gin

volv

ed

inth

eshapin

g

oftr

ee

oflif

ein

the

thre

edom

ain

sth

atw

eknow

pre

sently.

Inadditio

n,a

larg

eva

riety

of

5

vir

uses

has

been

recently

identified

inextr

em

eenvironm

ents

,hoste

dby

extr

em

ophili

c

mic

roorg

anis

ms,

inecosyste

ms

consid

ere

das

analo

gues

toth

ose

of

the

earl

yE

art

h.

The

earl

iesttr

aces

oflif

ew

ere

pre

serv

ed

by

the

pre

cip

itation

ofsili

ca

on

org

anic

str

uc-

ture

s.

The

stu

dy

of

the

insitu

and

experi

menta

lfo

ssili

sation

of

mic

roorg

anis

ms

allo

ws

better

unders

tandin

gof

the

fossili

sation

pro

cesses

and

help

sid

entification

of

traces

of

10

life

inancie

nt

rocks.

Ina

continuation

of

these

stu

die

s,

we

pre

sent

the

results

of

the

firs

texperi

menta

lfo

ssili

sation

by

sili

ca

ofvir

uses

from

extr

em

ophili

cA

rchaea

(SIR

V2

–

Sulfo

lobus

isla

ndic

us

Vir

us

2,

TP

V1

–T

herm

ococcus

pri

euri

ivir

us

1,

and

PA

V1

–P

y-

rococcus

abyssi

vir

us

1).

Our

results

confirm

that

vir

uses

can

be

fossili

sed,

with

sili

ca

pre

cip

itating

on

the

diff

ere

nt

viralstr

uctu

res

(pro

tein

s,

enve

lope)

ove

rseve

ralm

onth

s.

15

How

eve

rdiff

ere

nces

inth

esili

cifi

cation

pro

cess

were

noticed,

dependin

gon

the

viral

str

uctu

reand

com

positio

n.

The

fossili

sation

mechanis

mis

sim

ilar

toth

at

of

the

fossili

-

sation

of

mic

roorg

anis

ms.

This

stu

dy

thus

suggests

that

viralre

main

sor

traces

could

be

pre

serv

ed

inth

ero

ck

record

although

their

identification

may

be

challe

ngin

gdue

to

the

sm

all

siz

eofth

eviralpart

icle

s.

20

1In

tro

du

cti

on

The

role

of

vir

uses

inearl

yevo

lution

of

life

has

recently

been

reeva

luate

d.

For

alo

ng

tim

evir

uses

were

consid

ere

das

non-liv

ing

by-p

roducts

of

cellu

lar

activity

and

thus

unable

tohave

pla

yed

aro

lein

the

ori

gin

and

evo

lution

of

their

hosts

.O

ver

the

last

decade,

how

eve

r,our

vie

ws

on

the

ori

gin

and

natu

reof

vir

uses

have

dra

matically

25

2236

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


changed.

Str

uctu

ral

sim

ilari

ties

betw

een

vir

uses

infe

cting

phylo

genetically

dis

tant

hosts

suggest

that

vir

uses

were

already

active

at

the

tim

eof

the

Last

Univ

ers

alC

om

-

mon

Ancesto

r(L

UC

A)

and

pro

bably

befo

re(B

am

ford

et

al.,

2005;

Koonin

and

Dolja

,

2006;

Kru

povic

and

Bam

ford

,2008).

Com

para

tive

genom

ics

has

reve

ale

dth

at

most

genes

encoded

by

vir

uses

have

no

cellu

lar

hom

olo

gues,

testify

ing

the

cre

ativity

of

vi-

5

rocells

(Fort

err

e,

2010)

inin

venting

new

pro

tein

s.

Seve

ral

theori

es

and

dis

cove

ries

have

been

made

regard

ing

the

ancie

ntnatu

reofvir

uses

and

their

possib

lekey

role

sin

earl

yevo

lution

of

life

and

inth

eori

gin

of

modern

cells

(Fort

err

e,

2002;

Bam

ford

,2003;

Koonin

et

al.,

2006;

for

recent

revie

ws

and

altern

ative

vie

wpoin

ts,

see

More

ira

and

Lopez-G

arcıa

,2009;

Fort

err

eand

Pra

ngis

hvili

,2009a,

b).

For

insta

nce,

vir

uses

have

10

been

evo

ked

toexpla

inth

eori

gin

of

DN

Acells

.S

tart

ing

from

an

RN

Aw

orl

d(c

ells

with

RN

Agenom

es),

DN

Acould

have

appeare

din

vir

uses

(DN

Avir

uses)

and

then

have

been

transfe

rred

tocells

(Fort

err

e,

2002).

One

hypoth

esis

als

osuggests

that

inte

rac-

tions

betw

een

DN

Avir

uses

and

RN

Acells

(fusio

n,gene

transfe

rs)

could

have

resulted

inth

efo

rmation

ofth

eth

ree

dom

ain

s(E

ukary

ote

s,B

acte

ria,A

rchaea)

(Fort

err

e,2006).

15

Fin

ally

,th

edis

cove

ryofM

imiv

irus,a

gia

ntD

NA

vir

us

infe

cting

Am

oeba

(La

Scola

etal.,

2003),

perm

its

avir

us

genom

eto

be

inclu

ded

inth

euniv

ers

alt

ree

oflif

e.

This

suggests

that

gia

nt

vir

uses

could

be

inte

rmedia

tes

betw

een

cells

and

pre

vio

usly

know

nvir

uses

and

could

form

anew

dom

ain

oflif

e(R

aoult

etal.,2004).

Am

ajo

rdeve

lopm

ent

inre

cent

years

invirolo

gy

has

been

the

dis

cove

ryof

very

20

div

ers

enew

vir

uses

inextr

em

eenvironm

ents

.S

uch

environm

ents

are

part

icula

rly

in-

tere

sting

as

they

are

often

consid

ere

das

analo

gues

of

the

environm

ents

on

the

earl

y

Eart

hin

whic

hlif

ecould

have

ori

gin

ate

d.

Mic

roorg

anis

ms

found

inth

ese

environm

ents

are

als

oconsid

ere

das

possib

leanalo

gues

toth

eearl

iestfo

rms

oflif

eon

Eart

hand

pos-

sib

lyon

Mars

(Nis

betand

Sle

ep,2001;K

onhauser

etal.,2003).

Recognitio

nofth

ean-

25

cie

ntnatu

reofvir

uses

suggests

thatearl

ylif

efo

rms

could

have

,in

deed,hoste

dvir

uses.

The

stu

dy

ofvir

uses

ofextr

em

ophili

cm

icro

org

anis

ms

isa

rela

tive

lyre

centfield

.N

eve

r-

thele

ss,it

has

already

allo

wed

the

identification

ofseve

ralth

ousands

ofvir

uses

havin

g

diff

ere

ntm

orp

holo

gie

sand

chara

cte

ristics,and

als

oofnew

vir

us

fam

ilies

(see

revie

win

2237

https://www.researchgate.net/publication/249543839_Hot_spring_sinters_keys_to_understanding_Earth's_earliest_life_forms_Can_J_Earth_Sci?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Le

Rom

ancer

etal.,2007).

Vir

uses

have

been

identified

inall

know

nextr

em

eenviron-

ments

:hypers

alin

eenvironm

ents

(Ore

netal.,1997;D

yall-

Sm

ith

etal.,2003,P

agalin

g

et

al.,

2007;

Sim

e-N

gando

et

al.,

2010),

alk

alin

ela

kes

(Jia

ng

et

al.,

2004),

desert

s

(Pri

gentetal.,2005),

pola

rre

gio

ns

(Mara

nger

etal.,1994;K

epner

etal.,1998;B

orr

iss

et

al.,

2003;

Gow

ing,

2003),

indeep

subsurf

ace

sedim

ents

(Bird

et

al.,

2001),

and

in5

hydro

therm

alenvironm

ents

.T

he

searc

hfo

rnew

vir

uses

inth

ela

tter

environm

ent

has

been

especia

llyfr

uitfu

l,fo

llow

ing

the

pio

neeri

ng

work

ofW

olfra

mZ

illig

on

the

vir

uses

of

hypert

herm

ophili

cA

rchaea

(Mart

inet

al.,

1984;

Ric

eet

al.,

2001;

Rachelet

al.,

2002).

Many

new

viral

fam

ilies

have

been

identified

that

infe

ct

hypert

herm

ophili

cA

rchaea

in

terr

estr

ial

and

mari

ne

hot

spri

ngs

(Geslin

et

al.,

2003a,

2005;

Ort

mann

and

Suttle

,10

2005;

Pra

ngis

hvili

et

al.,

2006a;

Ort

mann

et

al.,

2006;

Le

Rom

ancer

et

al.,

2007).

Although

these

vir

uses

all

have

double

-str

and

DN

Agenom

es,

they

pro

duce

vir

ions

with

very

div

ers

em

orp

holo

gie

s(e

.g.

rod-s

haped,

fila

mento

us,

spin

dle

-shaped,

elli

p-

soid

,head

and

tail)

and

mostpro

tein

sencoded

inth

eir

genom

es

have

no

hom

olo

gues,

exceptsom

etim

es

inoth

er

virallin

eages

(Pra

ngis

hvili

etal.,2006b).

15

Although

str

uctu

raland

com

para

tive

genom

ics

poin

tto

the

antiquity

of

vir

uses,

their

fossil

rem

ain

shave

yet

tobe

dete

cte

din

the

rock

record

.A

lthough

challe

ngin

g,

con-

sid

eri

ng

the

siz

eof

viral

part

icle

s,

dete

ction

of

possib

lefo

ssili

sed

viral

rem

ain

sin

the

vic

inity

of

puta

tive

fossili

sed

mic

roorg

anis

ms

could

be

adirect

pro

of

of

the

antiquity

of

vir

uses

and

an

additio

nalclu

eto

assessin

gth

ebio

genic

ity

of

the

observ

ed

str

uctu

res.

20

The

old

est

know

nfo

ssil

mic

roorg

anis

ms,

dating

back

toalm

ost

3.5

bill

ion

years

(Ga)

ago,

were

pre

serv

ed

as

sili

cifi

ed

rem

ain

s(s

ee

revie

win

Westa

ll,2010).

The

pre

ser-

vation

by

sili

ca

was

due

toth

efa

ct

that

the

earl

yE

art

h’s

oceans

were

sili

ca

enri

ched

com

pare

dto

the

unders

atu

rate

doceans

of

the

pre

sent

Eart

hw

here

sili

ceous

org

an-

ism

sare

asin

kfo

rava

ilable

sili

ca.

An

additio

nal

sourc

eof

sili

ca

inth

eearl

yoceans

25

cam

efr

om

the

very

active

hydro

therm

al

pro

cesses

cyclin

gsili

ca

and

oth

er

ele

ments

from

the

cru

st

back

into

the

ocean

(see

revie

win

Westa

lland

South

am

,2006).

Insitu

and

experi

menta

lsili

cifi

cation

of

mic

roorg

anis

ms

inth

ese

hydro

therm

ally

-influenced

environm

ents

has

been

stu

die

din

depth

ove

rth

epast

two

decades

(Westa

llet

al.,

2238

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


1995;Topors

kie

tal.,2002;K

onhauser

etal.,2004;O

range

etal.,2009

and

refe

rences

there

in).

These

inve

stigations

have

pro

vid

ed

pre

cio

us

info

rmation

regard

ing

the

pro

-

cesses

invo

lved

and

help

ed

the

identification

of

sili

cifi

ed

traces

of

life

inancie

nt

rocks

(see

revie

win

Westa

ll,2010).

Recent

stu

die

shave

sta

rted

toexplo

reth

epossib

ility

thatvir

uses

could

be

pre

serv

ed

inth

efo

ssil

record

.P

oin

ar

and

Poin

ar

(2005)

report

ed

5

the

possib

lepre

sence

ofpre

serv

ed

vir

uses

in15

to100

Ma

insectpre

serv

ed

inam

ber.

Laid

ler

and

Ste

dm

an

(2010)

monitore

dth

eexperi

menta

lfo

ssili

sation

ofbacte

riophage

T4

for

afe

wdays

under

sim

ula

ted

hot

spri

ng

sili

cifyin

gconditio

ns

and

show

ed

that

sili

ca

could

pre

cip

itate

aro

und

viralstr

uctu

res

and

pre

serv

eth

em

.

Ourstu

dy

pre

sents

the

results

ofth

efirs

tlo

ng-t

erm

experi

menta

lfossili

sation

ofth

ree

10

vir

uses

and

the

firs

texperi

menta

lfo

ssili

sation

of

vir

uses

hoste

dby

hypert

herm

ophili

c

Arc

haea.

Am

ong

the

gre

at

vari

ety

of

vir

uses

inhypert

herm

ophili

cA

rchaea,

we

chose

for

this

stu

dy

vir

uses

pro

ducin

gvir

ions

(viral

part

icle

s)

with

very

diff

ere

nt

str

uctu

res

and

morp

holo

gie

s:

the

rod-s

haped

vir

us

SIR

V2

(Sulfo

lobus

isla

ndic

us

vir

us

2)

and

the

spin

dle

-shaped

TP

V1

(Therm

ococcus

pri

euri

ivir

us

1)

and

PA

V1

(Pyro

coccus

abyssi

15

vir

us

1)

vir

uses.

These

morp

holo

gie

sappear

tobe

specifi

cfo

rvir

uses

infe

cting

org

an-

ism

sliv

ing

inextr

em

eenvironm

ents

,either

hot

tem

pera

ture

environm

ent

(rod-s

haped,

spin

dle

-shaped)

or

hig

hsalt

(spin

dle

-shaped),

with

the

exception

ofsom

epla

ntvir

uses

whic

hals

opro

duce

rod-s

haped

part

icle

s.

Our

obje

ctive

sw

ere

(1)

todete

rmin

ew

heth

er

vir

uses

could

be

fossili

sed

by

sili

ca

and

20

wheth

er

they

could

be

pre

serv

ed

ove

rre

lative

lylo

ng

peri

ods

of

tim

e(s

eve

ralm

onth

s),

and

(2)

toeva

luate

wheth

er

recogniz

able

featu

res

inth

eviralstr

ain

sused

could

still

be

identified

once

sili

ca

had

pre

cip

itate

daro

und

them

.

2239

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


2M

ate

rials

an

dm

eth

od

s

2.1

Descri

pti

on

of

the

vir

uses

used

for

this

stu

dy

SIR

V2

(Sulfo

lobus

isla

ndic

us

rod-s

haped

vir

us

2)

belo

ngs

toth

eR

udiv

irid

ae

fam

ily

(Pra

ngis

hvili

et

al.,

1999;

Biz

eet

al.,

2009)

and

isa

lytic

vir

us

(i.e

.w

hic

hcauses

the

death

of

the

host

cell)

that

infe

cts

astr

ain

of

the

Cre

narc

haeota

Sulfo

lobus

isla

ndic

us,

5

an

acid

ophili

cand

hypert

herm

ophili

cA

rchaea

thatw

as

ori

gin

ally

isola

ted

from

sam

ple

s

taken

from

solfa

tari

cfield

sin

Icela

nd

(Zill

iget

al.,

1994).

SIR

V2

part

icle

sare

stiff

rods

up

to900

nm

long

and∼

20

nm

wid

e(F

ig.

1a;

Pra

ngis

hvili

et

al.,

1999)

and

consis

tof

asuperh

elix

inclu

din

ga

pro

tein

with

double

str

anded

linear

DN

A.

Incontr

ast

tooth

er

vir

uses,

such

as

rod-s

haped

mem

bers

of

the

Lip

oth

rixvir

idae,

SIR

V2

vir

ions

are

not

10

enve

loped.

TP

V1

(Therm

ococcus

pri

euri

ivir

us

1)

and

PA

V1

(Pyro

coccus

abyssi

vir

us

1)

were

isola

ted

from

the

Eury

arc

haeota

Therm

ococcus

pri

euri

istr

ain

Bio

-pl-0405IT

2and

Py-

rococcus

abyssi

str

ain

GE

23,

respective

ly.

The

latter

are

two

neutr

ophili

cand

hy-

pert

herm

ophili

cA

rchaea

of

the

Therm

ococcale

sord

er

(Geslin

et

al.,

2003b;

Gorl

as

15

et

al.,

2009,

2011).

TP

V1

and

PA

V1

are

non-lytic,

spin

dle

-shaped

vir

uses

(PA

V1:

120×

80

nm

;T

PV

1:

140×

80

nm

),chara

cte

rised

by

double

-str

anded

circula

rD

NA

,and

are

found

either

isola

ted

or

ingro

ups

(Fig

.1c,

d).

TP

V1

and

PA

V1

have

an

enve

lope

com

posed

of

viral

pro

tein

sand

lipid

sfr

om

the

host.

These

vir

uses

morp

holo

gic

ally

resem

ble

mem

bers

ofth

eF

usello

vir

idae

fam

ilybuthave

notyetbeen

cla

ssifi

ed.

20

2.2

Vir

us

iso

lati

on

The

vir

uses

used

inth

isstu

dy

were

harv

este

dfr

om

fresh

culture

softh

ehoststr

ain

s.

For

the

pro

duction

of

SIR

V2,

the

Sulfo

lobus

isla

ndic

us

str

ain

LA

L14/1

was

gro

wn

untilth

ela

teexponentialphase

was

reached,as

descri

bed

by

Zill

igetal.

(1994).

Cells

were

rem

ove

dby

low

-speed

centr

ifugation

(3500

gin

aS

orv

all

GS

3ro

tor)

.T

he

vir

uses

25

were

pre

cip

itate

dfr

om

the

supern

ata

ntby

the

additio

nof1

MN

aC

land

10%

PE

G6000

and

incubate

dove

rnig

ht

at

4◦

C.

Apelle

tof

vir

uses

was

colle

cte

dby

centr

ifugation

in

2240

https://www.researchgate.net/publication/12868332_A_Novel_Virus_Family_the_Rudiviridae_Structure_Virus-Host_Interactions_and_Genome_Variability_of_the_Sulfolobus_Viruses_SIRV1_and_SIRV2?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


aS

orv

all

GS

Aro

tor

at

23

000

gfo

r30

min

and

suspended

inTA

buffer

(20

mM

Tri

s-

acecate

,pH

6).

SIR

V2

were

puri

fied

by

centr

ifugation

ina

CsC

lbuoyant

density

gra

di-

ent

(0.4

5g

ml−

1)

ina

Beckm

an

SW

41

roto

rcentr

ifuge

at

250

000

gfo

r48

h.

Fra

ctions

conta

inin

gth

eviral

part

icle

sw

ere

colle

cte

dw

ith

asyri

nge

then

dia

lyzed

again

st

TA

buffer

for

furt

her

analy

sis

(Bettste

tter

et

al.,

2003).

The

vir

uses

were

sto

red

at

4◦

Cin

5

a20

mM

Tri

s-a

ceta

tebuffer

(pH

6)

untilused.

Therm

ococcus

pri

euri

istr

ain

Bio

-pl-0405IT

2(G

orl

as

et

al.,

2011)

and

Pyro

coccus

abyssi

str

ain

GE

23

(Era

uso

et

al.,

1993;

Mart

ein

sson

et

al.,

1995)

were

gro

wn

inth

e

mediu

mdescri

bed

by

Geslin

et

al.

(2003b),

at

80◦

Cand

85◦

C,

respective

ly,

up

toth

e

late

exponentialphase.

The

cells

were

pelle

ted

by

low

-speed

centr

ifugation

at

6000

g10

for

15

min

.T

PV

1and

PA

V1

were

pre

cip

itate

dfr

om

the

supern

ata

nt

in1

MN

aC

lw

ith

10%

PE

G6000

ove

rnig

ht

at

4◦

Cw

ith

gentle

stirr

ing.

The

pre

cip

itate

was

pelle

ted

by

centr

ifugation

at

13

000

gfo

r1

h15

min

,th

en

dra

ined

and

resuspended

ina

spe-

cifi

cbuffer

for

each

vir

us

(TP

V1:

TP

V1-b

uffer

–10

mM

Tri

s-H

CL,

100

mM

NaC

l,5

mM

CaC

l2;

PA

V1:

TE

buffer

–10

mM

Tri

s-H

Cland

1m

ME

DTA

,pH

8).

Asecond

pre

cip

ita-

15

tion

with

PE

G6000

(10%

)and

1M

NaC

lwas

made

duri

ng

1.5

hand

the

pre

cip

itate

was

colle

cte

das

descri

bed

above

.A

fter

centr

ifugation

(5000

gfo

r10

min

),th

esupern

ata

nt

was

kept

and

the

pelle

tw

as

extr

acte

dtw

om

ore

tim

es

under

the

sam

econditio

ns

with

reduced

volu

mes

ofT

PV

1or

TE

buffer.

The

vir

us-c

onta

inin

gsupern

ata

nts

were

poole

d

and

sto

red

for

one

nig

htat4◦

C.A

fter

centr

ifugation

(5000

gfo

r15

min

)to

rem

ove

resid

-20

ualcell

debri

s,

the

supern

ata

nt

was

concentr

ate

dby

ultra

centr

ifugation

at

33

000

gfo

r

1h

45

min

(Beckm

an

Optim

aLE

-80K

70.1

Tiro

tor)

and

the

pelle

tw

as

resuspended

in

TP

V1

or

TE

buffer.

Vir

uses

were

puri

fied

from

these

suspensio

ns

by

centr

ifugation

in

aC

sC

lbuoyant

density

gra

die

nt

(TP

V1:

1.3

2g

mL−

1;

PA

V1:

1.2

98

gm

L−

1)

ina

Beck-

man

Optim

aLE

-80K

70.1

Ti

centr

ifuge

roto

rat

180

000

gfo

r6

h(P

AV

1:

220

000

gfo

r25

24

h).

Fra

ctions

conta

inin

gth

enucle

icacid

sw

ere

dete

cte

dat

254

nm

and

colle

cte

d

usin

ga

density

gra

die

nt

fractionato

r(m

odel

185,

ISC

O).

These

fractions

were

then

dia

lyzed

again

st

ala

rge

volu

me

of

TP

V1

or

TE

buffer

and

were

sto

red

at

4◦

Cuntil

used.

2241

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


2.3

Silic

ificati

on

pro

ced

ure

The

fossili

sation

experi

ments

were

made

directly

inth

ebufferi

ng

media

of

the

vir

uses.

This

was

done

for

pra

cticalre

asons

as

well

as

tom

ain

tain

the

vir

uses

ina

favo

ura

ble

environm

ent

so

that

their

evo

lution

could

be

follo

wed

duri

ng

fossili

sation

ove

rseve

ral

month

s.

Although

itw

as

not

possib

leto

ascert

ain

the

accura

cy

of

the

finalsili

ca

con-

5

centr

ations

due

toth

esm

all

volu

mes

invo

lved,

fossili

sin

gm

edia

were

satu

rate

dw

ith

respectto

sili

ca.

Asto

ck

sili

ca

solu

tion

(3200

ppm

Si)

was

pre

pare

dfr

om

apure

sodiu

msili

cate

solu

tion

(Rie

del

de

Haen)

conta

inin

g∼

27%

SiO

2and

∼10%

NaO

H(N

a2S

i 3O

7,

M=

242

gm

ol−

1).

Its

pH

was

adju

ste

dto

8befo

rein

jection

into

the

vir

us

suspensio

n.

10

19

µL

of

asuspensio

nof

puri

fied

SIR

V2

were

mix

ed

with

1µL

of

the

sto

ck

sili

ca

so-

lution

ina

seale

dgla

ss

via

lto

obta

ina

finalconcentr

ation

of∼

160

ppm

Si.

For

TP

V1

and

PA

V1,180

µL

ofsuspensio

ns

ofpuri

fied

vir

uses

were

mix

ed

with

20

µL

ofth

esto

ck

sili

ca

solu

tion

toobta

ina

finalconcentr

ation

of∼

320

ppm

Si.

The

SIR

V2

fossili

sation

experi

ment

via

lsw

ere

sto

red

at

room

tem

pera

ture

(due

toth

esm

all

volu

me,

hig

hte

m-

15

pera

ture

sle

dto

eva

pora

tion).

The

TP

V1

and

PA

V1

fossili

sation

experi

ment

via

lsw

ere

pla

ced

inan

ove

nat60◦

C.

2.4

Ele

ctr

on

mic

rosco

py

The

via

lsw

ere

sam

ple

datdiff

ere

nttim

es

(betw

een

2and

60

days

for

the

SIR

V2

exper-

iment

and

betw

een

1and

180

days

for

the

TP

V1

and

PA

V1

experi

ments

)by

colle

cting

20

∼1

µL

of

the

sam

ple

and

imm

edia

tely

pre

pari

ng

itfo

rnegative

ly-s

tain

ed

transm

issio

n

ele

ctr

on

mic

roscopy

(Geslin

etal.,2003b).

For

negative

sta

inin

g,a

dro

ple

tofsam

ple

(either

unsili

cifi

ed

or

sili

cifi

ced)

was

pla

ced

on

acarb

on-c

oate

dcopper

TE

Mgri

d.

The

sam

ple

was

allo

wed

toabsorb

toth

ecarb

on

layer

for

2m

inbefo

rere

movin

gth

eexcess

liquid

with

apie

ce

of

filter

paper.

Som

e25

sam

ple

sw

ere

sta

ined

toin

cre

ased

contr

ast

by

pla

cin

ga

dro

ple

tof

satu

rate

dura

nyl

aceta

teeth

anolic

solu

tion

for

40

sand

then

rem

ovin

gth

eexcess

liquid

.S

om

egri

ds

2242

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


were

als

oobserv

ed

unsta

ined

tobetter

dis

tinguis

hsili

ca

depositio

non

the

viralstr

uc-

ture

s.

The

pre

pare

dsam

ple

sw

ere

then

air

dri

ed.

They

were

observ

ed

and

analy

sed

with

aP

hili

ps

CM

20

Tra

nsm

issio

nE

lectr

on

Mic

roscope

(Centr

ede

Mic

roscopie

Ele

c-

troniq

ue,U

niv

ers

ity

ofO

rleans),

equip

ped

with

an

ED

Xdete

cto

r(O

xfo

rdIn

str

um

ents

).

3R

esu

lts

5

3.1

Exp

eri

men

talfo

ssilis

ati

on

of

SIR

V2

SIR

V2

part

icle

sappear

as

stiff

rods

of

seve

ralhundre

ds

of

nanom

ete

rsin

length

and

∼20

nm

inw

idth

and

conta

ina

centr

al

cavity

(Fig

.1a),

not

vis

ible

on

unsta

ined

gri

ds

(Fig

.1b).

This

vir

ion

isnatu

rally

fragile

and

readily

bre

aks

up

into

fragm

ents

that

may

be

linked

toeach

oth

er

by

DN

Astr

ands

(Fig

.1b,

arr

ow

).A

part

from

fragm

enta

tion,

10

the

pre

serv

ation

of

SIR

V2

fragm

ents

duri

ng

fossili

sation

was

genera

llygood.

Inth

e

firs

tdays

of

the

fossili

sation

experi

ment,

agra

nula

rand

alv

eola

r-te

xtu

red

sili

ca

pre

cip

-

itate

form

ed

sponta

neously

(Fig

.2a,

b)

inboth

the

contr

olexperi

ment

without

vir

uses

and

inth

eexperi

ment

with

the

vir

uses,

indic

ating

that

neither

vir

us

part

icle

snor

the

bufferi

ng

mediu

mhad

any

influence

on

pre

cip

itation

or

on

the

str

uctu

reof

the

sili

ca.

15

Initia

lly,

after

one

week

of

fossili

sation

only

afe

wS

IRV

2part

icle

sw

ere

observ

ed

in

direct

conta

ct

with,

and

som

etim

es

trapped

within

,th

esili

ca

pre

cip

itate

(Fig

.2a)

but

most

were

not

(Fig

.2b).

Where

em

bedded

inth

esili

ca

deposit,

the

viral

helix

ap-

peare

dto

have

been

com

pacte

dor

decayed

and

the

vir

ion

was

only

vis

ible

as

afa

int,

dark

outlin

e(F

ig.2a,arr

ow

s)

thatpro

bably

repre

sents

degra

ded

DN

Aand

possib

lypro

-20

tein

s.

Sili

ca

nucle

ation

on

the

vir

ion

was

observ

ed

after

one

week

(Fig

.2c,arr

ow

)and,

after

30

days,

nanom

etr

icdark

part

icle

shad

form

ed

inth

ecentr

al

cavity

of

som

eof

the

vir

ions

(Fig

.2d).

This

phenom

enon

was

more

evid

ent

after

60

days

of

fossili

sation

with

vir

ions

show

ing,

on

unsta

ined

gri

ds,

astr

ongly

contr

aste

dcentr

alcavity

(Fig

.2e),

suggesting

thatth

ein

div

idualpart

icle

sobserv

ed

pre

vio

usly

had

continued

their

gro

wth

25

and

eve

ntu

ally

merg

ed.

More

pro

nounced

sili

ca

nucle

ation

on

the

outs

ide

of

the

viral

2243

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


part

icle

sw

as

als

oobserv

ed.

ED

Xanaly

ses

made

on

these

part

icle

sshow

ed

asig

nal

slig

htly

str

onger

sili

ciu

mth

an

the

sig

nalfr

om

backgro

und

nois

e(F

ig.

2f)

.F

ine

chain

s

of

sili

ca

part

icle

s(F

ig.

2g)

may

repre

sent

sili

ca

directly

pre

cip

itate

don

isola

ted

DN

A

str

ands,or

the

rem

nants

ofvir

ion

fragm

ents

com

ple

tely

cove

red

by

sili

ca.

3.2

Exp

eri

men

talfo

ssilis

ati

on

of

TP

V1

an

dP

AV

15

Although

agra

nula

r,alv

eola

rsili

ca

pre

cip

itate

form

ed

sponta

neously

within

one

day

of

the

experi

ment

(Fig

.3a),

itw

as

only

after

30

days

that

the

TP

V1

part

icle

sw

ere

observ

ed

indirect

conta

ct

with

the

pre

cip

itate

(Fig

.3b).

No

vir

ions

trapped

inth

e

sili

ca

pre

cip

itate

were

observ

ed.

The

firs

tsig

ns

of

possib

lesili

ca

pre

cip

itation

on

the

viralstr

uctu

res

occurr

ed

only

after

75

days

when

nanom

ete

rsiz

ed,dark

part

icle

sw

ere

10

observ

ed

within

(Fig

.3c)

and

on

the

enve

lope

of

the

vir

us

part

icle

s(F

ig.

3d).

Inth

e

latter

case,

ED

Xanaly

ses

(Fig

.3e)

indic

ate

that

this

pre

cip

itate

consis

tsof

both

sili

ca

and

cesiu

m,

the

latter

bein

gderi

ved

from

the

puri

fication

buffer.

Such

apre

cip

itate

had

not

been

observ

ed

inearl

ier

ste

ps

of

the

experi

ment

nor

on

unfo

ssili

sed

vir

uses

part

icle

s.

After

180

days,

the

vir

ions

were

sig

nifi

cantly

degra

ded

(Fig

.3f)

.S

om

eof

15

the

viral

part

icle

sconta

ined

only

afe

wnanopart

icle

sof

sili

ca

within

their

str

uctu

re

(Fig

.3f)

while

som

eoth

ers

were

heavily

coate

dand

impre

gnate

dw

ith

the

appare

ntly-

pro

gre

ssin

gS

iO2-C

spre

cip

itate

(Fig

.3g).

Monitori

ng

of

the

PA

V1

fossili

sation

experi

ment

was

com

plic

ate

dby

the

pre

sence

of

num

ero

us

fragm

ents

ofP.abyssi

flagella

as

well

as

num

ero

us

art

efa

cts

inth

epre

para

-20

tion

of

puri

fied

PA

V1

(Fig

.1d).

PA

V1

part

icle

scould

be

observ

ed

indirect

conta

ct

with

the

sili

ca

pre

cip

itate

after

only

one

day

(Fig

.4a).

This

pre

cip

itate

was

diff

ere

nt

from

that

form

ed

inth

eS

IRV

2and

TP

V1

experi

ments

and

consis

ted

of

very

fine

part

icle

s

(com

pare

Fig

s.

4c

and

3b),

possib

lyas

aconsequence

of

the

diff

ere

nt

com

positio

nof

the

PA

V1

bufferi

ng

mediu

m(w

hic

hin

clu

des

ED

TA

).A

lthough

no

pre

cip

itate

sfo

rmed

25

insid

eor

on

PA

V1

after

60

days

(Fig

.4b),

seve

ral

vir

us

part

icle

sw

ere

trapped

inth

e

sili

ca

pre

cip

itate

(Fig

.4c).

2244

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


4D

iscu

ssio

n

The

vir

uses

used

inth

isexperi

mentbelo

ng

todiff

ere

ntvir

us

fam

ilies

havin

gcom

ple

tely

diff

ere

nt

morp

holo

gie

sand

str

uctu

res.

This

may

expla

inw

hy

the

results

of

the

fossili

-

sation

experi

ments

were

diff

ere

nt.

Inth

ecase

of

SIR

V2,

anon-e

nve

loped

rod-s

haped

vir

us,

sili

ca

bin

din

gon

the

viralpart

icle

sw

as

sig

nifi

cant

and

pro

gre

ssiv

ew

here

as

only

5

limited

sili

ca

bin

din

goccurr

ed

on

TP

V1,

an

enve

loped

spin

dle

-shaped

vir

us.

Incon-

trast,

no

sili

ca

bin

din

gw

as

observ

ed

on

the

second

enve

loped

and

spin

dle

-shaped

vir

us,PA

V1.

4.1

Fo

ssilis

ati

on

of

the

vir

uses

The

fact

that

the

sili

ca

pre

cip

itate

sin

the

SIR

V2

experi

ment

were

inva

riably

linked

to10

DN

Astr

ands

(Fig

.2b)

suggests

thatth

esponta

neously

-form

ed

min

era

ldeposit

pro

ba-

bly

rapid

lyentr

apped

num

ero

us

SIR

V2

part

icle

s,as

was

observ

ed

by

Laid

ler

and

Ste

d-

man

(2010).

Som

epart

icle

sw

ere

bare

lyobserv

able

as

dark

outlin

es

within

the

min

era

l

pre

cip

itate

(Fig

.2a).

The

viralpart

icle

sth

at

rem

ain

ed

free

inth

em

ediu

mshow

ed

pro

-

gre

ssiv

esili

ca

bin

din

gw

ith

tim

eon

their

oute

rsurf

ace

and

inth

eir

centr

alcavity.

Initia

l15

sili

ca

nucle

ation

within

the

cavity

of

the

SIR

V2

part

icle

sdid

not

appear

todam

age

the

body

(Fig

.2d,

e)

although,

at

late

rste

ps

of

the

fossili

sation

pro

cess,

str

ings

of

sili

ca

part

icle

sobserv

ed

on

DN

Astr

ands

(Fig

.2g)

suggest

that

viralpart

icle

sm

ay

be

heav-

ilydam

aged

by

the

gro

wth

of

sili

ca.

The

rari

tyof

sili

cifi

ed

DN

Afr

agm

ents

may

be

due

toth

efa

ctth

atD

NA

degra

des

rapid

lyw

hen

exposed

toth

eenvironm

ent(S

chelb

le20

etal.,2008),

while

the

rem

ain

sofD

NA

trapped

insili

ca

(Fig

.2a,g)

may

be

pre

serv

ed.

The

sili

ca

bin

din

gto

the

vir

ions

oute

rsurf

ace

appears

tobe

the

result

of

the

direct

nucle

ation

of

sili

ca

(Fig

.2c)

follo

wed

by

poly

meri

sation,

and

of

the

bin

din

gof

already

form

ed

sili

ca

part

icle

son

the

vir

ions,

or

on

pre

vio

usly

bound

sili

ca

part

icle

s(F

ig.

2e)

(Bir

nbaum

et

al.,

1989;

Schultze-L

am

et

al.,

1995;

Westa

llet

al.,

1995;

Topors

kiet

al.,

25

2002;

Ora

nge

et

al.,

2009).

Sili

ca

pro

bably

bin

ds

via

cova

lent

or

hydro

gen

bonds

be-

tween

the

sili

ca

part

icle

sand

the

hydro

xyland/o

rcarb

oxylfu

nctionalgro

ups

conta

ined

2245

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


inth

epro

tein

ofth

ehelix

(see

revie

wofth

efo

ssili

sation

pro

cesses

inK

onhauser

etal.,

2004).

Fossili

sation

of

the

vir

uses

TP

V1

and

PA

V1

diff

ere

dfr

om

that

of

SIR

V2

because

the

form

er

have

an

enve

lope

that

conta

ins

lipid

sth

at

deri

vefr

om

the

hosts

(the

Arc

haea

P.

abyssi

and

T.

pri

euri

i,re

spective

ly)

(Geslin

et

al.,

2003b;

Gorl

as

et

al.,

2011).

Pre

vi-

5

ous

results

from

the

experi

menta

lfo

ssili

sation

of

Arc

haea

show

ed

that

the

sim

ple

cell

wall

of

these

mic

roorg

anis

ms

(S-L

ayer+

pla

sm

icm

em

bra

ne)

has

only

alim

ited

abili

ty

tobin

dsili

ca

(Ora

nge

et

al.,

2009).

Itw

as

there

fore

tobe

expecte

dth

at,

havin

ga

sim

-

ilar

com

positio

n,

the

enve

lopes

of

TP

V1

and

PA

V1

would

not

bin

dsig

nifi

cant

am

ounts

of

sili

ca.

This

was

indeed

observ

ed,

although

som

evir

uses

occasio

nally

occurr

ed

in10

direct

conta

ct

with

sili

ca

(Fig

.3b),

and,

after

75

days

of

exposure

toth

esili

ca

solu

tion,

nanom

ete

r-siz

ed

part

icle

sw

ere

ubiq

uitously

observ

ed

insid

eT

PV

1(F

ig.

3c,

f).

Only

at

this

adva

nced

sta

ge

inth

efo

ssili

sation

pro

cedure

(75

days),

were

nanopart

icle

sof

aS

iO2-C

spre

cip

itate

firs

tobserv

ed

on

the

vir

ion

enve

lopes.

The

sourc

eofth

eC

sw

as

the

cesiu

mchlo

ride

used

for

harv

esting

puri

fied

TP

V1.

Chela

ted

meta

lions

may

actas

15

bri

dges

inth

efixation

ofm

inera

lions

toorg

anic

mate

rials

.T

his

has

been

dem

onstr

ate

d

with

Fe(I

II)

chela

ted

toB

acte

ria

and

Arc

haea

(Beve

ridge

and

Murr

ay,

1976;

Beve

rdig

e

and

Kova

l,1981;

Ora

nge

et

al.,

2011).

How

eve

r,as

am

onova

lent

cation,

Cs+

cannot

act

as

adirect

inte

rmedia

tebetw

een

sili

ca

and

the

vir

ion

enve

lope

or

inte

rnalm

ate

ri-

als

.S

ilica

bin

din

gto

the

TV

1part

icle

sm

ostlik

ely

occurr

ed

inth

esam

ew

ay

as

with

the

20

SIR

V2

part

icle

s,nam

ely

thro

ugh

cova

lentor

hydro

xylbin

din

gofth

esili

ca.

4.2

Imp

licati

on

for

the

pre

serv

ati

on

an

did

en

tifi

cati

on

of

vir

alre

main

sin

rocks

The

thre

evir

uses

used

inth

isart

ificia

lfo

ssili

sation

stu

dy

were

well

pre

serv

ed,

apart

from

aslig

ht

defo

rmation

possib

lydue

toth

ele

ngth

of

the

experi

ment

sto

rage

pri

or

toth

efo

ssili

sation

experi

ment

rath

er

than

toth

eexposure

tosili

ca.

The

fossili

sation

25

was

made

directly

on

vir

ions

sto

red

inbuffere

dsolu

tions

whic

his

obvio

usly

not

anat-

ura

lconditio

n.

The

rationale

for

makin

gth

eexperi

ments

on

the

mic

roorg

anis

ms

in

abuffere

dsolu

tion

isth

epre

vio

usly

note

dim

port

ance

for

the

cell

tore

main

aliv

e,

or

at

2246

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


leastnotto

lyse,duri

ng

the

tim

enecessary

for

sili

ca

tofo

rma

deposit

aro

und

the

cells

thic

kenough

toallo

wfo

rfo

ssili

sation

and

furt

her

pre

serv

ation

(Ora

nge

et

al.,

2009).

This

peri

od

of

tim

eva

ries

depends

on

the

str

ain

or

the

type

of

mic

roorg

anis

m.

Vi-

ral

part

icle

sobvio

usly

do

not

lyse

but

their

physic

al

inte

gri

tycan

quic

kly

be

affecte

d

due

toth

eir

sm

all

siz

eand

constitu

tion.

Our

results

show

that,

ifviralpart

icle

scan

be

5

conserv

ed

ove

rseve

ralm

onth

s,th

ey

can

theore

tically

becom

efo

ssili

sed.

The

fact

that

vir

uses

can

be

fossili

sed

suggests

that

natu

rally

fossili

sed

vir

uses

may

occur

inro

cks

inth

esam

ew

ay

as

fossili

sed

mic

robia

lcells

.T

he

question

is,

how

could

we

identify

them

?M

orp

holo

gy

isa

cri

teri

on

for

their

identification

but

their

sim

-

ple

nanom

etr

ic-s

ized

str

uctu

res

may

be

confu

sed

with

min

era

lsor

oth

er

art

ifacts

ofth

e10

sam

esiz

e(o

fth

eord

er

of∼

100

nm

).O

nth

eoth

er

hand,

itm

ay

be

possib

leto

iden-

tify

fila

mento

us

or

rod-s

haped

str

uctu

res,

such

as

the

about

500×

50

nm

-siz

ed

SIR

V2

part

icle

s,

inancie

nt

rocks.

These

vir

uses

are

about

the

sam

esiz

eand

shape

as

the

so-c

alle

d“n

anobacte

ria”

identified

in3.9

Ga

old

carb

onate

concre

tions

inth

eM

art

ian

mete

ori

teA

LH

84001

(McK

ay

et

al.,

1996

–note

that

itis

now

accepte

dth

at

the

latter

15

are

sim

ply

min

era

lpre

cip

itations,

Gib

son

et

al.,

2001).

Although

sili

ca

pre

cip

itate

din

the

centr

al

cavity

of

SIR

V2

(Fig

.2e)

as

astr

uctu

reof

less

than

10

nm

inw

idth

and

seve

ralhundre

ds

of

nanom

ete

rslo

ng,

itw

ould

be

diffi

cult

todis

tinguis

hsuch

afe

atu

re

from

poly

meri

cfibri

lsor

fila

ments

of

oth

er

bio

logic

alori

gin

that

had

been

sili

cifi

ed.

Vi-

ral

org

anic

com

pounds,

such

as

lipid

sand

pro

tein

s,

could

als

obe

pre

serv

ed

inve

ry20

sm

all

am

ounts

butth

ey

would

rapid

lybe

degra

ded,hence

renderi

ng

their

dete

ction

and

identification

as

viralbio

mark

ers

diffi

cult.

Thus,

eve

nth

ough

itis

theore

tically

feasib

le,

dete

ction

ofvir

us

rem

ain

sin

rocks

will

be

hig

hly

challe

ngin

g.

5C

on

clu

sio

ns

This

stu

dy

isone

of

the

firs

tattem

pts

tofo

ssili

se

vir

uses

and

the

firs

tm

onitori

ng

of

the

25

fossili

sation

of

vir

uses

of

extr

em

ophili

cA

rchaea

ove

ra

peri

od

of

seve

ralm

onth

s.

Our

results

show

thatth

evir

uses

ofextr

em

ophili

cm

icro

org

anis

ms

can

be

fossili

sed

by

sili

ca

2247

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


and

thatth

estr

uctu

res

have

achance

ofbein

gpre

serv

ed

ifth

epro

cess

can

take

pla

ce

ove

ra

peri

od

of

seve

ral

month

s.

The

vir

us

str

uctu

reand

the

mole

cula

rcom

positio

n

of

their

oute

rsurf

ace

(lip

idenve

lope,

pro

tein

)cle

arl

ypla

ya

cru

cia

lro

lein

the

rate

and

manner

of

fossili

sation.

Although

faithfu

lpre

serv

ation

of

the

viral

str

uctu

res

ove

r

geolo

gic

al

tim

eis

pro

bably

unlik

ely

,som

etr

aces

may

neve

rthele

ss

rem

ain

inro

cks.

5

Our

stu

dy

thus

contr

ibute

sim

port

ant

info

rmation

toth

isnew

field

of

researc

h.

Due

to

the

gre

at

div

ers

ity

of

vir

uses

inte

rms

of

morp

holo

gy

and

str

uctu

re,

additio

nalstu

die

s

are

needed

tocom

ple

teth

ese

results

with

the

fossili

sation

of

oth

er

kin

ds

of

vir

us

in

ord

er

tobetter

unders

tand

the

mechanis

ms

invo

lved.

Ackn

ow

led

ge

me

nts

.T

his

rese

arc

hw

as

fun

de

dby

the

CN

RS

–P

IDO

rig

ine

sd

es

Pla

ne

tes

et

10

de

laV

ie(2

00

7–

20

09

).A

uro

reG

orl

as

wa

sfu

nd

ed

thro

ug

ha

Ph

Dgra

nt

fro

mth

eM

inis

tere

de

l’En

se

ign

em

en

tS

upe

rie

ur

et

de

laR

ech

erc

he.

Dom

iniq

ue

Ja

lab

ert

isth

an

ke

dfo

rh

isa

idw

ith

the

tra

nsm

issio

ne

lectr

on

mic

rosco

pe.

15

Th

ep

ublic

atio

no

fth

isart

icle

isfin

an

ce

dby

CN

RS

-IN

SU

.

Refe

ren

ces

Ba

mfo

rd,D

.H

.:D

ovir

use

sfo

rmlin

ea

ge

sa

cro

ss

diff

ere

ntd

om

ain

so

flif

e?

,R

es.M

icro

bio

l.,1

54

,

23

1–

23

6,

do

i:1

0.1

01

6/S

09

23

-25

08

(03

)00

06

5-2

,2

00

3.

Ba

mfo

rd,

D.

H.,

Gri

me

s,

J.M

.,a

nd

Stu

art

,D

.I.:

Wh

at

do

es

str

uctu

rete

llu

sa

bo

ut

vir

us

evo

lu-

20

tio

n?

,C

urr

.O

pin

.S

truc.

Bio

l.,

15

,6

55

–6

63

,d

oi:1

0.1

01

6/j.s

bi.2

00

5.1

0.0

12

,2

00

5.

Be

ttste

tte

r,M

.,P

en

g,

X.,

Ga

rre

tt,

R.

A.,

an

dP

ran

gis

hvili

,D

.:A

FV

1,

an

ove

lvir

us

infe

ctin

g

hyp

ert

he

rmo

ph

ilic

arc

ha

ea

of

the

ge

nu

sA

cid

ianu

s,

Viro

log

y,3

15

,6

8–

79

,2

00

3.

2248

https://www.researchgate.net/publication/10715992_Bamford_DH_Do_viruses_form_lineages_across_different_domains_of_life_Res_Microbiol_154_231-236?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy

https://www.researchgate.net/publication/10715992_Bamford_DH_Do_viruses_form_lineages_across_different_domains_of_life_Res_Microbiol_154_231-236?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy

https://www.researchgate.net/publication/7497569_What_does_structure_tell_us_about_virus_evolution?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy





BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Beve

rid

ge,

T.

J.a

nd

Kova

l,S

.F.

:B

ind

ing

me

tals

toce

lle

nve

lop

es

of

Esch

eri

ch

iaco

li,A

pp

l.

Enviro

n.

Mic

rob.,

42

,3

25

–3

35

,1

98

1.

Beve

rid

ge,

T.

J.a

nd

Mu

rray,

R.

G.

E.:

Up

take

an

dre

ten

tio

no

fm

eta

lsby

ce

llw

alls

of

Ba

cill

us

su

btilis

,J.

Ba

cte

rio

l.,

12

7,

15

02–

15

18

,1

97

6.

Bird

,D

.F.

,Ju

nip

er,

S.

K.,

Ric

cia

rdi-R

iga

ult,

M.,

Ma

rtin

eu

,P.,

Pra

irie

,Y.

T.,

an

dC

alv

ert

,S

.E

.:5

Su

bsu

rfa

ce

vir

use

san

db

acte

ria

inH

olo

ce

ne

/La

teP

leis

tocen

ese

dim

en

tso

fS

aa

nic

hIn

let,

BC

:O

DP

Ho

les

10

33

Ba

nd

10

34

B,L

eg

16

9S

,M

ar.

Ge

ol.,1

74

,2

27

–2

39

,d

oi:1

0.1

01

6/S

00

25

-

32

27

(00

)00

15

2-3

,2

00

1.

Bir

nb

au

m,

S.

J.,

Wire

ma

n,

J.W

.,a

nd

Bo

row

ski,

R.:

Sili

ca

pre

cip

ita

tio

nby

the

an

ae

rob

icsu

l-

ph

ate

red

ucin

gb

acte

riu

mD

esu

lfovib

rio

de

su

lfu

rica

ns:

effe

cts

up

on

ce

llmo

rph

olo

gy

an

dim

-10

plic

atio

ns

for

pre

se

rva

tio

n,

in:

Ori

gin

,E

volu

tio

n,a

nd

Mo

de

rnA

sp

ects

of

Bio

min

era

liza

tio

nin

Pla

nts

an

dA

nim

als

,e

dite

dby:

Cri

ck,R

.E

.,P

lenu

mP

ress,N

ew

Yo

rk,U

SA

,5

07

–5

16

,1

98

9.

Biz

e,

A.,

Ka

rlsson

,E

.A

.,E

ke

fja

rd,

K.,

Qu

ax,

T.

E.

F.,

Pin

a,

M.,

Pre

vost,

M.

C.,

Fo

rte

rre,

P.,

Te

na

illo

n,

O.,

Be

rna

nd

er,

R.,

an

dP

ran

gis

hvili

,D

.:A

un

iqu

evir

us

rele

ase

inth

eA

rch

ae

a,

P.

Na

tl.

Aca

d.

Sci.

US

A,

10

6,

11

30

6–

11

31

1,

do

i:1

0.1

07

3/p

na

s.0

90

12

381

06

,2

00

9.

15

Bo

rris

s,

M.,

He

lmke,

E.,

Ha

nschke,

R.,

an

dS

ch

we

de

r,T.:

Iso

latio

na

nd

ch

ara

cte

riza

tio

no

f

ma

rin

ep

sych

rop

hili

cp

ha

ge

–h

ost

syste

ms

fro

mA

rctic

se

aic

e,

Extr

em

op

hile

s,

7,

37

7–

38

4,

do

i:1

0.1

00

7/s

00

79

2-0

03-0

33

4-7

,2

00

3.

Dya

ll-S

mith

,M

.,Ta

ng

.,S

.L

.,a

nd

Ba

th,

C.:

Ha

loa

rch

ae

alvir

use

s:

how

div

ers

ea

reth

ey?

,R

es.

Mic

rob

iol.,

15

4,

30

9–

313

,d

oi:1

0.1

01

6/S

09

23

-25

08

(03

)00

07

6-7

,2

00

3.

20

Era

uso,

G.,

Reyse

nb

ach

,A

.L

.,G

od

froy,

A.,

Me

un

ier,

J.R

.,C

rum

p,

B.,

Pa

rte

nsky,

F.,

Ba

ross,

J.A

.,M

art

ein

sso

n,

V.,

Ba

rbie

r,G

.,P

ace,

N.

R.,

and

Pri

eur,

D.:

Pyro

co

ccu

sa

byssi

sp.

nov.

,a

new

hyp

ert

he

rmo

ph

ilic

arc

ha

eo

nis

ola

ted

fro

ma

de

ep

-se

ahyd

roth

erm

al

ven

t,

Arc

h.

Mic

rob

iol.,

16

0,

33

8–

34

9,

19

93

.

Fo

rte

rre,

P.:

Ah

ot

sto

ryfr

om

co

mp

ara

tive

ge

no

mic

s:

reve

rse

gyra

se

isth

eo

nly

25

hyp

ert

he

rmo

ph

ile-s

pe

cifi

cp

rote

in,

Tre

nd

sG

en

et.

,1

8,

23

6–

23

8,

do

i:1

0.1

01

6/S

01

68

-

95

25

(02

)02

65

0-1

,2

00

2.

Fo

rte

rre,

P.:

Th

ree

RN

Ace

llsfo

rri

bo

so

ma

llin

ea

ge

sa

nd

thre

eD

NA

vir

use

sto

rep

lica

teth

eir

ge

no

me

s:

ahyp

oth

esis

for

the

ori

gin

ofce

llula

rd

om

ain

,P.N

atl.A

ca

d.S

ci.

US

A,1

03

,3

66

9–

36

74

,d

oi:1

0.1

07

3/p

nas.0

51

03

33

10

3,

20

06

.30

Fo

rte

rre,

P.:

Ma

nip

ula

tio

nof

ce

llula

rsyn

the

se

sa

nd

the

na

ture

of

vir

use

s:

the

viro

ce

llco

nce

pt,

C.

R.

Ch

im.,

inp

ress.,

do

i:1

0.1

01

6/j.c

rci.2

01

0.0

6.0

07

,2

01

0.

Fo

rte

rre,

P.

an

dP

ran

gis

hvili

,D

.:T

he

gre

at

bill

ion

-ye

ar

wa

rb

etw

ee

nri

bo

so

me

-a

nd

ca

psid

-

2249

https://www.researchgate.net/publication/225799053_Pyrococcus_abyssi_sp_nov_a_new_hyperthermophilic_archaeon_isolated_from_a_deep-sea_hydrothermal_vent?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy









https://www.researchgate.net/publication/10696624_Isolation_and_characterization_of_marine_psychrophilic_phage-host_systems_from_Arctic_sea-ice_Extremophiles_7377-384?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy



https://www.researchgate.net/publication/16172799_Binding_of_metals_to_cell_envelopes_of_Escherichia_coli_K-12_Appl_Environ_Microbiol?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy






https://www.researchgate.net/publication/10716003_Haloarchaeal_viruses_how_diverse_are_they_Res_Microbiol?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy



https://www.researchgate.net/publication/22332612_Uptake_and_retention_of_metals_by_cell_walls_of_Bacillus_subtilis_J_Bact?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy








BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


en

co

din

go

rga

nis

ms

(ce

llsa

nd

vir

use

s)

as

the

ma

jor

so

urc

eo

fevo

lutio

na

ryn

ove

ltie

s,

An

n.

NY.

Aca

d.

Sci.,

11

78

,65

–7

7,

do

i:1

0.1

11

1/j.1

74

9-6

63

2.2

00

9.0

49

93

.x,

20

09

a.

Fo

rte

rre,

P.

an

dP

ran

gis

hvili

,D

.:T

he

ori

gin

of

vir

use

s,

Re

s.

Mic

rob

iol.,

16

0,

46

6–

47

2,

do

i:1

0.1

01

6/j.r

esm

ic.2

009

.07

.00

8,

20

09

b.

Ge

slin

,C

.,L

eR

om

an

ce

r,M

.,G

aill

ard

,M

.,E

rau

so,

G.,

an

dP

rie

ur,

P.:

Ob

se

rva

tio

no

fvir

us-lik

e5

pa

rtic

les

inh

igh

tem

pe

ratu

ree

nri

ch

me

nt

cultu

res

fro

md

ee

p-s

ea

hyd

roth

erm

al

ven

ts,

Re

s.

Mic

rob

iol.,

15

4,

30

3–

307

,d

oi:1

0.1

01

6/S

09

23

-25

08

(03

)00

07

5-5

,2

00

3a

.

Ge

slin

,C

.,L

eR

om

an

ce

r,M

.,E

rau

so.,

G.,

Ga

illa

rd,

M.,

Pe

rro

t,G

.a

nd

Pri

eu

r,D

.:PA

V1

,th

e

firs

tvir

us-lik

ep

art

icle

iso

late

dfr

om

ahyp

ert

he

rmo

ph

ilic

eu

ryarc

ha

eo

te,

Pyro

co

ccu

sa

byssi,

J.B

acte

rio

l.,

18

5,

38

88–

38

94

,d

oi:1

0.1

12

8/J

B.1

85

.13

.38

88

-38

94

.20

03,

20

03

b.

10

Ge

slin

,C

.,L

eR

om

an

ce

r,M

.,G

aill

ard

,M

.,a

nd

Pri

eu

r,D

.:D

ive

rsite

vira

lea

sso

cie

ea

ux

eco

syste

me

shyd

roth

erm

au

xo

ce

an

iqu

es

pro

fon

ds

et

au

xso

urc

es

ch

au

de

ste

rre

str

es,

Vi-

rolo

gie

,9

,3

57

–3

66

,2

00

5.

Go

rla

s,

A.,

Ge

slin

,C

.,a

nd

Pri

eu

r,D

.:T

V1

,th

efirs

tvir

us-lik

ep

art

icle

of

Th

erm

oco

ccu

s,

ahy-

pe

rth

erm

op

hili

cA

rch

ae

ag

enu

s,

Ori

gin

sL

ifeE

vol.

B,

39

,6

2,

20

09

.15

Go

rla

s,

A.,

Ala

in,

K.,

Bie

nve

nu

,N

.,Is

aa

c,

S.,

an

dG

eslin

,C

.:T

he

rmo

co

ccu

sp

rie

uri

isp.

nov.

,

an

ove

lhyp

ert

he

rmo

ph

ilic

arc

ha

eo

nis

ola

ted

fro

ma

de

ep

-se

ahyd

roth

erm

alve

nt

at

the

Ea

st

Pa

cifi

cR

ise,

su

bm

itte

dto

Int.

J.S

yst.

Evo

l.M

icr.

,2

01

1.

Gow

ing

,M

.M

.:L

arg

evir

use

sa

nd

infe

cte

dm

icro

eu

ka

ryo

tes

inR

oss

Se

asu

mm

er

pa

ck

ice

ha

bita

ts,

Ma

r.B

iol.,

142

,1

02

9–

10

40

,d

oi:1

0.1

00

7/s

00

22

7-0

03

-10

15

-x,2

00

3.

20

Jia

ng

,S

.,S

tew

ard

,G

.,Jelli

so

n,

R.,

Ch

u,

W.,

an

dC

ho

i,S

.:A

bu

nd

an

ce,

dis

trib

utio

na

nd

di-

vers

ity

of

vir

use

sin

alk

alin

ehyp

ers

alin

eM

on

oL

ake,

Ca

lifo

rnia

,M

icro

b.

Eco

l.,

47

,9

–1

7,

do

i:1

0.1

00

7/s

00

24

8-0

03-1

02

3-x

,2

00

4.

Ke

pn

er,

R.

L.,

Wh

art

on

Jr.

,R

.A

.,a

nd

Su

ttle

,C

.A

.:V

iru

se

sin

An

tarc

tic

lake

s,

Lim

no

l.

Oce

an

ogr.

,4

3,

17

54

–17

61

,1

99

8.

25

Ko

nh

au

se

r,K

.O

.,Jo

ne

s,B

.,R

eyse

nb

ach

,A

.L

.,a

nd

Re

na

ut,

R.W

.:H

otsp

rin

gsin

ters

:keys

to

un

de

rsta

nd

ing

Ea

rth

’sea

rlie

st

life

form

s,

Ca

n.

J.E

art

hS

ci.,

40

,1

71

3–

17

24

,2

00

3.

Ko

nh

au

se

r,K

.O

.,Jo

ne

s,

B.,

Ph

oe

nix

,V.

R.,

Fe

rris

,G

.,a

nd

Re

na

ut,

R.

W.:

Th

em

icro

bia

lro

le

inh

ot

sp

rin

gsili

cifi

ca

tion

,A

mb

io,

33

,5

52

–5

58

,2

00

4.

Ko

on

in,

E.

V.

an

dD

olja

,V.

V.:

Evo

lutio

no

fco

mp

lexity

inth

evira

lw

orl

d:

the

daw

no

fa

new

30

vis

ion

,V

iru

sR

es.,

11

7,1

–4

,d

oi:1

0.1

01

6/j.v

iru

sre

s.2

00

6.0

1.0

18

,2

00

6.

Ko

on

in,

E.

V.,

Se

nkevic

h,

T.

G.,

an

dD

olja

,V.

V.:

Th

ea

ncie

nt

vir

us

wo

rld

an

devo

lutio

no

fce

lls,

Bio

l.D

ire

ct,

1,

29

,d

oi:1

0.1

18

6/1

74

5-6

15

0-1

-29

,2

00

6.

2250

https://www.researchgate.net/publication/8450902_Abundance_Distribution_and_Diversity_of_Viruses_in_Alkaline_Hypersaline_Mono_Lake_California?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy









https://www.researchgate.net/publication/8064377_The_Microbial_Role_in_Hot_Spring_Silicification?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy




BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Kru

povic

,M

.a

nd

Ba

mfo

rd,

D.

H.:

Vir

us

evo

lutio

n:

how

far

do

es

the

do

uble

be

ta-b

arr

el

vira

l

line

ag

eexte

nd

?,

Na

t.R

ev.

Mic

rob

iol.,

6,

94

1–

94

88

,d

oi:1

0.1

03

8/n

rmic

ro2

03

3,

20

08

.

La

Scola

,B

.,A

ud

ic,

S.,

Ro

be

rt,

C.,

Ju

ng

an

g,

L.,

de

La

mb

alle

rie,

X.,

Dra

nco

urt

,M

.,B

irtle

s,

R.,

Cla

veri

e,

J.M

.,a

nd

Ra

ou

lt,

D.:

Ag

ian

tvir

us

ina

mo

eb

ae,

Scie

nce,

29

9,

20

33

,2

00

3.

La

idle

r,J.

R.

an

dS

ted

ma

n,

K.

M.:

Vir

us

sili

cifi

ca

tio

nu

nd

er

sim

ula

ted

ho

tsp

rin

gco

nd

itio

ns,

5

Astr

ob

iolo

gy,

10

,5

69

–57

6,

do

i:1

0.1

08

9/a

st.

20

10

.04

63

,2

01

0.

Le

Ro

ma

ncer,

M.,

Ga

illa

rd,

M.,

Ge

slin

,C

.,a

nd

Pri

eu

r,D

.:V

iru

se

sin

extr

em

es

enviro

nm

en

ts,

Rev.

Enviro

n.

Sci.

Bio

tech

.,6

,1

7–

31

,d

oi:1

0.1

00

7/s

11

15

7-0

06

-00

11

-2,2

00

7.

Ma

ran

ge

r,R

.,B

ird

,D

.F.

,a

nd

Ju

nip

er,

S.

K.:

Vira

la

nd

ba

cte

ria

ld

yn

am

ics

inA

rctic

se

aic

e

du

rin

gth

esp

rin

ga

lga

lblo

om

ne

ar

Re

so

lute

,N

.W

.T.,

Ca

na

da

,M

ar.

Eco

l.-P

rog

.S

er.

,1

11

,10

12

1–

12

7,

19

94

.

Ma

rte

insso

n,

V.

T.,

Wa

trin

,L

.,P

rie

ur,

D.,

Cap

rais

,J.

C.,

Ra

gue

ne

s,

G.,

an

dE

rau

so,

G.:

Ph

e-

no

typ

icch

ara

cte

riza

tio

n,

DN

Asim

ilari

tie

s,

an

dp

rote

inp

rofile

so

ftw

en

tysu

lfu

r-m

eta

bo

lizin

g

hyp

ert

he

rmo

ph

ilic

an

ae

rob

icA

rch

ae

ais

ola

ted

fro

mhyd

roth

erm

alve

nts

inth

eS

ou

thw

este

rn

Pa

cifi

cO

ce

an

,In

t.J.

Syst.

Ba

cte

rio

l.,

45

,6

23

–6

32

,d

oi:1

0.1

09

9/0

02

07

71

3-4

5-4

-62

3,

19

95

.15

Ma

rtin

,A

.,Ye

ats

,S

.,Ja

ne

kovic

,D

.,R

eite

r,W

.D

.,A

ich

er,

W.,

an

dZ

illig

,W

.:S

AV

1,

ate

mp

er-

ate

u.v

.-in

du

cib

leD

NA

vir

us-lik

ep

art

icle

fro

mth

ea

rch

arb

acte

riu

mS

ulfo

lobu

sa

cid

oca

lda

riu

s

iso

late

B1

2,

EM

BO

J.,3

,2

16

5–

21

68

,1

98

4.

McK

ay,

D.

S.,

Gib

so

nJr.

,E

.K

.,T

ho

ma

s-K

ep

rta

,K

.L

.,V

ali,

H.,

Ro

ma

ne

k,

C.

S.,

Cle

me

tt,

S.

J.,

Ch

illie

r,X

.D

.F.

,M

ae

ch

ling

,C

.R

.,an

dZ

are

,R

.N

.:S

ea

rch

for

pa

st

life

on

Ma

rs:

20

po

ssib

lere

licb

iog

en

ica

ctivity

inm

art

ian

me

teo

rite

AL

H8

40

01

,S

cie

nce,

27

3,

92

4–

93

0,

do

i:1

0.1

12

6/s

cie

nce.2

73.5

27

7.9

24

,1

99

6.

Mo

reira

,D

.a

nd

Lo

pe

z-G

arcıa

,P.:

Te

nre

aso

ns

toexclu

de

vir

use

sfr

om

the

tre

eo

flif

e,N

at.

Rev.

Mic

rob

iol.,

7,

30

6–

31

1,d

oi:1

0.1

03

8/n

rmic

ro2

10

8,

20

09

.

Nis

be

t,E

.G

.a

nd

Sle

ep,

N.

H.:

Th

eh

ab

ita

ta

nd

na

ture

of

ea

rly

life,

Na

ture

,4

09

,1

08

3–

10

91

,25

do

i:1

0.1

03

8/3

50

59

21

0,2

00

1.

Ora

ng

e,

F.,

We

sta

ll,F.

,D

isn

ar,

J.R

.,P

rie

ur,

D.,

Bie

nve

nu

,N

.,L

eR

om

an

ce

r,M

.,a

nd

De

farg

e,

C.:

Exp

eri

me

nta

lsili

cifi

ca

tio

no

fth

eextr

em

op

hili

cA

rch

ae

aP

yro

co

ccu

sa

byssi

an

d

Me

tha

no

ca

ldo

co

ccu

sja

nn

asch

ii,A

pp

lica

tio

ns

inth

ese

arc

hfo

revid

en

ce

oflif

ein

ea

rly

Ea

rth

an

dextr

ate

rre

str

ial

rocks,

Ge

ob

iolo

gy,

7,

40

3–

41

8,

do

i:1

0.1

11

1/j.1

47

2-4

66

9.2

00

9.0

02

12

.x,

30

20

09

.

Ora

ng

e,

F.,

Dis

na

r,J.

R.,

We

sta

ll,F.

,P

rie

ur,

D.,

an

dB

aill

if,P.:

Me

tal

ca

tio

nb

ind

ing

by

the

hyp

ert

he

rmo

ph

ilic

mic

roo

rga

nis

m,

Arc

ha

ea

Me

tha

no

ca

ldo

co

ccu

sja

nn

asch

ii,a

nd

its

effe

cts

2251

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


on

sili

cifi

ca

tio

n,

su

bm

itte

dto

Pa

lae

on

tolo

gy,

20

11

.

Ore

n,

A.,

Bra

tba

k,

G.,

an

dH

eld

al,

M.:

Occu

rre

nce

of

vir

us

like

pa

rtic

les

inth

eD

ea

dS

ea

,

Extr

em

op

hile

s,

1,

14

3–

14

9,

do

i:1

0.1

00

7/s

00

79

20

05

00

27

,1

99

7.

Ort

ma

nn

,A

.C

.a

nd

Su

ttle

,C

.A

.:H

igh

abu

nd

an

ce

so

fvir

use

sin

de

ep

-se

ahydro

the

rma

lve

nt

syste

min

dic

ate

vira

lm

ed

iate

dm

icro

bia

lm

ort

alit

y,D

ee

p-S

ea

Re

s.

Pt.

I,5

2,

15

15

–1

52

7,

5

do

i:1

0.1

01

6/j.d

sr.

20

05

.04

.00

2,

20

05

.

Ort

ma

nn

,A

.C

.,W

ied

en

he

ft,B

.,D

ou

gla

s,T.,

an

dYo

un

g,M

.:H

otcre

na

rarc

ha

ea

lvir

use

sre

vea

l

de

ep

evo

lutio

na

ryco

nn

ectio

ns,

Na

t.R

ev.

Mic

rob

iol.,

4,

52

0–

528

,d

oi:1

0.1

03

8/n

rmic

ro1

44

4,

20

06

.

Pa

ga

ling

,E

.,H

aig

h,

R.

D.,

Gra

nt,

W.

D.,

Cow

an

,D

.A

.,Jo

ne

s,

B.

E.,

Ma

,Y.,

Ve

nto

sa

,A

.,a

nd

10

He

ap

hy,

S.:

Se

qu

en

ce

an

aly

sis

ofa

nA

rch

ae

alv

iru

sis

ola

ted

fro

ma

hyp

ers

alin

ela

ke

inIn

ne

r

Mo

ng

olia

,C

hin

a,

BM

CG

en

om

ics,

8,

41

0,

do

i:1

0.1

18

6/1

47

1-2

16

4-8

-41

0,

20

07

.

Po

ina

r,G

.P.

an

dP

oin

ar,

R.:

Fo

ssil

evid

en

ce

of

inse

ct

pa

tho

ge

ns,

J.In

vert

eb

r.P

ath

ol.,

89

,

24

3–

25

0,

do

i:1

0.1

01

6/j.jip

.20

05

.05

.00

7,

20

05

.

Pra

ng

ish

vili

,D

.,A

rno

ld,

H.

P.,

Go

tz,

D.,

Zie

se,

U.,

Ho

lz,

I.,

Kri

stja

nsso

n,

J.K

.,a

nd

Zill

ig,

W.:

15

An

ove

lvir

us

fam

ily,

the

Ru

div

irid

ae

:str

uctu

re,

vir

us-h

ost

like

inte

ractio

ns

an

dg

en

om

eva

ri-

ab

ility

of

the

Su

lfolo

bu

svir

use

sS

IRV

1a

nd

SIR

V2

,G

en

etics,

15

2,

13

87

–1

39

6,

19

99

.

Pra

ng

ish

vili

,D

.,F

ort

err

e,

P.,

an

dG

arr

ett

,R

.A

.:V

iru

se

so

fth

eA

rch

ae

a:

au

nifyin

gvie

w,

Na

t.

Rev.

Mic

rob

iol.,

4,

83

7–8

48

,d

oi:1

0.1

03

8/n

rmic

ro1

52

7,

20

06

a.

Pra

ng

ish

vili

,D

.,G

arr

ett

,R

.A

.,a

nd

Koo

nin

,E

.V.:

Evo

lutio

na

ryg

en

om

ics

of

arc

ha

ea

l20

vir

use

s:

un

iqu

evira

lg

en

om

es

inth

eth

ird

do

ma

ino

flif

e,

Vir

us

Re

s.,

11

7,

52

–6

7,

do

i:1

0.1

01

6/j.v

iru

sre

s.2

00

6.0

1.0

07

,2

00

6b.

Pri

ge

nt,

M.,

Le

roy,

M.,

Con

falo

nie

ri,

F.,

Du

tert

re,

M.,

an

dD

uB

ow

,M

.S

.:A

div

ers

ity

of

ba

cte

-

rio

ph

ag

efo

rms

an

dg

en

om

es

ca

nb

eis

ola

ted

fro

mth

esu

rfa

ce

sa

nd

so

fth

eS

ah

ara

De

se

rt,

Extr

em

op

hile

s,

9,

28

9–

29

6,

do

i:1

0.1

00

7/s

00

79

2-0

05

-04

44

-5,

20

05

.25

Ra

ch

el,

R.,

Be

ttste

tte

r,M

.,H

ed

lun

d,

B.

P.,

Ha

rin

g,

M.,

Ke

ssle

r,A

.,S

tett

er,

K.

O.,

an

d

Pra

ng

ish

vili

,D

.:R

em

ark

able

mo

rph

olo

gic

al

div

ers

ity

of

vir

use

sa

nd

vir

us-lik

ep

art

icle

sin

ho

tte

rre

str

iale

nviro

nm

en

ts,

Arc

h.

Viro

l.,

14

7,

24

19

–2

42

9,

do

i:1

0.1

00

7/s

00

70

5-0

02

-08

95

-2,

20

02

.

Ra

ou

lt,

D.,

Au

dic

,S

.,R

ob

ert

,C

.,A

be

rge

l,C

.,R

en

esto

,P.,

Og

ata

,H

.,L

aS

co

la,

B.,

Su

za

n,

M.,

30

an

dC

lave

rie,J.

M.:

Th

e1

.2-m

eg

ab

ase

ge

no

me

seq

ue

nce

ofM

imiv

iru

s,S

cie

nce,3

06

,1

34

4–

13

50

,d

oi:1

0.1

12

6/s

cie

nce.1

10

14

85

,2

00

4.

Ric

e,G

.,S

ted

ma

n,K

.,S

nyd

er,

J.,W

ied

enh

eft

,B

.,W

illits,D

.,B

rum

fie

ld,S

.,a

nd

McD

erm

ott

,T.:

2252


















https://www.researchgate.net/publication/7275023_Evolutionary_genomics_of_archaeal_viruses_unique_viral_genomes_in_the_third_domain_of_life_Virus_Res?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy








BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Vir

use

sfr

om

extr

em

eth

erm

al

enviro

nm

en

ts,

P.

Na

tl.

Aca

d.

Sci.

US

A,

98

,1

33

41

–1

33

45

,

do

i:1

0.1

07

3/p

na

s.2

31

17

01

98

,2

00

1.

Sch

elb

le,

R.

T.,

Ha

ll,J.

A.,

Nea

lso

n,

K.

H.,

an

dS

tee

le,

A.:

DN

Ap

ers

eve

ran

ce

of

mic

roo

rga

n-

ism

sexp

ose

dto

sili

ca

:a

nexp

eri

me

nta

lstu

dy,

Ge

ob

iolo

gy,

6,

50

3–

51

1,

do

i:1

0.1

11

1/j.1

47

2-

46

69

.20

08

.00

17

7.x

,2

008

.5

Sch

ultze

-La

m,

S.,

Fe

rris

,F.

G.,

Ko

hn

au

se

r,K

.O

.,a

nd

Wie

se,

R.

G.:

Insitu

sili

cifi

ca

tio

nof

an

Ice

lan

dic

mic

rob

ialm

at:

imp

lica

tio

ns

for

mic

rofo

ssil

form

atio

n,

Ca

n.

J.E

art

hS

ci.,

32

,2

02

1–

20

26

,1

99

5.

Sim

e-N

ga

nd

o,

T.,

Lu

ca

s,

S.,

Ro

bin

,A

.,P

au

se

Tu

cke

r,K

.,C

olo

mb

et,

J.,

Be

tta

rel,

Y.,

De

sm

on

d,

E.,

Gri

ba

ldo,

S.,

Fo

rte

rre,

P.,

Bre

itb

art

,M

.,a

nd

Pra

ng

ish

vili

,D

.:D

ive

rsity

of

10

vir

us–

ho

st

syste

ms

inhyp

ers

alin

eL

ake

Re

tba

,S

en

eg

al,

Enviro

n.

Mic

rob

iol.,

inp

ress.,

do

i:1

0.1

11

1/j.1

46

2-2

92

0.2

01

0.0

23

23

.x,

20

10

.

To

po

rski,

J.K

.W

.,S

tee

le,

A.,

We

sta

ll,F.

,T

ho

ma

s-K

ep

rta

,K

.L

.,a

nd

McK

ay,

D.

S.:

Th

esim

u-

late

dsili

cifi

ca

tio

no

fb

acte

ria

–n

ew

clu

es

toth

em

od

es

an

dtim

ing

of

ba

cte

ria

lp

rese

rva

tio

n

an

dim

plic

atio

ns

for

the

se

arc

hfo

rextr

ate

rre

str

ialm

icro

fossils

,A

str

ob

iolo

gy,

2,

1–

26

,2

00

2.

15

We

sta

ll,F.

:E

arl

ylif

e:

na

ture

,d

istr

ibu

tio

na

nd

evo

lutio

n,

in:

Ori

gin

sa

nd

Evo

lutio

no

fL

ife,

an

Astr

ob

iolo

gic

alP

ers

pe

ctive

,e

dite

dby:

Ga

rga

ud

,M

.,L

op

ez-G

arcıa

,P.,

an

dM

art

in,

H.,

Ca

m-

bri

dg

eU

niv

ers

ity

Pre

ss,C

am

bri

dg

e,

UK

,3

91

–4

13

,2

01

0.

We

sta

ll,F.

an

dS

ou

tha

m,

G.:

Th

ee

arl

yre

co

rdo

flif

e,

Ge

op

h.

Mo

no

g.

Se

rie

s,

16

4,

28

3–

30

4,

20

06

.20

We

sta

ll,F.

,B

on

i,L.,

an

dG

ue

rzo

ni,

E.:

Th

eexp

eri

me

nta

lsili

cifi

ca

tio

no

fm

icro

org

an

ism

s,

Pa

lae

on

tolo

gy,

38

,4

95

–5

28

,1

99

5.

Zill

ig,

W.,

Kle

tzin

,A

.,S

ch

lep

er,

C.,

Ho

lz,

I.,

Ja

ne

kovic

,D

.,H

ain

,J.

,L

an

ze

ndo

rfe

r,M

.,a

nd

Kri

stja

nso

on

,J.

K.:

Scre

en

ing

for

Su

lfolo

ba

les,

the

irp

lasm

ids,

an

dth

eir

vir

use

sin

Ice

lan

dic

so

lfata

ras,

Syst.

Ap

pl.

Mic

rob

iol.,

16

,6

09

–6

28

,1

99

4.

25

2253

https://www.researchgate.net/publication/11019235_The_Simulated_Silicification_of_Bacteria-_New_Clues_to_the_Modes_and_Timing_of_Bacterial_Preservation_and_Implications_for_the_Search_for_Extraterrestrial_Microfossils?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy














https://www.researchgate.net/publication/239949946_The_Early_Record_of_Life?el=1_x_8&enrichId=rgreq-94b972ea-f7ac-4058-b6fe-05a108c30dd0&enrichSource=Y292ZXJQYWdlOzIzMDg4MjI4NztBUzo5OTEzNTM4MjM2MDA4NkAxNDAwNjQ3MTI1OTgy



BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Fig

.1

.T

EM

mic

rogra

ph

ssh

ow

ing

exa

mp

les

of

the

vir

use

su

se

dfo

rth

isstu

dy.

(A)

SIR

V2

pa

rtic

le,

no

teth

ece

ntr

al

cavity,

so

me

tim

ed

iscon

tinu

ed

.(B

)F

rag

me

nts

of

SIR

V2

pa

rtic

leo

n

an

un

sta

ine

dgri

d.

Note

the

ab

se

nce

of

vis

ible

fea

ture

s,

an

dth

eD

NA

str

an

dth

at

links

the

two

fra

gm

en

ts(a

rrow

).(C

)A

ggre

ga

teo

fT

PV

1p

art

icle

s.

(D)

PA

V1

pa

rtic

les

(arr

ow

s),

no

teth

e

pre

se

nce

of

rem

na

nt

Pyro

co

ccu

sa

byssi

fla

ge

llas

inth

esa

mp

le.

All

TE

Mm

icro

gra

ph

sw

ere

ma

de

at

20

0kV

on

gri

ds

ne

ga

tive

lysta

ine

dw

ith

ura

nylace

tate

,u

nle

ss

oth

erw

ise

sta

ted

.

2254

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Fig

.2

.T

EM

mic

rogra

ph

ssh

ow

ing

pro

gre

ssiv

este

ps

of

the

exp

eri

me

nta

lfo

ssili

sa

tio

no

fS

IRV

2

ata∼

16

0p

pm

Sis

ilica

co

nce

ntr

atio

n.

(A)

48

h,S

IRV

2p

art

icle

pa

rtia

llytr

ap

pe

din

sid

eth

esili

ca

pre

cip

ita

te(S

i).

Th

ea

rrow

su

nd

erl

ine

the

sh

ap

eo

fth

evir

ion

insid

eth

ep

recip

ita

te.

(B)

7d

ays,

fra

gm

en

tso

fS

IRV

2pa

rtic

les

inth

evic

inity

ofa

sili

ca

pre

cip

ita

te(S

i).

Th

etw

ose

pa

rate

dstr

an

ds

of

DN

Aa

revis

ible

(arr

ow

).(C

)7

days,

un

sta

ine

dgri

d,

fra

gm

en

to

fS

IRV

2p

art

icle

.A

da

rk

∼1

0n

mp

art

icle

isse

en

att

ach

ed

on

the

vir

ion

(arr

ow

).(D

)3

0d

ays,

SIR

V2

pa

rtic

le,

no

teth

e

nu

me

rou

sd

ark

sili

ca

pa

rtic

les

inth

ece

ntr

al

cavity.

(E)

60

days,

un

sta

ine

dgri

d,

fra

gm

en

tso

f

SIR

V2

pa

rtic

le,

no

teth

ed

ark

sili

ca

pa

rtic

les

att

ach

ed

on

the

ou

ter

su

rfa

ce

of

the

vir

ion

,a

nd

filli

ng

the

ce

ntr

alcavity.

(F)

60

days,

ED

Xsp

ectr

am

ad

eo

nth

eS

IRV

2p

art

icle

so

f(E

),sh

ow

ing

aS

isig

na

lslig

htly

hig

he

rth

an

on

the

ba

ckgro

un

d.

(G)

60

days,

un

str

ain

ed

gri

d,

DN

Astr

an

d

cove

red

by

sili

ca

pa

rtic

les.

All

TE

Mm

icro

gra

ph

sw

ere

ma

de

at

20

0kV

on

gri

ds

ne

ga

tive

ly

sta

ine

dw

ith

ura

nyla

ce

tate

,u

nle

ss

oth

erw

ise

sta

ted

.

2255

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Fig

.3

.T

EM

mic

rogra

ph

ssh

ow

ing

pro

gre

ssiv

este

ps

of

the

exp

eri

me

nta

lfo

ssili

sa

tio

no

fT

PV

1

ata∼

32

0p

pm

Sisili

ca

co

nce

ntr

atio

n.

(A)

On

ew

ee

k,tw

oT

PV

1p

art

icle

s(a

rrow

s)

ne

ar

asili

ca

pre

cip

ita

te(S

i).

(B)

On

em

on

th,

TP

V1

pa

rtic

le(a

rrow

)a

tta

ch

ed

toth

esili

ca

pre

cip

ita

te(S

i).

(C)

75

days,

TP

V1

pa

rtic

les

with

da

rkn

an

om

etr

icp

art

icle

sw

ith

in.

(D)

75

days,

un

sta

ine

dgri

d,

TP

V1

pa

rtic

les

on

wh

ich

ad

ark

pre

cip

ita

teh

as

form

ed

.(E

)7

5d

ays,

ED

Xsp

ectr

ao

bta

ine

d

on

the

vir

ion

of

(D)

an

do

nth

eb

ackgro

un

d,

sh

ow

ing

Si

an

dC

ssig

na

lso

nth

ep

recip

ita

te.

(F)

18

0d

ays,d

efo

rme

dT

PV

1p

art

icle

ne

ar

asili

ca

pre

cip

itate

(Si),n

ote

the

da

rkp

art

icle

with

in.

(G)

18

0d

ays,

un

sta

ine

dgri

d,

TP

V1

pa

rtic

lecove

red

an

dfille

dw

ith

ad

ark

pre

cip

ita

te.

All

TE

M

mic

rogra

ph

sw

ere

ma

de

at

20

0kV

on

gri

ds

ne

ga

tive

lysta

ine

dw

ith

ura

nyla

ce

tate

,u

nle

ss

oth

-

erw

ise

sta

ted

.

2256

BG

D

8,

22

35

–2

25

7,

20

11

Ex

pe

rim

en

tal

fos

sil

isa

tio

no

f

vir

us

es

F.O

ran

ge

et

al.

Title

Page

Abstr

act

Intr

oduction

Conclu

sio

ns

Refe

rences

Table

sF

igure

s

◭◮

◭◮

Back

Clo

se

Full

Scre

en

/E

sc

Pri

nte

r-fr

iendly

Vers

ion

Inte

ractive

Dis

cussio

n


Fig

.4

.T

EM

mic

rogra

ph

ssh

ow

ing

pro

gre

ssiv

este

ps

of

the

exp

eri

me

nta

lfo

ssili

sa

tio

no

fPA

V1

ata∼

32

0p

pm

Sisili

ca

co

nce

ntr

atio

n.

(A)

On

ed

ay,

PA

V1

pa

rtic

le(a

rrow

)a

tta

ch

ed

toth

esili

ca

pre

cip

ita

te(S

i).

(B)

60

days,

nu

me

rous

PA

V1

pa

rtic

les

ne

ar

the

sili

ca

pre

cip

ita

te.

(C)

60

days,

ag

gre

ga

teo

fPA

V1

pa

rtic

les

tra

pp

ed

ina

fin

ely

gra

ine

dsili

ca

pre

cip

ita

te.

All

TE

Mm

icro

gra

ph

s

we

rem

ad

ea

t2

00

kV

on

gri

ds

ne

ga

tive

lysta

ine

dw

ith

ura

nyla

ce

tate

.

2257

Experimental fossilisation of viruses from extremophilic Archaea

Documents

Transcript of Experimental fossilisation of viruses from extremophilic Archaea