Wound

PathogenDevelopmental signal

VSP, Thi2.1JA COI1

Light

CEV1

Hook

formation

PDF1.2

ERF1

SAPR1,

defencePathogen

PHYA,PHYB

DAD1 OPR3

OPDA

Defence

Pathogen

Developmental signal

C2H4

ETR1 CTR1

NPR1

Light induced genes

DDE2

Defence

COI1 Defence,

growth inhibition,fertility

Growth

inhibition

AtMYC2

EIN2

WRKY70

EIN3

New frontiers between plant responses to stress and

development and applications for health and fuel production

Alessandra.Devoto@rhul.ac.uk

Plant Molecular Science

and for Systems and Synthetic Biology Centre

CAR

Research Area 1.

Investigation on the mechanism of action of jasmonates on growth and

metabolite production and their potential application

G1 S G2 M

JAs are distress signals reducing vegetative growth, during defence responses

Sandra Noir, Moritz Bömer et al 2013

JAs

1a. Establish the link between jasmonate signalling and the cell cycle.

A Devoto 2/15

1b. The Effect of Jasmonates on Chromatin Remodelling

COI1Skp1/Ask1

Cullin

HDA6

Devoto et al 2002 Plant J 2002

Important to regulate environmental response

genes

Genome-wide analysis of histone modifications

by chIP-CHIP and chIP-SEQ

Collaboration with Motaki Seki, Jong-Myong Kim, RIKEN CSRS, Japan

Research Area 1. ctd

Investigation on the mechanism of action of jasmonates on growth and

metabolite production and their potential application

A Devoto 3/15

1c. Testing the effectiveness of JAs on cancer cells and developing functional

bioassays

•JA also suppresses cell proliferation in human cancer cell lines, opening a door for

the potential use of JAs as therapeutics

•Identification of active genes and 2 metabolites.

Moritz Bömer et al in preparation

Collaboration with A Harvey, Brunel University; N Smirnoff, University of Exeter

Research Area 1. ctd

Investigation on the mechanism of action of jasmonates on growth and

metabolite production and their potential application

A Devoto 4/15

Research Area 2.

Engineering Plant Cell Walls for biofuel production and to improve

biomass production and release

Cook , Francocci et al in preparation

Collaboration with S Ferrari, F Cervone, Rome ‘La Sapienza’ and

the late Prof P Bolwell,

•Tobacco and Arabidopsis lines down-

regulated for carbon flux into the

phenylpropanoid pathway, xylan,

lignification-specific peroxidase, and pectins

•Fungal pretreatment with white rots

David Kretschmann Nature Materials 2, 775 - 776 (2003)

A Devoto 5/15

Jasmonates

Defence/Cell death Cell cycle

ROS production MAPKs induction

JAs mediate senescence and wound responses …

…and suppress cell proliferation in human cancer cell lines.

Balbi and Devoto 2008. New Phytol, 77: 301-318

The JAs effects on basic cellular machineries such as induction of

cell cycle arrest, activation of MAPK cascades, production of ROS

species in animal cancer cells are shared with plants

Effects of jasmonates on plant and cancer cells

Mechanisms

identified in both cell

types

Possible diversification

in the mechanisms

controlling the final

outputs in either animal

or plant cells

A Devoto 6/15

MeJA alters the evolution of

the leaf area by affecting cell

size and cell number in a

COI1-dependent manner

Noir, Moritz Bömer et al (2013) Plant

Physiology 161 (4) 1930-1951

A Devoto 7/15

Full genome transcript profiling upon

continuous MeJA treatment during leaf

development

Ribosomal genes

…40 array chips and n QRTPCR later…

Noir, Moritz Bömer et al (2013) Plant Physiology 161 (4) 1930-1951

Continuous MeJA stress delays the onset of endoreduplication

in a COI1-dependent manner during leaf development

Flow cytometry: Nuclear DNA content

coi1-16B

aos

WT

Noir, Moritz Bömer et al (2013) Plant Physiology 161 (4) 1930-1951

Key processes regulated by MeJA continuous treatment

to control leaf size

Continuous MeJA treatment keeps Arabidopsis seedlings in a ‘stand by/ready-

to-go’ mode, where plants are ready to go/grow again once the stress signal is

released

It is not yet known whether and how the JA/SA

pathways might be exploited by pathogens to

alter plant cell growth…

A Devoto 9/15

•putative berberine bridge enzyme

•berberine bridge enzyme-like protein

•strictosidine synthasecytochrome P450-like protein CYP79B2

phenylalanine ammonia lyase (PAL1)4-coumarate:CoA ligase 2

cinnamyl-alcohol dehydrogenase

putative cinnamyl alcohol dehydrogenase;

chalcone synthase

putative flavonol 3-O-glucosyltransferase dihydroflavonol 4-reductase

leucoanthocyanidin dioxygenase (LDOX)UDP rhamnose-anthocyanidin-3-glucoside

•putative limonene cyclase

•Myrcene/ocimene synthase (MOS)squalene epoxidase homologue

S-adenosyl-methionine-sterol-C-

methyltransferase

alk

alo

ids

ph

en

oli

cs

terp

en

oid

s

… for secondary product synthesis

25% of the MeJA-induced genes…

Devoto et al 2005 PMB 58:497–513WT coi1

0 0.5 6 1 6 0 0.5 6 1 6hrs

JA JA

A Devoto 10/15

Growth inhibition of cancer cells incubated with plant mutants: JA,

dependency and selectivity…targeted identification of metabolites in

progress…

MDA-MB-361

0.0

20.0

40.0

60.0

80.0

100.0

120.0

G Col-gl1 Col-gl1

+

coi1-16 coi1-16

+

aos aos + COV99 COV99

+

cev1 cev1 +

RC

N

MCF-10A

0.0

20.0

40.0

60.0

80.0

100.0

120.0

140.0

G Col-gl1 Col-gl1

+

coi1-16 coi1-16

+

aos aos + COV99 COV99

+

cev1 cev1 +

RC

N

MDA-MB-361

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

80.0

90.0

Col-gl1 ** coi1-16 ** aos *** COV99 *** cev1 ***

inhib

itio

n in

%

MCF-10A

-20.0

-10.0

0.0

10.0

20.0

30.0

40.0

50.0

60.0

70.0

Col-gl1 * coi1-16 aos COV99 cev1 **

inhib

itio

n in %

Cancer cells

non -tumorigenic cells

Cancer cells

non -tumorigenic cells

Novel bioassay…

to improve the understanding of jasmonate-regulated pathways

affecting the growth of cancer cells

Moritz Bömer et al in preparation

Collaboration with A Harvey, Brunel University; N Smirnoff, University of Exeter

Credit: Charis Cook

COOK AND DEVOTO A (2011). J Sci Food and Agricult, 91: 1729–1732

Cigarettes kindly provided by Dr A Daudi!

‘LIVING IN A BOX’: The substrate for second generation biofuels

is lignocellulosic material obtained from plant cell walls

David Kretschmann Nature Materials 2, 775 - 776 (2003)

After the cell has grown to its

full size, cells in some tissue

develop secondary cell wall

layers between the primary cell

wall and the lumen

L-Phenyl alanine

Cinnamic Acid

p-coumaric Acid

Hydroxycinnamoyl CoA Esters

Hydroxylcinnamaldehydes

Monolignols

Lignin

PAL

C4H

4CL

CCR

CAD

Peroxidases

CINNAMATE-4-HYDROXYLASE C4H controls

how much carbon is allocated to the pathway

(Blee et al. 2001).

CINNAMOYL COA REDUCTASE CCR regulates

carbon allocation to specific monolignol

synthesis pathways (O’Connell et al. 2002)

Peroxidases including TP60 carry out the final

polymerisation of the monolignols to lignin (Bleeet al. 2003).

… and a xylan biosynthesis gene

UDP-glucuronate decarboxylase synthesises UDP-

xylose, which make up xylan backbone (Bindschedler

et al. (2007)

UDP glucose → UDP glucuronic acid → UDP xylose

Tobacco lines down-regulated in lignin synthesis genes

A Devoto 13/15

Degradation of a dead tree by ''Phanerochaetechrysosporium''; Mag5x.

http://microbewiki.kenyon.edu/index.php/File:Whiterot.jpg

Pretreatment of cell wall material with white rot fungi to

remove lignin from cell walls to improve cellulose

accessibility

Processing of CWs carried out to determine saccharification using

Phanerochaete chrysosporium, Trametes cingulata (lignin removal),

Trichoderma reesei (xylan and cellulose digestibility)

•Depolymerise lignin and can metabolise lignin as a primary carbon source.

•Live on dead trees in the wild

•Can strip lignin off cellulose with no need for alkalis, high pressure or high

temperature.

A Devoto 14/15

JAs and cell cycle

Chromatin remodellingRHUL, UK

Dr Sandra Noir – Post DocMoritz Bömer - PhD Student

Tsui Tjir-Li – MSc student

Dr Zoltan Magyar – Post Doc

Dr Virginia Balbi – Post DocPaolo Bombelli– Res assistant

Safina Khan – Res assistantKlara Godo – Research

assistant

Dr Keiko Sugimoto, RIKEN CSRS, JapanDr Motaki Seki, Dr Jong-Myong

Kim, RIKEN CSRS, JapanDr Naoki Takahashi, NAIST,

Japan

Metabolomics

Dr. H. Florance Dr Venura Perera

Prof. Nick Smirnoff University of Exeter

CSSB, Systems and synthetic

biology centre, RHUL, UK

SBS

Prajwal Bhat- PhD StudentEmanuele Libertini - PhD Student

CS Dr Hugh ShanahanProf Alberto Paccanaro

University of Milan, ItalyMonica Bertossi - PhD Student

Prof Piero Morandini

Effect of Jasmonateson the

Biology of Normal and Tumour CellsDrs Amanda Harvey and Aican

CynarBrunel University, UK

Biofuel production - Pectin

modified linesProf Felice Cervone Prof Giulia De Lorenzo

Prof Daniela BellincampiDr Simone Ferrari

Dr Fedra Francocci, Università di Roma La Sapienza, Italy