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VII ECPGR Ba

The Collection of Dof Barley and their

B dBreed

Stanca AM, Pagani D, Alberici R,Terzi V, Tondelli A, and Lunquist U

arley WG Meeting

Developmental Mutants r Potential Use in Pre-

di W kding Work

, U Nicosia, 10-12 May 2011 CYPRUS

19At that time the Mutanand Genetically charaand Genetically charathem a relationship b

hormone biosynthehormone biosynthe20

The collection has bdeveloping double muta

onFor many of them todaFor many of them, todabeen sequenced and th

For many others weFor many others we

970ts were Morphologically

acterized and for few ofacterized and for few of between mutation and esis has been foundesis has been found010been implemented by ants and by isolating new nesay gene/s involved haveay gene/s involved have he function evidenciatede don't know anythinge don't know anything

The domestication Barley belongs to the represents of branch pPoaceae. Diploid (2n = 2

The Plant morphology very well characterize

of cultivated barley y

tribeae Triticeae whichin the grass familyg y

x = 14) has been

ed

Th M t tThe Mutants arfollowing categ

SP- SP- LE- S

GR- GR

di id d i thre divided in thegories/classes:g

PIKEPIKEEAFTEMRAINRAIN

EAR MEAR MMUTANTSMUTANTS

The development of controlled by a single for vulgare), that is recallele responsible for p(Vsr1).

a six-rowed spike isallele, vrs1 (formerly v

essive to the dominantthe two-rowed spikep

The wild type Vrs1 allele (forThe wild-type Vrs1 allele (fora transcription factor that ina closely linked leucine zippwas strictly localized in the immature spikes, suggestisuppresses development offunction of Vrs1 resulted in rudimentary lateral spikeletrudimentary lateral spikeletfully developed fertile spphenotype. Komatsuda et al.PNAS 2007

r two rowed barley) encodesr two-rowed barley) encodescludes a homeodomain wither motif. Expression of Vrs1

lateral-spikelet primordial ofng that the VRS1 proteinf the lateral rows. Loss ofcomplete conversion of the

ts in two-rowed barley intots in two rowed barley intopikelets in the six-rowed

This feature is an evolutiby the two-rowed spspontaneous six-rowedspontaneous six rowed naturally and rapidly frombecause they lack thisbecause they lack thisrowed barley occurs p

dweeds.

ionary advantage offeredpikes in nature, andmutants are eliminatedmutants are eliminated

m wild barley populationsadaptation Thus six adaptation. Thus, six-

rimarily as cultivars of

SIX-ROWED BARLEY OMUTATION IN A HOMUTATION IN A HOZIPPER I-CLASS HOMEO Komatsuda T., M. PourAzhaguvel, H. Kanamori,Graner T Wicker A TGraner, T. Wicker, A. TFujimura, M. Matsuoka, T. PNAS 104(4): 1424 1429PNAS, 104(4): 1424-1429,

ORIGINATED FROM AOMEODOMAIN LEUCINEOMEODOMAIN-LEUCINEOBOX GENE

rkheirandish, C. He, P. D. Perovic, N. Stein, A.Tagiri U Lundqvist TTagiri, U. Lundqvist, T. Matsumoto, M. Yano

20072007

CORONA GRAMI

Seconda guerra punica “CoroImperatore Galliano 268 d.C.

INEA

ona Obsidionalis” ORZO?

Awn MMutants

The barley Hooded mutation caused by a dupThe barley Hooded mutation caused by a dupKai J. Müller*, Nicoletta Romano*, Ortrud GersCarlo Pozzi*, Francesco Salamini* & Wolfgang R

The hooded mutation causes the appearance of an extThe hooded mutation causes the appearance of an ext

Single dominant genetic locus

Homeobox gene Knox3 (Bkn3)

305-base pair duplication in intron 4

plication in a homeobox gene intronplication in a homeobox gene intronstner*, Federico Garcia-Marotot†, Rohde*

tra flower of inverse polarity on the lemmatra flower of inverse polarity on the lemma

Nature 374: 727-730 (1995)

In Hooded plants,develops at thedevelops at the between lemma and

, an extra flowersite of transitionsite of transitionawn.

The hooded locufor a regulator activity.

s may be codingr of peroxidase

GENETICS OF BGENETICS OF BSUPPRESSIONSUPPRESSION Roigh C., C. Pozzi, L. SaM.R. Stile, L. Rossini , A.M Genetics, 167: 439-488, 20

BARLEY HOODEDBARLEY HOODED

anti, J. Müller, Y. Wang,M. Stanca, F. Salamini

004

TH E R ELATIO N B ETA C TIV ITY A N D THEXPR ESSIO N O F TH EB A R LEY Stebbins G .L., V .K . G upta PNAS, 64: 50-56, 1969(This inform ation is current as of M

TW EEN PERO XID ASEE M O R PH O LO G ICA L

E H O O D ED G EN E IN

ay 2007)

GENETICS OF MUTATIGENETICS OF MUTATIDEVELOPMENT OF BRACTBRACT Pozzi C., P. Faccioli, V. TerP. Castiglioni, R. Fink, Rg , ,Bossinger, W. Rohde, F. Sa Genetics, 154(3): 1335-1346

IONS AFFECTING THEIONS AFFECTING THEA BARLEY FLORAL

rzi, A.M. Stanca, S. Cerioli,R. Capone, K.J. Müller, G.p , ,alamini

6, 2000.

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Stem MMutants

Leaf MLeaf MMutantsMutants

GrainGrain MutantsMutants

N: GENOTIPO NAKED H: GEN

MARCATORI MAPPATI VICINO AL GEN

bpN: GENOTIPO NAKED H: GEN

750

500

250

H N

sKT2 (0.9cM da nud)sKT2 (0.9cM da nud)

bp

N H

250

500

750

sKT3 (0.9cM da nud)

250

NOTIPO HULLED (vestito)

NE NUDO (nud)

bpNOTIPO HULLED (vestito)

750

500N H

sKT7 (comappa con nud)

250

pp )Controllo positivo ICE1

750

bp

250

500

750NH

sJ14 (2.7cM da nud)

H A PLO TYPE STR U C TN U D LO C U S IN A SSO C IA TIO N W ITH RA SSO C IA TIO N W ITH RSTR IPE (PYR EN O PH O R B arabaschi D ., L . C am pani, EG .P . Valè, A . G ianinetti, GPecchioniPecchioni Plant B reeding, 126: 24-29, 20

TU R E A R O U N D TH EB A R LEY A N D ITS

R ESISTA N C E TO LEA FR ESISTA N C E TO LEA FR A G R A M IN EA )

E . Francia, H . Toubia-R ahm e,. D elogu, A .M . S tanca, N .

007

• TAKETA et al. PNAS 200

Barley grain with controlled by an efactor (ERF) genefactor (ERF) gene

biosynthesi

the gene expressiohthe te

08

adhering hulls is ethylene response

regulating a lipidregulating a lipid is pathway

on is localized to esta

THE LODICULES OF CLEISTOGAMOUS AN

(A)stamlemlem

(B)aclacl

Thclecle

Alcle

ThThorg

Sc

ND NON CLEISTOGAMOUS BARLEY

). The lodicule (lo) located at the base of themen (st)open the spikelet by pushing apart themam (le) and palea (pa)mam (le) and palea (pa).

). A non cleistogamous and (C)leistogamous barley spike at anthesisleistogamous barley spike at anthesis.

e lodicules of (D)a non cleistogamous and (E) a istogamous barleyistogamous barley.

section of the spikelet in (F) a noni t d (G) l i t b leistogamous and (G) a cleistogamous barley.

he carpel (cp) is surrounded by the other floralhe carpel (cp) is surrounded by the other floralgans.

cale bars: D and E, 800 µm; F and G, 200 µm.

“Cleistogamous flowerth i f ithe suppression of mic

mRNA

Nair S K  Wang N  TuruspeNair S.K., Wang N., TuruspeSinsuwongwat S., Guoxiong C.Watanabe Y., Kanamori H., W

Matsumoto T.

PNAS 2010 Vol. 107 PNAS 2010 Vol. 107 

ring in barley arises from RNA id d H AP2 croRNA-guided HvAP2

cleavage”

ekov Y  Pourkheirandish M  ekov Y., Pourkheirandish M., ., Sameri M., Tagiri A., Honda I., 

Wicker T., Stein N., Nagamura Y., ., Komatsuda T.,

 n.1: 490‐495 n.1: 490 495

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Double Mutants

PLANT FOR THPLANT FOR THHE FUTUREHE FUTURE

STONE- BREAKER STONE- DRESSER

M A N N E R I S T

YIELD ENHANCEMENYIELD ENHANCEMENNT GENESNT GENES

Double Mutants

D. Pagani, R. Alberici, F. RizzC Morcia Udda Lunquist MiC. Morcia, Udda Lunquist, Mi

CRA-GPG Fiorenzuola

za, A. Tondelli, C. Crosatti, ichele Stanca Valeria Terziichele Stanca, Valeria Terzi

Le

CaratterizzazioneCaratterizzazionemorfologica

Caratterizzazione genetica

eafy lemmay

A CONSENSUS FUNCTION MAPSTRES

18 Candidate genes either collected from ( C ) COhomology searches (TC-) are boxed. COR an

and RFLP markers showing sequence homoitalic. Blu and red boxes inside chromosomes

respec

P FOR THE STUDY OF BARLEY ABIOTIC-SS TOLERANCE

GenBank accessions (Hv-) or obtained by i i i ffnd other abiotic stress-induced effector genes,

ology to barley stress-induced TCs are in bold s represent cold and drought tolerance QTLs, ti lctively.

tmc-ap3: a chloroplast-

Sub-cellurar fractionsSub-cellurar fractions

-localised cor protein

Sub-chloroplats fractionsSub-chloroplats fractions

HYPOTHES

PROTEINSYNTHESIS

INCREASEDFLUX ?

HYDROXYL RADICAFORMATION

SIS OF FUNCTION

AMINO ACIDSSYNTHESIS

PROTEINDEGRADATION

ALS AMINO ACIDSACCUMULATION

THE OUTER PLASTID ENVELOAS PRECURSOR TRANSLPROTOCHLOROPHYLLIDE OXPROTOCHLOROPHYLLIDE OX Reinhothe S F Quigley A SReinhothe S., F. Quigley, A. SReinbothe PNAS, 101(7): 2203-2208, 2004

OPE PROTEIN OEP 16: ROLELOCASE IN IMPORT OFXIDOREDUCTASE AXIDOREDUCTASE A

Springer A Schemenewitz CSpringer, A. Schemenewitz, C.

4

DNA chip ttechnology

The hardening transcrip

Total cor genes up rTotal cor genes down

Total cor genes correctly up Total cor genes correctly down

ptome: ARRAY analysis

regulated in WT : 850 n regulated in WT : 720

regulated in the mutants: 98 wn regulated in the mutants: 36

B A R L E Y G E N O M E N E TB A R L E Y G E N O M E N E T A N A L Y S I S A N D E X P L O

D I V E RT h e c e n t r a l g o a l o f t h i s p r o j e c ta s s o c i a t i o n m a p p i n g a p p r o a c ht y p e s f o r t h e d i s c o v e r y o f n et y p e s f o r t h e d i s c o v e r y o f n e( H o r d e u m v u l g a r e s s p . s p o n t a n

f o r c r o p b r e e d i n g . O u r a p p r o ag e n o m i c s b a s e o f b a r l e y a n d wc o n c e p t s p i o n e e r e d i n h u m a ne f f i c i e n c y o f t h e a s s o c i a t ii d e n t i f y i n g g e n e a l l e l e s i n H o rb r e e d e r .

: G E N O M I C S A S S I S T E D: G E N O M I C S - A S S I S T E D O I T A T I O N O F B A R L E Y R S I T Y

i s t o e s t a b l i s h a n i n c r e m e n t a l b a s e d o n d i f f e r e n t p o p u l a t i o nw g e n e a l l e l e s i n w i l d b a r l e yw g e n e a l l e l e s i n w i l d b a r l e y

n e u m ) , w h i c h c a n b e e x p l o i t e dc h w i l l b u i l d u p o n t h e s t r o n g

w i l l a p p l y a s s o c i a t i o n g e n e t i c ss a n d A r a b i d o p s i s t o t e s t t h e o n g e n e t i c s a p p r o a c h f o r

r d e u m t h a t a r e n e e d e d b y t h e

Th d bj ti i tThe second objective is to alleles, which have been studies, into advanced ba

d i d fprograms derived from wspontaneum germplasm anallow us to determine the eand extraction of usefuland extraction of useful programs based upon wide

it th f lrecruit the new useful genediscovered in the above

ack-cross (ABC) breedingid b t Hwide crosses between H.

nd elite cultivars. This willefficiencies of identificationalleles in barley breedingalleles in barley breedingcrosses.

The third major projectThe third major project huge DNA and marker dgproject to determine genetic parameters for b

objective is to use theobjective is to use thedata set obtained in the

important populationbarley.

Müller K.J., Romano NMaroto F., Pozzi C., Sal(1995). The barley Hoo

by a duplication in a hby a duplication in a hNature, 374: 727-730.,

N., Gerstner O., Garcia-lamini F. and Rohde W.oded mutation causedhomeobox gene intronhomeobox gene intron.

A single flower may ariseA single flower may arise individual vegetative shoot apshoot may be transformed yinflorescence that contains flowers. All grass inflorescenc

each spikelet containing one spikelet develops as a short

hi h i tt h d t th iwhich is attached to the mainoverlapping sterile bracts (gluthird bract is the lemma whichthird bract is the lemma which

lemma terminates in a long considered to be a modified lea

through modification of anthrough modification of anpex or more often a vegetative

into a flowering shoot orgseveral or many individual

ces are organized in spikelets;

or many florets. The barleytened stem axis, the rachilla,

i hi d b t axis or rachis and bears twomes) in alternating order. The

h bears a flower in its axil Theh bears a flower in its axil. The

pointed extension, an awn,af blade.

The inflorescence architectcontinuous story of reduct“panicle” of spikelets (as seedof spikelets resulting in threeof spikelets, resulting in threebarley and single sessile spirye. In wild Hordeum species,slender awns form a light disanemochory and zoochory. temporal and spatial regulatioreduction and sterility of the la

ture in the Poaceae is ation from a more originald in rice and oats) to a “spike”e sessile spikelets per node ine sessile spikelets per node inkelet per node in wheat and, the three spikelets and theirspersal unit that permits both

In two-rowed barley, stricton of Vrs1 expression leads toateral spikelets.

We speculate that either sexpression or regulation of Vexpression or regulation of V

complete repression of lati fl d f thinflorescence nodes of other found in wheat and rye. A Poavariability and regulation of exciting and productive way tof plant development and the e

strong alleles or differentialVrs1 orthologs could lead toVrs1 orthologs could lead toteral spikelet formation at

i i th P ispecies in the Poaceae, as isaceae-wide assessment of theVrs1 orthologs would be an

to improve our understandingevolution of grass species.

As yet, very little irelationship between trelationship between tthose genes which characteristics of highexpression of such chaexpression of such cha

is known about thethe primary effects ofthe primary effects ofaffect morphological

er plants and the finalaractersaracters.

The most critical stepare the ones whichare the ones which enzyme activity tomorphological effects.

ps in these pathwayslink gene controlledlink gene-controlled

o histological and.

ChloroplaChloroplaast Mutantsast Mutants

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