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Symbiotic and phylogenetic diversity of rhizobia associated with native and introduced Acacias in Algeria

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session 6 • Functional and structural diversity • bacteria

l B

Lactiç

acid bacteria in org.nie agrfculttJral soils

SomersElen', AllamsAmke',

"'*'

Croonenborghs'.vanOverbeekLee'. Yaodedeydeo

Jœ'

1K.U.leuven Microbial and MoIeoJlar Systems Kasteelpar Arenbe!g 20 3001 Heve.1ee. Belgium 'P1aot Research International, wageningen,theNelhlldands

According

to

a oonœpl foonulaled by H.P. Rusch (1906·1977). soli fertility of organic agriculturel soils canberelated to the presence of lac-ticacid batteria. HO'NeVer. lhere is liltle or no evidence ln literallJr8

to

support lt1isronœpt.

We have sampled al dilferenl lime points oyer 8 2 year period (2005-2006) 15 organic farmslnF1anders. Belgium.Soilsusedforthe orga· nieproduction

01

cabbage and Ieak~$l'Impled. Direct platingof soil dilutions oneltherMRS(withglucose ascaJ'1)onSOUrat) or M17(with

lactose ascarbon),COlTI'IlOflIyused growlhmediaforlacIic

acid

bac1eria,revealed no signifiCant numbefs of IaclicacicIbacteria.Thesame

reUts

Mlf8obtainedwi1hthe aJlture independenl DGGE analysis. using nested PCR 8SdescribedbyHeitigelal. (2002).identificatiOnof colonies obtained by direct plating using l.iIctobaciIus lIfOUl>specific primers for 165 rONAsequences ~1eCI maîIiy BaciIus, PHnibadIusand~spedes.

lnilsecondround.an er."'ic:tmenl&teP lot IadicacidbacteriI

was

nctded.byncubating dilutecl sail samples anaerobicatyinMRSOfM17 medium. priorlapIatingorOGGE 1NlIysis.ilSdesc:riledbyChanetal(2005).ms revealedhigh~anddiYetsityoIladic aeid bac-teria.5equencingthe165rONA.te'INllId!hefoIIowWlg Iadic:aeid baderia:Lac:toooc:aIsgaMelle,LBctx:ocalS~.

LlK.QltlOS/Dc

me.sen-tfHOideJ, VNi$seh,l.Bctobat::iIusWld Etltatl:lCXJ(;CUS,Sorneofthese isoIateswetetestedanclbJndposiWeb biocontIolacrMtyandIor cfirecl: plant

wowth

promotion.Two

UdococaIiI8ctis

isolalesshowedsignificantplanlgrow\hpromotiflgaclMtyin greeMousetria\$with

"'--.

Il canb8eoncIudedlhallacticaadbacteria,althoughpresent.areJ'lOtOominanllnorganlcfarmingsoilsin

Randers.

Nevertheless,ifthey

canb8enrichedltlrough agriaJltural Pf3C\icas.sorne oflheseshowpotenlialforbioalnlroland/orplantgi'OWlhpromotion. Heilig el al(2002) Appl.Environm.Microbiol. 68.114·123.

Chenetal(2005) Lelt. Appl.MiC1Obioi.40. 195-200.

Rusch H.P. (1964) Bodenfruchbar1leij.Kart F. Hauh Vertag,ed.(Heidelberg).

am

Symbiotic and ph)40genetic diversity of rhizobia associated

with

native and introduced Acacias in Alg.na

BoukIIalemZinebFm,Dome~Ollie.GalianaAntoine.DeLajudiePhilippe.LaguerreGiMle. BekkiAbdIIbder.Ge!jlM

AntoDa

~clesSyntJio$esT~ et~UMR 113ClRADIJRD/INRAI SupAgroJUMlI, 34398MonlpeIierCedex5. Franœ

lnthearidzoneswtIete

aop

producü:lnIsredl.œddue

to

Qoouglltcombined

to

Iow soi!lertiity,theuse of~nativeIegumenes adapted

to

suctl haf5htXlidlîonsooukIenhanc::e thepmdtJdiyiyofagroIorestry systems.InNorthAfrica,Acade speciesare goocI c:andida-tes b thisplSpOSesinœtheycan

tFJW

onN-deficienl50ÎISand~lheirNbaJal'lOllduetetheirS)'ITlbiollcassociationwîlh rhizobia,

~soibadetîa.

ln Ofder10ldentifyandseled. ellicienlacaCia-rtlizcbia syntIioticassociations.soi! sarnplesandroolnodulesFrom seven Acaciaspecies,l1ve nativelAehtenbet'giBna,A

htet.,

A.niIotica,A. seyaf, A.tortis)andIwointroduœd (A.kanoo,A.SB6gna).were coIlected irl arid and seJri.

arid regionsofAIgeriafromOran

to

TamanrassetAroIIectionof22 bac1eria1strainsWBSobtained aftet trapping on

Ac.cia

seedlings inocu-laledwilhsolisamples originating from nine geographic siles.165tONAiequencing reveal8dlhal the

new

strains repteS8l1led dilfll1"enl spa-cles in Slnorfllzobium and Rhizobium. This genetic diversity WBS confirmedby phenotypicch&racterisation through biochemical assays and host spec\Nm.Somastrains were partieularty tolerantlo loWpHand high NaCI c:oncentralions. temperature and osmotic slress in in vitro condttions. In a neKt slep, selectionofelTicienl rtlizobia displaying a high nitrogen-fixing polential uOOer stress conditions willbecanied out

inconll'Olled conditions before lasting ltlem in fietd conditions.

116

liID

Genttlc:

diversfty of rhizobia that nodulate

Prosopis jullfkJnl

in the and 'Istem arel of Morocco

Btna!B

Hlrww·Ou~MoIammed.Bemc:hiAbdebasset,Abdel~menHanaiI,8ouktlal6mNoumIddine. Missbah ElkIriuît.\Wapl'la

~MchamedPremier, F... ôesac:iences. e>..;ciaBiokJgieL..alxnlonde BiOklgie ôesfltantesetdes~ s .FacU'.6deS

SCIen-ces ,Oujda60.000 OujclaMoroc:co

5ymb1cticnilrogenfixation

n

!he aridregions depends

on

!heIwopartnInofthesyrnbiO$iS:

Iegume -

Itizotlium andIheiri<ptitude10

aaapl

teprev;iIing ctmalicandsolCMStrMlts.Vieshowed!hal ProsopisjcJiItxaisa sallolefant~species, moait is

net

aftededby the sanc:oncenntiOnsas high as seawaterIevels.This plant repraserns a verygoodcandidateteoombatdesel1il'icalion.

we

lhusisolated214 bac:l&rial strainsfromtherootnodu6es

ri

PtosopisjuIifIora,euItivatedindilfetenl soils

ri

theEastem areaof

Motoeco.

Wefoundlhal 24 strains IoIerated NaCI cooœnlr.Jtions highef than 30~(w/v). AMthestrains re-oodulatedtheiroriginals hosl They aH pœ-sessedthenodCgane. This wascheckedbyamplifyingthenodC genewhichcodesfora NodC nodulalionfactorbyPCRspecifieprWners. Theassessmentofmetabolic and physiologie properties 01\he$eisolaleshoweda hlgh phenotypic diversity.thisIs an advantage forthe plant b&cause~can be always nodulaledbydiflerent types 01 rtlizobia even ifthesoil or climate conditions have to change.

The RFlPofthe165 fONA using Mspl. Haelll and Cioi enzymes oonfinnedthedivel'Sity ofthestrains!hat oodulate thls specias inthesame

solI.We Illus obtalned lhree difterenl profiles.Thedivefsity cithestrains was alsa assessedbytheBllpIoralionofthebaeterial chromosome eatried

out

bytepPCR usingBOXA1R.Rep (1andIl) and ERIC (1 and Il) primers. The Resultsobtainedshoweda highdiversityofltlestralns. Sornertlizotlial strains kindly provided byooIleaguesofthe"laboratoireCommundeMiaobioIogieofthe JRDIISRAAJCAO" in ()al(ar,

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