Screening and selection of growth-promoting bacteria for ıt Dunaliella cultures Myriam Le Chevanton, Matthieu Garnier, Gaël Bougaran, Nathalie Schreiber, Ewa Lukomska, Jean-Baptiste Bérard, Eric Fouilland, Olivier Bernard, Jean-Paul Cadoret To cite this version: Myriam Le Chevanton, Matthieu Garnier, Gaël Bougaran, Nathalie Schreiber, Ewa Lukomska, et al.. Screening and selection of growth-promoting bacteria for ıt Dunaliella cultures. Algal Research, Elsevier, 2013, 2, pp.212–222. <10.1016/j.algal.2013.05.003>. <hal-00852257> HAL Id: hal-00852257 https://hal.inria.fr/hal-00852257 Submitted on 20 Aug 2013 HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés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`!;;VKZ!:!<K!;?!<;!;>!/!! :K! F&]_!`;;VKZ:!<K!;?!<K!?"[!@&82*&12/*(!0(!A,B)/2$2%/(!(1!0(!C/21(+,.2$2%/(!0()!#$%9()D!EF'GHG'D! :"! *9(!0(!$IE$(!0IJ(9D!<<;KK!M#MNG-D!F*&.+([(( ::! ( :;! ! " :;! Abstract :<! A*(W/29)!)190/()!,&W(!0(72.)1*&1(0!1,&1!8&+1(*/&!/.3$9(.+(!7/+*2&$%&$!7(1&82$/)7D!)9%%()1/.%!1,&1!1,(! :=! )($(+1/2.!&.0!+,&*&+1(*/a&1/2.!23!%*241,56*2721/.%!8&+1(*/&!),29$0!233(*!&!.(4!)1*&1(%B!32*!/76*2W/.%! :>! /.09)1*/&$! &$%&$! +9$1/W&1/2.[! E.! 1,(! 6*()(.1! )190BD! <L! +9$1/W&8$(! 8&+1(*/&! 4(*(! /)2$&1(0! 3*27! 7&*/.(! :?! 7/+*2&$%&(!)6(+/()!&.0!/0(.1/3/(0!9)/.%!">-!*'M#!6,B$2%(.(1/+!&.&$B)/)[!N,(!*(+2W(*(0!8&+1(*/&!4(*(! :L! 329.0!12!8(!7(78(*)!23!1,(! cA!&.0! iAH50/'0;$:/'5&$D!,I/0L-$7$AD%$.0;$:/'5&">A6$:/'50&C'?!VRFCZ! :P! &.0! %*&7562)/1/W(! 72.26,B$(1/+! +$9)1(*)[! N2! &00*())! 1,(! 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N,()(! 8&+1(*/&! +29$0! 8(! +2.)/0(*(0! &)! ,($6(*)! 32*! M! ;P! &++979$&1/2.!/.!!"#$%&'%%$(?L*!+($$)[!! <K! Keywords: H/+*2&$%&(D!!"#$%&'%%$D!8&+1(*/&D!/.1(*&+1/2.D!8&+1(*/&$!0/W(*)/1B! <"! Abbreviations: N7&]D!7&]/7&$!%*241,!*&1(O!ÄO7&]D!7&]/7&$!8/27&))!/.+*(&)(!&1!)1&1/2.&*B!6,&)(O!R_MD! <:! +&*82._./1*2%(.!*&1/2O!R,$!$D!+,$2*26,B$$!$!O!'#AYD!*&.027!&76$/3/+&1/2.!23!62$B72*6,/+!YM#[ <;! ! : <;! 1. Introduction <<! N,(*(! /)! &! 0/W(*)(! &**&B! 23! +9**(.1! &.0! 621(.1/&$! &66$/+&1/2.)! 32*! 7/+*2&$%&(D! 4,/+,! /.+$90(! 3220D! <=! &./7&$! 3((0D! ,(&$1,+&*(D! (.(*%B! &.0! 6,B+2*(7(0/&1/2.! b"c;d[! N,(! 8227! 23! 7/+*2&$%&$! W&$9(5&00/.%! <>! 2W(*! *(+(.1! 0(+&0()! ,&)! 0*&4.! &11(.1/2.! 12! 1,(! )190B! 23! 8&+1(*/&57/+*2&$%&(! /.1(*&+1/2.)! /.! &66$/(0! <?! &$%&$!+9$1/W&1/2.!b<D=d[!C&+1(*/&!+&.!+276(1(!4/1,!7/+*2&$%&(!32*!1,(!$/7/1(0!*()29*+()! b>D?d!2*!(W(.! <L! 6*209+(! 12]/+! )98)1&.+()! &%&/.)1! 7/+*2&$%&(! bLdD! &$$! 23! 4,/+,! +&.! 0(+*(&)(! +9$19*(! 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a/.%! +9$1/W&8$(! ?L! 8&+1(*/&! 3*27! W&*/29)! 7/+*2&$%&$! +9$19*()! 32*! )98)(f9(.1! 1()1/.%! /.! &))2+/&1/2.! 4/1,! !"#$%&'%%$( ?L[! ?P! -#h!"P[;[!E.!&!,/%,51,*29%,691!(]6(*/7(.1!4(!3/*)1!)+*((.(0!&!$&*%(!.978(*!23!7/+*2&$%&(!5!8&+1(*/&! LK! &))2+/&1/2.)! 32*! 1,(/*! (33(+1! 2.! 7/+*2&$%&(! %*241,[! N,*((! )($(+1(0! 8&+1(*/&! )1*&/.)! 4/1,! 621(.1/&$! L"! %*241,!&$1(*/.%!(33(+1)!2.!!"#$%&'%%$!?L(4(*(!39*1,(*!1()1(0!/.!&!3$&)e!(]6(*/7(.1[!'()9$1)!,/%,$/%,1(0! L:! 1,(! %*241,! &$1(*/.%! (33(+1)! 2.! !"#$%&'%%$( ?L[! -#h! "P[;! &.0! /.3$9(.+(! 23! 1,()(! 8&+1(*/&! 2.! ./1*2%(.! L;! /.+2*62*&1/2.!/.!7/+*2&$%&([!! L<! 2. Materials and methods L=! 2.1. Algal strain, maintenance and purification L>! !"#$%&'%%$( ?L*! -#h! "P[;! 4&)! 281&/.(0! 3*27! 1,(! +9$19*(! +2$$(+1/2.! 23! &$%&(! &1! 1,(! T./W(*)/1B! 23! L?! h2(11/.%(.!V-#hZ!h(*7&.BD!&.0!7&/.1&/.(0!&1!:K!iR!9.0(*!+2.1/.929)!$/%,1!4/1,!0&B$/%,1!3$92*()+(.1! LL! 198()! V=K! j72$! 6,212.)! 75:)5"Z[! R9$19*()! 4(*(! 6(*32*7(0! /.! )1(*/$(! G*$(.7(B(*! 3$&)e)! 3/$$(0! 4/1,! LP! &*1/3/+/&$! )(&4&1(*! V#-kD! )&$/./1B! ;=Z! b:=d! 3/$1(*(0! &1! K[::! j7! &.0! (.*/+,(0! 4/1,! 720/3/(0! k&$.(I)! PK! 7(0/97!b:>d[!#772./97!4&)!9)(0!&)!&!./1*2%(.!)29*+(!V"["?!7HZ!&.0!W/1&7/.)!4(*(!27/11(0[!N,(! 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v! ::"! '#AY! #.&$B)/)! A*/7(*! "! V=w5! hhNhRhhh##5;wZ! &.0! <! V=w5! ##h#hRRRhN5;wZ[! N,(! +B+$/.%! :::! 6*2%*&7!4&)!&)!32$$24)_!=!7/.!&1!P<!iRD!<=!+B+$()!/.+$90/.%!"!7/.!&1!P<!iRD!"!7/.!&1!;>!iR!&.0!:!7/.! ::;! &1! ?:! iR[! AR'! 6*209+1)! 4(*(! )(6&*&1(0! 2.! :! m! N#G! &%&*2)(! %($)! &.0! 1,(! 6*23/$()! 281&/.(0! 4(*(! ::<! +276&*(0!12!1,(!2*/%/.&$!8&+1(*/&$!*(3(*(.+(!)1*&/.!3*27!1,(!+2$$(+1/2.[! ::=! 2.5. Estimation of growth and statistics ::>! H&]/7&$! %*241,! *&1(D! N7&]! V05"Z! 23! !"#$%&'%%$( ?L*! 4&)! +27691(0! &++2*0/.%! 12! (f9&1/2.! "! 3*27! 1,(! ::?! $/.(&*!6&*1!23!1,(!$.51*&.)32*7(0!%*241,!+9*W(_! ! P (N7&]!r! F$. O !!! F/ Gf9&1/2.!" ::L! 4,(*(! O! /)! (/1,(*! R,$! $! 3$92*()+(.+(! V(]6(*/7(.1! "Z! 2*!6&*1/+9$&1(! +&*82.! V(]6(*/7(.1! :Z!09*/.%! 1,(! ::P! (]62.(.1/&$! %*241,! 6,&)(! &.0! /! /)! 1/7(! /.! 0&B)[! -/.+(! 8&+1(*/&$! +&*82.! +2.1*/891(0! 12! &! $24! $(W($! 12! :;K! 121&$! 6&*1/+9$&1(! +&*82.! *(+2W(*(0! 2.! hFnR! 3/$1(*)D! 7/+*2&$%&(! N7&]! +29$0! 8(! +27691(0! 3*27! 121&$! :;"! 6&*1/+9$&1(!+&*82.!0&1&!/.!(]6(*/7(.1!:[! :;:! H&]/7&$! 8/27&))! /.+*(&)(! O7&]! 23! !"#$%&'%%$( ?L*! 4&)! +27691(0! &1! )1&1/2.&*B! 6,&)(! &++2*0/.%! 12! :;;! (f9&1/2.!:_!! Gf9&1/2.!: O7&]!r!OKA(O&(((! :;<! 4,(*(! OK( &.0! O&! &*(! (/1,(*! R,$! $! 3$92*()+(.+(! V(]6(*/7(.1! "Z! 2*! 6&*1/+9$&1(! +&*82.! V(]6(*/7(.1! :ZD! :;=! *()6(+1/W($B!&1!1,(!)1&1/2.&*B!6,&)(!&.0!&1!1,(!8(%/../.%!23!1,(!+9$19*([! :;>! -/.+(! O7&]!4&)!+27691(0!3*27!3$92*()+(.+(!*(&0/.%)!/.!(]6(*/7(.1!"!&.0!3*27!6&*1/+9$&1(!+&*82.!/.! :;?! (]6(*/7(.1!:D!+276&*/)2.!32*! O7&]!8(14((.!1,(!142!(]6(*/7(.1)!4&)!6(*32*7(0!&31(*!.2*7&$/a&1/2.! :;L! &++2*0/.%!12!(f9&1/2.!;_! FO .2*7 7&] !?! FO 7&] /FO 7&] ! u Gf9&1/2.!; :;P! k,(*(! FO 7&] /)!7(&.!7&]/7&$!8/27&))!/.+*(&)(!&.0!u(/)!)1&.0&*0!0(W/&1/2.!/.!(]6(*/7(.1[! :<K! -1&1/)1/+&$!&.&$B)()!4(*(!6(*32*7(0!4/1,!-1&1%*&6,/+)s!)2314&*(!32*!1,(!3&+12*/&$!0()/%.!&66*2&+,!&.0! :<"! '! )2314&*(! VhMT! 6*2g(+1Z! ($)(4,(*([! -/.+(! (]6(*/7(.1)! "! &.0! :! /.W2$W(0! 2.$B! 1*/6$/+&1(! +9$19*()D! :<:! *()9$1)!&*(!(]6*())(0!,(*(&31(*!&)!7(0/&.!&.0!/.1(*f9&*1/$(!*&.%(!VEu'Z!*&1,(*!1,&.!7(&.!&.0!)1&.0&*0! :<;! (**2*[! E.! (]6(*/7(.1! :D! (33(+1)! 23! 8&+1(*/&! 2.! 7/+*2&$%&$! 6&*&7(1(*)! 4(*(! 1()1(0! 9)/.%! 1,(! x*9)e&$! :<<! k&$$/)! 1()1! Vgr=mZ[!N,(!+276&*/)2.!23!%*241,!6&*&7(1(*)!+27691(0!3*27!1,(!142!(]6(*/7(.1)!4&)! :<=! +&**/(0!291!9)/.%!-6(&*7&.I)!*&.e!+2**($&1/2.!+2(33/+/(.1!Vgr=mZ[! ! "K :<>! 3. Results :<?! 3.1. Purification of Dunaliella sp. SAG 19.3 culture :<L! #.! &](./+! +9$19*(! 23! !"#$%&'%%$( ?L*! )1*&/.! -#h! "P[;! 4&)! 281&/.(0! )9++())/W($B! 12! 1,(! *(6(&1(0! :<P! &.1/8/21/+!1*(&17(.1[!M2!+9$1/W&8$(!8&+1(*/&!4(*(!28)(*W(0!2.!1,(!H&*/.(!#%&*!6$&1()!/.2+9$&1(0!4/1,! :=K! )&76$()!23!1,/)!&$%&$!+9$19*([!E.!&00/1/2.D!8(32*(!1,(!9)(!23!1,/)!!"#$%&'%%$(?L[!+9$19*(!32*!(]6(*/7(.1)D! :="! &8)(.+(! 23! 9.+9$1/W&8$(! )1*&/.)! 4&)! )B)1(7&1/+&$$B! W(*/3/(0! 8B! (6/3$92*()+(.+(! 7/+*2)+26B! &31(*! :=:! -JC'%*((.!)1&/./.%[!! :=;! 3.2. Bacterial collection :=<! F2*1B5(/%,1!)1*&/.)!23!8&+1(*/&!4(*(!/)2$&1(0!3*27!"P!7/+*2&$%&(!)6(+/()D!23!4,/+,!?"m!4(*(!&+f9/*(0! :==! 3*27! 0/&127)[! E.! 1,(! +2$$(+1/2.D! &.&$B)/)! 32*! 6&*1/&$! ">-! *'M#! *(W(&$(0! 1,&1! ;?! )1*&/.)! 4(*(! %*&75 :=>! .(%&1/W(D! 23! 4,/+,! L! 8($2.%! 12! ,I/0L-$7$AD%$.0;$:/'5&">A6$:/'50&C'?( VRFCZ! /.+$90/.%! :! :=?! )6,/.%28&+1(*/&! &.0! >! 3$&W28&+1(*/&D! "?! 12! M%L-$L50/'0;$:/'5&$! &.0! ":! 12! 4$>>$L50/'0;$:/'5&$[! :=L! G$(W(.!)1*&/.)!4(*(!%*&7562)/1/W(!/.+$90/.%!"K!M:/�$:/'5&$!&.0!2.(!6$:&%%&(VN&8$(!:Z*! :=P! 3.3. High-throughput selection (experiment 1) :>K! G*G*1*(0L/&:$%(>'$?"5'>'#/(K05(>&:50$%7$'(L0L"%$/&0#(( :>"! N,(! 3&+12*/&$! &66*2&+,! 9)(0! 12! +276&*(! UY>LK! 12! 3$92*()+(.+(! &)! 6*2]/()! 32*! 7/+*2&$%&(! 6269$&1/2.! :>:! *()9$1(0!/.!821,!720($)!(]6$&/./.%!72*(!1,&.!PPm!23!1,(!W&*/&8/$/1B!28)(*W(0!2.!0&1&[!C21,!UY>LK!&.0! :>;! 3$92*()+(.+(!4(*(!)/%./3/+&.1$B!VcrK[K"Z!&.0!62)/1/W($B!*($&1(0!12!7/+*2&$%&$!+2.+(.1*&1/2.!VN&8$(!;Z[! :><! F2*!crK[K"D!.(/1,(*!f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mZ!4&)!281&/.(0!32*!1,(!](./+!!"#$%&'%%$(?L*!+9$19*([!E.1(*()1/.%$BD!;!8&+1(*/&$!)1*&/.)!/)2$&1(0!3*27! :??! 1,/)!](./+!+9$19*(!VT-0C0:0::"?(K$?:&$#?!-JKK"D!-JKK:!&.0!!&'/P&$(?L*!-J:=KZ!*()9$1(0!/.!.(%&1/W(! :?L! (33(+1)!V5"L!mD!5""!m!&.0!5::!mD!*()6(+1/W($BZ!4,(.!1()1(0!/.0/W/09&$$B[!)"5&:$"C$(?L[!)1*&/.!-J:<<! :?P! /)2$&1(0! 3*27! U-$%$??&0?&5$( ?L*D( *()9$1(0! /.! &! ::! m! 0(+*(&)(! /.! !"#$%&'%%$! ?L[! %*241,! *&1([! #.21,(*! :LK! )"5&:$"C$!)1*&/.!V-J"L>Z!&$)2!,&0!&.!/.,/8/12*B!(33(+1!2.!N7&]!V5"L!mZ[!N,(!&00/1/2.!23!+(*1&/.!)1*&/.)! :L"! *()9$1(0!/.!$())(*!*(09+1/2.)!/.!N7&]D!)9+,!&)!4/1,!V$%0>0#$?(?L[!-JKK;!V5":!mZ(&.0!M%/'50>0#$?(?L[! :L:! -JKK?!V5?!mZ[! :L;! F29*1((.! )1*&/.)! (],/8/1(0!N7&]!+$2)(!12!1,(!+2.1*2$D!)9+,!&)!1,(!)1*&/.)!&33/$/&1(0!12!M5/-50;$:/'5(?L[! :L<! -JKK<!&.0!12!6$:&%%"?(K05$>&#&?!-JKP?D!32*!(]&76$(!VF/%9*(!"Z[!! :L=! G*G*G*(<KK':/(0K(;$:/'5&$(0#(>$R&>$%(;&0>$??(&#:5'$?'(( :L>! H&]/7&$!8/27&))!/.+*(&)(!V O7&]Z!7(&)9*(0!&1!)1&1/2.&*B!6,&)(!4&)!72*(!)1*2.%$B!&$1(*(0!8B!8&+1(*/&$! :L?! &00/1/2.! 1,&.! N7&]! VF/%9*(! :Z[! G33(+1)! *&.%(0! 3*27! 5=?! m! 12! `:>! m! &.0! 4(*(! 7&/.$B! .(%&1/W([! N,(! :LL! )1*2.%()1!.(%&1/W(!(33(+1!V5=?!mZ!4&)!28)(*W(0!32*!1,(!](./+!!"#$%&'%%$(?L[!+9$19*([!-1*&/.)!-JKK"!&.0! :LP! -JKK:!/)2$&1(0!3*27!1,(!](./+!+9$19*(!&.0!&33/$/&1(0!4/1,!T-0C0:0::"?(K$?:&$#?!&$)2!0(+*(&)(0! O7&]! :PK! 4/1,!)1*2.%!(33(+1)!V5<:!m!&.0!5<<!mZ[! :P"! N4(.1B52.(!8&+1(*/&$!)1*&/.)!*()9$1(0!/.! O7&]!+$2)(!12!1,&1!23!1,(!&](./+!+2.1*2$!VF/%9*(!:Z[!\24(W(*D! :P:! O7&]!4&)!(.,&.+(0!8B!::m!&.0!:>m!4,(.!!"#$%&'%%$(?L*!4&)!&))2+/&1(0!4/1,!8&+1(*/&!-JKK?!&.0! :P;! -J:<<! &33/$/&1(0! 12! M%/'50>0#$?( ?L*! &.0! )"5&:$"C$( ?L[D! *()6(+1/W($B[! N,()(! 8&+1(*/&! 4(*(! /)2$&1(0! :P<! 3*27! 0/&127! +9$19*()_! U-$%$??&0?&5$( ?L[! 32*! 1,(! M%/'50>0#$?( ?L[! -JKK?D! &.0! H-$'0C$:/I%">( :P=! /5&:05#"/">!32*!1,(!)"5&:$"C$(?L[!-J:<<[!! :P>! 3.4. Flask cultures (experiment 2) :P?! 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M%/'50>0#$?( ?L*! -JKK?D! M! /.+2*62*&1/2.! 4&)! )/%./3/+&.1$B! (.,&.+(0! 96! 12! =>! m! 23! 1,(! ;>K! /./1/&$!M!(.*/+,7(.1[! ;>"! 4. Discussion ;>:! 4.1. Microalgae culture-based bacterial collection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g5! &.0! 5! ;>P! H50/'0;$:/'5&$D! ,I/0L-$7$AD%$.0;$:/'5&">A6$:/'50&C'?! &.0! %*&7562)/1/W(! 7&/.$B! &33/$/&1(0! 12! ;?K! 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