Cultivo de Protoplastos
2014
UNIVERSIDAD NACIONAL AGRARIA AGRARIA LA MOLINA
Facultad de Ciencias Departamento de Biología
Curso: CULTIVO CULTIVOS S DE CÉLULS !
TE"IDOS VE#ETLES VE#ETLES
Tema: Culti$o de %rotoplastos: Aislamiento, Aislamiento, cultivo e importancia importancia en en el mejoramiento vegetal.
Profesora: Dra. Antonietta Gutiérrez Rosati
-8 de mayo del 2014-
0
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
TABLA DE CONTENIDO I.
GLOSARIO GLOSARIO.... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........... ........... ....10 10
II. I!ROD"##I$ I!ROD"##I$... ....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ......... ............ ............... ............... .............. ......... 10 III. III.
D%&II#I$ D%&II#I$.... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ..........11 ......11
I'. I'.
#ARA#!% #ARA#!%R(S!I R(S!I#AS #AS D% LOS LOS )RO!O)LAS!OS. )RO!O)LAS!OS..... ........ ........ ........ ........ ............ ............... .............. .............. .........12 ..12
'. AISLA*I%!O AISLA*I%!O D% )RO!O)LAS!O )RO!O)LAS!OS.... S........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........... ............... ...............12 .......12 1. *étodo *e+,ni+o... *e+,ni+o....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........... ............... ............... .............. ........... ....12 12 2. *étodo enzim,ti+o. enzim,ti+o..... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........... .............. ..............1 .......1 'I.
)"RI&I#A# )"RI&I#A#I$ I$ D% LOS LOS )RO!O)LAS!O )RO!O)LAS!OS.... S........ ........ ........ ........ ........ ........ ........ ........ ........ .......... ............. .............1 ......1
1. Re Remo+i/n mo+i/n de deris y enzimas.... enzimas........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ .......... ............. ..........1 ...1 2. La elimina+i/n elimina+i/n de rotolast rotolastos os rotos... rotos....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ......... ............ ...........1 ....1 . )rueas )rueas de de iailidad iailidad de rotol rotolastos astos y de la 3orma+ 3orma+i/n i/n de de ared.................1 ared.................1 4. 'iailidad 'iailidad de rotolast rotolastos... os....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ .......... ............. ............... ...........1 ...1 'II.
!#I#AS !#I#AS D% #"L!I'O #"L!I'O )ARA ARA )RO!O)LA )RO!O)LAS!OS S!OS AISLADOS....... AISLADOS.............. ............... ............... ..........20 ...20
1. *edio de +ultio... +ultio....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........... ............... ............... ........... ....20 20 2. Re5uladores........... Re5uladores...................... ..................... ..................... ..................... ..................... ..................... ..................... ...................... ................... ........ 20 . &uentes +aronadas +aronadas..... ......... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ .......... ............. ............... ............... .............21 ......21 4. Sistemas Sistemas e6eriment e6erimentales ales ara ara el +ultio +ultio de rotol rotolastos astos aislado aislados..... s......... ........ ........... ........21 .21 7. Densidad Densidad de rotolas rotolastos tos y +re+imiento +re+imiento del rotol rotolasto asto en el medio de +ultio +ultio 21 . Aan+es innoatio innoatioss ara los +ultios de rotola rotolastos stos...... .......... ........ ........ ........ ........ .......... .............22 .......22 •
•
%stimula+ion elé+tri+a de los rotolastos.................. rotolastos............................. ..................... ..................... ...................22 ........22 Sulementa+i/n del medio de +ultio +on sur3a+tantes9 antii/ti+os y oliamidas 22
'III. 'III. !R !RAS&O AS&OR*A#I R*A#I$ $ G%!I#A G%!I#A #O )RO!O)LAS!O )RO!O)LAS!OS..... S......... ........ ........ ........... ............... ...............2 .......2 1. !rans3orma+i rans3orma+i/n /n mediada mediada or A5roa+terium... A5roa+terium....... ........ ........ ........ ........ ........ ........ ........ ........ ........ ............ ..........2 ..2 2. !rans3orma+i rans3orma+i/n /n or medio :u;mi+o oli+ati/ni+ oli+ati/ni+o..... o......... ........ ........ ........ ........ ........ ........ ........ ........ ...........2 .......2 . !rans3orma+i rans3orma+i/n /n or medio-ele+tr medio-ele+troora+ oora+i/n.... i/n........ ........ ........ ........ ........ .......... ............. .............. ............... ............24 ....24 4. Soni<+a+i/n9 Soni<+a+i/n9 liosoma9 liosoma9 mi+ro-i mi+ro-inye++i nye++i/n9 /n9 otoo otoora+i/n ra+i/n99 y trans3 trans3orma+ orma+i/n i/n or or medio-laser........................ medio-laser................................... ..................... ..................... ..................... ..................... ..................... ..................... ........................ ............. 24 I=.
A)LI#A A)LI#A#I$ #I$ D% LA &"SI$ &"SI$ D% )RO!O)LAS )RO!O)LAS!OS. !OS.... ...... ...... ...... ..... ..... ...... ....... ....... ....... ........ ....... ....... .......27 ...27
=. A'A#%S % LA I'%S! I'%S!IGA IGA#I$ #I$ D% D% )RO!O)LAS! )RO!O)LAS!OS OS D% )LA! )LA!AS....................2 AS....................2 a+i/n de
/n....
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
•
#onstitu+i/n +romos/mi+a y estalidad.............................. estalidad............................................................. ............................... 2
•
#riado de 5enoma9 +itolasmoma9 trans+ritoma y roteoma.........................2
4. 'aloriza+i/n a5r;+ola...................... a5r;+ola................................. ...................... ..................... ..................... ..................... ..........................2 ................2 =I.
?I?LIOGR ?I?LIOGRA&(A. A&(A..... ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ ........ .......... ............. .............. .........
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
I.
2014
GLOSARIO •
•
•
•
•
•
•
•
•
•
Protoplasto: #omonentes ios de las +élulas e5etales :ue est,n rodeados s/lo or la memrana lasm,ti+a9 +onstituyen las +élulas desnudas de una lanta. Sspe!s"o!es #ellares: +élulas lires y a5re5ados +elulares distriuidos en un medio en moimiento. ei dermis del >az C$llas %el mes&'lo: son +élulas de arén:uima9 entre la eidermis y el enés9 :ue oseen una 5ran +antidad de +lorolastos9 +lorolastos9 relatiamente desor5anizados y +on mu+>os esa+ios de aire entre las +élulas9 lo l o +ual 3a+ilita el inter+amio de 5ases.
Nrse #ells@ +élulas :ue ayudan a estimular al +re+imiento a rotolastos aislados de interés. %stos ueden ser de la misma o de di3erente ese+ie Protoplast(to(pla!t Protoplast(to(pla!t s)stems: Sistema ara el estale+imiento del +ultio e indu++i/n de la re5enera+i/n de la lanta a artir de rotolastos aislados. Pol"*ala#tro!asa: %nzima >idrolasa9 endo o e6o5lu+anasa9 :ue ini+ia el +orte >idrol;ti+o or el e6tremo no redu+tor del ol;mero ol ;mero de e+tina. Pe#t"!a: #onunto +omleo de ol;meros rami<+ados del tio de los +aro>idratos. La e+tina no solo est, resente en la l a ared +elular rimaria9 sino tamién en la l,mina media entre las +élulas de la lanta9 donde ayuda a las +élulas a mantenerse untas. De+r"s: #élulas no di5eridas durante el ro+eso de aislamiento de rotolastos o rotolastos rotos. rotos. *aterial de dese+>o durante la uri<+a+i/n. me dio. Osmot"#m: %stailizador o re5ulador osm/ti+o del medio.
L)mp,oprep: *edio de 5radiente de densidad re+omendado ara el aislamiento de +élulas mononu+leares mediante la e6lota+i/n de las di3eren+ias en la densidad +elular. +elular.
II. INTROD-CCIN "na di3eren+ia 3undamental entre las +élulas de lantas y animales es la resen+ia de una ared +elular en lantas. %sta9 a esar de u5ar un ael +lae en los ro+esos rela+ionados +on la estru+tura y 3un+i/n de la lanta9 es un ost,+ulo imo imort rtan ante te en la e6l e6lot ota+ a+i/ i/n n de la tran trans3 s3er eren en+i +ia a dire dire+t +ta a de AD a las las +é +élu lulas las indiiduales y la rodu++i/n de >;ridos som,ti+os or 3usi/n +elular B#omton et al.9 al.9 2000 2000C C es de+i de+ir9 r9 la ar ared +e +elu lula larr +o +ons nsti titu tuye ye un se seri rio o imed imedime iment nto o en los los ro5r ro5ramas amas de meoram meoramient iento o de +ulti +ultios os a traés traés de la >irida >irida+i/ +i/n n B!oma B!omarr and Dantu9 2008C.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
al5o Eni+o en +ultio de +élulas e5etales ya :ue e6isten +omo +élulas searadas y sin +ontinuidad +itolasm,ti+a +on las +élulas e+inas B#omton et al.9 2000C. Los rotolastos de lanta rimero se aislaron or Fler+er en 182 or +orte de las es+alas es+alas de ulo de +eolla +on un +u+>illo oJani y Razdan9 18C. %l rendimiento de rotolastos 3ue ao ao y el ro+ ro+ed edim imie ient nto o se limi limita taa a a +é +élu lula lass muy muy a+u a+uol olad adas as99 +é +élu lula lass nonomeristem,ti+as. %l aislamiento de rotolastos se >izo m,s amliamente ra+ti+ado en la dé+ada de 10 +on la e6tra++i/n y uri<+a+i/n de las enzimas :ue odr;an ser utilizadas ara de5radar la ared +elular de la lanta. #o+in5 B10C des+uri/ :ue :ue los los rot roto olas lasto toss inta inta+t +tos os 3,+i 3,+ilm lmen ente te odr odr;a ;an n se serr ote oteni nido doss median mediante te la in+u in+ua a+i +i/n /n de los los tei teido doss e5e e5eta tales les en una una solu solu+i +i/n /n +o +on+ n+ent entra rada da de +e +elu lula lasa sa rearada a artir del >on5o Myrothecium verruatria B#omton et al.9 2000C. %l aislamiento de rotolastos es a>ora de rutina en una amlia 5ama de ese+ies los rotolastos iales son oten+ialmente totiotentes. )or lo tanto9 +uando se les da los est;mulos :u;mi+os y 3;si+os +orre+tos9 +ada uno es +aaz9 te/ri+amente9 de la re5e re5ener nera+ a+i/ i/n n de una una nuea nuea are ared d y de somet someter erse se a la dii diisi si/n /n mit/ mit/ti ti+a +a ara ara rodu+ir +élulas >ias de la :ue las lantas 3értiles se ueden re5enerar mediante el ro+eso de +ultio de teidos BDaey et al.9 2004C. o s/lo los rotolastos aislados9 ero su rodu+to de 3usi/n9 un >;rido som,ti+o9 tamién se ueden re5enerar en lantas enteras. Asimismo ueden +atar AD e6tranero9 a traés de su memrana las lasm, m,ti ti+a +a desn desnud uda9 a9 ao ao trat tratam amien iento toss ese ese+; +;<+ <+os os.. )ro )roor or+i +ion onan an adem, adem,ss un sistema e6erimental ara una amlia 5ama de estudios io:u;mi+os y mole+ulares :ue an desde las inesti5a+iones en las roiedades de +re+imiento de las +élulas indiiduales >asta el transorte de memrana B!omar B!omar and Dantu9 2008C. Sistemas de roto rotolas lastos tos-a- -a-lan lanta ta B)ro B)roto tolas last-t t-to-l o-lant ant system systemsC sC est,n est,n dison disonil iles es ara ara mu+>as mu+>as ese+i ese+ies es +on una e6tens e6tensa a litera literatur tura a sin emar5o emar5o99 los ro+e ro+edimi dimient entos os ,si+os ara su aislamiento >an su3rido o+os +amios desde la rimera ez :ue 3ueron reortados BDaey et al.9 2004C. %s as; :ue la resente mono5ra3;a tiene +omo oetio 5eneral reisar los ase+tos +la +lae e a+ a+er er+a +a del del +ult +ulti io o de rot roto ola last stos os99 in+l in+luy uyen endo do desd desde e su deast >asta a sus sus ali+ ali+a+ a+io ione ness en el me meo ora ramie mient nto o 5enéti+o e5etal. Se mantendr, una isi/n 5loal del tema9 de manera :ue ueda se serr Etil Etil ara ara die diers rsos os tio tioss de lan lanta tas s aun: aun:ue ue tam tamién ién se e6o e6ond ndr, r,n n +a +aso soss ese+;<+os de interés :ue serir,n ara ilustrar el uso de rotolastos a+tualmente.
III. DE/INICIN Los rotolastos son +élulas de lantas9 >on5os o a+terias en las :ue la ared +elular se >a eliminado9 ero la memrana lasm,ti+a est, inta+ta. Sin emar5o9 esta reisi/n de literatura se +entrar, ese+;<+amente en los rotolastos de ori5en
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
esar de ser searado +omletamente de su ared +elular. Desde un unto de ista oras desués de la liera+i/n9 y9 . 1M8C. Deido a la ausen+ia de la ared +elular9 los rotolastos son ade+uados ara diersas y di3erentes maniula+iones 5enéti+as :ue no ser;an osiles +on lantas o +élulas inta+tas. Los rotolastos siren adem,s +omo >erramientas e6erimentales Eni+as ara mu+>as inesti5a+iones
I.. CA I CARA RACT CTER ERS STI TICA CAS S DE LOS PR PROT OTOP OPLAS LASTO TOS S Las +a Las +ara+ ra+te ter;s r;sti ti+a +ass ese ese+; +;<+ <+as as de los los rot roto ola last stos os dee deend nden en de su ori5e ori5en n de e6tra++i/n9 ya sea la ese+ie o +élula ese+ializada y el resultado otenido desués de ser +ultiado9 del método usado. usado. %n mu+>os mu+>os +asos B%e+>aut B%e+>aut et al9 201C9 de a+uerdo a la ese+ie9 se +ontrola el tio de resultado ara un
PROTOPLASTO
CARACTERSTICA
es&'lo
#ontienen +lorolastos erdes y en luz muestran a+tiidad 3otosintéti+a
Sspe!s"o!es Sspe!s"o!es #ellares
#ontienen 5ranos de almid/n Se ued ueden en usar usar ara ara aisl aislam amie ient nto o de nE+leos +elulares9 as; +omo +romosomas mit/ti+os y mei/ti+os.
Alero!a %e sem"llas %e a3e!a
Se+retan amilasa y RAsa en resen+ia de ,+ido 5ieréli+o
N&%los %e ra#es %e le*m"!osas
)ueden redu+ir el a+etileno
L"+res
%l +>o:ue osm/ti+o o+asiona +amios en el atr/n de RA y s;ntesis de rote;nas
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
&uente@ )rotolastos. Aislamiento9 +ultio y re5enera+i/n de lantas BSzaados9 L. 11C
. AI AISL SLA AIE IENT NTO O DE PR PROT OTOP OPLA LAST STOS OS Antes de entrar en los detalles sore el +ultio de rotolastos9 es imortante aislar lo m, m,ss sua suaeme ement nte e y m, m,ss r,id r,idam amen ente te osi osil le e los los rot roto ola last stos os ia iales les y no lesion les ionado ados. s. Lo Loss dos métodos métodos ara el aislamien aislamiento to de rotol rotolast astos os son@ son@ método método me+,ni+o y método enzim,ti+o.
a& '(todo '(todo 'ec)nico 'ec)nico %l método me+,ni+o tiene9 ante todo9 un interés >ist/ri+o9 ya :ue su rendimiento es ao9 ao9 su ro+e ro+edimi dimient ento o tedios tedioso o y su ali+a ali+ail ilida idad d limita limitada da BSzaa BSzaados dos99 L. 11C. 11C. Fler+er B182C 3ue el rimero en aislar rotolastos a artir de teidos de ulos de +eolla. Las +aas 3ueron sumer5idas sumer5idas en sa+arosa al 1.0 * >asta :ue el rotolasto rotolasto se +ontrai5a de su ared +elular9 y lue5o el teido lasmolisado 3ue +ortado +on un +u+>illo aio >iot/ t/ni ni+a +a Ba Baa a +on+entra+i/n de sa+arosaC B!omar B!omar and Dantu9 2008C. Figura 1 Tejido Tejido cortado en la línea punteada (A), algunas células liberan protoplastos completos y el resto de células producen producen protoplastos rotos muertos como se muestra en (B) marcado con asteriscos.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
enzimas uri<+adas >a sido utilizado amliamente or otro 5ruo de traaadores B!omar and Dantu9 2008C. Los rin+iales 3a+tores :ue se deen +onsiderar ara el e l é6ito de este método son@
%l tio de enzimas :ue se usar,n La +omosi+i/n del medio en :ue ellas a+tuar,n *aterial de ori5en de los rotolastos.
a) %nzimas ara el aislamiento de rotolastos
ay dos en3o:ues ,si+os ara el aislamiento enzim,ti+o de rotolastos@ BaC el tratamiento de un teido e5etal +on una mez+la de e+tinasa y +elulasa as; al mismo tiemo se searan las +élulas y de5radan sus aredes B)oten+ia y #o+in59 1M0C y BC el método se+uen+ial Ben dos asosC :ue imli+a la rodu++i/n de +élulas aisladas9 a artir del +allo o teido mediante el uso de e+tinasa B!omar and Dantu9 2008C9 las +uales a su ez se +onierten en rotolastos or un tratamiento +on +elulasa :ue di5iere la ared +elular Ba5ata y !aee9 1M0 +itado or Smit>9 1M8C. Las enzimas utilizadas ara el aislamiento de rotolastos son e6tra+tos +rudos de ori5en 3En5i+o. Las e+tinasas son ri+as en a+tiidad oli5ala+turonidasa mientras :ue las +elula +elulasas sas +ontie +ontienen nen >emi+e >emi+elul lulosa osa99 -194-194- 5lu+an 5lu+anasa asa99 :uitin :uitinasa asa99 liasa liasa99 nu+leasas y e+tinasa BSmit>9 1M8C. )ara utilizar las reara+iones enzim,ti+as +ome +o mer+ r+ial iales es B!a B!ala la 1C en al5un al5unos os tei teido dos9 s9 se a+ a+on onse sea a elim elimin inar ar las las sales sales :ue :ue +ont +o ntien ienen en esto esto se ued uede e >a+e >a+err ade6 G27 o G70 BSzaados9 L. 11C.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Tabla Tabla 1 Enimas !ue se utilian para el aislamiento de protoplastos (Fuente" #abados, $. 1%%1)
b) %l medio de in+ua+i/n
Dado :ue la elimina+i/n de la ared +elular resulta en la érdida de resi/n de la +élu +é lula la99 los los rot roto ola last stos os son son aisl aislad ados os y ma mant nten enid idos os en un me medi dio o >iert >iert/n /ni+ i+o o lasmol;ti+o roor+ionado or una sal inor5,ni+a e:uilirada o una solu+i/n de azE+ar monosa+,rido BSmit>9 1M8C. As;9 el material de in+ua+i/n :ue se utiliza ara el aislamiento de rotolastos +ons +o nsta ta de los los si5u si5uien iente tess eleme element ntos os@@ enzim enzimas as disu disuelt eltas as en una una solu solu+i +i/n /n :ue :ue +onten +onten5a 5a al5una al5unass sal sales es B+omo B+omo #a#l #a#l 9 F2)O49 *5#l2C9 un medio de +ultio9 un amorti5uado amorti5uadorr Bsolu+i/n Bsolu+i/n tam/nC9 y un estailizad estailizador or o re5ulador re5ulador osm/ti+o. osm/ti+o. %l +al+io B1 a 10m*C es un in5rediente esen+ial y el ion 3os3ato B0.7 a 2.0 m*C es ené<+o
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
%n teor;a9 es osile aislar rotolastos de +ual:uier tio de lanta9 /r5ano o teido9 ero las 3uentes :ue se usan +orrientemente son mes/a+iénd >a+iéndolo olo menos menos sus+e sus+eti tile le a la de5rad de5rada+i a+i/n /n +omlet +omleta a de +elula +elulasa sa BSmit>9 1M8C.
El mes&'lo #omo fe!te %e protoplastos. %l estado umedad. "sualmente "sualmente se utiliza utiliza una luz de 109000 a 09000 lu6 y un 3otoeriodo de 18 / 1 >. %s esen+ial +ontar +on un uen suministro de a5ua y +on 3ertiliza+i/n 3ertili za+i/n re5ular BSzaados9 L. 11C. Al5unas e+es los rotolastos aislados resultan m,s estales si las lantas se +onseran reiamente en la os+uridad durante al5unos d;as. %n +iertos +asos9 ara meorar la iailidad de los rotolastos aislados9 es a+onseale el retratamiento del del tei teido do 3oli 3oliar ar en un me medi dio o +o +on n >or >ormona monass de +re+ +re+im imie ient nto9 o9 en di3e di3errente entess +ondi+iones de luz y temeratura BSzaados9 L. 11C.
Las sspe!s"o!es #ellares #omo fe!te %e protoplastos . Deido a :ue las susensiones son >etero5éneas y a menudo est,n +omuestas de a5re5ados de di3erentes tamaNos y de +élulas lires9 no todas las +élulas est,n en +ondi+iones /tima /timass ara ara la liera liera+i/ +i/n n de roto rotola lasto stos s la liera+ liera+i/n i/n en este este +as +aso o deende deende tamién de las +ondi+iones de +re+imiento de las +élulas +ultiadas. Se >a en+ontrado en+ontrado :ue +uando +uando se +ultian +ultian rotolastos rotolastos deriados de susension susensiones9 es9 el é6ito deende +laramente de la +alidad de la susensi/n +elular :ue se >a utiliza utilizado. do. Lo Loss roto rotola lasto stoss de susen susensio siones nes +elula +elulare ress :ue est,n est,n en +re+im +re+imient iento o lo5ar;tmi+o +omienzan a diidirse muy r,idamente9 mientras :ue los deriados de
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
totiotentes. De manera similar9 en otras mono+otiled/neas tales +omo l,tano9 susensiones de +élulas 3ueron el material 3uente re3erida deido a su totioten+ia9 ya :ue los de la >oa de mes/a en+ontrado :ue los rotolastos de las 5ram;neas son e6tremadamente re+al+itrantes al +ultio +on la Eni+a e6+e+i/n de los rotolastos roenientes de tallos de ma;z B)otryus et al.9 1MMC9 no se >an odido +ultiar e6itosamente rotolastos de +ereales aislados de lantas enteras. Los resultados ne5ati ne5atios os de numer numeroso ososs e6erim e6eriment entos os >an lleado lleado a la +on+lu +on+lusi/ si/n n de :ue las +élulas di3eren+iadas de los +ereales deen >aer erdido el oten+ial de diisi/n y la totio totioten ten+ia +ialida lidad d B)otr B)otry yus9 us9 180C 180C en +amio9 +amio9 los roto rotola lasto stoss ais aislado ladoss de susen susensio siones nes +elular +elulares es emrio5 emrio5éni éni+as +as se >an +ulti +ultiado ado +on é6ito é6ito BSzaa BSzaados dos99 L. 11C ese+ialmente en el +aso del arroz B!an5 B!an5 et al.9 2001 +itado or Daey et al.9 2004C. !ras !ras el in3orme de :ue las +élulas +él ulas 5uardianas son una 3uente Eni+a de rotolastos totiotentes en la remola+>a azu+arera Ball et al.9 1C9 otros inesti5adores >an real realiz izad ado o e6e e6erim rimen ento tos. s. )or eem eemlo lo99 )ande andey y et al. B200 B2002C 2C in3o in3orm rmar aron on de los los ro+edimientos ara aislar rotolastos de +élulas 5uardianas de A. thaliana ara su uso uso en estu estudi dios os an >an sido sido rearados rotolastos in+luyen las del teido +entral de n/dulos de la ra;z de !icia in3e+tadas +on a+teroide a+teroidess de #hi$obium se rodu+en unto "aba9 donde las +élulas in3e+tadas +on +o n las +é +élu lula lass no in3e+ in3e+ta tada dass B)eit B)eiter er et al.9 al.9 200 200C. C. %l ais aisla lamie mient nto o ino inolu lu+r +ra a la dise++i/n de n/dulos lue5o la di5esti/n de la ared en solu+i/n >iert/ni+a9 la liera+i/n de rotolastos en solu+i/n li5eramente >iot/ni+a9 y la seara+i/n de las 3ra++iones 3ra++iones de rotolas rotolastos tos or +entri3u5a+ +entri3u5a+i/n i/n iso;+ni+a iso;+ni+a en 5radiente 5radiente de densidad. densidad. !ales !ales rotolastos son Etiles en las inesti5a+iones
d) %s:uema de ali+a+i/n ara aislamiento de rotolastos
"n es:uema de ali+a+i/n 5eneral ara la rodu++i/n de rotolastos9 utilizando el ro+edimiento de mi6ed-enzyme9 se muestra en la <5ura 2. )ara :ue las enzimas
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Desués Desués de la in+ua+i/n de la enzima9 los rotolast rotolastos os es3éri+os B&i5ura B&i5ura C deen seararse de los dese+>os +elulares9 surotolastos surotolastos y a+uolas. Surotolastos se 3orman durante lasm/lisis temrana +uando la rotolastos se diide en dos o m,s suunidades9 al5unos de los +uales ser,n anu+leados y or lo tanto no iales. La seara+i/n se uede +onse5uir 3,+ilmente en una ariedad de maneras@ reetida resusensi/n y +entri3u5a+i/n de rotolastos en un medio de laado de Kota+i/n de rotolastos en una solu+i/n de sa+arosa >iert/ni+a B&i5ura C aso de la mez+la de in+ua+i/n a traés de un tamiz de nylon de tamaNo de oro ade+uado :ue ermite s/lo rotolastos ara asar a traés o or el uso de un sistema de dos 3ases9 tal +omo de6trano de6trano - )%G Bolietilen5li+ Bolietilen5li+olC9 olC9 :ue se asa en una di3eren+ia di3eren+ia de densidad entre los rotolastos y +élulas BFanai y %dJards 9 1M re3erido en Smit>9 1M8C. #on5elar los rotolastos9 re+o5erlos desués del laado y e6aminarlos en el mi+ros+oio ele+tr/ni+o o tratarlos +on arillantadores Kuores+entes :ue se unen ese+;<+amente a la +elulosa9 reela la ausen+ia total de <ras de +elulosa en la suer<+ie de la lasmalema. Figura & Es!uema de aplicaci'n general para el aislamiento, el laado y el recuento de protoplastos de mes'ilo de la *oja. $os protoplastos aislados a partir de callos o suspensiones de células se manejan de la misma manera. Fuente #mit*, 1%+
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
( Petunia hybrida). hybrida). (#) $os Figura - (a) - (a) Protoplastos de mesófilo recién aislados (40 micras diam.) en medio l!"ido ( Petunia protoplastos (60 micras diam.) li#eradas por las cél"las c"lti%adas. &l n'cleo en posición central central est rodeada por cordones citoplasmticos ( Parthenocissus tricuspidata). tricuspidata). (c) "sión ind"cida por nitrato nitrato de sodio entre "n protoplasto del mesófilo (m)* !"e contiene cloroplastos* + "n protoplasto incoloro (c) aislado a partir de "na "na s"spensión de cél"las. $os plastidios son %istos entrando en el citoplasma del protoplasto incoloro. (d) ,icrofoto-rafa electrónica de "n protoplasto mesófilo recién aislada (n n'cleo/ c cloroplasto/ c% %ac"ola central* p plasmalema). (e) $a captación de Rhizobium () Rhizobium () en %esc"las en el citoplasma de "n protoplasto (p plasmalema). (,icroscopio electrónico proporcionadas por el r. r. , a%e+.) a%e+.)
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
I.. PI P-RI RI/I /ICA CACI CIN N DE LOS PR PROT OTOP OPLA LAST STOS OS %l é6ito del +ultio de rotolastos re:uiere una ola+i/n ura de rotolastos inta+tos y iales +on un alto rendimiento. As; los rotolastos deen ser uri<+ados retirando el material no di5erido BderisC9 rotolastos reentados y enzimas.
,& -emoci.n de de*ris / en+imas Los deris Los deris ueden ueden ser retir retirado adoss de la susens susensi/n i/n de roto rotolas lastos tos median mediante te el idratado. La susensi/n se +entri3u5a de nueo ara resoler los rotolastos y el medio de laado se de+anta. Las >uellas de la enzima se eliminan or laado de los rotolastos dos o tres e+es +on el medio.
*& La eliminaci.n de protoplastos rotos Los rot roto ola last stos os inta inta+t +tos os se se sea ara ran n de los los rot roto ola last stos os rotos otos me media diant nte e la susensi/n de la reara+i/n de rotolastos en solu+i/n de sa+arosa al 20-40 U y la +entri3u5a+i/n a 70 rm durante tres minutos. )rotolastos inta+tos se a+umulan en la arte suerior de la solu+i/n de sa+arosa y se retiran +uidadosamente +on una ieta BGre5ory y #o+in59 17 ener5;a y #o+in59 1M0 %ans et al9 1M2C.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
mi+ro mi+ros+o s+oio io Kuores Kuores+en +ente. te. Solamen Solamente te los roto rotolas lastos tos iales iales ueden ueden ser istos istos B!omar and Dantu9 2008C. Figura Formaci'n de anillo de puriicaci'n de protoplastos y el material de desec*o (roto o células intactas) se asientan en el ondo. Fuente Tomar and /antu, &00
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
II. I I. T5CN T5CNIC ICAS AS DE DE C-L C-LTI TIO O PAR PARA A PROT PROTOP OPLA LAST STOS OS AISLADOS 1. *edi *edio o de de +ul +ulti tio o )ara oder lantear medios de +ultio e<+ientes ara rotolastos9 lo rimero :ue se dee tener en +uenta es la resion osmoti+a. )or lo 5eneral9 los rotolastos +omienzan a re5enerar la ared +elular en minutos9 ero este ro+eso re:uiere +uidados en la resi/n osm/ti+a >asta :ue la ared +elular rimaria ueda +ontrarrestar la resi/n de tur5en+ia eer+ida sore el +itolasma. )or otro lado9 se >a +omroado :ue la resi/n osm/ti+a tamién inKuye en la mitosis. %sta Eltima se mantiene +uando se diluye el medio de +ultio +on una solu+i/n de +omosi+i/n + omosi+i/n similar ero +on menos resi/n osm/ti+a. )rotolastos de una misma ese+ie ero de di3erentes teidos y de di3erentes ese+ies tendr,n re:uerimientos nutri+ionales di3erentes. %n +onse+uen+ia9 los medios de +ultio a lar5o lazo se determinaran em;ri+amente. "niersalmente9 y +omo >emos desarrollado en las ra+ti+as9 el medio de +ultio ase es del *S B*uras>i5e y So559 12C y ?7 BGamor5 et al9 al 9 18C9 +on la adi+ion de
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
La mayor;a de los l os sistemas e6erimentales est,n asados en medios li:uidos9 semi-solidos o la +omina+i/n de amos. Disensar susensiones de rotolasto en la+as )etri es la o+i/n mas simle9 ya :ue el medio uede ser 3,+ilmente reemlazado ara 5raduar la osmolaridad y or tanto9 mantener el +re+imiento del rotolasto. Susensiones de 5otas de rotolastos son Etiles +uando >ay un numero redu+ido de rotolastos disoniles9 mientras :ue las té+ni+as de drolet-array >an 3a+ilitado las +omina+iones de +on+entra+iones de los re5uladores de +re+imiento. Los rotolastos aislados ueden resistir estar emeidos en medios semis/lidos. %stos emlean9 or lo 5eneral9 a5ar +omo a5ente de soorte9 sin emar5o9 se re+ono+e :ue la a5arosa tiene m,s e<+ien+ia +omo a5ente de soorte or su neutralidad. %stos medios9 +onteniendo a los rotolastos9 ueden ser disensados +omo +aas o 5otas9 +on el osterior in+remento de 270 ul en olumen en la+as etri. Se++ionar las +aas en se+tores9 se5uidas de an>os de estos o de 5otas en medio li:uido de la misma +omosi+i/n9 romuee el +re+imiento rotolasm,ti+o. %se método tamién se emlea ara +amiar la osmolaridad. %l al5inato tamién se uede usar ara solidi<+ar el medio9 e6oniéndolo a iones
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
%n el +aso de :ue >uiera una densidad e6+esia los nutrientes se a+aar;an r,idamente y las +élulas no odr;an sostener una diisi/n +elular +ontinua. %n la +ontra arte9 si >ay un una +antidad de ino+ulo insu<+iente9 las +élulas no odr;an sostener la diisi/n +elular9 deido a :ue >ar;a una 3alta de 3a+tores estimuladores del +re+imiento Be6+retadas de +elula a +elulaC in+luyendo amino,+idos9 este ro+eso llea or nomre medio +ondi+ionante o nurse +ulture. )reiamente se uede tratar el medio de +ultio ara :ue ueda 3omentar el +re+imiento a+tio de los nueos rotolastos aislados mediante el +ultio de +élulas en3ermeras :ue ueden ser de di3erente o de la misma ese+ie de los rotolastos aislados9 +omo or eemlo9 Lolium multiKorum uede estimular a las +élulas emrionarias e mrionarias del arroz. A:uellas +élulas :ue >an sido tratadas +on rayos = ara in>iir la diisi/n +elular ueden rodu+ir ese e3e+to en las otros rotolastos9 a+tuando tamién +omo nurse +ells. %stas +élulas rearadoras o indu+toras ueden ser usadas en el mismo medio o en memranas. . Aan+es Aan+es innoa innoatios tios ara ara los +ultios +ultios de rotola rotolastos. stos. !odos !odos estos aan+es tienen +omo oetio ma6imizar el +re+imiento +elular y la di3eren+ia+i/n9 arti+ularmente en +élulas en las :ue >a >aido maniula+i/n 5enéti+a. Sin emar5o9 +ae resaltar :ue no e6isten arametros 3;si+os ni en
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
"n nueo método :ue romete in+rementar la diisi/n mit/ti+a es el uso de sur3a+tantes no i/ni+os9 ese+ialmente de luroni+os. Se su5iere9 en el +aso del )luroni+o R &-89 adem,s de romoer la mitosis9 eer+e la di3eren+ia+i/n9 romoiendo la toma de nutrientes9 re5uladores del +re+imiento y o6;5eno. Al5unos antii/ti+os estimulan la diisi/n de +élulas deriadas de rotolastos. )or eemlo9 la +e3alosorina9 +e3ota6ima romueen en Passi%ora edulis9 +uando se le a5re5a 270 ulWmL. %n amos +asos el modo de a++i/n es des+ono+ido9 ero se sose+>a :ue la +e3ota6ima odr;a ser metaolizada ara 3ormar arte de +omonentes re5uladores del +re+imiento. Las oliaminas re5ulan la reli+a+i/n9 trans+ri+i/n y tradu++i/n del AD son tratados +omo una nuea +lase de re5uladores en lantas y son una resera de +arono y nitr/5eno en los +ultios de teidos. %n el +aso de la aena9 a+tua +omo un estimulante de s;ntesis de AD y en almendra9 la ar5inin5a estimula la diisi/n de rotolastos. !odos !odos estos aan+es tienen +omo oetio ma6imizar el +re+imiento +elular y la di3eren+ia+i/n9 arti+ularmente en +élulas en las :ue >a >aido maniula+i/n 5enéti+a. Sin emar5o9 +ae resaltar :ue no e6isten ar,metros 3;si+os ni en +uanto a la +omosi+i/n del +ultio9 or lo :ue los roto+olos deen ser
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
)ara la trans3eren+ia del 5en A5roa+terium-mediado9 los rotolastos son aislados y +ultiados or 2 a d;as reio al +o-+ultio +on +élulas A5roa+terium. Los rotolastos son mez+lados +on A5roa+terium a una tasa de 10X4 a 20X +élulas de lantas +ontra 10X a 10X8 +élulas a+terianas or mililitro e in+uadas untas or 24 a 0 >oras. Desués de la +o-+ultia+i/n9 los antii/ti+os :ue in>ien el +re+imiento de las a+terias son aNadidos al medio +ultio. A Las lantas tratadas se les ermite +re+er +re+er a un estado multi+elular antes de la adi+i/n sele+tia de antii/ti+os Banami+ina9 >i5romi+ina9 et+.C :ue in>ien el +re+imiento de +élulas no trans3ormadas. Las +élulas trans3ormadas :ue +re+en en un medio sele+tio son +aa+es de re5enerar lantas +on el 5en introdu+ido. La 3re+uen+ia de trans3orma+i/n usualmente Ku+tEa entre 0.2 a 17.U. el uso de A5roa+terium ara trans3ormar los rotolastos de lanta >a erdido el 3aor en aNos re+ientes or:ue no todas las ese+ies9 en ese+ial +ereales y astos9 son sensiles a la trans3orma+i/n or medio-A5roa+terium y a e+es es di3;+il in>iir el +re+imiento de A5roa+terium :ue si5ue en el +o+ultio. Adi+ionalmente9 el uso de sistemas de +ultios re5eneratios m,s simles >a eliminado la ne+esidad de usar rotolastos ara trans3orma+i/n .
2. Tra!sforma#"&! por me%"o me%"o 6m"#o pol"#at"&!"#o pol"#at"&!"#o %l :u;mi+o oli+ati/ni+o usado m,s +omEnmente ara trans3orma+i/n de AD es
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
%n la ele+troora+i/n9 un ulso elé+tri+o es e s utilizado ara indu+ir la asor+i/n de AD or los rotolastos de lantas9 es a traés de esta asor+i/n de AD :ue o+urre a traés de los oros en la memrana 5enerada en resuesta al ulso elé+tri+o. %n 5eneral9 el AD trans3ormale es mez+lado +on un rotolastos de una lanta antes de uno o m,s ulsos elé+tri+os de 200 a 1000 'W+m y 1 a 1000 V& de +aa+itan+ia. )ulsos +ortos Bt;i+amente mi+rose5undosC son t;i+amente menos letales a los rotolastos :ue los ulsos lar5os Bmilise5undosC. Los ar,metros /timos de ele+troora+i/n ar;an y deen ser roados ara nueas ese+ies. Las 3re+uen+ias de trans3orma+i/n y la iailidad +elular son 5eneralmente
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
La >irida+i/n som,ti+a >a ermitido rodu+ir >;ridos :ue no se >a;an odido otener otener or +ruzam +ruzamien ientos tos se6ua se6uales9 les9 in+remen in+rementad tado o as; el Kuo Kuo de 5enes 5enes en los +ultios. "n 5ran nEmero de traaos >an des+rito la 5enera+i/n de lantas Eni+as a traés del ro+eso de >irida+i/n som,ti+a or 3usi/n de rotolastos. La ola+i/n de +é +élu lula lass res esul ulta tant ntes es de es esto toss ro+ ro+es esos os de 3usi 3usi/n /n a a se serr una una me mez+ z+la la de rotolastos arentales9 >omo+ariontes y >etero+ariontes +on dos o m,s nE+leos. La 3usi/n de +élulas som,ti+as uede ali+arse ara@ 1C La suera+i/n de la in+omatiilidad en +ru+es interese+;<+os
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
estéril. %n determinados +asos es osile mediante su+esias retro3usiones eliminar +asi todo el 5enoma de uno de los arentales Be. el >;rido resistente a atrazina entre Solanum ni5rum y Ly+oersi+on es+ulentum [ain et al.9 188C. Otro +aso mode modelo lo de +ir +irid ida+ a+i/ i/n n es el ?ras ?rassi si+a +a-R -Ra a>a >anu nuss B)el B)elle leti tier er et al.9 al.9 18 18C@ C@ el >etero+ari >etero+arionte onte +ontiene el nE+leo de ?rassi+a ?rassi+a naus9 naus9 los +lorolastos +lorolastos resistentes resistentes a atra atrazi zina na de ?ras ?rassi si+a +a +a +am mes estr tris is y las las mito mito+o +ond ndri rias as de Ra>a a>anu nuss sati satia a :ue :ue +on
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Inertos in itro 3ueron 3ueron ne+esarios ne+esarios ara mantener mantener la re5enera+i/n re5enera+i/n sinensisC. Inertos de meristemos. BLiu and Den59 2000C •
)rotolastos ele+tro3usionados ele+tro3usionados de +élulas susendidas de ?onnaza nael oran5e BC. sinensisC +on rotolastos rotolastos del mes/ 5rae3ruit BC. paradisiC sin semillas ara 5enerar lantas tetraloides y diloides. BGuo et al.9 2000C Al5unos de los tetraloides mostraron m ostraron Kora+i/n re+oz. Guo et al. roone :ue los >;ridos som,ti+os ueden ser e6+elentes donadores de olen ara la meora de +ultiares +ultiares de omelo BtoronasC BtoronasC mediante la
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
7. SOLA SOLANA NACE CEAS AS %se %se+i +ies es de Sola Solanu num9 m9 en es ese e+i +ial al la aa aa B 'olanum 'olanum tuberosum tuberosumC9 >an sido +ara+terizadas e6tensamente en ro5ramas de >irida+i/n som,ti+a ara amliar el néti néti la di idad idad néti+ néti+ di i+i/ i+i/ de lo
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
9. AAN ANCE CES S EN EN LA IN INE EST STIG IGAC ACI IN N DE
2014
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Otro unto a tener en +uenta9 son los l os sulementos adi+ionados a los medios
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
2014
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Cultivo de Protoplastos
9I
CONCL-SIONES
2014
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
Trusted by over 1 million members
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.