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Використання прийомів генетичної інженерії і клітинної біології для отримання нових форм у видів з родини бобових і деяких інших рослин: Автореф. дис.

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Iaoe?iiaeueia Aeaaeai?y Iaoe Oe?a?ie

?inoeooo ee?oeiii? a?ieia?? oa aaiaoe/ii? ?iaeaia???

Ia i?aaao ?oeiieno

EO*OE IEEIEA A?EOI?IAE*

OAeE 575.22:577.21:581.143

AEEI?ENOAIIss I?EEII?A AAIAOE*II? ?IAEAIA??? ? EE?OEIII? A?IEIA?? AeEss
IO?EIAIIss IIAEO OI?I O AEAe?A C ?IAeEIE AIAIAEO ? AeAssEEO ?IOEO
?INEEI.

03.00.22 “Ee?oeiia a?ieia?y” 03.00.15 “Aaiaoeea”

AAOI?AOA?AO

aeena?oaoe?? ia caeiaoooy iaoeiaiai nooiaiy aeieoi?a a?ieia?/ieo iaoe

Ee?a 1998

Aeena?oaoe??th ? ?oeiien.

?iaioo aeeiiaii o a?aeae?e? ee?oeiii? ?iaeaia??? oa oeeoio?c?ieia??
?ineei ?inoeoooo ee?oeiii? a?ieia?? oa aaiaoe/ii? ?iaeaia??? IAIO oa a
Aioai?/iiio ?inoeooo? Oi?aa?neoaoo i. Ithioaia, O?I.

Io?oe?ei? iiiiaioe aeieoi? a?ieia?/ieo iaoe, i?ioani?, /eai-ei?. IAIO
A.A.Eoiao, ?inoeooo iieaeoey?ii? a?ieia?? ? aaiaoeee IAIO, caa.
a?aeae?eii

aeieoi? a?ieia?/ieo iaoe

A.I.Aaee?i, ?inoeooo a?ii?aai?/ii? o?i?? oa iaooio?i?? IAIO, caa.
eaai?aoi???th

aeieoi? a?ieia?/ieo iaoe

I.A. I?eia, ?inoeooo oeeoieia?? ? aaiaoeee IAI Aaea?on?, no. iaoe.
ni?a.

I?ia?aeia i?aai?caoe?y ?inoeooo
aa?iaeieia?? ?

a?ioaoiieia?? OAAI

Caoeno a?aeaoaeaoueny “ 10 ” a?oaeiy ” 1998 ?. i 14 aiae. ia can?aeaii?
Niaoe?ae?ciaaii? A/aii? ?aaee Ae.26.202.01 i?e ?inoeooo? ee?oeiii?
a?ieia?? oa aaiaoe/ii? ?iaeaia??? IAI Oe?a?ie ca aae?anith 252143, Ee?a,
aoe. Caaieioiiai 148.

C aeena?oaoe??th iiaeia iciaeiieoenue a a?ae?ioaoe? ?inoeoooo ee?oeiii?
a?ieia?? oa aaiaoe/ii? ?iaeaia??? IAI Oe?a?ie.

Aaoi?aoa?ao ?ic?neaiee “ 30 ” aeiaoiy 1998 ?.

A/aiee nae?aoa?

Niaoe?ae?ciaaii? ?aaee,

eaiaeeaeao a?ieia?/ieo iaoe E.A.Oa?anaiei

CAAAEUeIA OA?AEOA?ENOEEA ?IAIOE.

Aeooaeuei?noue oaie. ?icaeoie aaiaoe/ii? iaoee i?eca?a aei aeieeiaiiy
iiai? noa?e ae?yeueiino? ethaeeie, yea iia`ycaia c? ci?iith aaiaoe/ii?
eiinoeoooe?? aeeaeo ?noio ca aeiiiiiaith iiaeo i?aeoiae?a oa oaoiieia?e,
ui ia`?aeiai? i?ae caaaeueiith iacaith aaiaoe/ia aai aaiia ?iaeaia??y.
Oaea ca?ac o i?e?ia?ieia?/i?e oa oa?iaoeaaoe/i?e i?iieneiaino?, a
n?euenueeiio ainiiaea?noa? aeei?enoiaothoue ooaie aaeoa??e, a?ea?a,
ae??aeaeae?a, ee?oeii? e?i?? ?ineei oa oaa?ei, ni?oe ?ineei, ye? aoei
io?eiaii ca aeiiiiiaith aaiii-?iaeaia?ieo i?eeii?a. Aeey aaaaoiee?oeiieo
?noio, ye? aoei ci?iaii oaeei /eiii, iaa?oue aeieeea ie?aia iacaa –
o?ainaaii? i?aai?cie. O?ainaaii? i?aai?cie – oea ?ineeie, ?eae,
oaa?eie, oa ?io? ?noioe, ui ianooue a nai?io aaiii? aaiaoe/io
?ioi?iaoe?th, yeo aoei aaaaeaii ca aeiiiiiaith iaoiae?a aaiaoe/ii?
o?ainoi?iaoe??. Aeieeiaiiy o?ainaaieo i?aai?ci?a iia`ycaii ye c
?icaeoeii iaoiae?a aeania aaiii? ?iaeaia???, oiaoi oaoiieia??
?aeiiaeiaioieo AeIE, oae ? ?ic?iaeith iaoiae?a oaaaeaiiy
?aeiino?oeiaaii? AeIE o aeea? ee?oeie oa ?aaaia?aoe?? c ieo o?c?ieia?/ii
ii?iaeueieo aaaaoiee?oeiieo ?noio.

Oi/a c iiiaioo iioae?eoaaiiy ia?oeo iia?aeiieaiue i?i io?eiaiiy
aaiaoe/ii iiaeeo?eiaaieo aoea??ioe/ieo i?aai?ci?a (aiae. GMO) i?ieoei ia
a?eueo i?ae i`yoiaaeoeyoue ?ie?a, ca oeae /an aeaiee iaoeia?e iai?yiie
aeaa iniiao aeey noai?aiiy oe?eiai ?yaeo eiia?oe?eieo o??i, iniiaiith
iaoith yeeo ? noai?aiiy o?ainaaieo ?noio, ye? iathoue oe?eee ?yae iiaeo
ei?enieo iciae. Aaiaoe/ii iiaeeo?eiaai? ni?oe ?ineei aaea
aeine?aeaeothoueny ia aenia?eiaioaeueieo iieyo, a aeaye? aaea
neeaaeathoue cia/io /anoeio o aaeiaiio n?euenueeiainiiaea?nueeiio
i?iaeoeo? oaei? e?a?ie ye NOA. Oae, a 1997?. o?ainaaii? aaaiaia, niy ?
eoeo?oaeca caeiyee 18%, 13% oa 9% a?aeiia?aeii an?o iin?aieo ieiu, ia
yeeo ae?iuoaaeeny oe? eoeueoo?e a NOA. No/aniee ?icaeoie oaoiieia?e
aaiaoe/ii? ?iaeaia??? aea? iiaeeea?noue i?eionoeoe, ui aaea aei 2005
?ieo na?oiaee ?eiie i?iaeaaeo o?eueee ian?iiy o?ainaaieo ?ineei aoaea
neeaaeaoe aeecueei 6.6 i?euey?ae?a aeiea??a aeey iaei??? eiiiai??
“Monsanto”. Aeaye? ?io? ioe?iee ua a?eueo iioei?noe/i?.

Io?eiaiiy aaiaoe/ii iiaeeo?eiaaieo ?ineei aeey aeae?a c ?iaeeie aiaiaeo
caaaeaee aoei a oeaio?? oaaae aeine?aeiee?a, ye? i?aoeththoue a aaeoc?
iieaeoey?ii? oa ee?oeiii? a?ieia?? ?ineei. Oea aecia/aoueny ?yaeii
i?e/ei, na?aae yeeo ne?ae cacia/eoe aaaeeea?noue oeeo aeae?a aeey
ae?iaieoeoaa ii??aiyii iaaei?iaiai ?ineeiiiai a?eea. Cao?aeia ?a?iia ca
?aooiie aeanieo ?ano?n?a iie?eaa? nai? iio?aae a i?iaeoeoao ?ineeiiiai
iioiaeaeaiiy, ui i?noyoue a?eie, eeoa ia 29%. ?ioa /anoeia ?iii?oo?oueny
c NOA, A?acee??, A?aaioeie o aeaeyae? ni?aeo aia?a. I?e/iio niiaeeaaiiy
oaeeo i?iaeoeo?a ca?eueoo?oueny a na?aaeiueiio ia 3% a ??e. Iii?oia
i?noea a iinoa/aii? a e?a?ie AN iaoa??ae?a ,ui i?noyoue a?eie,
caeiathoue Oe?a?ia oa ?in?y, /ee noia?iee aenii?o ca ioe?ieith cao?aeieo
aenia?o?a aeioiaeea aei 1.14 iei. oiii ca?ia ai?ioo. Aea ca inoaii? ?iee
oey oeeo?a iino?eii cieaeo?oueny aei 321 oen. oiii a 1997?. Ca?eueoaiiy
ae?iaieoeoaa aiaiaeo eoeueoo? caeaaeeoue a?ae ?yaeo oiia, iaei??th c
yeeo ? oe?iea aeei?enoaiiy a?ioaoiieia?/ieo i?aeoiae?a, cie?aia
aaiaoe/ii? ?iaeaia??? ?ineei aeey naeaeoe?? iiaeo aenieii?iaeoeoeaieo ?
no?eeeo aei oai?ia, oe?aeiee?a oa iani?eyoeeaeo /eiiee?a iaaeieeoiueiai
na?aaeiaeua ni?o?a ?ineei.

Oaeei /eiii, aeine?aeaeaiiy ieoaiue, ye? iia’ycai? c aea/aiiyi i?iaeai
ia?aiino aaiaoe/ii? ?ioi?iaoe?? o aeu? ?ineeie; iioei?caoe?y i?eeii?a ?
iaoiae?a io?eiaiiy o?ainaaieo ?ineei o aeae?a c ?iaeeie aiaiaeo ? a
ca’yceo c oeei ?ica’ycaiiy aeayeeo i?iaeai eoeueoeaoaaiiy i?ioiieano?a
oa ?aaaia?aoe?? ?ineei c ee?oeiieo eoeueoo? ? iaaii aeooaeueieie ?
iathoue aaaeeeaa cia/aiiy ye aeey e?auiai ?icoi?iiy ooieoe?iioaaiiy
aeeaeo i?aai?ci?a, oae ? aeey i?aeoe/iiai canoinoaaiiy.

Ca’ycie ?iaioe c iaoeiaeie i?ia?aiaie, ieaiaie, oaiaie. Iaoeiaa ?iaioa
aeeiioaaeanue a ?aieao athaeaeaoieo oai a?aeae?eo oeeoio?c?ieia?? oa
ee?oeiii? ?iaeaia??? ?EAA? IAIO “Aea/aiiy iieaeoey?ii-aaiaoe/ieo
i?ioean?a o ee?oeiieo nenoaiao ,ui ?aeiino?othaaeeny, oa o?ainaaieo
?ineeiao” ? “Aea/aiiy iieaeoey?ii- a?ieia?/ieo i?ioean?a o o?ainaaieo oa
o?ainaaiiiieo ee?oeiieo e?i?yo oa ?ineeiao”. Aeine?aeaeaiiy oaeiae
i?iaiaeeeeny a iaaeao iaoeiaeo i?iaeo?a i??i?eoaoiiai iai?yieo 01.11. 00
A?ioaoiieia?y, i?iaeo?a Aea?aeaaiiai oiiaeo iaoeiaeo aeine?aeaeaiue,
i?iaeoo i?ia?aie i?aeo?eiee iaoeiaeo aeine?aeaeaiue o e?a?iao
eieeoiueiai N?N? I?aeia?iaeiiai iaoeiaiai oiiaeo (ISF), i?iaeoo
?a?iiaenueeiai ni?aoiaa?enoaa INTAS, i?iaeoo ?a?iiaenueeiai
Ni?aoiaa?enoaa “COPERNICUS”.

Iaoa oa caaaeaiiy aeine?aeaeaiiy. Iniiaiith iaoith ?iaioe aoei io?eiaiiy
oa aiae?c aaiaoe/ii iiaeeo?eiaaieo ?ineei oeyoii aeei?enoaiiy iaoiae?a
aaiaoe/ii? ? ee?oeiii? ?iaeaia???.

Aeey aeinyaiaiiy iinoaaeaii? iaoe iaiao?aeii aoei ae??oeoe ianooii?
caaaeaiiy:

· Iioei?coaaoe iaoiaee ?ciethaaiiy oa eoeueoeaoaaiiy i?ioiieano?a ye
iaeaaaeeea?oi? oiiae io?eiaiiy o?ainaaieo oa o?ainaaiiiieo oi?i ?ineei.

· Aeine?aeeoe iiaeeea?noue i?aeaeuaiiy ?aaaia?aoe?eii? caeaoiino? o
aeayeeo aeae?a c ?iaeeie aiaiaeo.

· Iioei?coaaoe oiiae “i?yiiai ia?aiino aai?a” ? aaiaoe/ii?
o?ainoi?iaoe?? ca aeiiiiiaith Agrobacterium tumefaciens aeey aeae?a c
?iaeeie aiaia?eo.

· Iioei?coaaoe iaoiaee niiaoe/ii? a?a?eaeecaoe?? aeey aeae?a c ?iaeeie
aiaiaeo.

· Ca aeiiiiiaith iieaeoey?ii-a?ieia?/ieo ? a?io?i?/ieo iaoiae?a aeiaanoe
i?enooi?noue /oaeeiiiai aaiaoe/iiai iaoa??aeo a aaiiiao io?eiaieo
?ineei.

Iaoeiaa iiaecia io?eiaieo ?acoeueoao?a. A ?acoeueoao? i?iaaaeaii? ?iaioe
aoei cai?iiiiiaaii oa ai?iaiaaii i?ea?iaeuei? niiniae eoeueoeaoaaiiy
i?ioiieano?a oa i?ioieiee ?aaaia?aoe?? ?ineei c i?ioieeii?a eiia?oe?eieo
ni?o?a ethoea?ie oa eiithoeie. ?aaaia?aoe?y ?ineei o eiithoeie iiaea
aooe io?eiaia ye oeyoii i?yiiai, oae ? iai?yiiai aia??iaaiaco, ui
i?aeoaa?aeaeaii ?acoeueoaoaie a?noieia?/iiai aiae?co. Oe? ?iaioe
a?aee?eee iiaeeea?noue io?eiaiiy iiaeo ni?o?a ?ineei ca aeiiiiiaith
iaoiae?a niiaoe/ii? a?a?eaeecaoe?? oa i?yiiai ia?aiino aai?a a
i?ioiieanoe ?ineei.

Iacaeaaeii a?ae ca?oa?aeieo aeine?aeiee?a aoee ?ic?iaeai? iaoiaee
aaiaoe/ii? o?ainoi?iaoe?? oeyoii aeaeo?iii?aoe?? i?ioiieano?a oa
io?eiai? o?ainaaii? ?ineeie ethoea?ie oa ee?oeii? e?i?? ni?.
Iieaeoey?ii-a?ieia?/iee aiae?c oa aiae?c aeoeaiino? oa?iaioo
iaii?oeeioinoio?ainoa?ace i?aeoaa?aeea o?ainaaiio i?e?iaeo io?eiaieo
e?i?e.

Aia?oa aoa cai?iiiiiaaiee oa ai?iaiaaiee iaoiae, ui i?aeaeuo?
?aaaia?aoe?eio caeaoi?noue o ethoea?ie oa ai?ioo oeyoii aaiaoe/ii?
o?ainoi?iaoe?? “shooty”-iooaioii Agrobacterium tumefaciens pGV 2206. A
?acoeueoao? i?iaaaeaieo aenia?eiaio?a aoee io?eiai? o?ainaaii?
?aaaia?aoe?ei? e?i?? ethoea?ie oa ai?ioo, ui aeicaieeei cai?iiiioaaoe
i?ioeaaeo?o “iiaea?eii? o?ainoi?iaoe??”, ui aea? iiaeeea?noue io?eioaaoe
o?ainaaii? e?i?? oeeo aiaiaeo eoeueoo? c aaiaie, ye? iathoue i?aeoe/iee
?ioa?an. Oeae i?aeo?ae neeaaea?oueny c aeae?eueeio aoai?a. sse ia?oee
e?ie i?iiiio?oueny io?eiaoe e?i?? ?ineei, caeaoi? aei ?aaaia?aoe?? a
eoeueoo?? in vitro, ca ?aooiie o?ainoi?iaoe?? aaiaie O-AeIE A.
tumefaciens. Ae?oaee aoai – oea o?ainoi?iaoe?y io?eiaieo e?i?e, caeaoieo
aei ?aaaia?aoe??, iaaiei aaiii, aeei?enoiaoth/e iaoiaee ye
aa?iaaeoa??aeueii? o?ainoi?iaoe??, oae ? “i?yiiai ia?aiino aai?a”.
O?ao?e aoai – a?aeiiaeaiiy aeo?aeiiai aaiioeio, ui iana o?eueee iaaiee
aai, ui oe?eaaeoue, ca ?aooiie o?iiiniiii? naa?aaaoe?? a ianooiieo
?aia?aoe?yo. O?ao?e aoai aecia/a?oueny /enoi noaoenoe/iith iiaeeea?noth
?ioaa?aoe?? aai?a O-AeIE a ??ci? o?iiiniie, ye? iaoiiaeththoue
caeaoi?noue aei ?aaaia?aoe??, ? iaaieo aai?a, ui oe?eaaeyoue, i?ney
iiaoi?ii? o?ainoi?iaoe??. C aeei?enoaiiyi nenoaie “iiaea?eii?
o?ainoi?iaoe??” aoee io?eiai? o?ainaaii? oi?ie ai?ioo, ye? i?noyoue
Ds-aeaiaio eoeo?oaece, ui o?ainiico?oueny. Oe? ?acoeueoaoe
a?aee?eaathoue oe?eeii iia? ia?niaeoeae a eeane/i?e aaiaoeoe? oaeiai
ia`?eoo ye ai?io oa iaaeathoue iiaeeea?noue aeey eeiioaaiiy oi?eaeueieo
aai?a oe??? ?ineeie ca aeiiiiiaith ?ina?oe?eiiai iooaaaiaco.

Cai?iiiiiaaii oa ai?iaiaaii iaoiae aaiaoe/ii? o?ainoi?iaoe?? ai?ioo ca
aeiiiiiaith iaiieiaaiieo ooai?a Agrobacterium tumefaciens. Io?eiai?
o?ainaaii? e?i??, ye? ianooue aai no?eeino? aei aa?a?oeeaeo –
oino?iio??oeeio. Aeey noaa?eueiiai io?eiaiiy iauaaee?a o ?aaaia?aio?a
ai?ioo aoei ?ic?iaeaii iaoiae ?iaeoeoe?? oea?o?iiy oa ooai?aiiy ian?iiy
o iaaii?a ai?ioo a eoeueoo?? in vitro. O o?ainaaieo e?i?e ai?ioo oeei
oeyoii aoei io?eiaii iauaaee?a. Iieaeoey?ii-a?ieia?/iee aiae?c
i?aeoaa?aeea i?enooi?noue iine?aeiaiinoae AeIE ia?aianaiiai aaio a
?aia?aoe?yo aaiaoe/ii iiaeeo?eiaaiiai ai?ioo. Oaeei /eiii io?eiai? e?i??
ai?ioo, ye? iathoue no?ee?noue aei aa?a?oeeaeo, ui a?aee?eaa? iia?
iiaeeeaino? a naeaeoe?? oaei? eoeueoo?ii? ?ineeie ye ai?io. E??i oiai,
aeania iaoiae ?iciiiaeaiiy ai?ioo a eoeueoo?? in vitro iaaei?
iiaeeea?noue aeei?enoaiiy iaoiaeo i?e?ieeiiaeueiiai ?iciiiaeaiiy aeey
oe??? eoeueoo?e. Oaeee iaoiae oe?iei aeei?enoiao?oueny aeey ?iciiiaeaiiy
aeaei?aoeaieo oa aeayeeo n?euenueeiainiiaea?nueeeo ?ineei. Aeei?enoaiiy
oeueiai iaoiaeo aeey ai?ioo aeicaiey? iacaeaaeii a?ae ee?iaoe/ieo oiia
?iciiiaeeoe oi?eaeueiee naeaeoe?eiee iaoa??ae aac ci?ie eiai aaiioeio c?
cia/ii a?eueo aenieei eiao?oe??ioii ?iciiiaeaiiy. Oae, ?ic?aooiee
na?ae/oue, ui c iaeiiai iaaiio aei e?ioey ?ieo iiaeia io?eiaoe aei 500
000 aaiaoe/ii iaeii??aeieo ?ineei i?e uii?ny/iiio eiao?oe??io?
?iciiiaeaiiy 3.

Io?eiaii aneiao?e/i? niiaoe/i? a?a?eaee i?ae aeaeaie c ?iaeeie aiaiaeo.
Iieacaii iiaeeea?noue aeei?enoaiiy ia?aianaiiai aaio
iaii?oeeioinoi-o?ainoa?ace ye aaiaoe/iiai ia?ea?a aeey a?aeai?o
niiaoe/ieo a?a?eae?a o aeae?a c ?iaeeie aiaiaeo.

O noeoiiino? oe? ?iaioe, ye? i?iaaaeaii ia ??ai? na?oiai? iiaecie,
aia?oa aeicaieeee aeei?enoaoe a?ioaoiieia?/i? iaoiaee aeey iie?ioaiiy
iaeaaaeeea?oeo n?euenueeiainiiaea?nueeeo aeae?a c ?iaeeie aiaiaeo. E??i
oiai, oe? aeine?aeaeaiiy ? iaeieie c ia?oeo ii aaiaoe/ii? ?iaeaia???
?ineei, i?iaaaeaieo ye a Oe?a?i?, oae ? ia oa?eoi??? eieeoiueiai N?N?.

I?aeoe/ia cia/aiiy io?eiaieo ?acoeueoao?a. Caeaoia aei ?aaaia?aoe??
e?i?y ethoea?ie ia iniia? ni?oo “Aanaei-Iiaeieyinueea” aeei?enoiao?oueny
a iaoeiaeo aeine?aeaeaiiyo ? naeaeoe?ei?e i?aeoeoe?:

· ?inoeoooo oeeoieia?? ? aaiaoeee NI ?AI (i.Iiainea??nuee, ?in?y) –
aeia?aeea, i?aeienaia aee?aeoi?ii ?inoeoooo aeaae. Ooiiei A. E.

· Iieoaanueeei III “Ae?oa” (i.Iieoaaa) – aeia?aeea, i?aeienaia cai.
aee?aeoi?a A?aaeaiei ?. I. ? iaoeiaei ni?a?ia?oieeii Eiia A. A.

· AIAe? ei?i?a ?i. A. ?. A?eueyina (Iineianueea iae., ?in?y) – aeo
ia?aaea/? ca i?aeienaie Iac?ia A. A. – caa a?aeae?eii a?ioaoiieia?? ?
ee?oeiii? naeaeoe??, Aaaoiaei?iai? I. I., n.i.n. ? Nieiaeei? E. A.,
n.i.n.

A?a?eaei? ee?oeii? e?i?? aeei?enoaii a iaoeiaeo aeine?aeaeaiiyo:

· Naeaeoe?eii-aaiaoe/iiai ?inoeoooo (i.Iaeana) – aeia?aeea, i?aeienaia
cai. aee?aeoi?a Na?a??aei A. A., caa. a?aeae?eii a?ioaoiieia?? Eoe`yithe
N. O.

Iiaeeo?eiaai? e?i?? ai?ioo ia?aaeai? aeey aei?iaoaaiue a

· INEI “Ieaa” (i.Iieoaaa) – eeno, i?aeienaiee aee?aeoi?ii ae.a.i.
i?ioani?ii I. I. *aeae?iei.

· ?inoeooo ?ineeiieoeoaa ?i. TH?ueaaa (i.Oa?e?a) – eeno, i?aeienaiee
aee?aeoi?ii ?inoeoooo E. A. Aiiaea?aiei oa caa. a?aeae?eii naeaeoe??
ai?ioo I. I. *ae?ea?iei.

Iniaenoee aianie iiooea/a. Aaoi?ii aeeiiaii aenia?eiaioe ii iioei?caoe??
oiia eoeueoeaoaaiiy i?ioiieano?a, ?aaaia?aoe?? ?ineei, io?eiaiith
o?anaaiieo oa o?ainaaiiiieo e?i?e. Cai?iiiiiaaia ?aeay oa cae?eniaia
noaia “iiaea?eii? o?ainoi?iaoe??”. Iieaeoey?ii-a?ieia?/iee oa
oeeoieia?/iee aiae?ce i?iaaaeai? ?acii c ni?aaaoi?aie noaoae,
iaaaaeaieie a aaoi?aoa?ao?.

Ai?iaaoe?y ?acoeueoao?a ?iaioe. ?acoeueoaoe ?iaioe aeiiia?aeaeeny ia
ianooiieo i?aeia?iaeieo eiia?anao, neiiic?oiao ? eiioa?aioe?yo:

· YIIth International Congress on Plant Tissue and Cell Culture;
Amsterdam, The Netherlands, June 24-29, 1990 – noaiaeiaa iia?aeiieaiiy

· 14 International Conference on Plant Growth Substances; Amsterdam, The
Netherlands, July 21-26, 1991 – noaiaeiaa iia?aeiieaiiy

· All-Union Conference on Trends in Plant Biotechnology; Pushino,
Russia, November 20-22, 1991 – aeiiia?aeue

· 1st Conference on Grain Legumes; Anger, France, June 1-3, 1992-
noaiaeiaa iia?aeiieaiiy

· VIIIth International Congress of Plant Tissue and Cell Culture;
Firenze, Italy, June 12-17, 1994 – noaiaeiaa iia?aeiieaiiy

· IVth European Congress of Cell Biology, Prague, Czech Republic, June
26-July 1, 1994 – noaiaeiaa iia?aeiieaiiy

· Ith International Symposium on Plant Biotechnology and Genetic
Engineering, Kiev, Ukraine, October 3-6, 1994 – aeiiia?aeue

· Vth International Congress of Plant Molecular Biology, Singapore,
21-27 September, 1997 – noaiaeiaa iia?aeiieaiiy

· VII International Conference In Vitro Plant Cell Biology,
Biotechnology and Germplasm Preservation, Moscow, 25-28 November 1997 –
aeiiia?aeue

Ioae?eaoe??. Iaoeia? ?acoeueoaoe ii oai? aeena?oaoe?? iioae?eiaaii a 45
ae?oeiaaieo ?iaioao, ui aeeth/a? 1 iiiia?ao?th, 1 aaoi?nueea na?aeioeoai
N?N?, 1 a?ioo?a iaoiaee/ieo ?aeiiaiaeaoe?e ? 20 noaoae, aeaeaieo a
i?aeia?iaeieo, oe?a?inueeeo ? ?in?enueeeo (eieeoi?o oeaio?aeueieo N?N?)
aeo?iaeao. Iiiia?ao?y, yea ia? ia`?i 8.84 ae?oeiaaieo a?eooa, oa 3
noaoo? iaienai? aac ni?aaaoi??a.

Ia’?i oa no?oeoo?a ?iaioe. Aeena?oaoe?y, aeeeaaeaia ia 284 noi??ieao
iaoeiiieniiai oaenoo, neeaaea?oueny ?c anooio, aeaio aeaa iaeyaeo
e?oa?aoo?e, o?ueio aeaa aenia?eiaioaeueii? ?iaioe, caeeth/aiiy,
aeniiae?a, iaoa??ae?a ? iaoiae?a, nieneo aeei?enoaii? e?oa?aoo?e,
?ethno?aoeaiiai aeiaeaoeo. ?iaioa i?noeoue 6 oaaeeoeue, 29 eiia?iiaaieo
iaethie?a. A?ae?ia?ao?/iee nienie aeeth/a? 581 e?oa?aoo?ieo aeaea?ae, ?c
ieo 553 ia ?iicaiieo iiaao.

AAIAOE*IA O?AINOI?IAOe?ss ?INEEI – I?AAeNOAAIEE?A ?IAeEIE AIAIAEO.

„I?yiee ia?aiin aai?a“ oeyoii aeaeo?iii?aoe?? i?ioiieano?a o aeae?a c
?iaeeie aiaiaeo. A aenia?eiaioao ii aea/aiith ia?aiino aai?a oeyoii
aeaeo?iii?aoe?? aoee aeei?enoai? i?ioiieanoe, ye? aeae?eyee c iac??eeo
ca?iaee?a ni?. Aeaeo?iii?aoe?th i?iaiaeeee ca aeiiiiiaith
aaeineiiaeaiiai iaie iaoiaeo. Aeaeeaaiiy i?ioiieano?a i?ney
aeaeo?iii?aoe?? neeaei 50-60%. Eoeueoeaoaaiiy i?ioiieano?a i?ney
aeaeo?iii?aoe?? i?iaiaeeee ca ?ic?iaeaiith iaoiaeeeith (Eo/oe, 1989).
A?aea?? o?ainoi?iaio?a cae?enithaaee ia iiaeeaiiio na?aaeiaeu? A5, yea
i?noeei 1 ia/e e?iaoeio aai AAIo c aeiaeaaaiiyi 50 ia/e
eaiai?oeeinoeueoaoo. Ca oaeeo oiia aoei a?ae?a?aii 21 eieii?th, ye? aoee
caeaoi? aei iiaeaeueoiai ?inoo oa ni?iiiaei? caeai?oe i?e ina?oeaii?.
I?e iiaeaeueoeo ianaaeao ia na?ae? na?aaeiaeua oaeiai ae neeaaeo
caeaoi?noue aei ?inoo caa?aaee 5 eieii?e.

Aiae?c aeoeaiino? iaii?oeeioinoio?ainoa?ace iieacaa i?enooi?noue oeueiai
oa?iaioo o a?aenaeaeoiaaieo ee?oeiieo e?i?yo. Aeioeia a?a?eaeecaoe?y ca
Naoca?iii aeyaeea iayai?noue iine?aeiaiinoae aaio
iaii?oeeioinoio?ainoa?ace o ee?oeiieo e?i?yo, ye? aoei i?iaiae?ciaaii.
Aoaeoeai?noue aaiaoe/ii? o?ainoi?iaoe?? neeaea 1 o 10-5 a?ae /enea
i?ioiieano?a, ui aoei aeei?enoaii aeey aaiaoe/ii? o?ainoi?iaoe?? (Eo/oe
oa ?io., 1990).

A ?ioee na??? aenia?eiaio?a ii aeaeo?iii?aoe?? eaeonieo i?ioiieano?a
eiithoeie eoaiai? aoee aeei?enoai? aiaeia?/i? oiiae, ui ? aeey
i?ioiieano?a ni?. I?e iiaeaeueo?e naeaeoe?? aoee a?ae?a?ai? n?i
ee?oeiieo e?i?e, ye? caa?aaee aeoeaiee ??no a i?enooiino?
eaiai?oeeinoeueoaoo. Aiae?c aeoeaiino? iaii?oeeioinoi-o?ainoa?ace
iieacaa iayai?noue oeueiai oa?iaioo o a?ae?a?aieo ee?oeiieo e?i?yo.

I?iaaaeai? aenia?eiaioe aeiaaee i?eioeeiiao iiaeeea?noue io?eiaiiy
ia?aiino aai?a o aeae?a c ?iaeeie aiaiaeo oeyoii aeaeo?iii?aoe??
i?ioiieano?a. Iaaeae? iai aaeaeiny aeei?enoaoe aeae?iii?aoe?th
i?ioiieano?a aeey io?eiaiiy o?ainaaieo ?ineei ethoea?ie.

Ethoea?ia i?ai?/ia (Medicago borealis L.), iaeaeeae/ee ?iaee/ ethoea?ie
iin?aii?, ? iaeiei c eoeueoeaiaaieo i?aaenoaaiee?a oeueiai aeaeo,
iniaeeai a ciiao iii??iiai ee?iaoo. A iaeiiio aeine?ae? ii
aeaeo?iii?aoe?? aeei?enoiaoaaee 1-2 x 105 i?ioiieano?a ethoea?ie
i?ai?/ii?, aeae?eaieo c eenoy anaioe/ii ae?iuaieo ?ineei e?i?? 94, ui
aoea io?eiaia c AIAe? ei?i?a ?i. A. ?. A?eueyina, i/i Eoaiaa,
Iineianueea iae. I?ney ia?aiino ee?oeiieo eieii?e ethoea?ie ia
aaa?eciaaia iiaeeaia na?aaeiaeua, ui i?noeoue 50 ia/e
eaiai?oeeinoeueoaoo, aoei a?ae?a?aii 56 eeii?a, ye? aoee caeaoi? aei
iiaeaeueoiai ?inoo oa ni?iiiaei? caeai?oe ia na?oe?. Oe? eieii??
ia?aiineee ia na?aea iiaeeaia na?aaeiaeua oaeiai ae neeaaeo, ? o?eueee
15 eeii?a i?iaeiaaeoaaee aeoeaii ?inoe. O an?o i?iaiae?ciaaieo ee?oeiieo
eieii?e aoea aeyaeaia aeoeai?noue iaii?oeeioinoio?ainoa?ace. Iaaeae? o
i’yoe oeeo eeii?a ia iiaeeaiiio na?aaeiaeu? A5, ui i?noeoue 0.5 ia/e
e?iaoeio ? 50 ia/e eaiai?oeeinoeueoaoo, aoei ?iaeoeiaaii ooai?aiiy
aia??i?ae?a, a i?ney oeueiai ? ii?iaeueieo ?ineei (Kuchuk et al., 1990).

I?iaaaeai? iaie aenia?eiaioe aia?oa iieacaee iiaeeea?noue io?eiaiiy
o?ainaaieo ?ineei o ethoea?ie oeyoii aeaeo?iii?aoe?? i?ioiieano?a. Oey
?iaioa aein? caeeoa?oueny ?aeeiith, ui iieno? io?eiaiiy o?ainaaieo
?ineei oeueiai aeaeo ca aeiiiiiaith aeaeo?iii?aoe?? i?ioiieano?a.

Oe?eaai a?aecia/eoe, ui a on?o i?iaaaeaieo aenia?eiaioao ii
aeaeo?iii?aoe?? i?ioiieano?a, ?cieueiaaieo c ?ineei aeae?a c ?iaeeie
aiaiaeo, e?euee?noue eaiai?oeeino?eeeo ee?oeiieo eeii?a, ui
a?aeae?athoueny nii/aoeo, cia/ii ciaioo?oueny i?ney ia?aianaiiy ia
na?ae? iiaeeai? na?aaeiaeua oiai naiiai neeaaeo. Oe? ?acoeueoaoe
na?ae/aoue i?i iiaeeea?noue ianoaa?eueii? ?ioaa?aoe?? /oaeeiieo aai?a a
aaiii ?ineei aai ae i?i ?o ?iaeoeaaoe?th ca ?aooiie ??ciiiai?oieo
iaoai?ci?a, iaeiei c yeeo iiaea aooe iaoeethaaiiy AeIE.

I?iaaaeai? aenia?eiaioe ii aeaeo?iii?aoe?? i?ioiieano?a aeae?a c ?iaeeie
aiaiaeo na?ae/aoue i?i iiaeeea?noue canoinoaaiiy oeueiai iaoiaeo aeey
aaiaoe/ii? o?ainoi?iaoe?? oeeo aeae?a, ca oiiae iayaiino? ?ic?iaeaii?
iaoiaeeee eoeueoeaoaaiiy i?ioiieano?a. Iaoiae aeaeo?iii?aoe??
i?ioiieano?a yaey? niaith iaeei c i?aeoiae?a aaiaoe/ii? o?ainoi?iaoe??
?ineei ?, oi/a a?i ia? iaiaaeaia canoinoaaiiy, iaeii/anii o iueiai ? ?
?yae ia?aaaa ii??aiyii c o?ainoi?iaoe??th ca aeiiiiiaith Agrobacterium
tumefaciens. Oae, canoinoaaiiy oeueiai iaoiaeo ia ? aeaeiniaoeeo?/iei,
ia a?aei?io a?ae iaoiae?a, ui aacothoueny ia aeei?enoaii? “iaacca?i?ieo”
ooai?a A. tumefaciens, ye? aeineoue /anoi aoaathoue aa??oeaioieie ii
a?aeiioaiith aei ??ciiiai?oieo ni?o?a ? aeae?a ?ineei. Ie??i
o?ainoi?iaoe?? yae?a, iaoiae aeaeo?iii?aoe?? i?ioiieano?a iiaeia
canoiniaoaaoe aeey aaiaoe/ii? o?ainoi?iaoe?? oei?iieanoii? AeIE, ui
a?aee?eaa? oe?eeii iia? ia?niaeoeae a io?eiaii? aenieiai ??aiy aeni?an??
/oaeeiieo aai?a a ?ineei?, ? iaeii/anii iaiaaeo? aaceiio?ieueia
?iciianthaeaeaiiy oeeo aai?a c ieeeii.

Io?eiaiiy o?ainaaieo ?ineei ethoea?ie oa ai?ioo c i?aeaeuaiith
caeaoi?noth aei ?aaaia?aoe??. A ?acoeueoao? ia?oeo ia oa?eoi??? N?N?
aenia?eiaio?a ii aaiaoe/i?e o?ainoi?iaoe?? ?ineei, ye? aoee i?iaaaeai? a
1984-1985 ?ieao o a?aeae?e? oeeoio?c?ieia?? oa ee?oeiii? ?iaeaia???
?inoeoooo aioai?ee, aoee a?aei?aoeueiaai? iaoiaee aaiaoe/ii?
o?ainoi?iaoe?? othothio, iniiaai? ia aeei?enoaii? A. tumefaciens (Eo/oe
oa Eaiaaneee, 1987).

Iaaeae? aenia?eiaioaeueia ?iaioa aoea cina?aaeaeaia ia ?ic?iaoe?
iaoiae?a aaiaoe/ii? o?ainoi?iaoe?? ? io?eiaii? o?ainaaieo ?ineei o
aeae?a c ?iaeeie aiaiaeo. I?iaaaeai? aeine?aee ii ?aaaia?aoe?? ?ineei o
ee?oeiieo e?i?e Medicago sativa L. ni?oo “Aanaeiiiaeieyineay” ia
??ciiiai?oieo iiaeeaieo na?aaeiaeuao aoee aac?acoeueoaoieie. Aeey
?iaeoeoe?? caeaoiino? aei ?aaaia?aoe?? o oeueiai ni?oo ethoea?ie aoei
cai?iiiiiaaii ni?iaoaaoe aeei?enoaoe “shooty”-iooaio Agrobacterium
tumefaciens pGV 2206. Oeae iooaio aoa io?eiaiee iai?ee?ioe? 70-o ?ie?a a
?acoeueoao? ?ina?oe?? ?iaeoeaoth/i? iine?aeiaiino?, ui i?noeoue aai
aaeoa??aeueii? no?eeino? aei eaiai?oeeio, a ?aeii aai?a neioaco aoenei?a
O-AeIE ieoii?iiaiai ooaio B6 A. tumefaciens (?en. 1). ?ai?oa aoei
anoaiiaeaii, ui i?ney aaiaoe/ii? o?ainoi?iaoe?? othothio c aeei?enoaiiyi
oeueiai aa?i-aaeoa??aeueiiai ooaio, aaeaeiny io?eiaoe ee?oeii? e?i??, ui
aeoeaii ooai?thaaee iaaiie.

I?ney ni?eueiiai eoeueoeaoaaiiy aenieaio?a ethoea?ie c “shooty”-iooaioii
A. tumefaciens pGV 2206, aoee ?iaeoeiaai? ee?oeii? e?i?? ia iiaeeaiiio
na?aaeiaeu? A5, yea i?noeoue 0.2 ia/e 2.4 D ? 0.5 ia/e e?iaoeio, a
oaeiae aioea?ioeee – eeaoi?ai oa ea?aai?oeee?i – aeey ae?i?iaoe??
aa?iaaeoa??e. Ee?oeii? e?i??, ye? ooai?eeeny, ia?anaaeaeoaaee ia na?aea
iiaeeaia na?aaeiaeua oiai naiiai neeaaeo ? eoeueoeaoaaee i?ioyaii
i?nyoey, a i?ney oeueiai ia?aiineee ia aacai?iiiaeueia na?aaeiaeua B5.
*a?ac i?nyoeue ae?iuoaaiiy ia oeueiio na?aaeiaeu? ee?oeii? e?i??
ethoea?ie ii/eiaee ooai?thaaoe aia??iiiae?ai? no?oeoo?e oaiii-caeaiiai
eieuei?o. I?ney ia?anaaeee oeeo no?oeoo? ia na?aea iiaeeaia na?aaeiaeua
oiai naiiai neeaaeo c a?ae?a?aieo 42 e?i?e i?i?inee ? aeaee ii?iaeuei?
?ineeie eeoa o?noue. I?e ia?anaaeoe? ia na?ae? aacai?iiiaeuei? iiaeeai?
na?aaeiaeua aa?o?aee iaaii?a, ui i?noyoue aieia? a?oiueee, aoee caeaoi?
ooai?thaaoe ei?ai?, aea oeae i?ioean aoa a?eueo o?eaaeei, i?ae i?e
ia?anaaeoe? iaaii?a aeo?aeiiai aaiioeio. A eiio?ieueiiio aenia?eiaio?
i?e aeio?eiaii? an?o oiia ae?iuoaaiiy, ca aeiyoeii ni?eueiiai
eoeueoeaoaaiiy c “shooty”-iooaioii A. tumefaciens, ia aacai?iiiaeueiiio
iiaeeaiiio na?aaeiaeu? ee?oeii? e?i?? ethoea?ie aeiaeieo aia??i?ae?a ia
ooai?thaaee, eeoa o aeayeeo eeii?a niinoa??aaee ?icaeoie ei?ai?a.

Ee?oeii? eoeueoo?e, ?iaeoeiaai? c ?ineei ethoea?ie, io?eiaieo i?ney
?aaaia?aoe??, oaeiae caa??aaee caeaoi?noue aei aia??iaaiaco. Iaea?aeai?
?ineeie ia?aiineee a a?oio, aea aiie oea?ee oa ooai?thaaee aeeoo?caeaoia
ian?iiy, yea caa??aaei i?e ia?aiin? ?o a eoeueoo?o in vitro caeaoi?noue
aei ?aaaia?aoe?? ?ineei.

Aeey aeieaco i?enooiino? iine?aeiaiinoae O-AeIE ieaci?aee pGV 2206 a
?ineeiao ethoea?ie, ye? aoee io?eiai? i?ney ?aaaia?aoe??, aoea
i?iaaaeaia Naoca?i-aeio a?a?eaeecaoe?y. sse ciiae aeei?enoiaoaaee aai
no?eeino? aei eaiai?oeeio, no?oeoo?ia /anoeia yeiai aoea i?enooiy a
O-AeIE ieaci?aee PGV2206. A ?acoeueoao? i?iaaaeaieo aenia?eiaio?a aoei
iieacaii i?enooi?noue iine?aeiaiinoae O-AeIE ieaci?aee pGV2206 a aaiii?
a?ae?a?aieo ?ineei ethoea?ie, ui aeiaiaeeoue ?o o?ainaaiio i?e?iaeo
(Kuchuk et al., 1990).

I?iaaaeai? aenia?eiaioe aeicaieeee c?iaeoe aeniiaie, ui caeaoi?noue aei
?aaaia?aoe?? o o?ainaaieo ?ineei ethoea?ie iiaea aooe aeeeeeaia aaiaie
O-AeIE “shooty”-iooaioa A. tumefaciens pGV 2206.

Io?eiai? ?acoeueoaoe aeicaieeee noi?ioethaaoe iiaee i?aeo?ae aeey
iie?ioaiiy ?aaaia?aoe?eii? caeaoiino? o ?ineei c iecueeei
?aaaia?aoe?eiei iioaioe?aeiiiieyaa? a ianooiiiio. Cai?iiiiiaaiee iaoiae,
ye? aoei iacaaii “iiaea?eiith o?ainoi?iaoe??th”, aoa aeei?enoaiee aeey
o?ainoi?iaoe?? oaeiai aaaeeeaiai n?euenueeiainiiaea?nueeiai aeaeo ye
ai?io.

A aenia?eiaioao aeei?enoiaoaaee e?i?th ai?ioo 1288, ui iaea iaaiee
?aaaia?aoe?eiee iioaioe?ae, aea a nenoai? a?aeai?o ia aacai?iiiaeueiiio
iiaeeaiiio na?aaeiaeu? ia aeyaeyea aeiaeieo iciae ooai?aiiy iaaii?a.
I?ney ni?aeoeueoeaoaaiiy aenieaio?a c? noaaea oa eenoe?a anaioe/ii
ae?iuaieo ?ineei ai?ioo e?i?? 1288 c “shooty”-iooaioii A. tumefaciens
pGV 2206 ia iiaeeaiiio na?aaeiaeu? B5, ui i?noeoue o?oiai?iiie (2.4 Ae
oa AAI) aoei eiaeoeiaaii eaeon. *a?ac iaeei i?nyoeue eoeueoeaoaaiiy
eaeon, ui ooai?eany, aoa ia?anaaeaeaiee ia aacai?iiiaeueia iiaeeaia
na?aaeiaeua. A aeayeeo ae?eyieao eaeonii? oeaieie ii/aee ooai?thaaoeny
ina?aaeee ?aaaia?aoe?? o aeaeyae? oaiii-caeaieo e?aiie. I?e iiaeaeueo?e
ia?anaaeoe? oe? ina?aaeee ooai?eee ee?oeii? e?i??, ye? noaa?eueii
ooai?thaaee iaaiie. Oaeei /eiii aaeaeiny a?ae?a?aoe 12 e?i?e ai?ioo,
caeaoieo aei ?aaaia?aoe??, ye? iino?eii ooai?ththoue ?aaaia?aioe ia
i?ioyc? o?eaaeiai /ano. Oae, iaeia c oeeo ia?aeiii io?eiaieo e?i?e
i?aeo?eio?oueny aac ao?aoe ?aaaia?aoe?eii? caeaoiino? i?ioyaii ainuei?
?ie?a. Iieaeoey?ii-a?ieia?/iee aiae?c c aeei?enoaiiyi
aeio-a?a?eaeecaoe?? ca Naoca?iii iieacaa i?enooi?noue iine?aeiaiinoae
O-AeIE o aaiii? a?ae?a?aieo ?aaaia?aoe?eieo e?i?e ai?ioo (Coaei oa ?io.,
1990).

Aeine?aeaeoth/e iiaeeea?noue canoinoaaiiy cai?iiiiiaaii? iaie iaoiaeeee
ii ?iaeoeoe?? ?aaaia?aoe?eii? caeaoiino? o ai?ioo i?ney
ni?aeoeueoeaoaaiiy c “shooty”-iooaioii A. tumefaciens, a aenia?eiaioao
ii aaiaoe/i?e o?ainoi?iaoe?? aoee aeei?enoai? a?n?i ni?oiaeo e?i?e
aieeaiaenueei? naeaeoe??, io?eiaieo a?ae o??ie “Nunhems Zaden”
(I?aea?eaiaee). I?ney ni?aeoeueoeaoaaiiy ?ineeiieo aenieaio?a c
“shooty”-iooaioii A. tumefaciens pGV 2206 iai aaeaeiny o o?ueio c
ainueie c?ace?a io?eiaoe e?i?? ai?ioo c aenieith ?aaaia?aoe?eiith
aeoeai?noth. Oe? e?i?? aoee iacaai? 912097-2206, 912074-2206 ?
911136-2206.

Ia iao iiaeyae, iacaaaeath/e ia an? iaiaaeaiiy, ?ic?iaeaiee iaoiae
io?eiaiiy caeaoieo aei ?aaaia?aoe?? e?i?e o ai?ioo oa ethoea?ie,
aeicaiey? iiaeieaoe i?iaeaie, iia’ycai? c o?oaeiiuaie ?iaeoeoe??
ooai?aiiy ?ineei c aeaaeeoa?a-ioe?eiaaii? oeaieie a eoeueoo?? in vitro.
Oeae i?aeo?ae aoa oni?oii canoiniaaiee aeey io?eiaiiy o?ainaaiieo ?ineei
ai?ioo, ui ianooue ??ciiiai?oi? aaiaoe/i? eiino?oeoe??, ye?
i?aaenoaaeythoue ?ioa?an ye aeey ooiaeaiaioaeueii? iaoee, oae ? aeey
i?aeoe/iiai canoinoaaiiy.

Eiino?othaaiiy ?ineei ai?ioo, ui i?noyoue iia?eueiee Ds aeaiaio
eoeo?oaece. Iaeiei c iiaeeeaeo i?aeoiae?a aeey ?ciethaaiiy oi?eaeueieo
iine?aeiaiinoae AeIE ? iiooe ? eeiioaaiiy aai?a ca aeiiiiiaith
o?ainiicii?a – aaiaoe/ieo aeaiaio?a, ui o?ainiicothoueny (gene tagging).
Na?aae ?ineeiieo o?ainiicii?a iaea?eueo aea/aiith ? ?iaeeia iia?eueieo
Ac/Ds aeaiaio?a eoeo?oaece. Oiio, iae/ano?oa aiie aeei?enoiaothoueny ye
nenoaia aaoa?ieia?/ieo o?ainiicii?a a aenia?eiaioao ii eeiioaaiith
aai?a.

Iacaaaeath/e ia oa, ui eiei ?ineei o yeeo i?aoeth? aaoa?ieia?/ia nenoaia
o?ainiicii?a Ac-Ds eoeo?oaece, iino?eii c?inoa?, iaeii/anii ? aeioaia?
iaia? ioae?eaoe?e, ui iienothoue iiae?ai? aenia?eiaioe o ai?ioo.
Iaiao?aeii cacia/eoe, ui ai?io ? iaei??th c n?euenueeiainiiaea?nueeeo
ca?iiaiaiaeo eoeueoo?, ye? iaea?eueo oe?iei aeei?enoiaothoueny, c aeia?a
aea/aieie ? ea?oiaaieie o?iiiniiaie. Oeae aeae ? i?aaenoaaieeii ?iaeeie
aiaiaeo, ui iathoue iaeio c oi?eaeueieo aaiaoe/ieo nenoai neia?ioe/ii?
o?enaoe?? acioo. Iiooe ? eeiioaaiiy oi?eaeueieo aai?a ai?ioo, cie?aia
aai?a neia?ioe/ii? o?enaoe?? acioo, c aeei?enoaiiyi aaoa?ieia?/ii?
nenoaie o?ainiicii?a ? aeinoaoiuei i?eaaaeeaei ye c oi/ee ci?o
ooiaea-iaioaeueieo, oae ? i?eeeaaeieo aeine?aeaeaiue. E??i oiai, o
ai?ioo ? aeioaia? ia iienai? ooieoe?iioth/? o?ainiiciie, oi/a
o?ainiiciiiiae?ai? iine?aeiaiino? AeIE aoee aeyaeai? a eiai aaiii? i?e
aea/aii? “iaiaeaeaanueei?” iooaoe?? – cii?oeoaao?noue ian?iiy. sse
ia?oee e?ie i?e eeiioaaii? oi?eaeueieo aai?a c aeei?enoaiiyi iaoiaeo
“gene tagging” iaiao?aeii aoei io?eiaoe o?ainaaii? ?ineeie ai?ioo, ui
i?noyoue Ds-aeaiaio eoeo?oaece.

Aeey io?eiaiiy o?ainaaiieo ?ineei ai?ioo ie aeei?enoiaoaaee ??ciiiai?oi?
oeyoe aaaaeaiiy AeIE a ?ineeii? ee?oeie. Iaeiei c iaoiae?a aoea
aeaeo?iii?aoe?y i?ioiieano?a ai?ioo a i?enooiino? ieaci?aee pSK3 (?en.
2). Oey eiino?oeoe?y aoea etha`ycii iaaeaia Dr. J. Hille (A?eueiee
oi?aa?neoao, i. Ainoa?aeai). Nii/aoeo ia naeaeoeaiiio na?aaeiaeu?, ui
i?noeoue 50 ia/e eaiai?oeeinoeueoaoo, aoei a?ae?a?aii 42 eieii??, ye?
caeai?ee i?e ina?oeaii?. I?e iiaeaeueoiio ae?iuoaaii? ia iiaeeaiiio
na?aaeiaeu?, ui i?noeoue naeaeoeaiee aaaio, aoei a?ae?a?aii 7 noaa?eueii
no?eeeo aei eaiai?oeeinoeueoaoo eeii?a.

No?ee? aei eaiai?oeeinoeueoaoo eeiie ia?aa??yee ia iayai?noue
iaii?oeeioinoio?ainoa?ace ca aeiiiiiaith aioeo?e, niaoeeo?/ieo aei
oeueiai a?eeo, aeei?enoiaoth/e noaiaea?oiee iaa?? aeey ELISA (Enzyme
Linked ImmunoSorbant Assay), io?eiaiee a?ae eiiiai?? ‘3’-5′” (NOA).
Io?eiai? aeai? iieacaee iayai?noue a?eeo NPTII o eeii?a ai?ioo,
a?ae?a?aieo i?ney aeae?iii?aoe?? i?ioiieano?a.

Iiaeaeueoee aiae?c o?ainoi?iaio?a i?iaiaeeee ca aeiiiiiaith aiae?co
iie?ia?acii? eaioethaiai? ?aaeoe?? (IE? – PCR). An? aea/ai? o?ainaaii?
eeiie aeaaaee a ?acoeueoao? aiie?o?eaoe?? o?aaiaioe AeIE iaiao?aeiiai
?ici??o (189 i.i.), i?e aeei?enoaii? i?aeia??a, niaoeeo?/ieo aei 35S
i?iiioi?o.

Aeey iiaeaeueoiai aeieaco ?ioaa?aoe?? AeIE ieaci?aee pSK3 a aaiii ai?ioo
i?iaiaeeee aeioeia-a?a?eaeecaoe?th ca Naoca?iii. Aeai?, io?eiai? iaie,
neoaeaoue aeieacii ?ioaa?aoe?? a aaiii ai?ioo aaia NPTII, ui aoiaeeoue
aei neeaaeo ieaci?aee pSK3. Aei oiai o an?o caie a?ae?a?aieo eeii?a aai
aoiaeeoue aei neeaaeo o?aaiaioo pici??ii i?eaeecii 7 oen. i.i. AEiaeieo
nioa a?a?eaeecaoe?? ia aoei aeyaeaii o AeIE eiio?ieueii?
iao?ainoi?iiaaii? ?ineeie (?a/ae e ae?., 1994).

I?ioa iaeii/anii aiae?c aeoeaiino? GUS aaia o a?ae?a?aieo eeiiao ia
aeyaea i?enooiino? aeoeaiino? -aetheo?ii?aeace. Aeey ia?aa??ee
iiaeeeaiai ?ia?aoaaiiy aeoeaiino? oeueiai aaio ca ?aooiie iaoeethaaiiy
caeeoe?a oeeoic?io a AeIE o?ainoi?iiaaieo ee?oei, a?aenaeaeoiaai? eaeone
ae?iuoaaeeny i?ioyaii 2 oeaei?a ia iiaeeaiiio na?aaeiaeu?, ui i?noeoue
5`-acaoeeo?ae?i. I?ney oeueiai eaeon, yeee oi?ioaaany ia na?aaeiaeu? c
5`-acaoeeo?ae?iii, iaa GUS iiceoeaio ?aaeoe?th. Oaeei /eiii aoei
anoaiiaeaii, ui aeiaeaaaiiy a iiaeeaia na?aaeiaeua aeaiaoeethth/iai
aaaioa, yeei ? 5`-acaoeeo?ae?i, a?aeiiaeth? aeoeai?noue ia?aianaieo
aai?a. Oeae oaeo na?ae/eoue i?i iiaeeea?noue ?ia?aoaaiiy aeoeaiino?
ia?aianaieo aai?a ca ?aooiie iaoeethaaiiy caeeoe?a oeeoic?io a AeIE
yae?a.

Io?eiai? ?acoeueoaoe ii a?aeai?o o?ainaaieo e?i?e ai?ioo aia?oa iieacaee
iiaeeea?noue aeei?enoaiiy eaiai?oeeinoeueoaoo ye naeaeoeaiiai aaaioa
aeey a?aeai?o o?ainoi?iaio?a o oeueiai aeaeo.

O?ainaaii? iaaiie ai?ioo,ye? iaee aaoaeiaaiee Ds-aeaiaio eoeo?oaece,
aoee io?eiai? oeyoii aaiaoe/ii? o?ainoi?iaoe?? ca aeiiiiiaith A.
tumefaciens. Ieaci?aea pSK3 aoea ?iiia?e?ciaaia a A. tumefaciens, ui
iana ieaci?aeo pMP 90 c Vir – iaeanoth. Io?eiaiei aa?iaaeoa??aeueiei
ooaiii aoei o?ainoi?iiaaii e?i?th ai?ioo 1288-2206-12 c aenieei
?aaaia?aoe?eiei iioaioe?aeii, yea aoea ?cieueiaaia iaie ?ai?oa i?ney
aaiaoe/ii? o?ainoi?iaoe?? “shooty”-iooaioii A.tumefaciens pGV 2206
(Coaei oa ?io., 1990). I?ney o?ainoi?iaoe?? aoei a?ae?a?aii 23 ee?oeiieo
e?i??, no?eeeo aei eaiai?oeeio. Caiaeii ia aacai?iiiaeueiiio iiaeeaiiio
na?aaeiaeu? aoei io?eiaii 5 ?aaaia?aio?a.

I?iaaaeaiee iieaeoey?ii-a?ieia?/iee aiae?c c aeei?enoaiiyi iie?-ia?acii?
eaioethaiai? ?aaeoe??, Naoca?i-aeio a?a?eaeecaoe?? oa ?ioii-oa?iaioiiai
aiae?co iaeiie/aiiy a?eeo NPTII iiai?noth aeiaaee o?ainaaiio i?e?iaeo
io?eiaieo eaiai?oeeino?eeeo ?aaaia?aio?a ai?ioo. Oaeiae aoea iieacaia
i?enooi?noue iia?eueiiai Ds-aeaiaioo a aaiii? a?ae?a?aieo ?ineei oeyoii
Naoca?i-aeio a?a?eaeecaoe?? c aeei?enoaiiyi iine?aeiaiino? AeIE naiiai
Ds-aeaiaioo ye ciiaeo (?a/ae oa ?io., 1995).

I?aenoiiaoth/e aeeeaaeaia aeua, iiaeia noaa?aeaeoaaoe, ui aia?oa
aeiaaaeaia iiaeeea?noue io?eiaiiy o?ainaaieo iaaii?a ai?ioo c
Ds-aeaiaioii eoeo?oaece. Oaeiae, aoea iieacaia iiaeeea?noue aeei?enoaiiy
eaiai?oeeinoeueoaoo ye naeaeoeaiiai aaaioo aeey a?aeai?o o?ainaaieo
?ineei ai?ioo.

Cae?eniai? aenia?eiaioe a?aee?eaathoue iia? ia?niaeoeae a aaiaoeoe?
oaeiai ia`?eoo ye ai?io oa aeicaieythoue eeiioaaoe oi?eaeuei? aaie ca
?aooiie aeei?enoaiiy iaoiaeo “gene tagging”.

Io?eiaiiy o?ainaaiieo ?ineei ai?ioo, no?eeeo aei oino?iio??oeeio. Iaoith
aenia?eiaio?a aoei io?eiaiiy o?ainaaiieo ni?o?a ai?ioo, ui ianooue aai
no?eeino? aei oino?iio??oeeio – iiaiai aa?a?oeeaeo a?aeiiiai oaeiae i?ae
iacaith BASTA aai aethoic?iao aiii?th. Aeey oe??? ?iaioe aoee
aeei?enoai? caeaoi? aei ?aaaia?aoe?? e?i?? 912097-2206, 912074-2206 ?
911136-2206, io?eiai? ?ai?oa o ni?o?a aieeaiaenueei? naeaeoe?? oeyoii
o?ainoi?iaoe?? ooaiii A. tumefaciens pGV 2206. Ia iniia? aaeoi?o pGA472
aoea ceiinopoeiaaia ieaci?aea pK3, yea iana ye aai no?eeino? aei
eaiai?oeeinoeueoaoo, oae ? aai no?eeino? aei oino?iio??oeeio. E??i oiai,
iaaeae?, ie oaeiae aeei?enoiaoaaee eiino?oeoe?? A. tumefaciens pHoe
106/pEHA 101 ? A. tumefaciens pHoe200/pEHA101, ye? ianooue eeoa aai
no?eeino? aei oino?iio??oeeio. An? oe? eiino?oeoe?? aoee ia?aianai? a
ooai A. tumefaciens N58N1, yea i?noeea ieaci?aeo pMP 90. Io?eiaieie
a?ia?ieie aaeoi?aie aoee i?iaaaeai? aenia?eiaioe ii o?ainoi?iaoe??
aeuaiaaaaeaieo e?i?e ai?ioo.

I?ney aaiaoe/ii? o?ainoi?iaoe?? c aeei?enoaiiyi oeeo eiino?oeoe?e o
e?i?e ai?ioo 912074-2206 ? 912097-2206 aoee a?ae?a?ai? o?ainoi?iaioe
ai?ioo, ye? ?inoooue oa caeai?thoue a i?enooiino? 10ia/e
oino?iio??oeeio. Ca aeiiiiiaith iie?ia?acii? eaioethaiai? ?aaeoe??
(IE?), aea ye i?aeia?e aeei?enoiaoaaeenue ye iine?aeiaiino?, ye?
niaoeeo?/ii ai?ciathoue 35S i?iiioi?, yeee aoiaeeoue aei neeaaeo
oeia?iiai aaio no?eeino? aei oino?iio??oeeio, oae ? i?aeia?e,
niaoeeo?/i? aeania aei neioaoe/iiai aaio no?eeino? aei oino?iio??oeeio,
aoea iieacaia i?enooi?noue iine?aeiaiino? AeIE ia?aianaiiai aaia o
a?ae?a?aieo ee?oeiieo e?i?yo. C oeeo e?i?e aoee io?eiai? ?aaaia?aioe,
aea i?ioean ooai?aiiy ei?ai?a o ieo aoa oneeaaeiaiee, ui ia aeaaaei
iiaeeeaino? io?eiaoe ian?iiy oeyoii aenaaeee a ??oio a?ae?a?aieo ?ineei.

I?iaeaio io?eiaiiy iauaaee?a c ian?iiy o o?ainaaieo iaaii?a ai?ioo ie
ae??oeee oeyoii, a?aei?iiei a?ae iienaiiai aoaeue-aea. Aeyaeeiny, ui i?e
ae?iuoaaii? ?o ia aacai?iiiaeueiiio iiaeeaiiio na?aaeiaeu? a
eoeueoo?aeueieo aaieao ia`?iii 250 ie ca oiia aaciiai?io c iaaeieeoi?i
iia?o?yi, aiie ii/eiathoue oea?noe aaciina?aaeiuei a eoeueoo?? in vitro
aac ooai?aiiy ei?ai?a. O iiaeaeueoiio c oeeo ea?oie caa’yco?oueny
iaaaeeea, aea oe?eeii iiaiioe?iia ian?iiy, yea iiaeia ye ia?aiineoe a
??oio, oae ? aenaaeaeoaaoe ciia a anaioe/i?e eoeueoo?? ia iiaeeaieo
na?aaeiaeuao, ye? i?noyoue naeaeoeaiee aaaio aeey ia?aa??ee no?eeino?
aei oino?iio??oeeio. Ca aeiiiiiaith oeueiai iaoiaeo ie io?eiaee
iauaaee?a o o?ainoi?iiaaieo e?i?e ai?ioo. Aiae?c oniaaeeoaaiiy iciaee
no?eeino? aei oino?iio??oeeio iieacaa, ui /anoeia ian?iiy, yea aoei
io?eiaii o iaei??? c o?ainoi?iiaaieo e?i?e 912074-2206, noaa?eueii
caa??aaea eiai aei ae?oai? ?aia?aoe?? aeeth/ii (Oaae. 1). Oe?
?acoeueoaoe ia?aeiieeai aeiaiaeyoue iiaeeea?noue io?eiaiiy o?ainaaiiiai
ian?iiy o ai?ioo, yea iana iciaeo no?eeino? aei aa?a?oeeaeo. Oaeei /eiii
aoee io?eiai? aaiaoe/ii ci?iai? ?ineeie ai?ioo oa ?oi? iauaaeee c
aeei?enoaiiyi i?ea?iaeueii? nenoaie ia?aiino aai?a, ui aoea iacaaia iaie
“iiaea?eia o?ainoi?iaoe?y”.

Oaaeeoey 1. Oniaaeeoaaiiy iciaee no?eeino? aei oino?iio??oeeio o
o?ainaaiieo ?ineei ai?ioo.

T2 aaia?aoe?y, ia?aa??aii ian?iiyE?i?y

ai?ioo

T1 aaia?aoe?y,

e?euee?noue ian?iiyAenaaeaeaii ian?iiy c eiaeii? O1 ?ineeieI?i?inei ia
na?aaei-aeu? c BASTACaa??aaei

caeaoi?noue ?inoe ia na?aaeiaeu? c BASTA*E?euee?noue IE?-

iiceoeaieo

912074-2206

4 (1 ia i?i?inei)

12

23

12

10

19

7

3

6

5

2

6

3Anueiai:47361411*IE? aiae?c an?o ?ineei, ye? ao?aoeee caeaoi?noue ?inoe
a i?enooiino? oino?iio??oeeio, ia aeaa iiceoeaiiai neaiaeo o aeiaeii? c
ieo.

Cai?iiiiiaaiee iaoiae “iiaea?eii? o?ainoi?iaoe??” aeicaiey? c aenieei
nooiaiai aoaeoeaiino? io?eioaaoe aaiaoe/ii ci?iai? ?ineeie ai?ioo. E??i
oiai, a?ae?a?ai? o?ainaaii? ?ineeie caa??aathoue aenieo ?aaaia?aoe?eio
caeaoi?noue, ui aeicaiey? aeei?enoiaoaaoe ?o a iiaeaeueoeo
aaiii-?iaeaia?ieo aenia?eiaioao. Oaeei /eiii, i?ea?iaeuei? ?ic?iaee ii
aaiaoe/i?e o?ainoi?iaoe?? ai?ioo c aeinoaoiuei aaeeeei nooiaiai
iaae?eiino? aeicaieythoue i?iaiaeeoe aaiaoe/io o?ainoi?iaoe?th ai?ioo,
a io?eiai? no?ee? aei aa?a?oeeaeo ?ineeie iiaeooue aooe aeei?enoai? a
i?aeoeoe? n?euenueeiai ainiiaea?noaa.

Io?eiaiiy o?ainaaiieo e?i?e ei?ai?a o ai?ioo ? ethoea?ie oiaeaaeaeii? c
aeei?enoaiiyi Ri ieaci?aee c A. rhizogenes. C iaoith aea/aiiy aieeao
aai?a ieaci?aee Ri, aeae?eaii? c A. rhizogenes, ia i?aaiiaaiac o aeae?a
c ?iaeeie aiaiaeo aoa i?iaaaeaiee ?yae aenia?eiaio?a ii o?ainoi?iaoe??
ai?ioo ? ethoea?ie oiaeaaeaeii? ooaiii A. tumefaciens C58C1, ye?
i?noyoue ieaci?aeo pRiA4, a oaeiae ieaci?aeo pGA 472, ui caaacia/o?
no?ee?noue aei eaiai?oeeinoeueoaoo o io?eiaieo o?ainaaiieo ?ineeiieo
e?i?e.

Ethoea?ia oiaeaaeaeia (Medicago lupulina) – iaeei c aeeei?ineeo aeae?a
ethoea?i – i?aaenoaaey? iniaeeaee ?ioa?an ye iiaeeeaee aeiii? iciaee
naiicaieeaiiy, yea a?aenooiy o eoeueoo?ieo aeae?a ethoea?i. A oie aea
/an oeae aeae ia no?auo?oueny cae/aeiei noaoaaei oeyoii c eoeueoo?ieie
aeaeaie ethoea?ie. Noai?aiiy o?ainaaiieo e?i?e o oeueiai aeaeo –
ethoea?ie oiaeaaeaeii?, ui ianooue naeaeoeai? iciaee a eoeueoo?? in
vitro,? aacia?a/ii oe?eaaei aeey aenia?eiaio?a ii niiaoe/i?e
a?a?eaeecaoe?? c ?ioeie aeaeaie ethoea?ie, ui aeei?enoiaothoueny a
n?euenueeiio ainiiaea?noa?.

Oaaeeoey 2. O?ainaaii? ee?oeii? e?i?? ? ?ineeie aeae?a c ?iaeeie
aiaiaeo, io?eiai? a ?EAA?

?ineeiiee aeae- e?i?yAaeoi?Naeaeoeaia iciaeaNiin?a o?ainoi?iaoe??Oi?ia
o?ainaaiii? e?i??Medicago sativa-Aanaei-

Iiaeieyinueea Pisum sativum

-1288

-911136

-912074

-912097

Pisum sativum

-1288-2206-12

Pisum sativum

-912074-2206

-912097-2206

Medicago lupulina

Medicago borealis – 94

Glycine max

Pisum sativum

-1288-2206-12pGV2206

pGV2206

pGV2206

pGV2206

pGV2206

pSK3

Ds-element

pK3

pK3

pRiA4# pGA472

pGA472

pGA472

pSK3

Ds-elementCaeaoi?noue aei ?aaaia?aoe??

Caeaoi?noue aei ?aaaia?aoe?? Caeaoi?noue aei ?aaaia?aoe?? Caeaoi?noue
aei ?aaaia?aoe?? Caeaoi?noue aei ?aaaia?aoe??

No?ee?noue aei eaiai?oeeio

No?ee?noue aei BASTA

No?ee?noue aei BASTA

No?ee?noue aei eaiai?oeeio

No?ee?noue aei eaiai?oeeio No?ee?noue aei eaiai?oeeio No?ee?noue aei
eaiai?oeeioAgrobacterium tumefaciens

Agrobacterium tumefaciens Agrobacterium tumefaciens Agrobacterium
tumefaciens Agrobacterium tumefaciens

Agrobacterium tumefaciens

Agrobacterium tumefaciens

Agrobacterium tumefaciens

Agrobacterium tumefaciens

Aeaeo?iii?aoe?y i?ioiieano?a Aeaeo?iii?aoe?y i?ioiieano?a
Aeaeo?iii?aoe?y i?ioiieano?aOa?oeeuei? ?ineeie

Oa?oeeuei? ?ineeie

Oa?oeeuei? ?ineeie Oa?oeeuei? ?ineeie Oa?oeeuei? ?ineeie

Iaaiie

Oa?oeeuei? ?ineeie Oa?oeeuei? ?ineeie

Eoeueoo?a ei?ai?a

?ineeie

Eaeon

Eaeon

A ?acoeueoao? i?iaaaeaieo aenia?eiaio?a ii eieoeueoeaoaaiith aenieaio?a
ai?ioo ? ethoea?ie oiaeaaeaeii? c A. tumefaciens pRiA4 pGA 472 ia
aacai?iiiaeueiiio iiaeeaiiio na?aaeiaeu?, ui i?noeoue 50 ia/e
eaiai?oeeinoeueoaoo aoee a?ae?a?ai? ee?oeii? e?i??, ye? aeoeaii ?inee.
I?e iiaeaeueoeo ia?anaaeeao ia iiaeeai? na?aaeiaeua oiai aea neeaaeo
/anoeia oeeo e?i?e aeoeaii ooai?thaaea ei?ai?, oa?aeoa?i? ii ii?oieia??
aeey caoai?thaaiiy “ai?iaeaa/ano? ei?ai?”, ui cae/aeii aeeeeea?oueny A.
rhizogenes. *anoeia io?eiaieo eaiai?oeeino?eeeo eeii?a ethoea?ie ?
ai?ioo ?inee o aeaeyae? iai?aai?ciaaii? ee?oeiii? iane. A?ae?a?ai?
eoeueoo?e ei?ai?a o iaio aeae?a ia ooai?thaaee aia??i?ae?a aai iaaii?a
ye ia aacai?iiiaeueieo iiaeeaieo na?aaeiaeuao, oae ? i?e aeiaeaaaii?
oeeoie?i?i?a. Aiae?c aeoeaiino? iaii?oeei-oinoio?ainoa?ace iieacaa
i?enooi?noue aeoeaiino? oeueiai oa?iaioo a io?eiaieo ee?oeiieo e?i?yo.

Oe? aenia?eiaioe aeicaieeee iai anoaiiaeoe, ui aeaee c ?iaeeie aiaiaeo
oae? ye ai?io ? ethoea?ia oiaeaaeaeia i?ney o?ainoi?iaoe?? ieaci?aeith
RiA4 ooai?ththoue ei?ai?, ui aeoeaii ?inoooue ? ? oeiiaeie aeey
o?ainoi?iaio?a, io?eiaieo c aeei?enoaiiyi A. rhizogenes. A?ae?a?ai?
e?i?? eoeueoo?e ei?ai?a o iaio aeae?a iaee iaii?oeeioinoio?ainoa?acio
aeoeai?noue, ui i?aeoaa?aeaeo? ?o o?ain- aaiio i?e?iaeo. AEiaeii?
iciaee i?aaiiaaiaco aai aia??iaaiaco o io?eiaieo e?i?e aeyaeaii ia aoei.

EOEUeOEAOAAIIss I?IOIIEANO?A, EE?OEIIEO E?I?E ? ?AAAIA?AOe?ss ?INEEI O
AeAssEEO ?INEEIIEO AEAe?A.

Eoeueoeaoaaiiy i?ioiieano?a ?ineei. Aeae?eaiiy ? eoeueoeaoaaiiy
i?ioi-ieano?a iaeaaaeeea?oeo n?euenueeiainiiaea?nueeeo aeae?a, ui
iaeaaeaoue aei ?iaeeie aiaiaeo, caaaeaee aoei a oeaio?? oaaae
aeine?aeiee?a, ye? i?aoeththoue a aaeoc? ee?oeiii? a?ieia?? ?ineei.

A aenia?eiaioao, ui ?icii/aeeny a 1985 ?ioe?, iniiaio oaaao aoei
i?eae?eaii ?ic?iaoe? niinia?a eoeueoeaoaaiiy i?ioiieano?a, ?cieueiaaieo
c oeaiei ?ioaeoieo ?ineei ni?. Io?eiaiiy aeinoaoiuei aaeeei? e?eueeino?
i?ioiieano?a c ?ineeiieo oeaiei ? iaei??th c iniiaieo i?iaeai, c yeeie
noeeaeeny aeine?aeieee i?ae /an ?ic?iaee niinia?a eoeueoeaoaaiiy
i?ioiieano?a o ni?. Aeei?enoaiiy eenoe?a ?ineei, ui ae?inee a anaioe/ieo
oiiaao, ye aeaea?aea aeey aeae?eaiiy i?ioiieano?a, ia aeicaieyei
ae??oeoe oeth i?iaeaio. I?ioa oaea iiaeeea?noue ia? i?noea o aaaaoueio
aeae?a c ??ciiiai?oieo ?iaeei, na?aae yeeo ? ? aeaee c ?iaeeie aiaiaeo.
Iacaeaaeii a?ae aiaeia?/ieo iia?aeiieaiue ,ui c’yaeeeny caiaeii, aoa
?ic?iaeaiee iaoiae eoeueoeaoaaiiy i?ioiieano?a ni?, yeee iieyaa? a
aeei?enoaii? iac??eeo ca?iaee?a ye aenieaio?a. A aenia?eiaioao aoee
aeei?enoai? iac??e? ca?iaeee ni? ?ici??ii a?ae 2 aei 10 ii.

I?ney i/euaiiy i?ioiieanoe c iac??eeo ca?iaee?a ni? eoeueoeaoaaee a
iiaeeo?eiaaiiio iiaeeaiiio na?aaeiaeu? EI8?. Ia?oa ae?eaiiy i?ioiieano?a
a?aeaoaaeiny ia 2-3 aeiao eoeueoeaoaaiiy. Ia i’yoo aeiao eoeueoeaoaaiiy
noniaic?th i?ioiieano?a ?icaaaeyee ??aiei ia’?iii iiaeeaiiai na?aaeiaeua
A5 c 0.35 I iai?oieii ye iniioeea ? o?oiai?iiiaie: 0.2 ia/e 2.4 Ae ? 0.5
ia/e e?iaoeio aai AAIo. A oe?e noi?o? i?ioiieanoe i?iaeiaaeoaaee
ae?eeoeny ? ooai?thaaee i?e?ieieii?? c 5-10 ee?oei. *a?ac 2-3 oeaei?
i?ney ia?oiai ?icaaaeaiiy i?e?ieieii?? ni? ?icaiaeeee ??aiei ia’?iii
oiai ae na?aaeiaeua N c 0.25 I iai?oieii ? oaeeie ae naieie
o?oiai?iiiaie. A oeeo oiiaao i?e?ieieii?? i?iaeiaaeoaaee ?icaeaaoeny,
ooai?thth/e 50-100 ee?oeii? eieii??. Oe? eieii?? ca aeiiiiiaith
ianoa??anueei? i?iaoee iaoa?iaoaaee ia u?eueia iiaeeaia na?aaeiaeua N,
ui i?noeoue 1 ia/e AAIa aai e?iaoei, ? ia?aiineee ia na?oei (4000ee) c
16 aiaeeiiei na?oeiaei ia??iaeii. A oaeeo oiiaao ee?oeii? eieii?? ni?
i?iaeiaaeoaaee ?icaeaaoeny ? iaaoaaee oaiii-caeaiiai caaa?aeaiiy.
Io?eiaiee i?aia?ao i?ioiieano?a ni? a?ae??ciyany aenieith caeaoi?noth
aei iiae?eo, ui aeicaieeei aeei?enoaoe oe? i?ioiieanoe aeey
?iaeea?aeoaeueiiai eoeueoeaoaaiiy a i?e?ieaieyo ia?iii 100 ie, c
iaeaeuith a ii??aiyii? c ?ioeie aeaeaie, ui aeine?aeaeoaaeeny,
aoaeoeai?noth iiae?eo yea aeinyaaea 46% (Ee?e/aiei oa ?io., 1990)

Aeey aeae?eaiiy i?ioiieano?a o aeayeeo ?ioeo aeae?a c ?iaeeie aiaiaeo ye
aenieaioe a?aee eenoee ?ineei, ui ?inee a anaioe/ieo oiiaao. Aeey
oa?iaioaoe?? ye iniioee aeei?enoiaoaaee noi?o ??aieo ia’?i?a ?ic/eio W5
? iiaeeo?eiaaiiai iiaeeaiiai na?aaeiaeua EI 8? c oei aea neeaaeii iaeoi-
? oeaethe?oe/ieo oa?iaio?a, yea aeia?a naaa ca?aeiiaiaeoaaea aeey
aeae?eaiiy i?ioiieano?a ni?.

Eoeueoeaoaaiiy i?iaiaeeee ia oiio ae iiaeeaiiio na?aaeiaeu?, ui
aeei?enoiaoaaee ? aeey i?ioiieano?a ni?. Ia?oa ae?eaiiy i?ioiieano?a
ai?ioo, ethoea?ie ? eiithoeie a?aecia/aee ia 2-3 aeiao eoeueoeaoaaiiy. O
i?ioiieano?a ethieio ? “aieoaeiiai” ai?ioo Cajanus cajan ia?o? ae?eaiiy
niinoa??aaee ia 5-6 aeiao eoeueoeaoaaiiy.

Aoaeoeai?noue aen?ao i?ioiieano?a o ??cieo aeae?a c ?iaeeie aiaiaeo aoea
iaiaeiaeiaith. Iaea?eueo aeoeaii ae?eeeeny i?ioiieanoe ni?, ? c aeineoue
iecueeith aoaeoeai?noth i?ioiieanoe ethieio. I?e eoeueoeaoaaii? ia
iiaeeaieo na?aaeiaeuao c i?aeaeuaiei ai?noii oeeoie?i?i?a ee?oeii?
eoeueoo?e “aieoaeiiai” ai?ioo ? ethieio ooai?thaaee caeai? eieii??, c
yeeo ia ?icaeaaeeny iaaiie, a o ethieio niiioaiii aeieeaee no?oeoo?e,
noiae? ia ei?ai?. O eaeonieo e?i?e ai?ioo, io?eiaieo c i?ioiieano?a c
aeoaea iecueeith aoaeoeai?noth, aaeaaaeiny ?iaeoeoaaoe ooai?aiiy iaaii?a
(Eo/oe, 1989; Sarangi et al., 1992).

O aeae?a c ?iaeeie aiaiaeo, ui iaeaaeaoue aei a?oie oo?aaeieo aai
ianiaeuieo, aaeaeiny io?eiaoe noaa?eueio ?aaaia?aoe?th ?ineei c
i?ioiieano?a. Oae, o ethoea?ie ia iiaeeaiiio na?aaeiaeu? N c 0.5 ia/e
e?iaoeio o 20% aenaaeaeaieo eieii?e aoei ?iaeoeiaaii ooai?aiiy
aia??i?ae?a, ui a iiaeaeueoiio oi?ioaaee ii?iaeuei? ?ineeie ia
aacai?iiiaeueieo iiaeeaieo na?aaeiaeuao.

Oaeei /eiii, iai aaeaeiny ?ic?iaeoe iaoiaee aeae?eaiiy ? eoeueoeaoaaiiy
i?ioiieano?a aeey aeae?a c ?iaeeie aiaiaeo. Iiai?noth aoea ae??oaia
i?iaeaia io?eiaiiy aeinoaoiuei? e?eueeino? ee?oeiieo eieii?e. ?acii c
oei ieoaiiy ?iaeoeoe?? aeniei? aoaeoeaiino? ?aaaia?aoe?? ?ineei o
io?eiaieo eeii?a aeiaaa? aeiaeaoeiaeo coneeue.

A ni?a?ia?oieoeoa? c Aioai?/iei ?inoeoooii Oi?aa?neoaoo i. Ithioaia aoa
?ic?iaeaiee iaoiae eoeueoeaoaaiiy i?ioiieano?a ? ?aaaia?aoe?? ?ineei
aeey Oenothera hookeri (Kuchuk et al., 1998). Aeaee oeueiai ?iaeo
iathoue ?yae oi?eaeueieo aaiaoe/ieo iciae, na?aae yeeo aeaoaaouee?anueea
oniaaeeoaaiiy oeeoiieaciaoe/ieo aai?a, iiaeeea?noue io?eiaiiy
i?aeaeaeiaeo a?a?eae?a, oaiiiai eiiieaenii? aaoa?iceaioiino?. Nenoaia
eoeueoeaoaaiiy i?ioiieano?a Oenothera a?oioo?oueny ia oaeaeeiio
cieaeaii? eiioeaio?aoe?? iniioeea a iiaeeaiiio na?aaeiaeu?, aea
eoeueoeaoaaeeny 4-5 – aeaii? i?e?ieieii??. Oe? aenia?eiaioe a?aee?eee
iiaeeea?noue aeey aeine?aeaeaiiy yaea?ii-oeeoiieaciao?/ieo aca?iiae?e c
aeei?enoaiiyi iaoiae?a niiaoe/ii? a?a?eaeecaoe?? ? o?ainoi?iaoe??
oei?iieanoii? AeIE o oaeiai iiaeaeueiiai ia’?eoa ye Oenothera.

Niiaoe/iee aia??iaaiac o eiithoeie eoaiai?. Aeey aea/aiiy i?ioean?a
?aaaia?aoe?? ?ineei a eoeueoo?? in vitro o oaei? oe?iei
?iciianthaeaeaii? ei?iiai? ?ineeie c ?iaeeie aiaiaeo ye eiithoeia eoaiaa
aoei i?iaaaeaia a?noieia?/ia aea/aiiy ooai?aieo ?aaaia?aio?a.

A ?iaio? aoee aeei?enoai? iacio?euei? i?ioiieanoe eiithoeie ye ia?aeii?
iaeiiee?oeii? no?oeoo?e, ui aeicaieeei iiaeaethaaoe nenoaio ?aaaia?aoe??
?ineeie c iaei??? ee?oeie. A i?ioean? eoeueoeaoaaiiy iacio?eueieo
i?ioiieano?a, aeae?eaieo ca ?ic?iaeaiith iaie iaoiaeeeith, aoei
aeyaeaii, ui i?e aoaeoeaiino? ooai?aiiy ee?oeiieo eieii?e, ui aeinyaa?
30% a?ae e?eueeino? aeae?eaieo i?ioiieano?a, eeoa 1,5% a?ae ?o
e?eueeino? aoee caeaoi? aei iiaeaeueoiai ?icaeoeo c ooai?aiiyi
niiaoe/ieo aia??i?ae?a. Oe? no?oeoo?e ia ooai?thaaee aeiaeiiai
i?ii?aeiiai eaeona, ? aia??i?aee aaciina?aaeiuei ?icaeaaeeny a iaaiie.
Io?eiai? ?acoeueoaoe noaee na?aeioeoaii iiaeeeaino? ?iaeoeoe?? i?yiiai
aia??iaaiaco c i?ioiieano?a eiithoeie, ui i?aeoaa?aeaeo? a?iioaco i?i
iiaeeea?noue ?nioaaiiy i?iaia??iiaeueieo ee?oei a niiaoe/ieo oeaieiao
eenoa eiithoeie. Iaaeae?, i?yia io?eiaiiy iiaiioe?iieo ?ineei c
i?i?inoe?a noeeioeiny c oneeaaeiaiiyie, ? oeae i?ioean cae?enithaaany
oeyoii ?iaeoeoe?? aoi?eiiiai niiaoe/iiai aia??iaaiaco.

I?ioean ooai?aiiy aoi?eiieo aia??i?ae?a o eiithoeie aoa oaeiae aea/aiee
ca aeiiiiiaith a?noieia?/iiai aiae?co. I?iaaaeai? niinoa?aaeaiiy
iieacaee, ui ooai?aiith aia??i?ae?a ia?aaeo? aeieeiaiiy i?iaia??iaaiieo
a?oi ee?oei, ye? ?icaeaaeeny c iaei??? aai aaaaoueio non?aenueeeo
aaeeeeo, aeoaea aaeoie?ciaaieo ee?oei. I?iaia??iiaeuei? a?oie ee?oei
yaeyee niaith ae??aiiee?oeii? no?oeoo?e c u?eueiith oeeoiieaciith ?
aeineoue aaeeeeie yae?aie ? yae?eoeaie. Niiaoe/i? aia??i?aee aeieeaee c
iaei??? aai aeae?eueeio ee?oei oeeo no?oeoo?. ?i?oe?aeuei? ee?oeie
aia??i?ae?a a?ae??ciyeeny a?ae iaaeieeoi?o ee?oei iioiauaiith
iaieiieith, ui i?inoaaeoaaeiny o iiaeaeueoiio ia aeoaea ?aii?e noaae??
?oiueiai ?icaeoeo. Noaae?? ?icaeoeo niiaoe/ieo aia??i?ae?a eiithoeie
noiae? c eeane/ieie, iienaieie ye aeey ceaioe/iiai aia??iaaiaco, oae ?
aeey niiaoe/iiai aia??iaaiaco ii?eae. Oe?eeii /?oei aeae?eythoueny
aeiaoey?ia, na?oeaaeaeia, oi?iaaeiaeaeia noaae??, a oaeiae noaae?y
i?i?inoaiiy. A ?acoeueoao? ooai?ththoueny a?iiey?i? no?oeoo?e c?
noaaeiaei ? ei?aiaaeie aiaenaie.

?acii c oei a ?icaeoeo niiaoe/ieo aia??i?ae?a eiithoeie niinoa??aaany
?yae a?aei?iiinoae a?ae i?ioeano ooai?aiiy ceaioe/ieo aia??i?ae?a. Oae,
aia??i?aee, ui noi?ioaaeeny c iaei??? ee?oeie, iaee a?eueo aeoyaiooo
aacaeueio /anoeio, ui cae??ieth? ?o a iaoa?einuee?e oeaiei?
aia??iiaeueiiai eiiieaeno. Iiaeeeai, oey no?oeoo?a aeeiio? ?ieue
i?aea?nea, ? ?? aaaaoiee?oeiia no?oeoo?a aeieeea aaea i?ae /an iaeia?oeo
noaae?e niiaoe/iiai aia??iaaiaco eiithoeie. O ceaioe/ieo aia??i?ae?a
eiithoeie i?aea?nie oaeiae aaaaoiee?oeiiee, aea oi?io?oueny a?i ia
i?ci?o?e aeiaoey?i?e noaae??. A oie aea /an a oi?ioaaii? niiaoe/ieo
aia??i?ae?a niinoa??aaeany iiaia aeeoa?aioe?aoe?y ca /anii ooai?aiiy
i?aea?nea. *anoeia c ieo, oae ye ? ceaioe/i?, oi?ioaaea aaaaoiee?oeiiee
i?aea?nie ia i?ci?e aeiaoey?i?e noaae??, ? a?i i?inoaaeoaaany a
oi?iaaeiaeaei?e noaae?? ? aeae? a ia??iae i?i?inoaiiy. Ii i??? ?icaeoeo
aia??i?aea ee?oeie i?aea?nea caeeoaeeny ae??aieie ? u?eueiiieaciaiieie,
a oie /an ye ee?oeie aeania aia??i?aea noaaaee a?eueoeie ? aeaaeae?
neeuei?oa aaeoie?ciaaieie, ui oa?aeoa?ii aeey niiaoe/ieo aia??i?ae?a,
ye? ?icaeaathoueny a oiiaao in vitro. I?ae ciiith aaaaoiee?oeiiiai
i?aea?nea ? aeania aia??i?aeii aeieeaea iaaaeeea ?iaaa?iaoe?y,
oa?aeoa?ia ? aeey ceaioe/ieo ca?iaee?a.

Aeaye? niiaoe/i? aia??i?aee iaee oe?ieo ciio i?e?aeiaiiy aei
aia??iaaiiiai eiiieaeno, ui i?aeoaa?aeaeo? ?oi? aaaaoiee?oeiia
iioiaeaeaiiy. Oae? ca?iaeee aeieeaee nii/aoeo o aeaeyae? oae caaieo
aeieiaiue, ui neeaaeaeeny c ae??aieo ee?oei ia iiaa?oi? aia??iiaeueiiai
eiiieaeno, caiaeii aiie aeeoa?aioe?thaaeeny a a?iiey?i? no?oeoo?e.

I?i?inoaiiy (i?aaiiaaiac) o niiaoe/ieo aia??i?ae?a eiithoeie
a?aeaoaa-einy ia oae /?oei, ye o ceaioe/ieo. Iae/ano?oa cai?cithaaeiny
ooai?aiiy ei?aiy, oiae? ye n?i’yaeie? ?icaeaaeeny ii?iaeueii, ? a ieo
caeeaaeaeany i?ia?aeia nenoaia. ?ieiee ?icaeoie ei?aiaaiai aiaeno
i?eai?/oaaany iaeaea iiai?noth, i?i ui na?ae/ea aeineoue aocueeee
aacaeueiee e?iaoeue aia??i?aea. Aea a ?yae? aeiaaee?a, a?eueo
?icaeiaiith aeyaeyeany ei?aiaaa /anoeia aia??i?aea ? iaio ?icaeiaieie
n?i’yaeie?. Niinoa??aaeiny oaeiae ia??aiii??ia ooai?aiiy iaio
n?i’yaeieae. Aeineoue /anoi aeieeaee oanoeeeiaai? aia??i?aee, iaaii
caaaeyee u?eueiiio ?icoaooaaiith ?oi?o ?i?oe?aeae. Ia??aee?nieie aoee
aeiaaeee aoi?eiiiai aia??iaaiaco, eiee caeeaaeea iiaeo aia??i?ae?a
a?aeaoaaeany a oeaieiao iiia?aaei?o, ui ciaoiaeeeeny ia i?ci?oeo
noaae?yo ?icaeoeo, aea ca yeeiinue i?e/eiaie coieieaoeo na?e ??no.
Aoi?eii? aia??i?aee oaeiae iaee ye iaeii-, oae ? aaaaoiee?oeiia
iioiaeaeaiiy. A aaaaoueio aeiaaeeao aia??i?aee coieiyee na?e ?icaeoie ia
aeiaoey?i?e aai oi?iaaeiaeaei?e noaae?yo. ?o ee?oeie ca?eueooaaeeny,
aaeoie?coaaeeny ? aeaaaee ii/aoie iiaiio aia??iaaiiiio eaeono.

Iao? aeine?aeaeaiiy na?ae/aoue i?i cia/io aiaeia?th a noaae?yo ?icaeoeo
ceaioe/iiai ? niiaoe/iiai aia??iaaiaco eiithoeie. Io?eiai? c niiaoe/ieo
aia??i?ae?a ?ineeie eiithoeie oni?oii ae?iuoaaee ye a oiiaao oaieeoe?,
oae ? ?oi? iauaaeee – a oiiaao a?aee?eoiai ??oioo aac iii?oieo a?aei?i
a?ae ?ineei aeo?aeiiai aaiioeio (?aaeeiiaiei oa ?io., 1994; Radionenko
et al., 1994).

A?noieia?/ia aea/aiiy ?aaaia?aoe?eieo e?i?e ai?ioo. Aeey iiaeaeueoiai
aea/aiiy cania?a ?aaaia?aoe?? o aeae?a c ?iaeeie aiaiaeo aoea
aeei?enoaia ee?oeiia e?i?y ai?ioo, ui noaa?eueii ooai?th?
?aaaia?aoe?eiicaeaoiee eaeon. I?e a?noieia?/iiio aea/aii? i?ioeano
?aaaia?aoe?? o io?eiaieo e?i?e ai?ioo ia aoei aeyaeaii /?oeeo oac
?icaeoeo, ye oea ia? i?noea o aeiaaeeo niiaoe/iiai aia??iaaiaco. A oie
aea /an a?aecia/aii ?icaeoie i?iaia??iaaiieo ae?eyiie, ui neeaaeaeeny c
ae??aieo ee?oei c u?eueiith oeeoiieaciith, ui ioi/ai? eaeonieie
ee?oeiaie cia/ii a?eueoiai ?ici??o. Oe? i?iaia??iiaeuei? a?oie ee?oei
i?e eoeueoeaoaaii? ?o ia aacai?iiiaeueieo iiaeeaieo na?aaeiaeuao aeaaaee
ii/aoie ooai?aiith a?oiueie, ? iiaeaeueoee ?icaeoie a?aeaoaaany ii oeyoo
i?aaiiaaiaco c ooai?aiiyi iaaii?a. Iaeiae, yeui oe? ee?oeii? e?i?? ia
i?ioyc? ei?ioeiai /ano (3-5 aeiae) eoeueoeaoaaoe a i?enooiino? 2.4 Ae, a
i?ney oeueiai ia?aianoe ia aacai?iiiaeueia iiaeeaia na?aaeiaeua, oi
?ieiee a?aei?/aeiny ooai?aiiy no?oeoo?, noiaeeo c aeiaoey?ieie.

A?noieia?/ia aea/aiiy i?ioean?a ?aaaia?aoe?? o oeoe?iaiai ao?yeo.
Oeoe?iaee ao?ye – iaeia c iaeaaaeeea?oeo n?euenueeiainiiaea?nueeeo
eoeueoo?. Aea/aiiy i?ioean?a ?aaaia?aoe?? o oe??? eoeueoo?e ia? aaeeea
cia/aiiy aeey ?ic?iaee a?ioaoiieia?/ieo iaoiae?a ? canoinoaaiiy ?o aeey
iie?ioaiiy ?aeiiiaaieo a Oe?a?i? ni?o?a. sse aenieaioe aeei?enoiaoaaee
/a?aoee eenoe?a, ui eoeueoeaoaaee ia iiaeeaiiio na?aaeiaeu? IS, yea
i?noeoue 1ia/e AAI. I?ney ia?aaoaaiiy o oeeo oiiaao c ee?oei ia?aio?ie
cieeaee aeeth/aiiy. Ee?oeie ?icii/eiaee ae?eaiiy, a yeeo nii/aoeo a?aee
o/anoue o?eueee aeae?eueea oa??a ee?oei ia?aio?ie. Ae?eaiiy a?aeaoaaeiny
ii oeio ae?iaeaiiy, a ?acoeueoao? /iai ooai?th?oueny ciia ae??aieo
ee?oei, c a?eueo u?eueiith oeeoiieaciith ? a?eueoeie yae?aie.
Ooai?ththoueny ciie, iiae?ai? aei ?ai?oa iienaieo iaie aeey eiithoeie ?
ai?ioo. Iaaeae? i?ioean a?aeaoaa?oueny ii oeio i?aaiiaaiaco. A ie?aieo
ae?eyieao ia?enoaiaoe/ieo cii, ui ooai?ththoueny, ii/eiathoue aeieiaoeny
aoai?ee. Oeaio?aeueia /anoeia aoai?ea caiaeii aeeoa?aioe?th?oueny a
aiaen iaaiia, a a ia?eoa??ei?e cii? aiaena a ?acoeueoao? ia?eeeeiaeueieo
iiae?e?a ee?oei ? aeayeiai ?oiueiai ?icoyao ooai?ththoueny eenoia?
aoai?ee. Oaeei /eiii oi?iothoueny a?oiueee. Ii aeiaaeei? /a?aoea
aiaeii/an caeeaaea?oueny aeae?eueea a?oiueie (Aaiieeiaa oa ?io., 1995)

C i?iaaaeaieo aenia?eiaio?a iiaeia c?iaeoe aeniiaie, ui i?ioean
?aaaia?aoe?? ye oeyoii i?aaiiaaiaco o ai?ioo ? oeoe?iaiai ao?yea, oae ?
oeyoii niiaoe/iiai aia??iaaiaco o eiithoeie aa?a na?e ii/aoie ia noaae??
ooai?aiiy ae??aiiee?oeiieo no?oeoo?, ui aecia/athoueny ye
i?iaia??iiaeueiee eiiieaen, aea i/aaeaeii ? a?aeaoaa?oueny aeoea?caoe?y
iaaieo aai?a, ui a?aeiia?aeathoue ca iaoai?cie ii?oiaaiaco. Naia ia oe?e
oac? iiaeiii a?aeaoaaoeny iaeiie/aiiy niaoeeo?/ieo iao?e/ieo ?IE, ye?
iiaeooue aooe eeth/ai aeey iiooeo aai?a, caeo/aieo aei i?ioeano
?aaaia?aoe??. Oiaoi i?ioeane, ui i?eaiaeyoue o iiaeaeueoiio aei
ooai?aiiy ii?iaeueii? ?ineeie c niiaoe/ieo ee?oei, a?aeaoaathoueny
aieiaiei /eiii ia noaae?? ooai?aiiy i?iaia??iiaeueieo a?oi ee?oei.
Iaaeae?, ?icaeoie oeeo ee?oei iiaea eoe ye ii oeyoo niiaoe/iiai
aia??iaaiaco, oae ? ii oeyoo i?aaiiaaiaco a caeaaeiino? a?ae aaiaoe/ieo
? o?c?ieia?/ieo /eiiee?a.

ANEIAO?E*IA NIIAOE*IA A?A?EAeECAOe?ss ssE IAeEI ?C NIINIA?A IA?AIINO
AAIAOE*II? ?IOI?IAOe?? I?AE ?INEEIAIE.

Io?eiaiiy niiaoe/ieo a?a?eae?a i?ae aeaeaie c ?iaeeie aiaiaeo c
aeei-?enoaiiyi aaiia-ii?ii?iaiiy. Iacaaaeath/e ia oni?oe, yeeo
aeinyaioea a inoaii?e /an aaiaoe/ia ?iaeaia??y ye i?aeo?ae, caniiaaiee
ia iaoiaeao aaaaeaiiy a ?ineeiio ee?oeio niaoe?aeueii noai?aieo
aaeoi?ieo iieaeoe AeIE, ia iiaeia iaaeiioe?ithaaoe iiaeeeainoae
ee?oeiii? ?iaeaia??? ?ineei. ?acii c oei iaiao?aeii a?aecia/eoe, ui
iiae?e ia aaiaoe/io ? ee?oeiio ?iaeaia??th ?ineei ? aeineoue oiiaiei, ?
a na?oia?e iaoeia?e i?aeoeoe? aiie ?icaeyaeathoueny ye oe?e?nia nenoaia
iiaeo i?aeoiae?a i?ae caaaeueiith iacaith aaiaoe/ia ?iaeaia??y ?ineei.

A iaoeo aenia?eiaioao ii aneiao?e/i?e niiaoe/i?e a?a?eaeecaoe?? i?ae
aeaeaie c ?iaeeie aiaiaeo, ui ?icii/aeeny a 1987 ?ioe?, aoee ?ic?iaeai?
iaoiaee niiaoe/ii? a?a?eaeecaoe?? c aeei?enoaiiyi aaiia-ii?ii?iaiiy
iaeiiai c ia?oia??a, ? ye ?acoeueoao aoee a?ae?a?ai? a?a?eaei? eeiie ?
?aaaia?iaai? a?a?eaei? ?ineeie.

Aeey ?iaeoeoe?? ceeooy i?ioiieano?a i?ae ??ciiiai?oieie aeaeaie c ?iaeo
Medicago, a oaeiae i?ae eiithoeiith (Trifolium pratense L.) ?
ethoea?iith i?ieeaith (M. varia L.) ca iniiao aoei acyoi iaoiae,
?ic?iaeaiee aeey ceeooy iacio?eueieo i?ioiieano?a o N. plumbaginifolia
ca?aeii yeiio noi?o i?ioiieano?a ia?iaey?oueny ia i?ioyc? 20 oa ?ic/eiii
40% IAAa 4000, ui i?noeoue aenieo eiioeaio?aoe?th ?ii?a Na2+ ? ?I ??aiee
9.5-10. Canoinoaaiiy oeueiai iaoiaeo ni?eyei ye io?eiaiith a aeinoaoi?e
e?eueeino? i?iaeoeo?a ceeooy oae ? aeinoaoiueiai a?aenioeo aeaeeaaiiy
aaoa?iea??ii?a ? aaouee?anueeeo i?ioiieano?a.

A ?acoeueoao? i?iaaaeaieo aenia?eiaio?a, i?ney ceeooy iacio?eueieo
i?ioiieano?a M. sativa, ni?o “Aanaeiiiaeieyinueea” c ii?ii?iaieie
aeicith 300A? iacio?eueieie i?ioiieanoaie M. varia, ui ianooue aai
no?eeino? aei eaiai?oeeinoeueoaoo, aoee a?ae?a?ai? a?a?eaei? ee?oeii?
eeiie. O aeayeeo c oeeo eeii?a aaeaeiny ?iaeoeoaaoe ooai?aiiy
aia??iiiae?aieo no?oeoo?, a a iaeiiio aeiaaeeo aoee io?eiai?
?ineeie-?aaaia?aioe (Eo/oe oa ?io., 1991). Iiaeia i?eionoeoe, ui
io?eiai? a?a?eaei? eeiie i?noyoue a nai?io aaiii? aaie M. varia, ui
iaoiiaeththoue ?aaaia?aoe?eio caeaoi?noue, aaeaea aeey eieii?e,
io?eiaieo c i?ioiieano?a ethoea?ie ni?oo “Aanaeiiiaeieyinueea”, aeiaeii?
?aaaia?aoe?eii? iiae?? io?eiaoe ia aaeaeiny. I?iaaaeaiee o?iiiniiiee
aiae?c iieacaa, ui a?ae?a?aiee a?a?eae iana ieoiiei?aeiee iaa?? o?iiinii
c 3-4 aeiaeaoeiaeie o?iiiniiaie.

Iaaeaeue o ethoea?ie aaaeei aeeoa?aioe?thaaoe o?iiiniie aaouee?a, iaeiae
?acoeueoaoe, iioae?eiaai? noiniaii aneiao?e/ii? niiaoe/ii?
a?a?eaeecaoe?? o aeae?a ?iaeo Nicotiana, a oaeiae i?ae Nicotiana ?
Atropa, na?ae/aoue, ui a oaeeo eiia?iaoe?yo caeaa?eueoiai
a?aeae?athoueny a?a?eaee c iiaeai?iei iaai?ii o?iiinii ?aoeei??ioa ?
aeayeei /eneii o?iiinii aeiii?a. A iaoeo aenia?eiaioao ie aeei?enoaee
aaouee?anuee? aeaee c oao?aiei?aeiei /eneii o?iiinii 2n=32, ui i?ecaaei
aei io?eiaiiy a?a?eaeo c ieoiiei?aeiei iaai?ii c aeiaeaoeiaith
e?euee?noth o?iiinii aeiii?a.

Ii?oieia?y io?eiaieo ?ineei ?noioii a?ae??ciyeany a?ae iaio aaouee?a.
Oae, a?eueo?noue eenoe?a aoea iaii?iaeueii? ?aaeoeiaaii? oi?ie. ?ineeie
iaee neeueii oei?i/ai? i?aeaocey, ? o ieo niinoa??aaeiny oneeaaeiaiiy
ei?aiaooai?aiiy a eoeueoo?? in vitro. Oaeee oaiioei i?a aooe
?acoeueoaoii ye iaaii? ianoi?niino? aaiii?a ?aoeei??ioa ? ii?ii?iaiiai
aeiii?a, oae, iiaeeeai, ? ne?aenoaii o?eaaeiai ae?iuoaaiiy a?a?eaeieo
ee?oeiieo eoeueoo? ia?o i?ae aaeaeiny ?iaeoeoaaoe ?aaaia?aoe?th o
io?eiaiiai a?a?eaeiiai ee?oeiiiai eeiio.

C i?aeoe/ii? oi/ee ci?o a?eueo oe?eaaith aoea ae?oaa a?a?eaeia
eiia?iaoe?y i?ae ?ioeie aeaeaie c ?iaeo Medicago. Iaeei c i?aaenoaaiee?a
oeueiai ?iaeo – ethoea?ia oiaeaaeaeia (M. lupulina) caeaoia aei
naiicaieeaiiy, ui ? aeoaea ei?eniith iciaeith c oi/ee ci?o naeaeoe??
ethoea?ie ca i?aeaeuaiith i?iaeoeoeai?noth ian?iiy. A oie aea /an
ethoea?ia oiaeaaeaeia acaaae? ia caeaoia aei noaoaaiai no?auoaaiiy c
ieoeueoo?aieie aeaeaie ethoea?ie, ui ia aeicaiey? caeo/aoe ?? aei
cae/aeieo naeaeoe?eieo i?ia?ai.

A i?iaaaeaieo aenia?eiaioao i?ney ceeooy iacio?eueieo i?ioiieano?a M.
borealis e?i?y 94 c ii?ii?iaieie i?ioiieanoaie M. lupulina RiA4,
?cieueiaaieie c io?eiaii? ?ai?oa o?ainaaiii? eaiai?oeeino?eei? ee?oeiii?
e?i??, aaeaeiny a?ae?a?aoe ee?oeii? e?i?? a aeai?e a?a?eaei?e
eiia?iaoe??. Iaiao?aeii a?aecia/eoe, ui ii?oieia?y iaea?aeaieo eeii?a
aoea noiaea nei??oa ia ii?oieia?th ee?oeiieo e?i?e M. borealis. Io?eiai?
aneiao?e/i? niiaoe/i? a?a?eaee ?icaeaaeeny o aeaeyae? caeaieo ee?oeiieo
e?i?e aac aoaeue-yeeo iciae ei?aiaooai?aiy. ?aaaia?aoe?y ?ineei o
a?a?eaeieo eieii?e aoea neeueii oneeaaeiaia ? iai ia aaeaeiny io?eiaoe o
ieo ii?iaeuei? iaaiie.

Aea/aiiy ca aeiiiiiaith aeioeia a?a?eaeecaoe?? ca Naoca?iii
oniaaeeoaaiiy ?eainiiaeuei?o aai?a o io?eiaieo a?a?eaeieo ee?oeiieo
e?i?e i?ae M. borealis ? M. lupulina iieacaei, ui o aneiao?e/ieo
niiaoe/ieo a?a?eae?a oe? aaie i?enooi? a?ae iaio ia?oia??a.

Aiaeia?/iee aiae?c aoa i?iaaaeaiee aeey eiia?iaoe?? M. sativa+ M. varia.
O aeiaaeeo aeei?enoaiiy aiaeiioeeaace ?ano?eeoe?? EcoRV ia aoei
aecia/aii aeiaeieo a?aei?iiinoae a niaeo?ao a?a?eaeecaoe?? o oeeo aeaio
aeae?a. I?ioa o a?a?eaeieo eeii?a a?aei?/aeiny aeieeiaiiy iiaeo nioa
a?a?eaeecaoe??. Oea yaeua iiaea aooe iiyniaia aeieeiaiiyi iiaeo eean?a
?eainiiaeueieo iiaoi??a o niiaoe/ieo a?a?eae?a, ui aaea a?aecia/aeiny
?ai?oa o niiaoe/ieo a?a?eae?a i?ae N. plumbaginifolia ? Atropa
belladonna.

I?eiouaiiy i?i iiaeeea?noue ia?aianaiiy aai?a, ui ?aaoeththoue
caeaoi?noue aei ?aaaia?aoe?? a eoeueoo?? in vitro, oeyoii niiaoe/ii?
a?a?eaeecaoe?? i?ae ??cii-iai?oieie aeaeaie c ?iaeo Medicago
i?aeoaa?aeaeo?oueny oaeiae ?acoeueoaoaie, io?eiaieie i?e a?a?eaeecaoe??
i?ioiieano?a eiithoeie eoaiai?, ?cieueiaaieo c o?eaaei eoeueoeaiaaieo
ee?oeiieo eoeueoo?, ui ao?aoeee ye caeaoi?noue aei ?aaaia?aoe??, oae ?
aei neioaco oei?io?eo, c ii?ii?iaieie iacio?eueieie i?ioiieanoaie M.
varia, ye? iathoue iciaeo no?eeino? aei eaiai?oeeinoeueoaoo, aai aac
oe??? iciaee. O a?a?eaeieo e?i?e, iicia/aieo iiia?aie 3 ? 4, ui aoee
io?eiai? i?ney ceeooy i?ioiieano?a eiithoeie c ii?ii?iaieie
iacio?eueieie i?ioiieanoaie ethoea?ie, ye? ia ianooue naeaeoeaieo iciae,
aoei ?iaeoeiaaii ooai?aiiy aia??iiiiiae?aieo no?oeoo?, a i?ney oeueiai ?
iaaii?a.

Iaaii aaie, ui aecia/athoue caeaoi?noue aei ?aaaia?aoe?? o ethoea?ie, ia
c/aieai? c aaiaie no?eeino? aei eaiai?oeeinoeueoaoo, oiio ui i?ney
a?a?eaeecaoe?? i?ioiieano?a eiithoeie c iacio?eueieie eaiai?oeeino?eeeie
i?ioiieanoaie ethoea?ie aoee io?eiai? ye caeai?, oae ? oei?io?eaeaoaeoi?
eeiie,no?ee? aei eaiai?oeeinoeueoaoo. A oie aea /an iaia? aeaieo i?i
aieea aeice ii?ii?iaiiy ia naa?aaaoe?th oeeo iciae. sse i?e aeic?
ii?ii?iaiiy iioiieano?a ethoea?ie a 1000 A?, oae ? i?e aeic? a 25 A?
aoee io?eiai? eaiai?oeeino?ee? a?a?eaei? eeiie i?ae eiithoeiith ?
ethoea?iith, caeaoi? aei iicaeai?iiy.

Oaaeeoey 3. Aneiao?e/i? niiaoe/i? a?a?eaee, io?eiai? a ?EAA? i?ae
aeaeaie ?iae?a Medicago ? Trifolium.

?ineeia-?aoeei??ioIi?ii?iaiee aeiii?Naeaeoeaia iciaeaOei a?a?eaeieo
e?i?eMedicago sativa

(iacio?e)

Medicago borealis

(iacio?e)

Medicago borealis

(iacio?e)

Trifolium pratense

(a?eee eaeon)

Trifolium pratense

(a?eee eaeon)M. varia

(iacio?e)

M. lupulina

(eaeon)

M. varia

(iacio?e)

M. varia

(iacio?e)

M. varia

(iacio?e)No?ee?noue aei eaiai?oeeio

No?ee?noue aei eaiai?oeeio

No?ee?noue aei eaiai?oeeio

Caeaoi?noue aei iicaeai?iiy

No?ee?noue aei eaiai?oeeioIaaiie c ei?aiyie

Caeaiee eaeon

Caeaiee eaeon

Iaaiie ? aia??i-iiae?ai? no?oeoo?e

Caeaiee ? a?eee

eaeon

Iaiao?aeii a?aecia/eoe, ui caeaoi?noue aei ?aaaia?aoe?? o aneiao?e/ieo
niiaoe/ieo a?a?eae?a a cia/i?e i??? aoea cieaeaia o ii??aiyii? c
ee?oeiaie aeiii?a aai ?aoeei??ioa. Oae, a aenia?eiaioao i?ney ceeooy
iacio?eueieo i?ioiieano?a M. borealis, e?i?y 94, c ii?ii?iaieie
iacio?eueieie i?ioiieanoaie M. varia, ui ianooue aai no?eeino? aei
eaiai?oeeinoeueoaoo, a?ae?a?ai? a?a?eaei? eeiie ia ooai?thaaee
aia??iiiae?aieo no?oeoo?. A oie aea /an io?eiai? c i?ioiieano?a aaouee?a
ee?oeii? e?i?? c aeinoaoiuei aenieith aoaeoeai?noth oi?ioaaee oae?
no?oeoo?e ? ooai?thaaee ii?iaeuei? ?ineeie.

A ?acoeueoao? i?iaaaeaieo aenia?eiaio?a aoee ?ic?iaeai? iaoiaee ceeooy
i?ioiieano?a o aeae?a Medicago ? Trifolium c canoinoaaiiyi ?iaeoeaaoe??
iaeiiai c ia?oia??a ca aeiiiiiaith aaiia-ii?ii?iaiiy. Oaeiae aoea
iieacaia iiaeeea?noue ia?aiino i?e aneiao?e/i?e niiaoe/i?e
a?a?eaeecaoe?? o aeae?a Medicago ? Trifolium iciaee no?eeino? aei
eaiai?oeeinoeueoaoo, aaoaeiaaiiai a aaiii aeiii?a oeyoii aaiaoe/ii?
?iaeaia???. E??i oiai, aoea iieacaia iiaeeea?noue ia?aiino iciaee
?aaaia?aoe?eii? caeaoiino? i?e aneiao?e/i?e niiaoe/i?e a?a?eaeecaoe?? o
oeeo aeae?a.

Io?eiai? ?acoeueoaoe ii aneiao?e/i?e niiaoe/i?e a?a?eaeecaoe?? o aeae?a
c ?iae?a Medicago ? Trifolium a?aee?eaathoue iia? ia?niaeoeae a
naeaeoe?? oeeo aaaeeeaeo n?euenueeiainiiaea?nueeeo eoeueoo?. Aiie
aeicaieythoue iaea?aeoaaoe iiaee aeo?aeiee naeaeoe?eiee iaoa??ae c
ei?enieie aaiaie, ia?aianaieie c ?iae?a ? aeae?a, o yeeo cae/aeia
no?auoaaiiy iaiaaeaia aai acaaae? iaiiaeeeai. E??i oiai, a?aee?eaa?oueny
iiaeeea?noue aeei?enoaoe aeai? i?aeoiaee aeey io?eiaiiy oeea??ae?a o
oeeo aeae?a.

AENIIAEE

1. Ai?iaiaaii nenoaio aaiaoe/ii? o?ainoi?iaoe?? oeyoii aeaeo?iii?aoe??
i?ioiieano?a. Io?eiaii eaiai?oeeino?ee? ?ineeie ethoea?ie ? ee?oeii?
e?i?? ni? oa ai?ioo. Iieaeoey?ii-a?ieia?/iee aiae?c ? aiae?c aeoeaiino?
oa?iaioo iaii?oeeioinoio?ainoa?ace i?aeoaa?aeea o?ainaaiio i?e?iaeo
io?eiaieo e?i?e.

2. Cai?iiiiiaaii ? ai?iaiaaii iaoiae, ui i?aeaeuo? ?aaaia?aoe?eio
caeaoi?noue o ethoea?ie ? ai?ioo oeyoii aaiaoe/ii? o?ainoi?iaoe??
iooaioii “shooty” Agrobacterium tumefaciens pGV 2206. Io?eiaii
?aaaia?aoe?ei? e?i?? ethoea?ie ? ai?ioo, ui aeicaiey? aeei?enoiaoaaoe ?o
aeey aaiaoe/ii? o?ainoi?iaoe?? ?ioeie aaiaie.

3. E?i?y o?ainaaiiiai ai?ioo c i?aeaeuaiith caeaoi?noth aei ?aaaia?aoe??
aoea aeei?enoaia aeey ianooiii? o?ainoi?iaoe?? aaeoi?ieie
eiino?oeoe?yie, ui ianooue naeaeoeaiee aai no?eeino? aei
eaiai?oeeinoeueoaoo. Io?eiaii o?ainaaii? iaaiie ai?ioo, ui ianooue
iia?eueiee Ds-aeaiaio eoeo?oaece. Iieacaii iiaeeea?noue aeei?enoaiiy
“iiaea?eii? o?ainoi?iaoe??” aeey io?eiaiiy o?ainaaiieo e?i?e ai?ioo c
aaiaie ,ui oe?eaaeyoue.

4. Cai?iiiiiaaii ? ai?iaiaaii iaoiae aaiaoe/ii? o?ainoi?iaoe?? ai?ioo ca
aeiiiiiaith iaiieiaaiieo ooai?a Agrobacterium tumefaciens. Io?eiaii
o?ainaaii? e?i??, ui ianooue aai no?eeino? aei oino?iio??oeeio.

5. ?ic?iaeaii niin?a ooai?aiiy ian?iiy o ai?ioo a anaioe/i?e eoeueoo??.
Cai?iiiiiaaii iaoiaeeeo i?e?ieeiiaeueiiai ?iciiiaeaiiy ai?ioo, ui
aeicaieeoue eeiioaaoe aei 500 000 aaiaoe/ii iaeii??aeieo iniaei a ??e.

6. O o?ainaaiieo e?i?e ai?ioo oeyoii ?iaeoeoe?? ooai?aiiiy ian?iiy a
eoeueoo?? in vitro io?eiaii iioiinoai. Iieaeoey?ii-a?ieia?/iee aiae?c
i?aeoaa?aeea i?enooi?noue iine?aeiaiinoae AeIE ia?aianaiiai aaia a
iieie?iiyo aaiaoe/ii iiaeeo?eiaaiiai ai?ioo.

7. I?ioiieanoe aeae?a ?ineei c ?iaeeie aiaiaeo, oaeeo ye eiithoeia,
ethoea?ia, niy, ai?io, ethiei, aieoaeiee ai?io, i?e aeei?enoaii?
?ic?iaeaieo iaoiae?a eoeueoeaoaaiiy caeaoi? aei ooai?aiiy ee?oeiieo
eieii?e c aenieei nooiaiai aoaeoeaiino?.

8. Cai?iiiiiaaii ? ai?iaiaaii i?ioieiee ?aaaia?aoe?? ?ineei c ee?oeiieo
e?i?e, io?eiaieo c i?ioiieano?a, ui aoee aeae?eai? c eiia?oe?eieo ni?o?a
ethoea?ie ? eiithoeie. ?aaaia?aoe?y ?ineei o eiithoeie iiaea aooe
io?eiaia ye oeyoii i?yiiai, oae ? iai?yiiai aia??iaaiaco, ui
i?aeoaa?aeaeaii ?acoeueoaoaie a?noieia?/iiai aiae?co.

9. ?ic?iaeaii iaoiaeeeo eoeueoeaoaaiiy i?ioiieano?a ? ?aaaia?aoe??
?ineei o Oenothera hookeri – iiaeaeueiiai ia’?eoo aeey
iieaeoey?ii-aaiaoe/ieo aeine?aeaeaiue.

10. Io?eiaii aneiao?e/i? niiaoe/i? a?a?eaee i?ae aeaeaie c ?iaeeie
aiaiaeo. Iieacaii iiaeeea?noue aeei?enoaiiy ia?aianaiiai aaia
iaii?oeeioinoi-o?ainoa?ace ye aaiaoe/iiai ia?ea?a aeey a?aeai?o
niiaoe/ieo a?a?eae?a o aeae?a c ?iaeeie aiaiaeo.

CIENIE IOAE?EAOe?E

1. Eo/oe I.A. Aaiaoe/aneay eiaeaia?ey aunoeo ?anoaiee.// Ee?a “Iaoeiaa
aeoiea”.-1997.-152n.

2. Aaiieeiaa I.A., Aieiaei A.Y. Oaaaeuie/ I.A., Eo/oe I.A. ?aaaia?aoeey
?anoaiee o naoa?iie naaeeu (Beta vulgaris) a eoeueoo?a in vitro,
aenoieiae/aneia eco/aiea i?ioeannia ?aaaia?aoeee.// Oeeoieiaey e
aaiaoeea.-1995.-29, N 6.-N. 14-22.

3. Coaei A.E., Eo/oe I.A., Ooiaiiaa E.A., Aeeiineay I.A, Aeaaa TH.TH.
Aaiaoe/aneay o?ainoi?iaoeey ai?ioa, iiin?aaeiaaiiay Agrobacterium
tumefaciens// Aeiiieeia?u e eeaoea.-1990.-6, N 3.-P.77-80.

4. Ee?e/aiei E.A., Eo/oe I.A., Naoii E.A., Aeaaa TH.TH.
Eiaeeaeaeoaeueiia eoeueoeae?iaaiea ?anoeoaeueiuo i?ioiieanoia e
eeaoie.// Aeiee. AI Oe?aeiu.-1990.- N 3.-C. 63-65.

5. Eo/oe I.A., Eaiaaneee E.O. Iaoiaeu aaiaoe/aneie o?ainoi?iaoeee
?anoaiee.// Aeioaoiieiaey.- 1987.- 3, N 3. – N. 365-369.

6. Eo/oe I.A. Auaeaeaiea e eoeueoeae?iaaiea i?ioiieanoia iyoe aeaeia
naiaenoaa aiaiauo// Oeceieiaey ?anoaiee.-1989.-3, N 4. -N.821-824.

7. Eo/oe I.A., Oaoianeee A.I., Eiia?ieoeeee E.E., Aeaaa TH.TH. Iieo/aiea
aaiaoe/anee o?ainoi?ie?iaaiiuo eeaoi/iuo eeiiia nie iooai
yeaeo?iii?aoeee i?ioiieanoia.// Aeioaoiieiaey.- 1990.- N 5.- C. 30-31.

8. Eo/oe I.A., Oaoianeee A.I., Ai?iiei A.E., Aeaaa TH.TH. Iieo/aiea
aneiiao?e/iuo niiaoe/aneeo aea?eaeia a ?iaea Medicago.// Aeiiieeia?u e
eeaoea.-1991.-7, N 4.- N. 54-60.

9. Eo/oe I.A., Aeaaa TH.TH. Oeeoio?c?ieia?y oa ee?oeiia ?iaeaia??y
?ineei.// Oe?. aio. aeo?iae.- 1992.-49, N 2.- N. 89-92.

10. Eo/oe I.A. Aaiaoe/aneay o?ainoi?iaoeey aunoeo ?anoaiee,
iiin?aaeiaaiiay aaeoa?eyie ec ?iaea Agrobacterium. //Oni. nia?.
aeie.-1997. – 117, N 6-C. 645-658.

11. Eo/oe I.A., Aeaaa TH.TH. I?eiaiaiea o?ainaaiiuo ?anoaiee aeey oeaeae
i?aeoe/aneie naeaeoeee.// Oeeoieiaey e aaiaoeea.-1997.- 31, N 4.-N.
102-114.

12. Eo/oe I. Aaiaoe/ia ?iaeaia??y – aoiaeaeaiiy a a?ieia?/io a?o.//
A?ni. IAI Oe?a?ie.-1998.-N 3-4.- N. 28-34.

13. Ni?i/eineee A.A., I?ioiaaneee A.E, A?iaeceineee Ae.I., Eo/oe I.A.
Eiioiioethi?anoeaioiay iee?ineiiey eiiiiiaioia oeeoineaeaoa a eeaoeao
?anoaiee.// Oeeoieiaey e aaiaoeea.-1990.-24, N 3. -C. 64-65.

14. ?aaeeiiaiei I.A., Oaaaeuie/ I.A., Aoaecue A.I., Aaiieeiaa A.I.,
Eo/oe I.A. ?acaeoea niiaoe/aneeo capiaeuoae a eaeeoniie eoeueoopa
eeaaapa eoaiaiai (Trifolium pratense L.)// Oeeoieiaey e aaiaoeea.-1994.-
28, N 5.- N. 15-20.

15. ?a/ae E.E., Noipiaeaiei N.A., Eo/oe I.A. Aaiaoe/aneay o?ainoipiaoeey
aipioa eiinopoeoeeae, niaeapaeauae Ds-yeaiaio eoeopocu, iaoiaeii
yeaeopiiipaoeee.// Oeeoieiaey e aaiaoeea.-1994.-28, N 6.- N. 49-54.

16. ?a/ae E.E., Noipiaeaiei N.A., Eo/oe I.A. Iieo/aiea e
iieaeoey?ii-aeieiae/aneee aiaeec o?ainaaiiuo ?anoaiee ai?ioa (Pisum
sativum L.), niaea?aeaueo Ds-yeaiaio eoeo?ocu.// Aeiiieeia?u e
eeaoea.-1995.-11, N 3-4.- N. 82-87.

17. Kuchuk N., Komarnitski I., Shakhovsky A., Gleba Y. Genetic
transformation of Medicago species by Agrobacterium tumefaciens and
electroporation of protoplasts.// Plant Cell Rep.-1990.- 8.-P. 660-663.

18. Kuchuk N.V., Gleba Y.Y. The development of biotechnological
approaches for improvement of grain legume species.// In: Proc. of 1
Eur. Conference on Grain Legumes.- France, Anger, 1-3 June 1992.- P.
121-122.

19. Kuchuk N., Herrmann R.G., Koop H.U. Plant regeneration from leaf
protoplasts of evening primrose (Oenothera hookeri).//Plant Cell Rep. –
1998.- 17, No. 8. – P. 601-604.

20. Radionenko M. A., Kuchuk N.V., Khvedynich O.A., Gleba Y.Y. Direct
somatic embryogenesis and plant regeneration from protoplasts of red
clover (Trifolium pratense L.)// Plant Sci.-1994.- 97. – P. 75-81.

21. Sarangi B.K., Kuchuk N.V., Gleba Y.Y. Isolation and culture of
protoplasts of pigeonpea (Cajanus cajan L.).// Plant Cell Rep.
-1992.-11.-P. 462-465.

22. A.n. 1549995 NNN? io 15.11.89. Eo/oe I.A., Aeaaa TH.TH. Niinia
auaeaeaiey e eoeueoeae?iaaiey i?ioiieanoia ec ?anoeoaeueiuo oeaiae nie.

23. Iiiio A.I., Eo/oe I.A., Ianoa?iae O.I. Iaoiaeu eeaoi/iie eiaia?ee
?anoaiee. – I?ai?eio 88.2, a. Eeaa “Eiienney ONN? ii aeaeai THIANEI”,
1988, 23n.

Eo/oe I.A. Aeei?enoaiiy i?eeii?a aaiaoe/ii? ?iaeaia??? ? ee?oeiii?
a?ieia?? aeey io?eiaiiy iiaeo oi?i o aeae?a c ?iaeeie aiaiaeo ?
aeayeeo ?ioeo ?ineei.

Aeena?oaoe?y o aeaeyae? ?oeiieno ia caeiaoooy iaoeiaiai nooiaiy aeieoi?a
a?ieia?/ieo iaoe ca niaoe?aeueiinoyie 03.00.22 “Ee?oeiia a?ieia?y”,
03.00.15 “Aaiaoeea”. ?inoeooo ee?oeiii? a?ieia?? oa aaiaoe/ii?
?iaeaia???, Ee?a 1998.

?ic?iaeaii iaoiaeeeo eoeueoeaoaaiiy i?ioiieano?a ? ?aaaia?aoe?? ?ineei o
ethoea?-ie, eiithoeie, ai?ioo ? o Oenothera hookeri. I?ioiieanoe ni?,
ethieio, aieo-aeiiai ai?ioo i?e aeei?enoaii? ?ic?iaeaieo iaoiae?a
eoeueoeaoaaiiy caeaoi? aei ooai?aiiy ee?oeiieo eieii?e c aenieei
nooiaiai aoaeoeaiino?. Io?eiaii eaiai?oeeino?ee? ?ineeie ethoea?ie ?
ee?oeii? e?i?? ni? oa ai?ioo oeyoii aeaeo?i-ii?aoe?? i?ioiieano?a.
Cai?iiiiiaaii iaoiae, ui i?aeaeuo? aoaeoeai?noue o?ain-oi?iaoe?? o
ai?ioo oa ethoea?ie – “iiaea?eia o?ainoi?iaoe?y”, ui aeeth/a? 1)
io?e-iaiiy e?i?? ?ineei, ye? caeaoi? aei ?aaaia?aoe?? a eoeueoo?? in
vitro, ca ?aooiie o?ain-oi?iaoe?? iooaioii “shooty” A. tumefaciens
pGV2206; 2) – o?ainoi?iaoe?y e?i?e, ye? caeaoi? aei ?aaaia?aoe??,
naeaeoeaiei aaiii. Oaeei oeyoii aoei io?eiaii ?aaaia-?aoe?ei? e?i??
ethoea?ie ? ai?ioo ? aeae? o?ainaaii? ?ineeie ai?ioo c Ds o?ainii-ciiii
eoeo?oaece oa c aaiii no?eeino? aei oino?iio?eoeeio. I?enooi?noue aai?a
a aaiii? ?ineei aoei aeieacaii ca aeiiiiiaith iaoiae?a iie?ia?acii?
eaioethaiai? ?aaeoe??, aeio- a?a?eaeecaoe?? ca Naoca?iii, NPT II
aiae?co, aiae?co GUS aeoeaiino?, ?ioiioa?iaioiiai aiae?co (ELISA).
I?aeaeaeia? niiaoe/i? a?a?eaee Medicago sativa +M.varia ? M.borealis+
M.lupulina aoee io?eiai? ca aeiiiiiaith aneiao?e/ii? a?a-?eaeecaoe??.
O?iiiniiiee aiae?c oa aeio-a?a?eaeecaoe?y ?eainiiaeueii? AeIE
i?aeo-aa?aeeee a?a?eaeio i?e?iaeo io?eiaieo e?i?e. Aoei i?iaaaeaii
a?noieia?/iee aiae?c i?ioeano ?aaaia?aoe?? o eiithoeie, ai?ioo oa
oeoe?iaiai ao?yea.

Eeth/ia? neiaa: aiaia?, oeoe?iaee ao?ye, Oenothera, i?ioiieano,
?aaaia?aoe?y ?ineei, aaiaoe/ia o?ainoi?iaoe?y, niiaoe/ia a?a?eaeecaoe?y.

Eo/oe I.A. Eniieueciaaiea i?eaiia aaiaoe/aneie eiaeaia?ee e eeaoi/iie
aeieiaee aeey iieo/aiey iiauo oi?i o aeaeia ec naiaenoaa aiaiauo e o
iaeioi?uo ae?oaeo ?anoaiee.

Aeenna?oaoeey a aeaea ?oeiiene ia nieneaiea o/aiie noaiaie aeieoi?a
aeieiae/aneeo iaoe ii niaoeeaeueiinoyi 03.00.22 “Eeaoi/iay aeieiaey” e
03.00.15 “Aaiaoeea”, Einoeooo eeaoi/iie aeieiaee e aaiaoe/aneie
eiaeaia?ee, Eeaa 1998.

?ac?aaioaiu iaoiaeu eoeueoeae?iaaiey i?ioiieanoia e ?aaaia?aoeee
?anoaiee o ethoea?iu, eeaaa?a, ai?ioa e o Oenothera hookeri.I?ioiieanou
nie, ethieia, aieoaeiiai ai?ioa niiniaiu n aunieie yooaeoeaiinoueth
ia?aciauaaoue eeaoi/iua eieiiee i?e eniieueciaaiee ?ac?aaioaiiuo
iaoiaeia eoeueoe-ae?iaaiey. Iooai yeaeo?iii?aoeee i?ioiieanoia iieo/aiu
eaiaieoeei-onoie/eaua ?anoaiey ethoea?iu e eeaoi/iua eeiee nie e ai?ioa.
I?aaeeiaeai iaoiae, iiauoathuee yooaeoeaiinoue o?ainoi?iaoeee o ai?ioa e
ethoea?iu – aeaieiay o?ainoi?iaoeey. Nooue iaoiaea caeeth/aaony a
neaaeothuai 1) iieo/aiea ?aaaia?aoeeiiiiniiniaiuo eeiee iooai
o?ainoi?iaoeee iooaioii “shooty” A. tumefaciens; 2) o?ainoi?iaoeey
?aaaia?aoeeiiiuo eeiee naeaeoeaiui aaiii. Oaeei ia?acii auee iieo/aiu
?aaaia?aoeeiiiua eeiee ethoea?iu e ai?ioa e a aeaeueiaeoai o?ainaaiua
?anoaiey ai?ioa, eae n Ds o?ainiiciiii eoeo?ocu, oae e n aaiii
onoie/eainoe e oinoeiio?eoeeio. I?enoonoaea aaiia a aaiiia ?anoaiee auei
aeieacaii n eniieueciaaieai iaoiaeia iieeia?aciie oeaiiie ?aaeoeee,
aeio-aea?eaeecaoeee ii Naoca?io, aiaeeca aeoeaiinoe NPTII, aiaeeca GUS
aeoeaiinoe, eiioiioa?iaioiiai aiaeeca (ELISA). Iaaeaeaeiaua niiaoe/aneea
aea?eaeu Medicago sativa+M.varia e M.borealis+ M.lupulina auee iieo/aiu
n iiiiuueth aneiiao?e/iie aea?eaeecaoeee. O?iiiniiiue aiaeec e
aeio-aea?eaeecaoeey ?eai-niiaeueiie AeIE iiaeoaa?aeeee aea?eaeioth
i?e?iaeo iieo/aiiuo eeiee. Aue i?iaaaeai aenoieiae/aneee aiaeec
i?ioeanna ?aaaia?aoeee o eeaaa?a, ai?ioa e naoa?iie naaeeu.

Eeth/aaua neiaa: aiaiaua, naoa?iay naaeea, Oenothera, i?ioiieano,
?aaa-ia?aoeey ?anoaiee, aaiaoe/aneay o?ainoi?iaoeey, niiaoe/aneay
aea?eaeecaoeey.

Kuchuk N.V. “Application of genetic engineering and cell biology
approaches to produce new forms of species from Leguminosae family as
well as of some other plants.”

Dissertation thesis as manuscript is presented for academic degree the
doctor of biological sciences in fields 03.00.22 “Cell biology” and
03.00.15 “ Genetics”.

Institute of Cell Biology and Genetic Engineering, Kiev, 1998.

Methods of protoplast cultivation and plant regeneration have been
developed for alfalfa, clover and pea and for Oenothera hookeri.
Protoplasts isolated from soybean, lupine and “pigeonpea” have been
cultivated with high efficiency. But, only callus has been obtained.
Protoplast electroporation has been successfully applied to produce
transgenic alfalfa plants as well as soybean and pea calli by using
kanamycine as selectable agent. To produce transgenic alfalfa and pea
plants double transformation system has been developed. The adopted
approach involves two transformation steps: 1) production of
regenerable legume lines through transformation with “shooty” mutant of
A.tumefaciens pGV2206; 2) transformation of these regenerable legume
lines with the gene construct containing selectable genes. Regenerable
transgenic alfalfa and pea lines have been obtained using the developed
approach. Transgenic pea plants carried of maize Ds trasposon as well as
phosphinotricine resistant plants have been produced. The presence of
transferred genes in the genomes of produced plants has been confirmed
by PCR, Southern blot hybridization, NPTII activity analysis, ELISA
analysis, GUS activity analysis. Interspecific hybrid Medicago
sativa+M.varia plants and M.borealis+ M.lupulina calli have been
developed using the asymmetric somatic hybridization. Chromosome number
analysis and Southern blot hybridization of ribosomal DNA have confirmed
the hybrid nature of lines developed. Processes of plant regeneration of
clover, pea and sugar beet have been studied with histological analysis.

Key words: legumes, sugarbeet, Oenothera, protoplast, plant
regeneration, genetic transformation, somatic hybridization,

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