CONCEPT
(https://mrsmaths!"#$!r%prss#&!m/')'/))/*/h+,hr-,"t+&-",+"r+",-a"%-s!mat+&-.s+!"/0(https://mrsmaths!"#$!r%prss#&!m/')'/))/*/h+,hr-,"t+&-",+"r+",-a"%-s!mat+&-.s+!"/0
ABSTRACTABSTRACT
T!%a12s S&"ar+! +s m!r !r 3ss 3+4 th+s#T!%a12s S&"ar+! +s m!r !r 3ss 3+4 th+s#
)# A,r+&3tra3 3a"%s ar s% .!r +"%str+a3 !r m+"+", !r)# A,r+&3tra3 3a"%s ar s% .!r +"%str+a3 !r m+"+", !r .!r 3a"% 533+", !. S!3+% Wasts !r .!r %$33+", ths.!r 3a"% 533+", !. S!3+% Wasts !r .!r %$33+", ths &as+", %+m+"+sh+", a,r+&3tra3 3a"%s a6a+3a73#&as+", %+m+"+sh+", a,r+&3tra3 3a"%s a6a+3a73#
'# La7!r +s 7&!m+", s&ar& a"% 8p"s+6'# La7!r +s 7&!m+", s&ar& a"% 8p"s+6 %+s&!ra,+", .armrs t! ,r!$#%+s&!ra,+", .armrs t! ,r!$#
*# W+3% A"+ma3s9 Rats a"% "t+m31 ra+"s9 hrr+&a"s9*# W+3% A"+ma3s9 Rats a"% "t+m31 ra+"s9 hrr+&a"s9 &1&3!"s t& ar %ama,+", a,r+&3tra3 53%s a"% ths&1&3!"s t& ar %ama,+", a,r+&3tra3 53%s a"% ths r%&+", t!ta3 a6a+3a73 1+3%s#r%&+", t!ta3 a6a+3a73 1+3%s#
# Psts a"% +"s&ts ar 7&!m+", m!r a"% m!r# Psts a"% +"s&ts ar 7&!m+", m!r a"% m!r pst+&+% rs+sta"t a"% r%&+", th 1+3%s#pst+&+% rs+sta"t a"% r%&+", th 1+3%s#
 
=# Y!",r ,"rat+!"s ar attra&t% t! m!r=# Y!",r ,"rat+!"s ar attra&t% t! m!r rm"rat+6 !&&pat+!"s#rm"rat+6 !&&pat+!"s#
># O6r ,"rat+!"s +"hr+t% 3a"%s ,!t %+6+%% a"%># O6r ,"rat+!"s +"hr+t% 3a"%s ,!t %+6+%% a"% 7&am sma33r $+th 3ssr &3t+6a73 3a"% a.tr7&am sma33r $+th 3ssr &3t+6a73 3a"% a.tr 6!3ms !. th 7!"%ar+s ar +"&ras%#6!3ms !. th 7!"%ar+s ar +"&ras%#
I" th+s &!"t8t a "% ar+ss t! s!36 a33 ths pr!73ms#I" th+s &!"t8t a "% ar+ss t! s!36 a33 ths pr!73ms# A 7ttr pr!p!s+t+!" +s t! "a73 a .armr t! s !"A 7ttr pr!p!s+t+!" +s t! "a73 a .armr t! s !" s% $h+&h &a" 1+3% !" &r!p a7!6 th ,r!"% a"%s% $h+&h &a" 1+3% !" &r!p a7!6 th ,r!"% a"% s+m3ta"!s31 a"!thr &r!p "%r th ,r!"% 3+4s+m3ta"!s31 a"!thr &r!p "%r th ,r!"% 3+4 POMATO $h+&h $as %s+,"% t! pr!%& T!mat!sPOMATO $h+&h $as %s+,"% t! pr!%& T!mat!s a7!6 th ,r!"% a"% P!tat!s "%r th ,r!"%?a7!6 th ,r!"% a"% P!tat!s "%r th ,r!"%? thr71 .armr $+33 7 ha6+", a &ha"& t! ha6 a 7ttrthr71 .armr $+33 7 ha6+", a &ha"& t! ha6 a 7ttr pr+& at3ast t! !" !. thm#pr+& at3ast t! !" !. thm#
I" th+s art+&3 @!rts ar ma%I" th+s art+&3 @!rts ar ma% )#)# t! &!mp+3 th a6a+3a73 t&h"!3!,+st! &!mp+3 th a6a+3a73 t&h"!3!,+s '#'# t! 7ttr "%rsta"% th pr!t!&!3s +"6!36% +"t! 7ttr "%rsta"% th pr!t!&!3s +"6!36% +"
s+mp3r trmss+mp3r trms 3.3. t! s +mpr!6+s% 6rs+!"s !. th mth!%!3!,+s t!t! s +mpr!6+s% 6rs+!"s !. th mth!%!3!,+s t!
a,m"t ssta+"% $!r3% .!!% pr!%&t+!" +" tha,m"t ssta+"% $!r3% .!!% pr!%&t+!" +" th &!m+", Yars&!m+", Yars..
MISSION:MISSION: I am looking for associates who can involve with us in technology,I am looking for associates who can involve with us in technology, production and marketing on contract asis.production and marketing on contract asis.
PROLOGUEPROLOGUE !!% pr!%&t+!" mst %!73 71 '; t! mt th %ma"% !. th $!r3%2s ,r!$+", p!p3at+!" a"% +""!6at+6 strat,+s ar "%% t! h3p &!m7at h",r9 $h+&h a3ra%1 a@&ts m!r tha" ) 7+33+!" p!p3 +" th $!r3%9 s6ra3 8prts t!%a1 t!3% th S&!"% C!mm+tt (E&!"!m+& a"% +"a"&+a30 %r+", a pa"3 %+s&ss+!" !" N$ &!!prat+!" .!r ,3!7a3 .!!% s&r+t1#
 
!. th .!!% pr+& &r+s+s9 th 5"a"&+a3 a"% &!"!m+& &r+s+s9 a"% &3+mat &ha",#
Ra3 Pa"%1a-L!r&h9 Ch+. !. Sta@ a"% Ha% !. th '' V+s+!" .!r !!%9 A,r+&3tr9 a"% th E"6+r!"m"t I"+t+at+69 I"tr"at+!"a3 !!% P!3+&1 a"% Rsar&h I"st+tt (IPRI09 sa+% that9 %sp+t @!rts t! +mpr!6 .!!% s&r+t1 71 th +"tr"at+!"a3 &!mm"+t19 th "m7r !. p!p3 ,!+", h",r1 has rma+"% r3at+631 &!"sta"t !6r th 3ast ha3. &"tr19 $h+&h .!r&% th st+!": What %! $ "% t! %! %+@r"t31
O" a" +"+r1 a7!t th t"s+!" 7t$" spp!rt+", sma33h!3%r .armrs a"% th "% .!r s7sta"t+a39 3ar,r-s&a3 a,r+&3tra3 +"6stm"t t! +mpr!6 .!!% s&r+t19 h sa+% +t $as a" +ss !. ma8+mm &+"&19 a"% that !" $a1 t! "&!ra, that &!3% 7 thr!,h spp!rt !. &!!prat+6s# Or ma+" !7 "!$ +s t! sh!$ that9 $+th th a6a+3a73 rs!r&s9 $ &a" ,t ,!!% rs3ts %r+", th "8t .$ 1ars9 h sa+%#
(http://$$$#"#!r,/prss/"/'J/,a.*''#%!&#htm0
HISTORY HISTORY 
Th &!"&pt !. ,ra.t+", r3at% p!tat!s a"% t!mat!s s! that 7!th ar pr!%&% !" th sam p3a"t $as !r+,+"a331 %63!p% +" )J>> at th Ma8 P3a"&4 I"st+tt .!r D63!pm"ta3 B+!3!,1 +" TK7+","9 Grma"19 a"% a3th!,h ha3th19 th p3a"t pr!%&% "+thr p!tat!s "!r t!mat!s#' Th Ma8 P3a"&4 I"st+tt .!r P3a"t Br%+", Rsar&h +" 3" pr!%&% a p3a"t $+th .r+t +" )JJ#'
As $+th a33 ,ra.ts9 th+s p3a"t $+33 "!t !&&r +" "atr a"% &a""!t 7 ,r!$" .r!m s%9 7&as th t$! parts !. th p3a"t rma+" ,"t+&a331 sparat9 a"% !"31 r31 !" a&h !thr .!r "!r+shm"t a"% ,r!$th# L+4 m!st sta"%ar% t1ps !. p3a"t ,ra.t+",9 a sma33 +"&+s+!" +s ma% +" th stm !. 7!th p3a"ts a"% th1 ar strapp% t!,thr# O"& th &ts ha6 ha3% a"% th p3a"ts ar !+"%9 th 3a.1 t!p !. th p!tat! p3a"t &a" 7 &t a$a1 a"% th r!!ts !. th t!mat! &a" 7 rm!6%9 3a6+", th 3a6s !. th t!mat! p3a"t t! "!r+sh th r!!ts !. th p!tat! p3a"t#* Th r!!tst!&4 (p!tat!0 a&ts as a sta73 a"% ha3th1 r!!t s1stm a"% th s&+!"s (t!mat!0 ar &h!s" .!r th+r .r+t9 <!$rs !r 3a6s# Th t!mat!s sh!3% 7 ra%1 t! har6st a.tr a7!t )' $4s %r+", th smmr m!"ths9 th p!tat!s sh!3% 7 ra%1 a.tr th t!mat! 3a6s 7,+" t! %+ 7a&49 "!rma331 +" ar31 atm"#  Gra.t+", +" th+s $a1 &a" 7 s% t! pr!%& ma"1 %+@r"t r3at% &r!ps .r!m th sam p3a"t9 .!r 8amp3 th ,r!$+", p!p3ar+t1 !. r+t sa3a% trs9 $h+&h +s a s+",3 tr that pr!%&s m3t+p3 t1ps !. &+trs .r+ts9 !r a tr $+th a 6ar+t1 !.  .r+ts $+th st!"s (pa&h9 p3m t&#0# 
 
I" )J>'9 Car3s!" a"% h+s ass!&+ats pr!%&% th 5rst +"tr-sp&+5& s!mat+& h17r+% 7t$" N+&!t+a"a ,3a&a a"% N# 3a",s%!r+# I" )J>9 M3&hrs a"% h+s &!-$!r4rs %63!p% th 5rst +"tr-,"t+& s!mat+& h17r+%s 7t$" S!3a"m t7r!sm (p!tat!0 a"% L1&!prs+&!" s&3"tm (t!mat!0# Th h17r+%s ar 4"!$" as QP!mat!s !r T!pat!s2 (http://www.biotechnology4u.com/plant_biotechnology_applications_cell_tissue_c ulture.html)
 
 
 
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Pr6+!s31 NONGWOO BIO %63!p% a "$ CMS 3+" (NWB-CMS0 .!r ra%+sh a"% th NWB-CMS 3+" +"%% pr!6+%% 3!ts !. s7-3+"s !. Ra%+sh# A"% th B+!t&h"!3!,1 I"st+tt has %63!p% th "$ Cr&+.ra p3a"ts 71 .s+", th NWB-CMS &33s !. Ra%+sh t! th !r%+"ar1 &33s !. Ch+"s Ca77a, a"% Ca77a,#
(http://$$$#"!",$!!7+!#&!m/"!",$!!/htm3/t&h'#htm3 0
SCIENCESCIENCE Th s% +s th matr9 .rt+3+F% !63# A.tr .rt+3+Fat+!"9 th hap3!+% &33s !. th m7r1! sa& %+s+"t,rat# Th matr"a331 %r+6% %+p3!+% &33s !. th !63 %63!p +"t! th har%9 $atr-rs+sta"t !tr &!6r+", !. th s%9 &a33% th tsta9 !r s% &!at# Th %+p3!+% F1,!t %63!ps +"t! th m7r1!9 a"% th tr+p3!+% "%!sprm &33s m3t+p31 a"% pr!6+% "tr+t+!"# Th tsta sa331 sh!$s a s&ar &a33% th h+3m $hr th !63 $as !r+,+"a331 atta&h% t! th ."+&3# I" s!m s%s a r+%, a3!", th tsta &a33% th raph sh!$s $hr th ."+&3 !r+,+"a331 $as prss% a,a+"st th !63# Th m+&r!p13 !. th !63 sa331 sr6+6s as a sma33 p!r +" th s% &!at that a33!$s passa, !. $atr %r+", ,rm+"at+!" !. th s%#
I" s!m sp&+s9 th ."+&3 %63!ps +"t! a 3ar,r str&tr !" th s% &a33% a" ar+39 $h+&h +s !.t" 7r+,ht31 &!3!r%9 +&19 a"% &!"ta+"s s,ars that ar &!"sm% 71 a"+ma3s that ma1 a3s! %+sprs th s% (as +" "tm,9 arr!$r!!t9 !8a3+s9 a"% &ast!r 7a"0# Th+s +s %+st+"&t .r!m th .r+t9 $h+&h .!rms .r!m th !6ar1 +ts3.#
Th m7r1! &!"s+sts !. th &!t13%!"(s0 9 p+&!t139 a"% h1p!&!t13# Th &!t13%!"s rsm73 sma33 3a6s9 a"% ar sa331 th 5rst ph!t!s1"tht+& !r,a"s !. th p3a"t# Th p!rt+!" !. th m7r1! a7!6 th &!t13%!"s +s th p+&!t139 a"% th p!rt+!" 73!$ +s th h1p!&!t13# Th p+&!t13 +s a" ap+&a3 mr+stm that pr!%&s th sh!!t !. th ,r!$+", p3a"t a"% th 5rst tr 3a6s a.tr ,rm+"at+!"# Th h1p!&!t13 %63!ps +"t! th r!!t# O.t" th t+p !. th h1p!&!t139 th ra%+&39 +s th 5rst +"%+&at+!" !. ,rm+"at+!" as +t 7ra4s !t !. th s%# 3!$r+", p3a"ts ar &3ass+5% as m!"!&!t13%!"s !r %+&!t13%!"s (m!st ar "!$ &a33% %+&!ts 0 7as% !" th "m7r !. &!t13%!"s pr!%&% +" th m7r1!# C!mm!" m!"!&!t13%!"s +"&3% ,rasss9 s%,s9 3+3+s9 +r+ss9 a"% !r&h+%s? &!mm!" %+&!t13%!"s +"&3% s"<!$rs9 r!ss9 3,ms9 s"ap%ra,!"s9 a"% a33 "!"&!"+.r!s trs#
 
rsr6# I" ,ra+"s s&h as &!r" a"% $hat9 th !tr 3a1r !. th "%!sprm &!"s+sts !. th+&4-$a33% &33s &a33% a3r!"9 $h+&h ar h+,h +" pr!t+"#
 
A"at!m1 !. th rpr!%&t+6 !r,a"s +" a",+!sprms#  (http://$$$#7+!3!,1r.r"&#&!m/R-S/Rpr!%&t+!"-+"- P3a"ts#htm3X+8FF*T'J1*"0
SEARCH O LITERATURESEARCH O LITERATURE
 
APPLICATIONS O SOMATIC HYBRIDIATION 
a) Crat+!" !. h17r+%s $+th %+sas rs+sta"& - Many disease resistance genes (e.g. tobacco mosaic virus, potato virus , club rot disease) could be success!ully trans!erred !rom one species to another. ".g resistance has been introduced in tomato against diseases such as #M$, spotted wilt virus and insect pests. b) E"6+r!"m"ta3 t!3ra"& - using somatic hybridi%ation the genes con!erring tolerance !or cold, !rost and salt were introduced in e.g. in tomato. c) C1t!p3asm+& ma3 str+3+t1 - using cybridi%ation method, it was possible to trans!er cytoplasmic male sterility. d) Za3+t1 &hara&trs - somatic hybrids with selective characteristics have been developed e.g. the production o! high nicotine content.
CHROMOSOME NUMBER IN SOMATIC HYBRIDS  #he chromosome number in the somatic hybrids is generally more than the total number o! both o! the parental protoplasts. &! the chromosome number in the hybrid is the sum o! the chromosomes o! the two parental protoplasts, the hybrid is said to be symmetric hybrid. 'symmetric hybrids have abnormal or wide variations in the chromosome number than the eact total o! two species.
(http://www.biotechnology4u.com/plant_biotechnology_applications_cell_tissue_c ulture.html)
 
Gra.t+", !r ,ra.ta,) +s a h!rt+&3tra3 t&h"+ $hr71 t+sss .r!m !"p3a"t ar +"srt% +"t! th!s !. a"!thr s! that th t$! sts !. 6as&3ar t+sss ma1 !+" t!,thr# Th+s 6as&3ar !+"+", +s &a33% +"!s&3at+!"# Th t&h"+ +s m!st &!mm!"31 s% +" as8a3 pr!pa,at+!" !. &!mmr&+a331 ,r!$" p3a"ts .!r th h!rt+&3tra3 a"% a,r+&3tra3 tra%s# I" m!st &ass9 !" p3a"t +s s3&t% .!r +ts r!!ts a"% th+s +s &a33% th st!&4 !rr!!tst!&4 # Th !thr p3a"t +s s3&t% .!r +ts stms9 3a6s9 <!$rs9 !r .r+ts a"% +s &a33% th s&+!" !r &+!"#) Th s&+!" &!"ta+"s th %s+r% ,"s t! 7 %p3+&at% +" .tr pr!%&t+!" 71 th st!&4/s&+!" p3a"t#
I" stm ,ra.t+",9 a &!mm!" ,ra.t+", mth!%9 a sh!!t !. a s3&t%9 %s+r% p3a"t&3t+6ar +s ,ra.t% !"t! th st!&4 !. a"!thr t1p# I" a"!thr &!mm!" .!rm &a33% 7% ,ra.t+",9 a %!rma"t s+% 7% +s ,ra.t% !"t! th stm !. a"!thr st!&4 p3a"t9 a"% $h" +t has +"!s&3at% s&&ss.3319 +t +s "&!ra,% t! ,r!$ 71 pr"+", !@ th stm !. th st!&4 p3a"t st a7!6 th "$31 ,ra.t% 7%#
!r s&&ss.3 ,ra.t+", t! ta4 p3a&9 th 6as&3ar &am7+m t+sss !. th st!&4 a"% s&+!" p3a"ts mst 7 p3a&% +" &!"ta&t $+th a&h !thr# B!th t+sss mst 7 4pt a3+6 "t+3 th ,ra.t has ta4"9 sa331 a pr+!% !. a .$ $4s# S&&ss.3 ,ra.t+", !"31 r+rs that a 6as&3ar &!""&t+!" ta4 p3a& 7t$" th ,ra.t% t+sss# [!+"ts .!rm% 71 ,ra.t+", ar "!t as str!", as "atra331 .!rm% !+"ts9 s! a ph1s+&a3 $a4  p!+"t !.t" st+33 !&&rs at th ,ra.t 7&as !"31 th "$31 .!rm% t+sss +"!s&3at $+th a&h !thr# Th 8+st+", str&tra3 t+ss (!r $!!%0 !. th st!&4 p3a"t %!s "!t .s# (http://"#$+4+p%+a#!r,/$+4+/Gra.t+",0
NOVEL ATERNATIVE- SOMATIC USIONNOVEL ATERNATIVE- SOMATIC USION S8a3 h17r+%+Fat+!" s+"& t+m +mmm!r+a3 has 7" s% as a mth!% .!r &r!p +mpr!6m"t 7t +t has +ts !$" 3+m+tat+!"s as +t &a" !"31 7 s% $+th+" mm7rs !. sam sp&+s !r &3!s31 r3at% $+3% sp&+s# Ths9 th+s 3+m+ts th s !. s8a3 h17r+%+Fat+!" as a ma"s !.  pr!%&+", 7ttr 6ar+t+s# D63!pm"t !. 6+a73 &33 h17r+%s 71 s!mat+& h17r+%+Fat+!"9 thr.!r9 has 7" &!"s+%r% as a" a3tr"at+6 appr!a&h .!r th pr!%&t+!" !. spr+!r h17r+%s !6r&!m+", th sp&+s 7arr+r# Th t&h"+ &a" .a&+3+tat 7r%+", a"% ," tra"s.r9 71pass+", pr!73ms ass!&+at% $+th &!"6"t+!"a3 s8a3 &r!ss+", s&h as9 +"trsp&+5&9 +"tr,"r+& +"&!mpat+7+3+t1# Th+s t&h"+ !. h17r+% pr!%&t+!" 6+a pr!t!p3ast .s+!" a33!$s &!m7+"+", s!mat+& &33s ($h!3 !r part+a30 .r!m %+@r"t &3t+6ars9 sp&+s !r ,"ra rs3t+", +" "!63 ,"t+& &!m7+"at+!"s +"&3%+", s1mmtr+& s!mat+& h17r+%s9 as1mmtr+& s!mat+& h17r+%s !r s!mat+& &17r+%s#
Th m!st &!mm!" tar,t s+", s!mat+& h17r+%+Fat+!" +s th ," !.  s1mmtr+& h17r+%s that &!"ta+" th &!mp3t "&3ar ,"!ms a3!",
 
$+th &1t!p3asm+& !r,a"33s !. 7!th par"ts# Th+s +s "3+4 s8a3 rpr!%&t+!" +" $h+&h !r,a"33 ,"!ms ar ,"ra331 &!"tr+7t% 71 th matr"a3 par"t# O" th !thr ha"%9 s!mat+& &17r+%+Fat+!" +s th pr!&ss !. &!m7+"+", th "&3ar ,"!m !. !" par"t $+th th m+t!&h!"%r+a3 a"%/!r &h3!r!p3ast ,"!m !. a s&!"% par"t# C17r+%s &a" 7 pr!%&% 71 %!"!r-r&+p+"t mth!% !r 71 &1t!p3ast-pr!t!p3ast .s+!"# I"&!mp3t as1mmtr+& s!mat+& h17r+%+Fat+!" a3s! pr!6+%s !pp!rt"+t+s .!r tra"s.r !. .ra,m"ts !. th "&3ar ,"!m9 +"&3%+", !" !r m!r +"ta&t &hr!m!s!ms .r!m !" par"t (%!"!r0 +"t! th +"ta&t ,"!m !. a s&!"% par"t (r&+p+"t0# Ths mth!%s pr!6+% ,"t+& ma"+p3at+!" !. p3a"ts !6r&!m+", hr%3 !. s8a3 +"&!mpat+7+3+t19 thr719 sr6+", as a mth!% !.  7r+",+", t!,thr 7"5&+a3 tra+ts .r!m ta8!"!m+&a331 %+st+"&t sp&+s $h+&h &a""!t 7 a&h+6% 71 s8a3 &r!sss# S6ra3 paramtrs s&h as9 s!r& t+ss9 &3tr m%+m a"% "6+r!"m"ta3 .a&t!rs +"<"& a7+3+t1 !. a pr!t!p3ast %r+6% h17r+% &33s t! %63!p +"t! a .rt+3 p3a"t# Th ,"ra3 stps +"6!36% +" s!mat+& h17r+%+Fat+!" a"% &17r+%+Fat+!" mth!%s ar 3a7!rat% +" +,r )*#) a"% )*#' #
Steps involved in somatic hybridization
Th s!mat+& .s+!" pr!&ss !&&rs +" .!r stps:
Th rm!6a3 !. th &33 $a33 !. !" &33 !. a&h t1p !. p3a"t s+", &333as "F1m t! pr!%& a s!mat+& &33 &a33% a pr!t!p3ast
)# Th &33s ar th" .s% s+", 3&tr+& sh!&4 (3&tr!.s+!"0 !r &hm+&a3 tratm"t t! !+" th &33s a"% .s t!,thr th "&3+# Th rs3t+", .s% "&3s +s &a33% htr!4ar1!"#
'# Th s!mat+& h17r+% &33 th" has +ts &33 $a33 +"%&% t! .!rm s+", h!rm!"s
*# Th &33s ar th" ,r!$" +"t! &a33ss $h+&h th" ar .rthr ,r!$" t! p3a"t3ts a"% 5"a331 t! a .33 p3a"t9 4"!$" as a s!mat+& h17r+%#
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( #orrence, ames (*++). igher iology (*nd ed.). odder 0ibson1 Mahesh. 2lant Molecular biotechnology. *++3. oo.)
 
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LIMITATIONS AND SOLUTIONSLIMITATIONS AND SOLUTIONS
 
 
 
 
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PLAYERS IN THE IELDPLAYERS IN THE IELD Ph1t!$3t (Head ofce: Kölsumer Weg 33 D-41334 Nettetal P +49.2162.!"9 # +49.2162.!921" $$$.%hyt&$elt.c&m) has sta73+sh% !pt+ma3 s1stms .!r ma"1 p3a"t sp&+s9 +"&3%+", 6" %+&3t !"s 3+4 ,rasss9 &ra3s a"% 3,ms# Ph1t!$3t a3ra%1 app3+% th SH t&h"!3!,1 .!r ma"1 %+@r"t app3+&at+!"s#
 
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PHASES O THE STUDIESPHASES O THE STUDIES
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IDENTIICATION O PLANTS TO BE MERGEDIDENTIICATION O PLANTS TO BE MERGED
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PROBABLE COMBINATIONS THAT MAY BE CHOOSENPROBABLE COMBINATIONS THAT MAY BE CHOOSEN
)# B3a&4,ram ()# B3a&4,ram (Vigna MungoVigna Mungo00 ^ Pa"t (^ Pa"t ( Arachis hypogaea Arachis hypogaea00
'# S!1 ('# S!1 (Glycine max Glycine max  0 ^ O"+!" (0 ^ O"+!" ( Allium Allium cepacepa00
*# P%+"a (*# P%+"a (Mentha ArvensisMentha Arvensis 0 ^ Carr!t (0 ^ Carr!t (aucus carotaaucus carota00
 
sativus!sativus!
;# C!r+a"%r (;# C!r+a"%r (#oriandrum sativum#oriandrum sativum 0 ^ Carr!t (0 ^ Carr!t (aucus carotaaucus carota00
=# Cha"a(=# Cha"a(#icer arietinum#icer arietinum00 ^ Carr!t (^ Carr!t (aucus carotaaucus carota00
># Gar (># Gar (#yamopsis tetragonoloba#yamopsis tetragonoloba 0 ^ Pa"t (0 ^ Pa"t ( Arachis hypogaea Arachis hypogaea00
PROTOCOLSPROTOCOLS Pr!t!p3ast Is!3at+!":
&solation o! protoplasts (5ig. .3) is readily achieved by treating cells/tissues with a suitable miture o! cell wall degrading en%ymes. 6sually, a miture o!
 
pectinase or macero%yme (+.7-7.38) and cellulose (7-*8) is appropriate !or most plant materials. emicellulase may be necessary !or some tissues. 0enerally crude commercial preparations o! en%ymes arc used.
 #he p o! en%yme solution is ad9usted between 4. and .+ and the temperature is ept at *;-<+=>. #he osmotic concentration o! en%yme miture and o! subse?uent media is elevated (usually, by adding ;++-++ m mol l-7 sorbitol or mannitol) to stabili%e the protoplasts and to prevent them !rom bursting. 6sually, ;+-7++ m mol l-7 >a>l* is added to the osmoticum as it improves plasma membrane stability. #he cells and tissues are incubated in the en%yme miture !or !ew to several (generally, 7@-7) hours1 naed protoplasts devoid o! cell wall are gradually released in the en%yme miture.
2rotoplasts have been isolated !rom virtually all plant parts, but lea! mesophyll is the most pre!erred tissue, at least in case o! dicots, !or this purpose. &n general, !ully epanded leaves are sur!ace-sterili%ed, their lower epidermis is peeled oA with a pair o! !orceps and the peeled areas are cut into small (>a. 7 cm*) pieces with a scalpel and suspended in the en%yme miture.
Bhen epidermis cannot be peeled, lea! may be cut into >a. 7 mm* pieces and treated with the en%yme miture1 vacuum inCltration may be used to !acilitate the entry o! en%ymes into the tissues. '!ter the period o! incubation, protoplasts are washed with a suitable washing medium in order to remove the en%ymes and the debris.
 
0enerally, MF and ; media, and their modiCcations are used !or protoplast culture. #he media are supplemented with a suitable osmoticum and, almost always, with an auin and a cytoinin, their types and concentrations depending mainly on the plant species.
'!ter -7+ days o! culture, protoplasts regenerate cell wall, and the osmolarity o! medium is gradually reduced to that o! normal medium. #he macroscopic colonies are trans!erred onto normal tissue culture media. 2rotoplasts are very sensitive to light1 there!ore, they are cultured in diAuse light or dar !or the Crst 4- days.
'# Pr!t!p3ast s+!":
 #he techni?ues !or protoplast !usion are pretty well reCned and highly eAective !or almost all the systems. ' number o! strategies have been used to induce !usion between protoplasts o! diAerent strains/species1 o! these the !ollowing three (5ig. .7+) have been relatively more success!ul.
2rotoplasts o! desired strains/species are mied in almost e?ual proportion1 generally, they are mied while still suspended in the en%yme miture. #he protoplast miture is then sub9ected to high p (7+.;) and high >a*G concentration (;+ m mol l-7) at <=> !or about <+ min (high p-high >a*G treatment). #his techni?ue is ?uite suitable !or some species, while !or some others it may be toic.
 
 #he washing medium may be alaline (p 3-7+) and contain a high >a*G ion concentration (;+ m mol l-7)1 this approach is a combination o! 2"0 and high p- high >a*G treatments, and is usually more eAective than either treatment alone.
2"0 is negatively charged and may bind to cation lie >a*G, which, in turn, may bind to the negatively charged molecules present in plasma lemma1 they can also bind to cationic molecules o! plasma membrane.
Huring the washing process, 2"0 molecules may pull out the plasma lemma components bound to them. #his would disturb plasma lemma organisation and may lead to the !usion o! protoplasts located close to each other (5ig. .77).
 #he above !usion techni?ues are nonselective in that they induce !usion between any two or more protoplasts. ' more selective and less drastic approach is the electro!usion techni?ue, which utili%es low voltage nonuni!orm alternating electric current pulses to bring the protoplasts in close contact (5ig. .7*). 5usion o! protoplasts is brought about by a short pulse o! high voltage.
 #he duration o! high voltage is a !ew microseconds, and the voltages ranges !rom ;++ to 7,+++ $/cm. #he high voltage creates transient disturbances in the organisation o! plasma lemma, which leads to the !usion o! neighbouring protoplasts. #he entire operation is carried out manually in specially designed e?uipment, called electroporator.
"lectro!usion has been mostly used with the members o! Folanaceae o!ten with very high rate (over ;+8) o! !usion. #his approach induces general !usion among protoplats and there is no control on the type o! protoplasts entering !usion. &n a modiCcation o! electro!usion, protoplast pairs are individually trans!erred into micro!usion chambers with the help o! a micromanipulator set up.
 
microseconds I a!ter mutual dielectrophoresis (7 M%1 @;-+ $/cm). #his techni?ue is called microelectro!usion1 it leads to highly speciCc and nearly 7++8 pairwise !usion. Many worers !eel that this !usion techni?ue is more desirable than the others !or a number o! important reasons.
*# S3&t+!" !. H17r+% C33s:
 
&n somatic hybridi%ation eperiments, only the heteroaryotic or JhybridD protoplasts, particularly those resulting !rom !usion between one protoplast o! each o! the two !usion partners, are o! interest.
owever, they !orm only a small proportion o! the population (usually +.;-7+8).  #here!ore, an eAective strategy has to be employed !or their identiCcation and isolation. #his step is called the selection o! hybrid cells, is the most critical, and is still an active area o! investigation.
' number o! strategies have been used !or the selection o! hybrid protoplasts. (7) Fome visual marers, e.g., pigmentation, o! the parental protoplasts may be used !or the identiCcation o! hybrid cells under a microscope1 these are then mechanically isolated and cultured.
5or eample, the protoplasts o! one species may be green and vacuolated (derived !rom mesophyll cells), while those o! the other may be non-vacuolated and non-green (obtained !rom cell cultures). Bhere such !eatures are not available, the protoplasts o! two parental species may be separately labelled with diAerent Kuorescent agents.
 #his would allow the microscopic identiCcation o! hybrid protoplasts !rom the parental ones. #his approach can even be adapted to automatic cell sorting permitting the recovery o! large numbers o! heteroaryons in a very short time.  #his approach is, however, time consuming, and re?uires considerable sill and eAort.
 
deCciencies) o! the parental species, which are not epressed in the hybrid cells due to complementation between their genetic systems.
 #hese properties may be sensitivity to culture medium constituents, antimetabolites, temperature, etc., inability to produce an essential biochemical (auotrophic mutants), chlorophyll or some other pigmentation, etc. #hese properties may be naturally present in the parental species or may be artiCcially induced through mutagenesis.
 #he !ollowing eample should be enough to clearly bring out the essential !eatures o! the complementation approach. 2rotoplasts o! 2etunia hybrida !orm calli on the MF medium, while those o! 2. parodii produce only small colonies.
5urther, actinomycin H (7 Eg ml-7) inhibits cell division o! 2. hybrida protoplasts, but it has no eAect on those o! 2. parodii. #hus protoplasts o! both these 2etunia species !ail to produce macroscopic colonies (calli) on MF medium supplemented with 7 Eg ml-7 actinomycin H.
owever, their hybrid cells (2. hybrida G 2. parodii1 it may be noted that somatic hybrids are denoted by a G sign as against the seual hybrids being designated by a symbol) divide normally on this medium to produce macroscopic colonies.
 #his selection strategy eploits those natural properties o! the two parental species, which show complementation in the hybrid cells and, at the same time, permit their selection. #hese strategies are simple, highly eAective and the least demanding, but their application is drastically limited by the nonavailability o! suitable properties (both natural and induced) in most o! the parental species o! interest to the eperimenters.
' variation o! the complementation approach uses selectable marers. &n this approach, (<) diAerent selectable marers lie antibiotic and herbicide resistance are engineered into the two !usion partners. #he hybrid cells, in such cases, are resistant to both the concerned selection agents, while the parental cells will be sensitive to one and the other selection agent.
' more general and widely applicable strategy, but demanding more wor than the previous approach, is (4) to culture the entire protoplast population without applying any selection !or the hybrid cells. 'll the types o! protoplasts !orm calli1 the hybrid calli are later identiCed on the basis o! callus morphology, chromosome constitution, protein and en%yme banding patterns, etc.
 
densities since neighbouring colonies are liely to !use at higher densities1 ideally, they should be cultured in microdrops, each drop containing but a single cell. Many worers tend to !avour this approach since it does not depend the presence o! appropriate but diLcult to Cnd marers in the parental species.
# R,"rat+!" !. H17r+% P3a"ts:
nce hybrid calli are obtained, plants are induced to regenerate !rom them since this is a prere?uisite !or their eploitation in crop improvement. 5urther, the hybrid plants must be at least partially !ertile, in addition to having some use!ul property, to be o! any use in breeding schemes. #he culture techni?ues have been reCned to a state where plant regeneration has been obtained in a number o! somatic hybrids (#able .;).
Ta73 #;# A 3+st !. s!m %+sta"t s!mat+& h17r+% p3a"ts:
ut even today, it has not been possible to recover hybrid plants and/or calli !rom a number o! somatic combinations1 this phenomenon is called Jsomatic incompatibilityD. #he reasons !or somatic incompatibility are not clearly understood. #he somatic hybrids are o! the !ollowing two types: (7) symmetric and (*) asymmetric hybrids.
Symmetric Hybrids:
Fome somatic hybrid plants retain the !ull or nearly !ull somatic complements o! the two parental species1 these are called symmetric hybrids. Fuch hybrids provide uni?ue opportunities !or synthesi%ing novel species, which may be o! theoretical and/or practical interest. 5re?uently, somatic hybrids between distantly related seually incompatible species are sterile, precluding their incorporation into a breeding programme.
 
plants may be epected to be partially !ertile. #hese somatic hybrids can now be used in breeding programmes !or limited gene/chromosome introgression !rom the species contributing the haploid protoplast.
' possible approach to the improvement o! apparently useless somatic hybrids, e.g., nonKowering somatic hybrid Haucus carota G 'egopodium podagraria, is to !use protoplasts !rom the hybrid with those o! one o! the parental species. #he !usion o! a somatic hybrid protoplast with that !rom one o! its parents is called somatic bac hybridi%ation.
Bhen protoplasts !rom the above somatic hybrid were !used with carrot protoplasts, the resulting somatic hybrid produced Kowers. Fuch hybrids can now be ordered into breeding programmes with the aim o! gene/chromosome introgression.
 Asymmetric Hybrids:
Many somatic hybrids ehibit the !ull somatic complement o! one parental species, while all or nearly all chromosomes o! the other parental species are lost during the preceding mitotic divisions1 such hybrids are re!erred to as asymmetric hybrids.
 #he available evidence suggests that such hybrids are liely to show a limited introgression o! chromosome segments !rom the eliminated genome(s) due to drastically enhanced chromosomal aberrations and/or mitotic crossing over in vitro.
'symmetric hybrids can be obtained even !rom those combinations, which normally produce symmetric hybrids by the !ollowing approach: protoplasts o! one o! the parental species are irradiated with a suitable dose o! -rays or gamma-rays to induce etensive chromosome breaage.
&n such cases, chromosome segment introgression may be maredly enhanced. &t may be pointed out that asymmetric hybrids are essentially cytoplasmic hybrids or cybrids, ecept !or the introgressed genes.
$ate o% &lasma'genes:
 
 #he cytoplasmic genes (generally studied in terms o! chloroplast types or chloroplast HN', cp-HN' mitochondrial HN', mtHN') appear to be distributed randomly during the mitotic cell divisions. 's a result, some cells receive chloroplasts o! one parental species, some others o! the other species and a small proportion retains the chloroplasts o! both the species.
 #his is reKected in the plants regenerated !rom these cells. #he same applies to mitochondria as well. &n addition, the distribution o! chloroplasts is independent !rom that o! mitochondria.
 #here!ore, a somatic hybrid plant may contain chloroplasts !rom one parental species and mitochondria !rom the other !usion parent. #here is considerable evidence that the genomes o! both chloroplasts and mitochondria, particularly the latter, undergo recombination in the hybrid cells1 this produces recombinant organelles in the progeny.
;# C17r+%s:
>ybrids or cytoplasmic hybrids are cells or plants containing nucleus o! one species but cytoplasm !rom both the parental species. #hey are produced in variable !re?uencies in normal protoplast !usion eperiments due to one o! the !ollowing: (i) !usion o! a normal protoplast o! one species with an enucleate protoplast or a protoplast having an inactivated nucleus o! the other species, (ii) elimination o! the nucleus o! one species !rom a normal heteroaryon, or (iii) gradual elimination o! the chromosomes o! one species !rom a hybrid cell during the subse?uent mitotic divisions. >ybrids may be produced in relatively high !re?uency by (i ) irradiating (with -rays or gamma-rays) the protoplasts o! one species prior to !usion in order to inactivate their nuclei, or (ii) by preparing enucleate protoplasts (cytoplasts) o! one species and !using them with normal protoplasts o! the other species.
 #he ob9ective o! cybrid production is to combine the cytoplasmic genes o! one species with the nuclear and cytoplasmic genes o! another species. ut the mitotic segregation o! plasmagenes, as evidenced by the distribution o! chloroplasts, leads to the recovery o! plants having plasmagenes o! one or the other species only1 only a small proportion o! the plants remain JcybridD, which would !urther segregate into the two parental types.
 
cytoplasmic male sterility !rom N. tabacum to N. sylvestris, !rom 2. hybrida to 2. aillaris, etc. (vi) &n addition, mitochondria !rom one parental species may be combined with the chloroplasts o! the other parental species.
(http://www.yourarticlelibrary.com/biotechnology/plant-tissues/somatic- hybridi%ation-o!-hybrid-plants-eplained-with-diagram/<<**@/
RESULTSRESULTS
DISCUSSIONDISCUSSION
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EPLORING MARETEERSEPLORING MARETEERS
STATUTORY REGULATIONS TO BE TAENSTATUTORY REGULATIONS TO BE TAEN CARECARE
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R,3at+!" !. GMOs  #he overseeing o! testing, use and commercialisation o! 0Ms I whether plants, animals or microorganisms I re?uires a special regulation system. #his system establishes the legal and institutional !ramewor !or the control o! potential negative eAects o! 0Ms on the environment or human and animal health (Obeanu, *++<). &n the 6F, transgenic plants are only introduced into the environment or on the maret !ollowing approval !rom the !ollowing governmental agencies responsible !or environmental, and human and animal health protection: 7. 6F Hepartment !or 'griculture (6FH'), *. "nvironmental 2rotection 'gency ("2'), <. 5ood and Hrug 'dministration (5H'). &n the 6F and >anada, transgenic plants are grown and used !or human and animal !ood, and separate storage and labelling are not mandatory. Fince 733+, in the "uropean 6nion, special legislation has been drawn up, enhanced and etended, with the purpose o! providing environmental and human health protection, and creating a common maret in the Celd o! biotechnology. #hus: - "6 Hirective No. *73/733+ (amended by Hirective No. 7/733) regulates the contained use o! genetically modiCed microorganisms (!or research and commercialisation), - "6 Hirective No. **+/733+, concerning the deliberate release o! genetically modiCed organisms into the environment, was the main initiative taen by the "6, and was subse?uently supplemented by several >ommission Hecisions (@*<, 77, 7*, 7</*++<), - "6 Hirective No. 7/*++7 regulates the deliberate release o! genetically modiCed organisms into the environment. #his Hirective repealed Hirective No. **+/733+. aving come into !orce on 7 ctober *++*, Hirective 7/*++7 both updates and consolidates 7; eisting regulations. #his Hirective also deals with mandatory in!ormation to the public, the long-term monitoring o! eAects, labelling and traceability, in all stages o! 0M introduction on the maret. #wo other acts have been adopted and published in the Lcial ournal o! the "uropean >ommunities, with respect to the >ommunity system o! 0M traceability, the labelling o! genetically modiCed !ood and !odder, and the continuous procedure o! authorisation or introduction o! 0Ms in the environment as !ood or !odder: - "> Pegulation No. 7*3/*++< (** Feptember *++<) on genetically modiCed !ood and !odder, and - "> Pegulation No. 7<+/*++< (** Feptember *++<) on the traceability and labelling o! genetically modiCed organisms, and 0M-based !ood and !odder.  #hese regulations amended "6 Hirective No. 7/*++7.
WEB REERENCESWEB REERENCES
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