Journal of Genetic Engineering and Biotechnology (2012) 10, 4753

cade!y of "cientific
#e$earch % &echnology
and
'ational #e$earch (enter, Egy)t
Journal of Genetic Engineering and Biotechnology
***+el$e,ier+co!-locate-.ge/
#&0(1E
2or)hological and !olecular characteri3ation of $o!aclonal
,ariation$ in ti$$ue culture4deri,ed /anana )lant$
5a!al 6+ /dellatif
7
, del E+ 8ega3y, 8aron 2+ /o$ha!a, 8a!dy + E!ara,
/delha!eed + El4"hahed
9lant Biotechnology :e)art!ent, Genetic Engineering and Biotechnology #e$earch 0n$titute (GEB#0), "adat
(ity, 2inoufiya ;ni,er$ity, Egy)t
#ecei,ed 11 :ece!/er 2011< re,i$ed 20 )ril 2012< acce)ted 5 2ay 2012
,aila/le online = June 2012
5E>?@#:"
"o!aclonal ,ariation< #9: !arAer< (lu$ter analy$i$< 9(@@#:<
2or)h
ologica
l trait$
/$tract 0n thi$ $tudy, 40000 ti$$ue culture4
deri,ed /anana )lant$ (,itro)lant$) at different
gro*th $tage$, i+e+ accli!ati3ation, nur$ery and
o)en field of /anana (2u$a $))+) culti,ar BGrand 'aineC *ere $creened
for $o!aclonal ,ariation$ u$ing !or)hological in,e$tigation$ and
!olecular charac4teri3ation+ &he total detected ,ariant$ *ere grou)ed
into 25 off4ty)e$ (t*o of the! died) in addition to the nor!al )lant+
#ando! !)lified 9oly!or)hic :' (#9:) *a$ carried out to $tudy
the difference$ a!ong the nor!al culti,ar BGrand 'aineC and it$ 23
,ariant$ u$ing 17 ar/itrary )ri!er$+ (lu$ter analy$i$ re$ult$ re,ealed
that B*inged )etioleC and Bdefor!ed la!inaC *ere !ore related to the
nor!al )lant+ 8o*e,er, BGiant )lantC and B*eaA )lantC related to each
other
and
clu$ter
ed
*ith
nor!a
l )lant+
ccor
ding to
)rinci)al coordinate analy$i$, !o$t of the
,ariant$ *ere aggregated nearly, *herea$
B,ariegated )lantC *a$ $e)arated a)art fro! the
other ,ariant$+ &hi$ !ay reflect the genetic
difference /et*een B,ariegated )lantC and the
other ,ariant$+ &he re$ult$ o/tained fro! /oth
!olecular and !or)hological analy$e$ *ere in
contiguou$ *ith /etter re$olution *hen u$ing
7
(orre$)onding author+ ddre$$D 9lant Biotechnology
:e)art!ent, Genetic Engineering and Biotechnology
#e$earch 0n$titute (GEB#0), 2inoufiya ;ni,er$ity, = El4
8a$$an Ben ly "t+, 1$t Eone, "adat (ity 7F, Egy)t+ &el+D
G20 04H2=012=4+
E4!ail addre$$D Aa!al2004grIyahoo+co! (5+6+
/dellatif)+
1=H74157J K 2012 cade!y of "cientific #e$earch %
&echnology+ 9roduction and ho$ting /y El$e,ier B+L+ ll
right$ re$er,ed+
9eer re,ie* under 'ational #e$earch
(enter, Egy)t+
htt)D--dM+doi+org-10+101=-.+.ge/+2012+05+
002
9roduction
and ho$ting
/y El$e,ier
4H 5+6+ /dellatif et al+
the 9(@@#: analy$i$ than clu$ter analy$i$+ &hu$, it can /e $aid that !olecular !arAer$ can
/e u$ed to eli!inate the unde$ira/le $o!aclonal ,ariant$ fro! the la/ *ithout additional culture
of the ,itro)lant$ in the field in order to $a,e ti!e and effort$+
K 2012 cade!y of "cientific #e$earch % &echnology+ 9roduction and ho$ting /y El$e,ier B+L+ ll
right$ re$er,ed+
1+ 0ntroduction
Banana (2u$a $))+) i$ one of the !o$t i!)ortant !e!/er$ of the
2u$aceae fa!ily+ Banana$ are gro*n in 12H countrie$ *ith a total
culti,ated area of 4+F2 !illion hectare$ and total *orld )roduction
of F7+3H !illion !etric ton$+ 0ndia ranAed fir$t all o,er the *orld in
/anana )roduction, *hich )roduce$ 27 !illion !etric ton$ N20O +
9lant$ *hich ha,e /een )ro)agated /y in ,itro ti$$ue culture
techniPue$ are Ano*n to eMhi/it a *ide array of genetic and
e)igenetic ,ariation *hich i$ Ano*n a$ $o!aclonal ,ariation N7O +
lthough the cau$e$ of genetic in$ta/ility are )oorly under$tood,
chro!o$o!e in$ta/ility i$ /elie,ed to /e one of the !o$t co!!on
cau$e$ of ti$$ue cul4ture4induced ,ariation N15O +
(haracteri3ation of induced !utation$ and $o!aclonal
,ariation$ /et*een induced !utant BG'=0C and it$ original
,ariety BGrand 'aineC of geno!ic :' u$ing ar/itrary )ri!4er$
*a$ )erfor!ed /y 6ernande3 et al+ N4O + 0n addition 9ancholi et al+
N12O $tated that a #ando! !)lified 9oly!or)hic :' (#9:)
!arAer /a$ed )rotocol *a$ de,elo)ed to $creen for $o!aclonal
,ariation in /anana$ in ti$$ue culture, u$ing (ach4aco Enano
(B), >anga!/i () and 9i$ang *aA (BB) )lant$+ &hey
re)orted that 17Q of the )lant$ *ere found to /e ,ariant$ and the
,ariation *a$ genoty)e4de)endent+ &hey found al$o that
,aria/ility increa$ed *ith an increa$e in the co)y nu!/er of
geno!e , /ut it decrea$ed *ith an increa$e in the co)y nu!/er
of geno!e B+ &heir re$ult$ indicated that #9: !arAer$ could /e
u$ed to !onitor the le,el$ of $o!acl4onal ,ariation+
#a.a!anicAa! and #a.!ohan N13O re)orted that, out of the 414
deca!er )ri!er$ $creened for /anana #9: analy$i$, 34 could
)roduce a!)lification+ &*enty4fi,e )ri!er$ $ho*ed high le,el of
)oly!or)hi$! and $iM of the !o$t )ro!i$ing )ri!er$ (@9401,
@9403, @9413, @9B404, @9B410 and @9B412) *ere u$ed for
#9: analy$i$+ #ecently, "aifullah et al+ N17O re)orted that 13
,arietie$ of the culti,ated /anana, )rocured fro! 0'0B9,
Belgiu!, *ere $creened u$ing #9::' !arAer$+ @nly three
#9: )ri!er$ (a!ong 20 te$ted) *ere cho$en a$ )roducing
)oly!or4)hic :' /and$ differentiating the in,e$tigated culti,ar$+
Ba$ed on tho$e identity !arAer$, the genetic fidelity /et*een
,ariou$ $u/culture le,el$ *ere deter!ined+
619 !arAer$ *ere u$ed in con.unction *ith !or)hologi4
cal de$cri)tor$, i$oe3y!e$, agrono!ic trait$ and #ando!
!)lified 9oly!or)hic :' (#9:) !arAer$ to characteri3e
the 2u$a acce$$ion$ in the gene /anA N1HO + 2icro$atellite
!arA4er$ *ere al$o u$ed to characteri3e /anana genoty)e$
NH,2O+ (re$te et al+ N2O re)orted that )henetic analy$i$ of
!icro$atellite !arAer /a$ed on Jaccard $i!ilarity indeM
deri,ed fro! )re$4ence or a/$ence of the allele$ agreed *ith
the !or)hological cla$$ification+
&he !ain o/.ecti,e of thi$ $tudy *a$ to characteri3e the
)roduced /anana ,itro)lant$ for /oth !or)hological and
!olecular (#9:) !arAer$
and to co!)are the re$ult$
of /oth !arAer ty)e$+
2+ 2aterial and !ethod$
2+1+ 9lant !aterial and
!or)hological trait$
8ealthy and unifor! /anana
off$hoot$ of culti,ar BGrand
'aineC *ere $elected fro! a
far! at h!ed @ra/y
Lillage, Badr (ity, Beheira
Go,ernorate, Egy)t in
ugu$t 200H+ &he off$hoot$
*ere )roliferated at the
9lant &i$$ue (ulture 1a/o4
ratory, 9lant Biotechnology
:e)t+, Genetic Engineering
and Biotechnology
#e$earch 0n$titute (GEB#0),
"adat (ity, 2inu4fiya
;ni,er$ity, Egy)t+ &*enty4
fi,e off ty)e$ *ere )roduced
throughout the )roliferation
and the e,aluation of the
)ro4duced ,itro)lant$+ &he
off ty)e$ *ere con$idered a$
$o!aclonal ,ariant$ N5O + ll
)roduced /anana
$o!aclonal ,ariant$ and
nor!al ,itro)lant$ *ere
gro*n in greenhou$e$ and
o)en field during the )eriod
fro! 200F until 2011 in a
far! at h!ed @r4a/y
Lillage, Badr (ity, Beheira
Go,ernorate, Egy)t in order
to /e e,aluated+
&a/le 1 &he /anana culti,ar
BGrand 'ainC $o!aclonal
,ariant$ and their
a))earance $tage$+
'o
+ 9henoty)e ca$e ))earance $tage
ccli!ati3ation 'ur$ery
6ield
1 'or!al G
2 ")ear $ha)e la!ina G
3 1eathery la!ina G
4 ?inged )etiole G
5 $y!!etric la!ina G
= 1a!ina defor!ation G
7
8alf ,ariegated
la!ina G
H Lariegated )lant G
F "tri))ed la!ina G
10 2alfor!ed )lant G
11 6an $ha)e )lant G
12 :*arf )lant G
13 ")rocAet la!ina
14 :efault la!ina
15 #eddi$h la!ina
1= 1ong )etiole
17
9ale green )$eudo
$te!
1H Ele)hant ear $ha)e
1F Erected leaf
20
BlacAened )$eudo
$te!
21 "hattered )unch
22 Giant )lant
23 Ligor )lant
24 ?eaA )lant
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