NHN NHANH in vitro GING HOA CC NHT RIVALRY

Study on Micropropagation of Japan chrysanthemum Rivalry

Nguyn Th L Anh, Nguyn Quang Thch

Summmary
The experiments ware carried out on new variety of chrysanthemum from Japan: Rivalry.
The research results showed that: HgCl
2
0,2% in 7 min. was the best for sterization treatment of the
stem and leaf tissue of chrysanthemum;Stem tissue was better than leaf one for explants. The initial
and multipropagation medium was MS+1ppm BA.The MS medium without growth regulations was
the best for rooting of shoot.The mix of soil, sand and burned rice husk (1:1:1) was suitable for
transfering the plantlets to ambient condition.Fourteen thousand plantlets ware produced by tissue
culture due to apply above research results. Those plantlets grew and gave their flowers.
Key words: Rivalry, tissue culture , multipropagation


1. m u
Trong cc loi hoa ct hin nay, ch ng sau hoa hng, hoa cc l mt trong nhng loi hoa
c a chung v trng ph bin nc ta (Linh Xuan Nguyen, 1998). Hoa cc c a chung
bi s a dng v chng loi v mu sc; phong ph v kiu dng, kh bn v ph hp vi nhiu
mc ch s dng. Hn th, cy cc c iu kin ra hoa to ngun sn phm hng ho lin tc
v n nh quanh nm (John, Harold, 1999). Chnh v vy, nhu cu c c cc ging cc mi thng
qua chn to v nhp ni nhm p ng c nhu cu ca th trng lun c ngi sn xut quan
tm, ch trng. gp phn lm a dng v ph bin ging mi, ng thi p ng mt phn nhu
cu cy ging cho ngnh sn xut hoa cc nc ta, ging cc Nht Rivalry c nghin cu
nhn nhanh bng k thut nui cy m.
2. Vt liu, phng php nghin cu
Nguyn liu cho cc th nghim in vitro l l v on thn ca cnh hoa cc Rivalry
(Dendranthema indicum Gr.). y l ging cc c kch thc hoa ln (ng knh hoa gp i cc
i o), mu sc mi l, ngun gc t Kyushu Nht Bn, c Vin Sinh hc Nng nghip du
nhp vo Vit Nam nm 2000. Nguyn liu cho cc th nghim ex vitro l cy con hon chnh nhn
c t nui cy in vitro.
Th nghim c tin hnh theo phng php nghin cu nui cy m hin hnh (Robert,
Dennis, 2000), trn nn mi trng c bn MS (Murashige,Skoog, 1962). iu kin nui cy: nhit
phng nui 25- 27
o
C, m phng nui 70%, cng chiu sng: 3000lux, thi gian chiu
sng: 16 gi/ngy. Th nghim c b tr ngu nhin, 3 ln lp li, mi cng thc 30 c th.Cc ch
tiu th nghim c quan st thng xuyn vi 7-10 ngy 1 ln (tu theo yu cu ca th nghim)
S liu thc nghim c x l thng k sinh hc theo chng trnh IRRISTAT. Cc th nghim
c tin hnh ti phng th nghim nui cy m, t bo thc vt v vn thc nghim ca Vin
sinh hc nng nghip, trng i hc nng nghip I.


3. Kt qu v tho lun
3.1 Giai on to ngun mu khi u
To c ngun mu sch c kh nng sng cao l iu kin tin quyt cho s thnh cng ca
vic nui cy in vitro. Trn c s ca nghin cu cng b trc y (Nguyn Th L Anh v cng
s, 1999), kt qu kh trng mu cy bng HgCl
2
nng 0,2% trong thi gian 7 pht cho t l mu
sch, sng i vi m l l 76,7% v m thn l 86,7%. Cc mu ny c nui cy ti sinh chi.

Bng 1. nh hng ca BA n s ti sinh chi t m l (sau 4 tun)
Cng thc
T l mu to
chi (%)
S chi/mu
cy (chi)
Chiu cao chi
(cm)
S l/chi
(l)
/C (MS) 0,0 0,0 0,0 0,0
MS + 0,5ppm BA 40,0 1,5 1,3 2,5
MS + 1,0ppm BA 43,3 1,6 1,5 4,4
MS + 1,5ppm BA 76,6 3,1 1,6 5,8
MS + 2,0ppm BA 66,7 2,5 1,4 3,2
MS + 2,5ppm BA 53,3 2,0 1,3 2,5
MS + 3,0ppm BA 50,0 1,2 1,2 1,8
LSD (0,05) 3,9 0,1

S c mt ca n cht BA vi cc nng khc nhau trong mi trng nui cy c kh
nng kch thch kh nng ti sinh chi t m l (bng 1). Ch cc cng thc b sung BA mi c kh
nng ti sinh chi. Tuy nhin, hiu qu kch thch ti sinh chi s b gim i khi b sung BA nng
cao. Khi b sung BA nng 1,5ppm cho hiu qu ti sinh chi cao nht. Bn cnh , cht
lng chi ti sinh th hin qua cc ch tiu sinh trng cng tt nht.

Bng 2. nh hng ca BA n s ti sinh chi t on thn mang 1 mt ng (sau 4 tun)
Cng thc
T l mu
to chi (%)
S chi/mu
cy (chi)
Chiu cao chi
(cm/chi)
S l/chi
(l/chi)
/C (MS) 100 1,0 6,9 2,1
MS + 0,5ppm BA 100 3,4 6,3 1,6
MS + 1,0ppm BA 100 4,6 6,6 1,5
MS + 1,5ppm BA 100 4,1 6,4 1,3
MS + 2,0ppm BA 100 3,8 6,7 1,2
MS + 2,5ppm BA 100 3,3 6,4 1,1
MS + 3,0ppm BA 100 3,1 7,7 1,0
LSD (0,05) 0,1 0,1 0,4 0,1

on thn mang mt ng cho t l ti sinh t 100% trn tt c cc mi trng (bng 2), do
kh nng bt chi ca mt ng t b nh hng bi cht iu tit sinh trng (trong gii hn nghin
cu). Tuy nhin, mu cy ch to cm chi khi c cy trn mi trng c b xung BA. iu ny
chng t BA kch thch kh nng ti sinh chi ca mu cy v b xung BA nng 1,5mg/lt
cho t l ti sinh t 100%, s chi ti sinh cao nht (t 4,6 chi/mu cy), chi ti sinh sinh
trng mnh (ch km cng thc i chng).
3.2. Giai on nhn nhanh
Giai on ny c thc hin bng phng thc to cm chi t cc chi ti sinh thu nhn c
trong giai on trn.


Bng 3. nh hng ca BA n h s nhn chi trn hai ngun mu (sau 4 tun)
Chi ti sinh t m l Chi ti sinh t m thn Cng thc
H s
nhn (ln)
Chiu cao
chi (cm)
S l/
chi (l)
H s
nhn (ln)
Chiu cao
chi (cm)
S l/ chi
(l)
/C (MS) 1,0 2,5 6,3 1,0 4,4 14,5
MS + 0,5 ppm BA 3,6 1,7 5,9 4,7 3,4 6,4
MS + 1,0 ppm BA 6,0 2,0 6,0 7,1 3,2 6,3
MS + 1,5 ppm BA 4,1 1,5 5,2 5,4 3,0 6,5
LSD (0,05) 0,3 0,1 0,2 0,3 0,4 0,6
Sau 4 tun nui cy cho thy mi trng c b sung BA c tc dng kch thch kh nng
chi ca cy hoa cc (ch cc cng thc b xung BA mi c kh nng chi) trn c hai ngun
mu l chi ti sinh t m l v chi ti sinh t m thn (bng 3). Tuy nhin, b sung BA nng
cao s lm gim kh nng chi, h s nhn gim 1,7 ln (t 7,1 xung 5,4) khi tng nng BA
t 1,0ppm ln 1,5ppm. H s nhn i vi ngun mu l cy ti sinh t on thn cao hn so vi
chi ti sinh t m l (7,1 ln so vi 6,0). Mi trng ti u nhn nhanh cy hoa cc trn c hai
loi ngun mu l: MS + 1mg/lt BA. Trn mi trng ny cho h s nhn l 7,1 i vi chi ti
sinh t m thn v 6,0 i vi chi ti sinh t m l.
3.3. Giai on to cy in vitro hon chnh

Bng 4. nh hng ca -NAA n sinh trng v kh nng ra r ca chi cc
ti sinh t cc ngun mu khc nhau (sau 2 tun)
Chi c ngun gc t m l (chi ti sinh t m l)
Cng thc T l ra
r (%)
S r/cy
(r)
Chiu di r
(cm)
S l/cy
(l)
chiu cao
cy (cm)
/C (MS) 80,0 9,1 3,8 7,3 2,3
MS+0,25pp mNAA
86,7 13,5 3,0 6,5 1,9
MS+0,5ppm NAA
100,0 15,8 3,0 6,9 2,0
LSD 2,8 2,0 0,1 0,1
Chi c ngun gc t m thn (chi ti sinh t m thn)
Cng thc T l ra
r (%)
S r/cy
(r)
Chiu di r
(cm)
S l/cy
(l)
chiu cao
cy (cm)
/C (MS) 100 9,6 5,0 11,6 2,8
MS+0,25ppm NAA
100 12,0 2,8 10,3 2,7
MS+0,5ppm NAA
100 14,8 2,2 10,5 2,8
LSD 0,6 0,1 1,0 0,03

B xung -NAA lm tng s r/cy nhng li lm gim cht lng ca b r khi so vi mi
trng khng b sung -NAA (/C). Mt khc, chi ti sinh t m thn c kh nng ra r mnh
hn so vi chi ti sinh t m l. iu ny chng t c mt s thay i ln v sinh l ca chi
cc ti sinh t m l, do m l phi qua giai on to chi bt nh trong khi chi ti sinh t m
thn thc cht l chi c nhn nhanh theo phng php vi gim cnh nn khng phi qua giai
on ti sinh chi bt nh.
Cy cc c kh nng t ra r rt mnh, t l ra r trn mi trng khng b sung cht iu tit sinh
trng t 80,0% (i vi chi ti sinh t m l) v 100% (i vi chi ti sinh t m thn). iu
chng t: chi cc c kh nng tng hp v t iu chnh lng cht iu tit sinh trng ni sinh
theo hng to r bt nh.
3.4 Giai on vn m
Bng 5. nh hng ca gi th n sinh trng, pht trin ca cy cc cy m (sau 4 tun)
Gi th
T l sng
(%)
tng
chiu cao
(cm)
S l mi/ cy
(l)
Quan st
Tru hun 100 3,1 4,9 Cy mp, l to, xanh u
Ct 100 2,2 4,3 L to, xanh, thn mp
t 36,7 2,9 4,4
Cy to, mp, kho, l to v dy, mu
xanh m
X than 53,3 1,5 4,2 Cy yu, chm pht trin, l vng mng
t+ct (1:1) 80,0 3,0 4,6 Cy kho, l to xanh
t+ct+tru
hun (1:1:1)
100 3,4 5,0
Cy mp, kho, mc u, thng cng l
to xanh thm dy bn

Trong giai on vn m, gi th c nh hng quyt nh n kh nng sng, sinh trng
ca cy cc cy m. Nhn chung, cc gi th c b xung tru hun s cho t l sng, sinh trng,
pht trin ca cy cc cy m tt (gi th tru hun, v t + ct +tru hun theo t l 1:1:1 cho t l
sng t 100%, cy sinh trng mnh). Gi th x than v t t ra khng thch hp vi cy cc cy
m. iu ny c th do gi th x than c cc c tnh l ho khng ph hp vi s pht trin ca b
r cn gi th t li b trong thi gian u tin khi ra cy v vy lm hn ch kh nng sinh
trng cng nh tng kh nng nhim bnh ca cy.

3.5 S sinh trng, pht trin ca cy cc cy m trong sn xut

Bng 6. S sinh trng, pht trin ca cy hoa cc Nht cy m v
hoa cc vng i a (trng v ng 2002 ti Hi Phng)
Ch tiu theo di Cc vng i a Cc Nht Rivalry
Thi gian t trng n ra hoa
( )
77 83
Chiu cao cy (cm) 48,2 86,5
ng knh thn lc thu hi (cm) 0,60 1,10
T l ra hoa (%) 100 100
ng knh bng hoa (cm) 6,7 13,3
ng knh cung hoa (cm) 0,3 0,8
Chiu cao bng hoa (cm) 12,3 14,2
Mu sc hoa Vng m Vng ti
c im hnh thi
Cy c thn l
cn i
Cy c thn l to, nng,
d
p dng cc kt qu nghin cu nu trn, Trung tm ng dng khoa hc k thut cng ngh
- S khoa hc Cng ngh Hi Phng sn xut c 14.200 cy cc Nht ging Rivalryv trng
th ti ng Hi - Hi Phng. Cc ch tiu sinh trng ca ging cc ny nh chiu cao cy, ng
knh thn lc thu hi, ng knh bng hoa, ng knh cung hoa, chiu cao bng u cao hn cc
i a. T l ra hoa 100% (bng 6).
Kt qu ny khng nh quy trnh nhn ging c th vn hnh sn xut ng thi cng gii
thiu cho sn xut mt ging hoa cc mi l. Do c kch thc hoa ln, chiu cao v ng knh
bng hoa gn tng ng nhau nn nng dn gi l hoa cc n lng. Ging hoa ny c th s
dng tt vo cc mc tiu trang tr.
4. Kt lun
Quy trnh nhn ging cy hoa cc Nht Rivalry bng k thut nui cy m vi ch kh
trng thch hp cho m thn v m l cc l: HgCl
2
0,2% trong 7 pht, ch kh trng ny m l
cho t l mu sch, sng t 76,7% v m thn t 86,7%. M thn mang mt ng c s dng
lm vt liu nui cy khi ng v cy ti sinh t m thn s cho cht lng cy con tt hn, kh
nng nhn nhanh cng nh ra r mnh hn. Mi trng MS + 1ppm BA c chn nui cy khi
ng, nhn nhanh in vitro. Mi trng MS khng b xung cht iu tit sinh trng c chn lm
mi trng to cy hon chnh, trn mi trng ny chi ra r 100% sau 2 tun nui cy, kh nng
sinh trng v pht trin ca cy con tt. Gi th thch hp cho cy cc cy m ngoi vn m l:
t+ ct + tru hun (1:1:1), gi th ny cho t l sng t 100%, cy con sinh trng mnh nht.
Vi qui trnh ny, sn xut c hng chc ngn cy ging trng th nghim Hi Phng, H
Ni, Thi Bnh.

Ti liu tham kho
Nguyn Th L Anh, Nguyn Quang Thch, T Th Hoi Thu (1999). Nghin cu cc phng php
nhn nhanh mt s ging hoa cc chm H lan. Bo co khoa hc Hi ngh Quc gia v Cng
ngh sinh hc, tr. 859-865
John M. D., Harold F. W., (1999). Floriculture Principles and Species, Prentice Hall, Inc., p. 292-
303
Linh Xuan Nguyen, (1998). Cut Flower Production in ASIA. FAO, Bangkok, Thailand, p.63-67
Murashige T. and Skoog F., (1962). A revised medium for rapid growth and bioassay with tobacco
tissue culture. Physiol. Plant. 15. pp: 473- 497.
Robert N. Trigiano, Dennis J. G., (2000). Plant Tissue Culture Concepts and Laboratory Excercises,
CRC Press LLC, p. 75-85