KHOA HpC C 6 M 0 NOHi
IVHAIXI GidlVG IM VITRO C A Y DLTA S A P
{Makapuna cacanut^
Tniong Qudc Anh\ Urong Thd MinhS Truong TTiJ T^ AIlh^ Throng Vinh Hii' T6MTAT
Cay dira {Cocos nucifera L) lA mOt trong nhOng c4y Idy dAu da nlfin chu ydu ciia vung nhl^t ddi, dugc trdng rfit phd bidn d nhidu nudc. Ngo^ v i ^ trich dflu tir cam dira, c4c ph4n phy khic nhu xcr, gio, nudc dira cQng duiTc Sli dyng d^ ch^ bi^n t h ^ h cdc silii phSm c6 gii tri cao nhu thim m difa, than ho^t tinh, th?ch diia.
Tinh TVd Vinh c6n c6 gidng dira d^c ru^t, mOt gi^ng dira bSn dja djc tnmg ntfi ti^ng. Vdi cic t h ^ h phin gdm mdi trudng ca bSn Y3, dudng, aga, than ho^t tinh vi ch^t di^u hda sinh trudng NAA. tir 250 quii thu th§p ban ddu da thu dugc 235 phdi, sau 6 thAng tao ra 177 diy, dgt tj 1^ 71% so vdi s6 phdi thu thftp ban diu.
K^t qua thi nghi$m Anh hudng ciia ndng dO dudng d^n khd nSng niy mim ctia phdi cho thiy, trftn mfii trudng nudi c^y c6 b5 sung 60 g/L sucroza, ty 1$ phdi niy mim cao nhit (77,5%), sv khic bi^t niy c6 y nghia v^ rn^lt thdng V.i. Thi nghidm v^ chit di^u hda ainh trudng NAA 16n si; hinh thinh rh trong nudi ciy phdi dira Sip vdi cic ndng dd 0,5, 1 vd 1,5 mg/L cho k^t qui nhu sau; cic chdi con tgo rk trftn mdi tnrdng Y3 c6 chua 1 mg/L NAA IA tdt nhit. Sau 6 tuin nudi ciy. 9096 chdi t?o 'M ri, dii 2-3 cm. Hi thdng ri phit tri^ii tdt vi phin nhinh nlii^u r^ ngang khic bi^l cd y nghla so vdi cic nghidm thuc khic. Sy thinh cdng trong phuong phip nudi ciy in vitro cudi ciing phg thudc vio ty 1$ ciy sdng d giai do^n vudn uom. K^t qui thi nghidm cho thiy, gii th^ cit c6 dd gift nudc k^m nftn h?n chd sy ngip ung cua bd ri trong giai do^n diu nin cho ty 1^ sdng.
Tu khda: Makapuno, phdi soma, dua SAp, Y3.
LMOBMI
Cay dira {Cocos nucifera L.) la mdt trong nhiing cay lay dau da nien chii y^u cua viing nhi^t ddi, dirpc trong rat pho bien b nhi^u nudc. Cay dira tir Idu da dugc xem la cay ciia cudc sdng (tree of life), Ciy dira cd rat nhi^u cdng dyng so vdi cac lo&i c^y trong khac vi h4u h^t cac phan cua qua dira, 1^ dira v4 thSn dira deu cd the dugc sir dung phuc vy cho nhu diu cua ddi song con ngudi.
O Vi^t Nam cd nhi^u giong dira, nhdm dira l^y dau g6m cd dira Ta, dira Dau, dira Dia, dira Bj...
ngoai vi^c l^y d^u tir cam dira, c^c p h ^ phu khic nhu xa, gao, nudc dCra ciing diig«c sii dung d^ ch^
bi^n thinh cac san p h i n c6 gi^ tri cao nhu tham s a dira, than hoat tinh, thach dira. Ngo^ cic giong dira chu y^u ndu tr&n, Vi^t Nam cdn cd gidng dira die rupt, cdn dupe gpi la dira Sip. Dira S^p cd nguon gdc tu Philippin, la hidn tugmg dot bi^n gien cua gidng dira cao Laguna, chi phdi bdi mot gien l$n duy nh^t (RiUo, E. P. va M. B. F. Paloma, 1992). Dira Sip phSn bd b Viet Nam chu y^u b tinh Tr^ Vinh, die bidt d huydn Cau Ke, noi dupe eho la viing diit tdt nhit d^
trong dira Sip. Cac loai trii dira ndi chung thudng trai qua vii giai doan: khi dira cdn non, com m^m
'Vifn Khoa hpc KJ thu(it Ndng nghidp mien Nam
d^o, nudc ngpt, khi g i i thi com dira ciing lai, nudc nhgt d ^ v i cd th^ I6n men. Ri6ng dira S ^ thi sau khi trii qua giai doan cdn non vdi cam dira vi nudc dira, sg ti^p tv>. phit tri^n day d4n ph4n com diia 1^, Xip gdn ddy khoang trdng cua gio dira, chi d^ 1^
m$t khdng gian nhd chinh giua vdi chit ldng s$t, cd miii thom d$c tnmg; com dira dgng xdp, m ^ vi deo ehii khdng cd.i ciing nhu com dira cua cic qui dua khic (Vd Van Long vi ctv. 2005).
Dira Sip khic c i c gidng dua cao d chd m6i budng thudng cd 2-3 trii d$c rudt, sd cdn lai binh thudng. V^ phirong p h ^ nhin gidng, ddi vdi cic gidng dira thudng chi cdn chpn cSy m? khde m?nh, trii sai, chit lupng com (dira khd), nudc (dira non) vira y, chd trai gii (khd) thu hii trii d^ gidng. E)di vdi dira Sip, chi nhiing trii khdng cd siy> mdi cd kha nang tao phdi, t<io m<>ng, mdm, t^o ra cay dira Sip gidng; nhiing ti-ii cd s i p khdng th^ d^ gidng. Do thij phdn chto, th^ he cSy con khd x i c dinh v^ tinh tr?iiig vi chdt lupng u ^ . Do dd, mudn nhin gidng dira Sip, ngudi ta irom trii binh thudng tr^n cay dira Sip va th^ h$ ti^p theo ciing cho ti 1$ trii dac mdt tuong duong 25%. Vi v^y, d^ khdc phyc han ch^ niy, cic n h i khoa hpc. da nghi6n ciiu s i n xudt ra gidng dto Sip bdng phuong phip nudi cdy phoi d^ nhin gidng vi gia tang ti 1$ trii s ^ . K^t q u i nghifin ciiu nhin
12 NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 10/2012
KHOA H p c CdNO NOH$
gidng in vitro tr6n dira da chi ra rdng, ung dung phuong phap nhin gidng in vitro la kha thi cho vi&c tai t^o ciy. BSn canh dd, nhd sir ling dung dia chdt di^u hda sinh hudng ciing da lim tang ty 1^ hinh thanh phdi v i ldm tang ty 1$ ciy con. Vi v^y, trudc y6u cdu cua thuc ti^n san xudt can phii nghidn ciiu
"Nhan gidng in vitro ciy dira Sip {Makapuno coconut" nhdm muc dich ho trg cho viec hoin thidn quy trinh nhin gidng dira Sip, giiip cho cdng tic lai tao v i bao ton gidng dira Sip hi^n nay.
L VAT L P Vii PHUONG PHAP l.V$tli§u
Dira Sip thu tai xa Hda Tan, huy^n Cdu K^, tinh T r i Vmh d^ lay phdi dya vao cic y^u td sau:
- Qua dira Sip chpn tir cdc cSy m? da dupe tuy^n chpn va dinh ddu trudc dd, qua cd kich thudc trung binh va ning cin, qua dupe chpn phai d^u d^n, khdng di dang va bi siu b^nh phi hoai. Qui dupe chpn sau khi dupe thu hoach d^ trong bdng mit 2 - 4 tudn d^ hoan tdt qua trinh chin sinh ly.
- Mdi trudng ca bin Y3 vdi cac Ihanh phdn gdm:
Khoing da lupng: NH4CI, KNO3, KCl, MgzSO^, KH2P04. Cic mudi trung, vi lupng: MnS04.4H20, ZnSOi.THaO, CuSO^.SHaO, KI, CoClz.eHjO, H3BO3, Na2.EDTA.2H2O, FeS04.7H20, NiCl2.6H20, NaH2P04.2H20, CaCl2.2H20. Cac mudi him ca v i vitamin: Myo-hiositol, nicotinic axit, pyridoxin HCl, thiamin HCl, glyxin, Ca-D-pantothenat, biotin.
- Phdi diia 10-12 thing tudi, dudng sucroza, aga, than hoat tinh, NAA, BA, IBA, canxi hypoclorit, con, NaOH, HCL, nudc cat ti^t trung, cac dung cij trong phdng.
2. Vd triing mdu nudi cdy
Dua Sip thu v^ phdng thi nghidm, bd ddi trii ra lam hai phdn theo chi^u ngang, sau dd diing dung cu lay mdu (stain steel cork borer) dn mgnh vio phdn trung tam phia b^n cd vdt ldm d^ lay dupe mot vimg com dira chiia phdi, mdu sau khi dirpc Idy ra bao quan trong nude cdt vd trung. Rua mdu bdng cdn 70° trong 1 phiit Rua lai 3 lan bang nudc cat vo trung (5 phiit/ldn). Tich lay phdi rdi ngam trong canxi hypoclorit 5% trong 20 phiit, riia lai bdng nude cdt vd triing 3 ldn, mdi lan 5 phut rdi cdy vao mdi trudng da chuan bi sdn.
3. Bd tri thi nghidm
Di6u ki§n phdng nudi: inh sing den huynh quang, cudng dd inh sing 1800 lux, thdi gian chidu sing 16 gid/ngiy, nhi^t dO phdng nudi 25''C (± 2°C), am dp phdng 40%.
Hdp khii triing d nhi^t dd 121°C, hdp 20 phiit ddi vdi mdi tnrdng nudi cdy vi 30 phiit ddi vdi dung cu.
Thi nghidm dupe bd tri hoan toin ngau nhi^n 4 ldn nhdc 1^. Sd mdu trdn mdi nghidm thiic la 10 mdu.
4. Xii 1]^ sdlieu
Cic kdt qud thi nghidm xir ly thdng ke theo phuong phip phin tinh phuong sai theo bang ANOVA So sinh kdt qud vi xdp hang theo phuong phip Duncan. Su dung phdn m^m xii ly thdng ke MSTATC v i Microsoft Excel.
•.KETQUAvAniAOLUAN
1. Nhin gidng diia Sip bdng phuong phip nudi cdy phdi
Nhin gidng dira Sip bdng phuong phip uam truydn thdng cua nong dan cd ty Id qua sap/buong chi dat 20-25%. Theo Torres (1937) vi Zuniga (1953), mpt trong nhimg bidn phip lam ting ty Id sap/buong
>75% l i sir dung nhung cay tir nudi cdy phdi. Tir nhung thanh tuu v^ nudi ciy phoi dua Sip cda Philippin, d^ tai cung d i nghi&n ciiu quy trinh nudi cdy phdi gdm 5 giai doan sau day:
- Giai doan chuan bi lay phdi tir qua diia Sip.
- Giai do?ii khic triing phdi dira.
- Giai doan nay mam ctia phdi.
- Giai dogn phat tri^n cua phdi thinh ciy dira.
- Giai doan thich nghi ciy cdy phdi tnrde khi dua ra vudn uom.
Vdi quy trinh nudi cay phdi dira Sip nhu tr^n, ttx 250 qua thu thip ban ddu, thu dupe 235 phdi, sau 6 thing tao ra 177 ciy dat t? 1§ 71% so vdi sd phdi thu th$p ban ddu (bang 1).
Bing 1. Ty 1$ phit triin cua phdi b cic giai doan trong phdng Ihi nghidm Stjqui
ban ddu
250
Tich phdi
S6 ph6i
235
«)
94
l y i ^ p h a i niy mdm (3
thing tu6i) S6 phdi
207 TylS
(%)
83
T}'!^ phdi phit tri& (6 thing
tudi) So ciy 177
Tyl«(%) 71
NONG NGHlfP VA PHAT TRIEN NONG THON - KY 2 - THANG 10/2012
KHOA HQC C 6 N 0 NCHt
2. V6 trung mdu nu6i cdy
Bing 2. Kit qud xii 1^ hypoclorit canxi v6i thM gian Nghidm
thiic (NT) 1 2 3 4 5
:vw
^D„„,N6ngd0 hypoclorit
canxi W 5 5 5 5 5
IhMgian xiil^
(phiit) 5 10 15 20 25
Mdu sdng 2,43 c 5,11 be 6,89 ab 8 ^ a 6,44 ab 12,05 3,98
Tyl«ph6i s6neOi)
24,33 51,10 68,90 82,30 64,43
Anh hudng eua thdi gian xii \^ hypoclorit canxi ddn vi^c vd trimg mdu phdi dira Sip.
Biiu dd 1. Anh hirdng ciia thdi gian xii l^ hypoclorit camd vdi ndng dd 5% ddn v i ^ vd triing mdu phdi dira
S ^ sau 30 ngiy nudi cdy
Ki^t qui thi nghidm anh huong eiia thdi gian xii ly hypoclorit eanxi vdi nong dd 5% ddn v i ^ vd triing mdu phdi dira sip sau 30 ngay nudi cdy dupe trinh bay b bang 2 vi bidu dd 1. Kdt qud eho thdy, xu ly hypoclorit canxi 5% trong thdi gian 20 phiit eho kdt qua cd ty 1§
mdu sdng cao nhdt la 82,3%, khic bi^t cd y nghia so vdi nghidm thiic xii ^ vdi thdi gian 5 va 10 phiit Kdt qua so vdi nghidm thuc xtr ly d 15 phiit va 25 phiit thi khdng khic bi^t ed y nghia. Tuy nhidn, ty Id mdu sdng cd xu hudng giam ddn theo Ihdi gian (xu ly sau 20 phiit), cd Ihd do do ddc ciia hypoclorit canxi anh hudng den phdi ndu xii ly d thdi gian lau hon 20 phiit
Cay dira sap Id mdt loai ciy rat khd khii trimg, ciy cd dd nhi&m tap rdt cao vdi moi chat khtr trimg;
ndu ndng dp ehdt khii trimg thdp, thdi gian khii trimg ngdn thi ty Id mdu cdy bi nhi^m cao, nhung chi cdn tang ndng dp hoac thdi gian khtr trimg I6n la ^ 1$ ciy chdt da tang ldn mdt cich dang kd. Khi khu
trimg bang hypoclorit canxi vdi ndng dd thdp, thdi gian khu triing ngdn t^ 1$ mdu cdy hi ahikm cao. KA qud eiia thi nghidm khu triing phdi dira Sip dupe su dving cho eie thi nghidm tidp theo vd nghifen cuu i n h hudng eiia ndng dd dudng vd chdt di^u hda sinh trudng ddn si; niy mdm ctia phdi dira Sip.
3. Anh hudng ciia ndng dO dudng ddn si; phit tridn ciia phdi diia Sip
Bdng 3. Anh hudng ctia ndng dfy dtrdng ddn t^ 1(
sdng cua phdi sau 30 ngdy nudi cdy Nghidm
thilc (NT) 1 2 3 4 CV(%) LSD„,„
Dining (g/L)
0 30 60 90
StiphAi sdng 5,44 b 7,44 ab
9,11a 7,89 ab
6,44 3,54
Tyl*ph«
8dng(X) 54,43 74,43 91,10 78,9
Dd ddnh gii dnh hudng ciia ndng dd dudng d^a k h i ning niy mdm v i tang trudng cua phdi, nghi^
ciiu da thuc hi$n thi nghidm vdi eie ndng dp dudng khdc nhau. Kdt q u i thu dupe dupe trinh bay d bing 3 v i bidu dd 2. Kdt q u i thi nghidm cho thay, 6 nghidm thiic 3 vdi ndng dd dudng 60 g/L cho ty le phdi sdng cao nhdt (91,1%) sau 30 ngiy nudi cdy. G nghidm thiic 1 khdng cd bd sung dudng, ty 1^ phoi sdng thdp nhdt (54,43%), khic bidt ed y nghla so voi cic nghidm thuc khic vd cdc phdi phdt tridn c h ^ hon. Ndng dO dudng trong mdi trudng nudi cdy c6 anh hudng ldn ddn su phdt sinh phdi. Kdt qui cho thdy sd lupng phdi sdng sdt ti 1$ thudn vdi ndng do dudng. Tuy nhi€n ndu ndng dd dudng virpt qui gioi h ^ cQng lim ti 1$ chdt phdi tang cao do ndng do dudng cao gSy iic chd si; tgo chdi va rfe bdt djnh.
f r
Bidu dd 2. Anh hu6ng ciia ndng dd dudng ddn t^ 1$
sdng cua phdi diia S ^ sau 30 ngdy nudi cdy Thi nghidm ciing tidn hdnh dinh gid anh huong eua ndng dO dudng ddn khdi lupng cua phdi dira S ^ d giai do?n 60 ngay sau khi cdy. Kdt qua dupe trinh
14
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 10/2012
KHOA HQC C 6 N 0 NGHt
bay a bang 4 cho thiy, a cic nghi$m thiic, cic ph5i dCia Sap duoc ciy t r t a m6i truemg c6 nong dp duemg tang dan, thi c i c chi ti6u v^ sinh truong nhir kich thuoc, khii luong phdi d^u tSng. Tuy nhiSn, cic chi tieu tang trudng iia m0t miic n h i t dinh. 6 nghidm thiic 3 vol nong do du6ng 60 g/L, ph6i dira cho khoi luong cao nhat (2,72 g), khic bi^t rat c6 y nghla so voi cac nhijm thiic con lai. Nghidm thiic 1 c6 khdi luong phoi khong dat ti6u chuin (0,57 g).
Bang 4. Anh htr6ng cua ndng dO du6ng d^n khdi lupng ph6i dira Sip sau 60 ngiy nudi c^y Nghiem
thiic (NT) 1 2 3 4 CV(S6) LSD„m
Dirong (g/L)
0 30 60 90
Khdi linpng phdi (g) 0,57 d 1,49 b 2,72 a 1,11c 5,10 0,21 Dua vao bang tide nghidm phin h?uig d bdng 5 va bidu dd 3 cho thdy, nong dp dudng c6 anh hudng ddn chidu cao dira Sip in vitro a giai doan 150 ngay sau cay. Mdi trudng vdi ndng dO dudng 60 g/L cho ket qua tdt nhdt (9,57 cm), khic bifit rat ed y nghla ve mat thdng kd so vdi cac nghidm thiic khic. Chidu cao dat thdp nhdt d nghidm thuc khdng bd sung nong dd dudng (chidu eao chi dat 2,83 cm).
Bdng 5. Anh hudng ciia ndng dd dudng ddn chidu cao dira Sip in vitro sau 150 ngiy nudi cdy Nghiem
thiic (NTI 1 2 3 4 CV(9S) LSD„„,
Duong (g/L)
0 30 60 90
Khdi luong phdi (g) 2,83 d 7,53 b 9,57 a 5,88 c 3,58 0,63
Chi4u cao (cm)
15 10 5 0
2.83
1 7,53
•
2 9,57
•
3 5,88
•
4— N g h i ^ H i - t h u e —
Bidu dd 3. Anh hirdng ciia ndng dO dudng ddn su phdt tridn chidu cao dira Sdp in vitro sau 150 ngiy
nudi cdy
4. Anh hudng ciia ndng dd BA ddn si; phit tridn phdi dua Sdp in vitro
Kdt qua ciia vide bd sung ndng dd BA vio mdi trudng nudi eay ddn su phdt tridn ciia cay dira Sip in vitro dupe trinh biy d bdng 6, bidu dd 4.
Bing 6. Anh hudng cua ndng dd BA ddn ty Id phdi sdng sau 60 ngiy nudi cdy Nghidm thiic
(NT) 1 2 3 4 CV(%) LSD„,
BA (mg/U
0 0,5
1 1,5
Sdphdi sdng 6,11b 8,89 a 8,87 a 7,89 ab
7,14 2,31
Tyl$ phdi sdng 61,10 88,90 88,67 78,90
Vdi mue tifiu xie dinh ndng dd ehdt didu hda sinh trudng BA thich hpp ddn sir phit tridn cua phdi dira Sip in vitro kdt qua eho thay; Sau 60 ngay nudi cdy, cic nghidm thiic ddu cho kdt qua tdt. Su bo sung ndng do BA tir 0,5 ddn 1,5 mg/L cd ty Id phdi sdng trdn 78%, su khac bifet khdng cd y^ nghia vd mat thdng kd. Chi khic bidt cd y nghla so vdi nghidm thvic khong bd sung BA. Ty 1^ phdi sdng ciia nghidm thirc 2 la 88,90%, nghidm thiic 3 la 88,67%, nghidm thtic 4 la 78,90% so vdi nghidm thiic khdng bd sung BA (nghiem thirc 1), ty Id sdng chi dat 61,10%.
I !
i
l y
78,90
Nghidm thtrc Bidu dd 4. Anh hudng ciia ndng dO BA ddn ty Id
phdi sdng sau 60 ngiy nudi cdy Sd hdu vd su phat trien ciia la cung dugc thu thap (bang 7, bieu do 5, hinh 1). Mdi trudng nudi cay ed bd sung ndng dp BA = 0,5 mg/L cho kdt qua tdt nhdt ddi vdi su phat tridn cua l i 13,07 cm sau 150 NONG NGHIEP VA PHAT TRIEN N 6 N G THON - KY 2 - THANG 10/2012 15
KHOA HQC C d N G N G H l
iiK'iiy nuoi cay va kliac bii''l laf lo v iij^'lua ^<i viVi i:'ic iijjhiem tliirc con l.ii. cac !:i i:iy 1<i li.iii v.i x.uili Iv'i
\ur\) danh gia vc ty 1^ plioi ^"n^!. va sir pli.il li icii la cho thay niing do BA dmn- hu MIIIJ; la i).','> IHK/I ' h"
hicu qua l(")t nhal
'Illl llK'hit'ni i'un^ Iicii Iianh i]n chiru can . av (hni Sap III \ilji> Ket qua ihi iu;hiiiii i\\un liiiih hav iMiiij^ baiin S va hicu th\ (M IK. Ihav, iiioi liinriii; niioi
<,\\ ^'i^ h o ^ u u j , ' \\\ -- 0,r. iiiK'/l . i i o kri qu.l It,I Illlal (ten sir phai liicii uia ihtfii cati cav {V\.i\i .in) TlDntr nioi ti"ir(niK' klioiiK' co hn sunt; I'.A llii rav v.'iii phai ini'ii, iihini>,' sir ph.il Incii < hu ii i aulhaii ktin 1 uy ilhicn. ho snuj; liA (|ua nhicu Ihi sr ir, . ho r|oii sir iihal liion c\\.\ cav din S.ip ui \ilin. vi \\\ ii iiniin Miih InroiiK' HA CO t.K duuK ihiu dav sir phai Incii la di'i \oi ca\ dini Saji nuoi c.iv Ur plaa Iron iiioi li-innu;
V:i Co Ihc ho MiiiK chal di('u ho:i sndi Inr.nij; UA
\'ao iron.i; (|uy li-|iih iili.tii Ki'nik' dira S;i|) //; vHra voi noii^ do O.fi ni^' I. iV diiic d.iy ^ir |)hal liirn < ii.a chuni cao thau va chicii dai la KV smii co cay < on hoan cliinh dira ra vuon innn
Bang 7. Anh hmrng ciia ndng dc) BA d<^n sir phat tndn la diia sip m vitro sau 150 ngiy nudi cdy Nghiem thirc
(.N"n 1 2 3 4 CV(«) USD„„,
(mg/L) 0 0.5 1,0 L5
Chiru dai l.i (era) 4.87 c 13.07 a 8,63 b 1.33 d 3.61 0.69
1 3 , 0 7
u
S.(. J
Nghidm thU'c
A 1 nii;/l.
(linh 1. Aiili liuf'nig nia nc^ng d(> liA ddn suphSl
\n6n l/i <iir;i S/ip in vitn> s;iu 150 n^ay nudi cdy Bang 8. Anil hirc'nig ciia nimg d() BA ddn suphSt tni^ri <lii(^ii cao tii^ii dua S<''ip in v;£/r;sau 150 ng^y
nudi ciy NKIUCIIIIIIIIC-
(NO 1 3 4 CV(%) LSD„„i
BA (mg/L)
0 0,5 1,0 1,5
Cliiou dai la (cm) 4,47 c 7.13 a 5.87 b 2.03 d 4,15 0.55
a
iidu do 5. Anh hudng cua nong dy BA ddn su phit tridn Ii diia Sip in vitro sau 150 ngiy nudi cdy
Bii^u do (>. Anh hirc'nig ciia ndng dO BA ddn syphit tndn cliiiHi cao cay dira Sap in \itro sau 150 ngay
nudi cdy
5. Aidi humig ciia ndng dO NAA ddn qui trinJi hiiih thaiih va phat tridn cdy dira Sip in vitro
Ket (jua till ngliicm dui.»c tlie hien a bang9va bidu dd 7 Chat dicu hoa sinh tnnVng NAA phii \m clio qua Irinh tao rd cay dira sap, n^u sir dung NAA CO ndng do tliich h(,Tp (Jii ty le cay ra re cao, chi^udai r^ vua phai, cay phat tnen toL Ket qua tlii nghiem chi) thay, (V nghidm thuc 3 (ndng dd NAA = 1 mg/U so Immg rh nhidu nhdt (6,73 re). O nghidm thitc 1 kliong CO bd sung NAA thi s6 luong r^ it nhat {IJ'J d'). Nghiem thuc 2 va ngliiem tliiic 4 khdng co sii
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 2 - THANG 1O/2012
KHOA Hpc C 6 N G N G H |
i4c bi$t. Vi^ b6 sung NAA v4o m6i tniimg nuoi iy 14m c4y phat tri& nhanh v^ s6 lui?ng ri, tao si/
in doi cho cSy.
Thi nghiSm cung ti& hinh xac djnh n6ng dp AA thich hpp d& chi^u da rS diia Sip nu6i ciy i6i. Sau 150 ng4y nu6i cSy giua cic nghidm thiic c6 r khic bi^t rit co y nghia, a nghi6m thuc 3 v6i n6ng J NAA . 1 mg/l chi^u dii rS dii nhiit (6,37 cm), ghi^m thiic 1 cho chi^u dii rS thip nhit (3,47 cm).
Jing 9. Anh huong cua n4ng dS NAA iia s6 lupng 4 v4 chiSu dii rS cua dira Sip in Klro sau 150 ngiy
nu6i ciy Nghi$m thuc
(ND 1 2 3 4 CV(%) LSDo.,,
I - 4 .
NAA ( m g / q
0 0,5
1 1,5
Chi^u dii r§
(cm) 3,47 d 4,83 b 6,37 a 4,27 c 4,40 0,55 .^-cUfi!
,<t^ - ^ ^
^^-^ ^<:z -<-v /
S6 rS/ciy
1,70 c 5,30 b 6.73 a 4,93 b 6,22 0,79
= . «
Biiu d6 7. Anh hudng ciia n6ng dd NAA din sd uigng rlvcLchilu dii FI ciia dira S ^ 10 vitro^xx 150
ngiy nudi c ^
6. Anh huong cua giA thi l€n ty 1§ sdng cua ciy I vitro sau 180 ngiy nudi c^y
Su thinh c6ng trong phuong phap nu6i c^y mo ay trong cu6i cimg phu thupc vio ty 1§ cay sdng b iai dogn vuon uom. O mpt s6 loai thi hifen sy kh6 han b giai doan vucm uom hon nhung loii khic i6ng qua ly le sdng r^t thap. Co nhiiu nguyfin nhin htr Ty-1& tong quang hpp tiiap, cirong dO inh sing lu trong phong nuoi c^y (Lees, 1994
). Kit qua da xic dinh dupe r ^ g , ciy nudi d(y 1 vitm trong dilu ki|n cudng do anh sing dupe ing cudng se giiip cai thi|n ty 1| cay sdng esr vitro,
»di voi dira Sap, theo Assy-Bah vi ctv (1986) va Rillo 1998), t^ le sdng ciia ciy con b giai doan esr vitm hoing 40%. Trong sudt qui trinh nay, dilu ki|n moi Tidng ciia cay con se din d ^ thay ddi theo thdi Lan; thdi gian ban ddu li g ^ lien vdi dilu ki|n in itro vi kit thiic la gan vdi dilu kidn ciia nhi ludi. Do 6 viec nghien ciru cac dilu ki|n m vitro va ear vitro
phii hpp li cki thilt n h ^ giiip ting ty le ciy sdng.
Theo kit qui nghien ciiu ciia Dr Aurora G. Del Rosario (1997) chuyin ciy dira Sip nudi c^y phdi tu binh tam giie ra nhi ludi cho th^y: Khi ciy b binh tam giie dupe 3 - 4 li thi mang vio nhi ludi vi hu&i luy|n dudi inh sing tu nhidn vi nhi|t dO phdng Sau 2 tu^, ciy dupe l^y ra tu binh tam giie, riia sach vdi nudc dl loai bd mdi trudng nudi cdy. Sau dd diuig thudc trir ndm Dithane M45 vdi ning dd 2 g/L dl xir \f nim rdi cdy vio cit da khu trimg.
Kit qui d bing 10 cho thdy, d gii thi cit ty 1|
sdng eao nhdt (52,34%), vi gii dil eit ed dd giu nudc kdm nin han ehl su ngip ling eiia bd rl trong giai d o ^ ddu. Gii thi cit pha xa dira cQng cho miic dO thoing khi tdt, tuy nhidn kit cdu long leo khdng tgo dupe tinh dn dinh cho ciy vung, lim bd rl hd h ^ kIm, rl bj thdi tao dilu ki|n cho ndm mdc phit triin lie ehl s\f phit triin ciia ciy. Gii thi ddt pha xo dira cung cd do thoing khi, tuy nhidn ddt ciing cd kit cdu ch^t ndn dl lim bp rl ngip ling.
Bing 10. Anh hudng ciia gii thi ldn ^ 1$ sdng dia ciy in vitro sau 180 ngiy nudi cdy Nghidm
thiic F l F2 F3 F4 CV(«) L S D „ ,
Giith*
Xadira Cit C i t : x a dira D i t : xo dira
Tyl&s^ng Slit ciia cay con (%)
33,42 b 52,34 a 46,32 ab 34,16 b 5,80 7,60 N.KfrLUAN
- Trong qui trinh khir trimg m^, sir dung chdt khir triing hypoclorit eanxi vdi ndng dp 5% trong thdi gian xir ly 20 phut li thich hpp nhdt T^ 1| sdng dat 823-
- Ndng dp dudng sucroza thich hpp nhdt cho st^
sinh trudng va phat triin ciia tir phdi diia din ciy hoin chinh li 60 g/L
- Ndng do BA thich hpp nhdt trong qui trinh nudi cdy dira Sip in vitro li 0,5 mg/L
- Ndng do NAA thich hpp nhdt trong qui trinh tao rl ciy dira Sip in vitro li 1 mg/L
- Gii thi cit eho ty le sdng cao nhat (52,34%).
Ldi dm aa: Nghidn cihi nay rfupc s(/ hd tr</ tai chinh tir BQL Du in Ndng nghi€p - Chuong trinh nghidn cihi Ndng nghidp hudng tdi khach hang, Bd Ndng nghidp va Phat triin ndng thdn.
Till i|u HUM KHAO
1. Assy-Bah B., Durand-Gasselin T. & Pannetier C. (1987). Use of zygotic embryo culture to collect
| 6 N G NGHIEP V A PHAT TRIEN N O N G THON - KY 2 - THANG 10/2012 17
KHOA Hpc C 6 N 0 NOHt <
germlasm of coconut (Cocos nucifera L ) . Genet. 9. M. T. Perez Numez et al., 2007. ImproV Resour. Newslet. 71:4-10. somatic embryogenesis from Cocos nucifera
2. Assy-Bah, B. F. Engelmann. T. Durand- plumule explants.
Gasselin, 1986. Culture in vitro coconut embryoes. 10. Maximova et al., 2002. Efficiency, genoty 3. Azpeitia et al., 2003. Effect of 22(S). 23(S)- variability, and cellular origin of primary and secondj homobrassinolide on somatic embryogenesis in somatic embryogenesis of Theobroma cacao.
plumule explants of Cocos/7(/r;/e/-a culture in vitro. 11. Perera et al., 1999. Identification a 4. Blake J. etal.. 1995. Somatic embryogenesis in characterization of microsatellite loci in cocon coconut. {Cocos nucifera L.) and analysis of cocon
5. De Kler, G. J. - Plant homones in tissue populations in Srilanka.
culture. Catalogue 2000-2001 Duchefa Biochemie 12. Raemakers et al., 1995. Secondary somal BV. p. 19 -25. embryogenesis and applications in plant breeding.
6. Eeuwens,C. J., 1976. Mmeral requirements for 13. Rillo, E. P. and M. B. F. Paloma, 1992. J growth and callus initiation of tissue explants excised vitro culture of Macapuno coconut embiyoi from mature coconut palms {Cocos nucifera L) and Coconuts Today 9(1):90-101.
culture in vitix). Physiologia Plantarum 36:23-28. 14. Rillo E. P. (1998). PCA's embryo cultur 7. Elida P., Rillo, 1997. Coconut embryo culture, technique in the mass production of Makapim International Symposium on Coconut Biotechnology, coconuts. In: Batugal P & Engelmann F (eds) Coconi
8. Lees R. P. (1994). Effects of light environment on Embryo Cultiire 27-31 October (1997). IPGRI. Baaat photosynthesis and growth in vitro. Physiology. Pp 31-46. Guinobatan, Albay, Philippines ^ p 69-78).
IN VITRO PROPAGATION OF MAKAPUNO COCONUT
Truong Quoc Anh, Luong The Minh, Truong Thi TU Anh and lYuong Vinh Hai lai la Summary
The coconut {Cocos nucifera L.) is very important crop in tropical areas. It is called "the Tree of Life"
because of many usea one can derive from it such as roofing matenals, timber, fibres, nut water as drinks and oil and providing cash and subsistance to smallholders. Makapuno coconut is an expensive delicacy and Its planting material is highly priced in Vietnam. Instead of the usual formation of a solid endosperm and cocounut water, the solid endosperm of the Makapuno coconut remains as a jelly, sometimes filling the entire cavity. The Makapuno coconuts are found as a small proportion of normal fruit on a fem palms, generally from the "Laguna Tall" ecotype. Although the Makapuno coconut contains an apparently normal embryo, it fails to germinate properly as the endosperm contains substances which are obviously lethal. A protocol was developed for Makapuno coconut regeneration using plumules from mature zygodc embryos.
Embryos were sterilized with calcium hypochloride 556 in 5-25 minutes. Results indicated that highly effective sterilization occurred vnih calcium hypochloride in 20 minutes. The highest rate of living of embryos was 82.3%. Germination of Makapuno coconut embryo was tested in Y3 medium (Eeuwens) with BA, activated charcoal supplemetation. Percent gemination, embryo weight was greater when embryos were cultured in Y3 medium with 60 g/L sucrose is 91% compare to other treatment of significant diference following statistic. Expenment of suitable NAA concentration for root creation and root lenghL The average number of root per explants after 150 days of culture in 1 mg/L freatment was 6.73 for the highest root regeneration of compare to 0 mg/L treatment of 1.7, 0.5 mg/L of 5.3 and 1.5 mg/L treatment of 4.93. The success of any in vitro culture is measured not only in tiie number of plants regenerated but more importantly, the number of plants that have been tiansplanted and established in the field and grown to maturity. Result has indicated that sand medium shown the highest living rate of Makapuno.
Key words: Makapuno, somatic embryo, Y3 medium. ^ Ngudi phan bi|n: GS.TSKH. Trdn Duy Qu^
Ngiy nhin bii: 4/9/2012 Ngiy thdng qua phan bl|n: 3/10/2012 Ngiy duylt ding: 8/10/2012
