Nội Dung 1: Nghiên Cứu Kỹ Thuật Tạo Hạt Nhân Tạo Từ Phôi Cây Lan Vanda Nội Dung 2: Khảo Sát Sự Hiện Diện Của Mycorrhiza Trên Một Số Giống Lan

B GIO DC V O TO
TRNG I HC NNG LM THNH PH H CH MINH

B MN CNG NGH SINH HC
NI DUNG 1: NGHIN CU K THUT TO HT NHN

TO T PHI CY LAN VANDA
NI
DUNG
2:
KHO
ST
HIN
MYCORRHIZA TRN MT S GING LAN
Ngnh hc: CNG NGH SINH HC

Nin kho: 2003-2007
Sinh vin thc hin: NGUYN HONG QUN
Thnh ph H Ch Minh
-2007-
DIN
CA
B GIO DC V O TO
TRNG I HC NNG LM THNH PH H CH MINH
B MN CNG NGH SINH HC
NI DUNG 1: NGHIN CU K THUT TO HT NHN

TO T PHI CY LAN VANDA
NI
DUNG
2:
KHO
ST
HIN
DIN
Gio vin hng dn

TS.TRN TH DUNG
Sinh vin thc hin

NGUYN HONG QUN
ThS.T TH M THUN
Thnh ph H Ch Minh
-2007-
CA
LI CM N
Thnh knh khc ghi cng n cha m tn ty sut i v con con
c c ngy hm nay.
Xin t lng bit n su sc n TS. Trn Th Dung v ThS. T Th M
Thun tn tnh hng dn v gip em trong sut thi gian thc hin
v hon thnh lun vn tt nghip ny.
Chn thnh cm n:
- Ban Gim Hiu trng i Hc Nng Lm
- Qu thy c trng i Hc Nng Lm
- Qu thy c B Mn Cng Ngh Sinh Hc.
tn tnh truyn t kin thc cho ti trong sut qu trnh hc tp
cng nh thc hin lun vn ny.
Xin gi li cm n n:
KS Nguyn Th Thu Hng, KS L Hng Thu Tin cng tp th cn b
nhn vin Trung Tm Cng Ngh Sinh Hc v Phng Thc Tp Bnh Cy
trng i Hc Nng Lm, cng tt c bn b trong v ngoi lp Cng
Ngh Sinh Hc 29 lun ng vin, gip ti trong thi gian hc tp v
thc hin ti.
TP. H Ch Minh, Ngy 25 thng 8 nm 2007
Sinh vin
Nguyn Hong Qun
ii
TM TT
NGUYN HONG QUN, sinh vin b mn Cng Ngh Sinh Hc kho
29, i Hc Nng Lm Thnh Ph H Ch Minh, thng 8 nm 2007
TI:
NI DUNG 1: NGHIN CU K THUT TO HT NHN TO T
PHI CY LAN VANDA.
NI DUNG 2: KHO ST S HIN DIN CA MYCORRHIZA
TRN MT S GING LAN
Ging vin hng dn: TS. TRN TH DUNG
ThS. T TH M THUN
ti c tin hnh ti Trung Tm Cng Ngh Sinh Hc- Trng i Hc
Nng Lm Tp H Ch Minh, thi gian thc hin t thng 3/2007 n thng 8/2007
Ni dung 1: Nghin cu k thut to ht nhn to t phi cy lan Vanda
Th nghim 1: Kho st nh hng ca nng sodium alginate n s hnh thnh
ht nhn to.
Th nghim c b tr theo kiu hon ton ngu nhin 1 yu t, gm 5
nghim thc, lp li 3 ln.
Kt qu thu c nng 30 g/l cho v ht nhn to trn, u, p v t l
ny mm cao.
Th nghim 2: Kho st nh hng ca mi trng to ht nhn to
nghim thc, lp li 3 ln
Kt qu thu c mi trng dinh dng MS, ht nhn to c a phi lan
Vanda cho t l sng st v ny mm cao.
iii
Th nghim 3: Kho st mi trng bo qun ht nhn to

nghim thc, 3 ln lp li.
Mi trng MS bo qun ht tt nht, cho ht nhn to c t l ny mm cao
nht sau khi bo qun.
Th nghim 4: Kho st s ny mm ca ht nhn to trn cc gi th khc nhau
nghim thc, 3 ln lp li.
T l ht nhn to ny mm trn bng gn cao nht (khong 100%)
Ni dung 2: Kho st s hin din ca mycorrhiza trn mt s ging lan
Kt qu kho st cho thy mycorrhiza c cc dng sau:
-Si nm n mu xanh, phn nhiu nhnh n xuyn qua nhiu lp t bo
-Si nm cun trn trong 1 t bo
-Si nm l cc cun trn nm cc t bo, ni vi nhau bng si nm
Gia cc ging lan trng, t l hin din ca mycorrhiza c s khc nhau do
ngun gc ca ging
iv
SUMMARY
NGUYEN HOANG QUAN, student of Biotechnological faculty chronology
29, Nong Lam University of Ho Chi Minh City, 8 -2007.
TOPIC:
CONTENT 1: RESEACHING TECHNOLOGY DOES ARTIFICIAL
SEED FROM SOMATIC EMBRYONIC VANDA ORCHID.
CONTENT 2: EXAMINING PRESENCE OF MYCORRHIZA ON
SOME ORCHID GENUS
Supervisor: TRAN THI DUNG, ph.D

TU THI MY THUAN, M.D
Topic was doen at Biotechnological Centre- Nong Lam University of Ho Chi
Minh City, doen from 3-2007 to 8-2007.
CONTENT 1: RESEACHING TECHNOLOGY DOES ARTIFICIAL SEED
FROM SOMATIC EMBRYONIC VANDA ORCHID.
Experiment 1: Examining sodium alginate strength influenced artificial seed
formation.
Experimentation was set quite accident one factor, include 5 assays, 3 times
repetition.
Resulting in 30 g/l strength gave full, nice artificial seed shell and ratio of
germination highly
Experiment 2: Examining influence of nutritional environment do artificial seed.
repetition.
Resulting in MS nutritional environment gave ratio of life and germination

highly, consist with artificial seed from embryonic Vanda.
Experiment 3: Examining maintainable environment arfiticial seed.
repetition.
MS environment maintained seed better, gave germinant ratio higher after
maintenance.
Experiment 4: Examining germination of arfiticial seed on different substances.
repetition.
Ratio of germinant arfiticial seed on the silk- coton was highest.
CONTENT 2: EXAMINING PRESENCE OF MYCORRHIZA ON SOME
ORCHID GENUS.
Resulting in mycorrhiza had forms:
-
The hyphae was dyed with blue, divided many branches across root cells.
The hyphae scrolled in one cell.
Hypha cirumvolutions entered into cells, allyed each other by the hyphae.
The present ratio of mycorrhiza had difference among orchid genus because of
growing simulant origin.
vi
MC LC
Trang
Trang ta ..........................................................................................................i
Li cm n .......................................................................................................ii
Tm tt .............................................................................................................iii
Summary ..........................................................................................................v
Mc lc .............................................................................................................vii
Danh sch cc bng .......................................................................................... x
Danh sch cc biu ......................................................................................xi
Danh sch cc hnh........................................................................................... xii
1 GII THIU .................................................................................................1
1.1 t vn ..................................................................................................1
1.2 Mc tiu nghin cu ...................................................................................2
1.2.1 Mc ch..................................................................................................2
1.2.2 Yu cu....................................................................................................3
2 TNG QUAN TI LIU .............................................................................4
2.1 Khi qut v cy lan ...................................................................................4
2.1.1 Phn loi thc vt hc .............................................................................4
2.1.2 Lch s cy lan .......................................................................................4
2.1.3 Tnh hnh sn xut lan .............................................................................5
2.1.4 c im thc vt hc ca cy lan.......................................................... 6
2.1.4.1 C quan dinh dng .............................................................................6
2.1.4.2 C quan sinh dc ca lan- t chc hoa ................................................7
2.1.5 Cc iu kin c bn cho cy lan ............................................................ 8
2.1.6 Cc phng php nhn ging lan ............................................................ 8
2.1.6.1 Gieo ht ................................................................................................ 8
2.1.6.2 Tch chit ............................................................................................. 9
vii
2.2 Phng php nui cy m lan ...10

2.2.1 Khi qut v lch s nui cy m ............................................................ 10
2.2.2 Cc phng php nui cy ......................................................................10
2.2.3 Cht dinh dng trong mi trng nui cy in vitro .............................. 12
2.2.4 Cc bc nhn ging in vitro ..................................................................14
2.2.5 Cc yu t nh hng nhn ging in vitro ..............................................15
2.3 Cng ngh to ht nhn to..
16
2.3.1 Khi nim ................................................................................................ 16
2.3.2 Mc ch..................................................................................................16
2.3.3 Cc nhn t to ht nhn to ...................................................................17
2.3.3.1 To phi v tnh ...................................................................................17
2.3.3.2 C ch pht sinh phi soma .................................................................18
2.3.3.3 To v bc bng sodium alginate ........................................................19
2.3.3.4 Quy trnh to ht nhn to ....................................................................21
2.4 Nm r cng sinh- mycorrhiza.. 22
2.4.1 Khi nim ................................................................................................ 22
2.4.2 Cc dng mycorrhiza...............................................................................23
2.4.2.1 Ectomycorrhiza ...................................................................................23
2.4.2.2 Endomycorrhiza ...................................................................................23
2.4.3 Tc ng c ch ca mycorrhiza ............................................................. 24
2.4.3.1 M rng din tch hp thu ca r cy ...................................................24
2.4.3.2 S trao i dinh dng .........................................................................24
2.4.3.3 S hnh thnh cht kch thch sinh trng mycorrhiza ........................24
2.4.3.4 Nng cao sc chng chu ca cy ........................................................25
2.4.3.5 Tng kh nng khng bnh ca cy trng ............................................25
2.4.4 S xm nhp ca mycorrhiza ..................................................................25
3 VT LIU V PHNG PHP NGHIN CU ....................................... 26
3.1 i tng th nghim .................................................................................. 26
3.2 Thi gian thc hin ..................................................................................... 26
3.3 Ni dung nghin cu ................................................................................... 26
3.4 Vt liu nghin cu ..................................................................................... 27
viii
3.4.1 Trang thit b v dng c nghin cu.
27
3.4.2 Mu s dng v iu kin nui cy.. 27

3.4.3 Mi trng s dng
27
3.5 Phng php nghin cu ............................................................................. 28

3.5.1 Chun b mi trng..
3.5.2 B tr th nghim
3.6 X l s liu
28
29
..32
4 KT QU V THO LUN ....................................................................... 33

4.1 Ni dung 1: Nghin cu k thut to ht nhn to t phi cy lan ............ 33
4.1.1 Th nghim 1: Kho st nh hng ca nng sodium alginate n
s hnh thnh ht nhn to ..............................................................................36
4.1.2 Th nghim 2: Kho st nh hng ca mi trng to ht nhn to 39
4.1.3 Th nghim 3: Kho st mi trng bo qun.
41
4.1.4 Th nghim 4: Kho st kh nng ny mm ca ht.
43
4.2 Ni dung 2: Kho st s hin din ca mycorrhiza cc ging lan........... 45

5 KT LUN V NGH ...........................................................................49
5.1 Kt lun ......................................................................................................49
5.2 ngh .......................................................................................................50
6 TI LIU THAM KHO .............................................................................51
7 PH LC ......................................................................................................53
ix
DANH SCH CC BNG

Bng 2.1: S pht trin ca m trn mi trng nui cy ....................................... 17
Bng 4.1: Nng alginate nh hng ln hnh thi v ht nhn to ..................... 36
Bng 4.2: T l sng ca ht nhn to cc nng alginate ................................ 38
Bng 4.3: T l ny mm ca ht nhn to cc nng alginate khcnhau .... 39
Bng 4.4: T l sng ca ht nhn to cc mi trng dinh dng khc nhau 40
Bng 4.5: T l ny mm ca ht nhn to cc mi trng dinh dng .............. 42
Bng 4.6: Kho st nh hng ca mi trng bo qun ht nhn to ................... 42
Bng 4.7: T l ny mm ca ht nhn to sau khi bo qun .................................. 43
Bng 4.8: T l ht nhn to ny mm trn cc gi th ........................................... 44
Bng 4.9: S hin din ca mycorrhiza trn cc ging lan..................................... 45
DANH SCH CC BIU

Biu 4-1: T l sng ca ht nhn to cc nng alginate khc nhau . 38
Biu 4-2: T l sng st ca ht nhn to mi trng khc nhau .......... 40
Biu 4-3: S hin din ca mycorrhiza trn cc ging lan ........................ 46
xi
DANH SCH CC HNH

Hnh 2-1: Cu to hoa lan ............................................................................... 8
Hnh 2-2: Cu to ht nhn to ....................................................................... 16
Hnh 2-3: Cc giai on pht sinh phi soma ................................................. 19
Hnh 2-4: Cng thc ho hc ca alginate ...................................................... 20
Hnh 2-5: Cc dng phi soma ............................................................................................ 21
Hnh 2-6: Quy trnh to ht nhn to .............................................................. 22
Hnh 4-1: Cc dng phi v tnh ..................................................................... 33
Hnh 4-2: Cu trc ca phi v tnh .................................................................34
Hnh 4-3: Mi trng to ht nhn to ............................................................ 34
Hnh 4-4: Cc bc to ht nhn to................................................................ 35
Hnh 4-5: S hnh thnh ht nhn to ............................................................. 35
Hnh 4-6: Cu to vi th ca ht nhn to ........................................................36
Hnh 4-7: Ht nhn to cc nng alginate ............................................... 37
Hnh 4-8: Ht nhn to ny mm .................................................................... 39
Hnh 4-9: Ht nhn to trn cc mi trng bo qun .................................... 42
Hnh 4-10: Ht nhn to ny mm trn cc gi th ......................................... 44
Hnh 4-11: Vng m c mycorrhiza ................................................................ 47
Hnh 4-13: Cc dng mycorrhiza .................................................................... 47
Hnh 4-12: Dng mycorrhiza cun trn .......................................................... 48
xii
CHNG I
GII THIU
1.1 t vn
Hin nay, x hi chng ta ngy cng pht trin v hin i. Cuc sng thng b
p lc nng n t cng vic.V vy nhu cu gii tr v vui chi lnh mnh l rt cn
thit i vi mi ngi. Trong chi hoa, cy cnh cng l mt th vui tao nh,
gip chng ta ho quyn vo thin nhin, nng cao cht lng i sng, gip ta
sng lu sng kho.
Hoa l biu tng cho v p v iu thnh thin. Mi loi hoa c mu sc,
hng thm c trng cho tng loi, cho tng vng, tng min.Tng loi hoa li
tng trng cho tnh cch ca mi ngi, mi biu tng trong cuc sng, i din
cho tnh yu, tnh bn, tnh cm con ngi. Hin nay hoa tr nn ph bin rng
ri i vi tt c mi ngi, l mn qu, l thng ip trao nhau trong nhng ngy
l, ngy k nim, ngy hp mt. Hoa cng c xem nh mt vt trang tr trong nh
tng v m quan, gn gi vi thin nhin. Nhiu l hi v hoa c t chc cc
quc gia.
Trong , hoa lan hin c nhiu ngi rt quan tm v khng nhng cm
mn trc v p ca hoa m cn thc mc, tm hiu cch sinh tn ca lan trong t
nhin. Lan rt a dng phong ph v mu sc, hng thm, chng loi v c bit
l phn b rng khp th gii. Ngi trng lan thng c v nh mt ngh nhn,
cn ngi thng thc v p ca chng c xem nh l ngh s.
V mt kinh t, trng v kinh doanh hoa lan l mt ngnh em li li nhun
cao. Gi tr sn lng hoa lan th gii ngy nay ln ti hng t la. V vy cc
nc H Lan, c, Thi Lan, Philippin, Malaysia vn ni ting v ngh trng
lan th nay h pht trin hn na quy trnh sn xut lan nhanh hiu qu hn. ng
thi, y mnh lai to ging chn ra nhng ging mi l, kh nng chu ng vi

hon cnh khc nghit ca t nhin.Vit Nam chng ta cng nghin cu rt nhiu
v lan, c bit l k thut nhn ging nhanh bng k thut nui cy m.
c bit, cy lan rt kh gieo ht ngoi t nhin, a phn mun nhn ging
nhanh thng p dng tch chit hoc nui cy in vitro. Vi s pht trin ca k
thut to ht nhn to, chng ta c th ng dng ly phi soma cho bc v nhn to
c cha mi trng dinh dng, nhm bo qun phi v to iu kin cho ht nhn
to ny mm. Vi k thut ny, th khc phc hn ch ny mm ca ht lan trong t
nhin, v gip nhn nhanh cc ging lan trng v cc loi thc vt kh ny mm
trong t nhin.
Trong h sinh thi nng nghip, rt nhiu nm c li cho cy trng, nm cng
sinh vi r l rt ph bin, chng gip cy nhn c mt s cht khong t t,
nht l tng cng hp th photpho d dng, gip cy tng trng nhanh, ng thi
cy li cung cp ngun cacbon cho nm, thng c gi l mycorrhiza. Nm bt
c mi quan h , chng ta c th ng dng vo lnh vc sn xut cy in vitro.
V vy, cn phi c nhiu nghin cu hn na v mycorrhiza, c bit l trn r lan.
c s ng ca b mn Cng Ngh Sinh Hc cng vi s hng dn tn tnh
ca TS Trn Th Dung, ThS T Th M Thun, chng ti tin hnh nghin cu
ti ny gm hai ni dung:
NI DUNG 1: NGHIN CU K THUT TO HT NHN TO T PHI
CY LAN VANDA
NI DUNG 2: BC U KHO ST S HIN DIN CA
1.2 Mc tiu nghin cu
1.2.1 Mc ch
-Bo qun phi soma hiu qu v lu tr ging d dng hn.
-To c s cho vic phn lp mycorrhiza trn cy lan ng thi th nghim
trong nui cy in vitro.
1.2.2 Yu cu
Theo di t l sng, t l ny mm ca ht nhn to.
Thu thp mu r lan, nhum v quan st mycorrhiza
CHNG 2
TNG QUAN TI LIU
2.1.Khi qut v cy lan
2.1.1 Phn loi thc vt hc
Ngnh: Angiospermae (ht kn)
Lp:Monocotyledonae
B: Orchidales
H: Orchidaceae
H ph: Orchidoideae
2.1.2 Lch s cy lan
Ngi ta tng rng cy lan c bit n trc tin chu u qua bn vit tay
bng ting Hy Lp Xem Xt Cy C (Enquiry intoPlants) ca Theophrastus
(khong nm 370-285 TCN). K thc th cy lan c bit n u tin
phng ng: Khng T (551-479 TCN) sau khi i chu du thin h, khng c
nc no s dng, nn v li nc L. Trn ng v, ng thy hoa lan mc chen
vi cy c rng su bn than rng: i, hoa lan c mi thm vng gi, nay ti
tt mt mnh chn sn lm, mc xen ln vi loi c hoang di, chng khc no
bc hin gi khng gp thi, ng chung vi bn b phu, m t cc bi th, ph,
vnh v hoa lan sau ny ca cc thi nhn Trung Hoa u b t nhiu nh hng.
phng ng, lan c ch n v p duyn dng ca l v hng thm
tuyt diu ca hoa. V vy trn thc t lan ch yu chim ngng trc tin l l
ch khng phi mu sc ca hoa. phng Ty, lan c bit n trc ht l
cng dng v dc liu v sau l sc hp dn ca hoa cng cc c tnh v thc
vt ca n.Theophrastus c xem l ng t ca thc vt hc v l cha ca
ngnh hc v lan v n nm 1836 John Lindley t tn cho h lan l Orchidaceae.
Vit Nam, nm 1789 Joanis Loureiro, nh truyn gio ngi B o Nha ln

u tin m t trong cunFlora Cochinchinesis v ch n khi ngi Php n
mi c nhiu cng trnh cng b. H.Lecomte m t 101 ging gm 750 loi lan cho
c 3 nc ng Dng. Nm 1993 Phm Hong H pht hin v m t khong 653
loi.
2.1.3 Tnh hnh sn xut lan
Trn th gii
Chu u, H Lan l quc gia duy nht c cng ngh trng lan xut khu sm,
ng dng trng trong nh knh nn xut khu c quanh nm, c bit l lan
Cymbidium. Cn Italia l quc gia nhp khu lan nhiu nht Chu u, ch yu t
cc nc: Thi Lan 64 triu cnh, H Lan 10 triu cnh vo nm 1993. M nhp t
Thi Lan 16,4 triu cnh, Singapore 289 ngn cnh Dendrobium vo nm 1994.
Chu , ng u th gii v nhp khu lan vn l Nht, ch yu l
Dendrobium, Oncidium, Cymbidium, Phalaenopsis t Malaysia, Singapore v Thi
Lan l nhiu nht
Vit Nam
Do chng ta vng c kh hu nhit i gi ma, v vy phong lan rt a dng.
Bn cnh cc loi lan rng ni ting v mu sc v hng thm cn c cc ging
lan nhp ngoi nh Dendrobium, Vanda, Mokara, Phalaenopsis, Cattleya cn
phn b lan rng th khp c nc:
Vng kh hu lnh: Cao Nguyn v cc tnh pha Bc c th trng c cc loi
lan nh Dendrobium, Cymbidium, Phalaenopsis, Cattleya, Paphiopedilum)
Vng kh hu nng m nh Nam v Trung Trung B, nht l min ng
Nam B c cc loi lan nh: Cattleya, Vanda, Mokara, Cymbidium, Ngc im
Tuy nhin c nhng kh khn ca ngi trng lan nc ta:
-Nng dn trng lan cn nh l, thiu quy hoch chin lc, k thut so vi
nc ngoi
-Cha to c ging lan ring bit cho Vit Nam c th cnh tranh vi cc
ging lan nhp ngoi, nht l Thi Lan.
-Th trng tiu th cn nhiu kh khn, cha n nh kinh t lu di cho

ngi trng lan.
Tuy nhin, thnh ph H Ch Minh c nhiu chnh sch, ngun vn u t
xy dng v m rng khu nng nghip cao, chuyn i i c cu nng nghip
cho ph hp vi tnh hnh v xu hng pht trin ca thnh ph, trong cy lan
cng l cy ch lc.
2.1.4 c im thc vt hc ca cy lan
2.1.4.1 C quan dinh dng
a) S pht trin cy a thn
Trng hp lan Cau dip (Spathoglottis plicata Bl.)
y l loi a lan, gi hnh l mt thn ngn ph mp. Mi gi hnh c mt s
l bao che v mang r y. y l to ra to thnh b bao quanh gi hnh. nch
ca mi b l c th c cc chi m mi chi c th pht trin thnh cnh mang hoa
(pht hoa) hay to ra c quan dinh dng mi (gi hnh mi).
Cc chi nch l y c che ch bi nhng l ngn khng c phin l bn
trn. Khi chi bt u ph to c dng mt gi hnh s khi th cc l l ra v pha
nh, sau cc r mi xut hin v pha y. Cui cng cc l pht trin ln ln,
tch tr dng cht v pha y lm cho gi hnh ln dn ra.
Trng hp lan Bch cu (Dendrobium crumenatum Sw.)
Xut hin t ng bng n ni cao, ph sinh sng bm trn cy hay trn hc
, r chng khng lun b m t nh trong t nh lan Cau dip.Cnh lan mi
sinh ra t gc ca cnh c: Gm gc nh mang r, tip l on ph mp khng r,
khng l, k trn l on di lng khng mang l mc xen hai hng, trn cng l
on di nh lng y nh vy nhng khng c l, tt c gi l gi hnh. Nh vy
Bch cu sng nhiu nm (cy a nin) nh cc n v ni tip nhau, chi mi sinh
ra t chi bn gc ca n v trc v c th tip tc pht trin theo chiu
ngang.
b) S pht trin cy n thn

kiu pht trin c tr, nu ta ct mt on pha ngn ca thn th on b
ct ri ny vn pht trin v pha nh. Cn nu ct on thn ca cy (nh gi
hnh) th on ct ri y khng th pht trin v pha nh.V d nh cy Vanda,
Ngc im (Rhynchostys gigantea), H ip (Phalaenopsis) L c phin hnh
tr di v c b bc kn thn. Thn lun lun mc cao ln v pha nh. S mc di
ca nh khng c gii hn nn cy ch c mt thn pht trin v hn. S pht trin
ngng li khi nh b tn thng, lc chi bn s x rch b l mc di ra
thnh nhnh, nhnh ny cng sinh trng khng gii hn nh nh ban u.
2.1.4.2 C quan sinh dc ca lan- t chc hoa
Hoa lan kiu tam phn (chia 3): 3 l i, 3 cnh hoa, 3 tm b
Phn hoa dnh li thnh phn khi, t khi ri tng ht nh cc loi hoa khc
S hin din ca tr l phn hp nht ca b phn sinh dc c v b phn
sinh dc ci. l c im quan trng ch c hoa lan.
Hnh dng, kch thc ca mi, cng thm v tr ca nhu c lm cho
hoa lan khng u, c s i xng hai bn r rt.
Tuy cu trc chung nh vy nhng trong chi tit tng bng hoa lan cng c
nhng khc bit m t ta chia chng ra nhng ging loi khc nhau, ch cn
cp n cnh mi thi cng rt phong ph. Nhng 99% cc cy lan c 1 nhu
c th m thi s cn li c 2-3 nhu c th (Nguyn Thin Tch,1996). V phi
tnh th i a s cy lan l hoa lng tnh, nhng ging Catasetum v Cycnoches
hoa li l n tnh.
Cnh mi: mt s loi th rt ln so vi hai bn cnh, mt s loi c cnh
mi rt nh.
Hnh dng: Mi c th nguyn hay c thu, c ra, c tua, cun hay nhnc
cu trc rt k l nh c sc, c sng, c gai, c lng, trn lng hay nhn..c ca,
mc, ti ng sau..
Mu sc: Thng khc hai bn, hoc c sc, im, nhng mu khc bit
Hnh 2-1: Cu to hoa lan

2.1.5 Cc iu kin c bn cho cy lan
m : Thng thng ti thiu l 70% thch hp cho s tng trng ca
nhiu loi. Tuy nhin m l tng vn l m ca vng bn x
pH ca nc: pH ca nc ph hp cho lan tng trng l 5-6, nu ln 7
hoc trn 7 th khng nn ti nc cho cy.
Nhit : Tu thuc vo nhit ca iu kin a l, vng cao th s c
s phn b ging lan ph hp.
nh sng: Yu t quyt nh cho s quang hp, h hp v nht l cho s tr
hoa.
Vanda l trn cn 100% nh sng
Vanda l sp thnh hng cn 70% nh sng
Dendrobium cn 70% nh sng
Cattleya cn 50% nh sng
Phalaenopsis cn khong 30% nh sng
thng gi: Lan c 2 nhm chnh l phong lan v a lan, tc gi
khong 10-15 km/gi, ngha l gi lm l v cc cnh nh hi rung ng, nu trng
ni thng gi v mt m, cy lan s tng trng nhanh v t b bnh hn (Nguyn
Minh Trc, 1996).
2.1.6 Cc phng php nhn ging lan
2.1.6.1 Gieo ht
Trong thin nhin, s th phn lan do cn trng thc hin, do cu trc ca
hoa lan thch ng vi iu ny. C 2 phng cch th phn:
- S t th phn: Khi phn hoa hoa ny c ri vo num ca chnh hoa

y, iu ny khng th xy ra ngoi t nhin i vi hoa lan c.
- S th phn cho: Khi phn hoa hoa ny c vo num ca hoa khc
trn cng cy hay cng mt loi, hay khc loi, khc ging (ch con ngi mi lm
c).
Ht lan qu nh, khng cha cht d tr v ch c mt phi cha phn ho nn
khng th pht trin theo cch bnh thng m vic lm cho ht lan ny mm thnh
cy lan trng thnh l vn kh khn.
Nm 1844, Neumann- ngi Php- cho ny mm thnh cng ht lan khi ng
ling ht lan quanh gc cy lan ln, nhng khng hiu v sao.
Nm 1899, Noel Bernard- ngi Php- khm ph b mt , v nhng ht
lan ny mm u b nhim nm. Nm 1904, ng c lp cc nm r lan v cho nhim
ln ht lan th 100% ht ny mm.
Ngi ta khm ph ra 3 loi nm gip ht lan ny mm:
-
Rhizoctonia repens gip ht lan Cattleya, Laelia ny mm.
Rhizoctonia mucoroides gip ht lan Vanda, Phalaenopsis ny mm.
Rhizoctonia lanugiosa gip ht lan Oncidium, Miltonia ny mm.

Nm 1922, Knudson M, thnh cng khi gieo ht lan trn mi trng
thch c b sung dinh dng (thm 2% ng )
2.1.6.2 Tch chit

a) Tch bi
Dng cho cc ging lan a thn nh Cattleya, Dendrobium. gc ca mi
gi hnh c t nht mt mt ng nn c th tch mi hng gi hnh thnh mt n v
trng, nhng cy sau khi tch kho hn th tt nht mi n v c t nht hai
gi hnh, v c mt ng.
b) Tch cy con
Dendrobium, Thunia thng to ra cy con trn gi hnh, khi cy con ny
kho mnh, c r tt, c th tch ra khi gi hnh trng.
Phalaenopsis c lc cy con mc trn cng pht hoa n khi c r tt th tch

ra trng.
Vi nhng loi n thn nh Vanda, H ip, Ngc im kho mnh, hoc
b tn thng nh ngn, chng s sinh ra cy con t chi bn gn gc, cc chi
ny c r di ra, c th tch ra trng ring.
2.2 Phng php nui cy m lan

2.2.1 Khi qut v lch s nui cy m
Nm 1838, hai nh sinh vt hc ngi c, Schleiden v Schwann a ra
thuyt t bo v nu r mi c th sinh vt c phc tp ti u th u c cu to
n v rt nh l t bo.
Nm 1902, Haberlandt thc hin nui cy t bo thc vt u tin t l mt s
cy mt l mm nh: Errythronium, Orrnithogalum
Nm 1922, Kotte v Robins lp li th nghim ca Haberlandt nhng trn nh
sinh trng ca r mt cy ho tho, trn mi trng lng c mui khong v
glucose. Tuy nhin s sinh trng ny ch ko di trong thi gian ngn.
Nm 1934, White v Gautheret nui cy thnh cng r c chua trn mi
trng mui khong v dich chit nm men.
Nm 1934, Nobecourt v Gautheret duy tr s sinh trng ca m so c rt.
Nm 1955, Cc cht kch thch s phn bo c Skoog t tn l kinetin, v
gp vi cc cht kch thch phn bo t nhin gi l cytokinin.
Nm 1956, Morel, hc tr ca Gautheret, p dng thnh cng nui cy m vo
cy lan (Cymbidium) to ra cc protocorm.
n nay, hu ht cc ging lan c nhn ging nhanh bng phng php
nui cy m nh: Cattleya, H ip, Ngc im, ngay c lan Hi ni ting ca Vit
Nam (Cu lc b hc vin hoa lan, 1999) .
2.2.2 Cc phng php nui cy
a) Nui cy nt n thn
S dng mu cy l chi ngn hoc chi bn c mang mt on thn ngn.
Chi ny c kch thch tng trng, ra r to cy nguyn vn, nhiu chi v l
10
c hnh thnh, tip tc cy chuyn trn mi trng dinh dng thch hp, phng
php ny dng nhn ging v tnh in vitro v c th p dng c in vivo (Nguyn
c Lng, 2002).
b) Nhn chi bn
Ging nh nui cy nt n thn v nguyn tc nhng khc l nui cy nt n
thn c s ko di chi, thn v thng khng cn n cytokinin pht trin. Cn
phng php nhn chi bn, chi ngn c c lp trn mi trng dinh dng v
cc chi bn t cc nch l pht trin di tc dng ca cytokinin nng cao.
Cytokinin c tc dng hn ch u tnh ngn cc chi bn pht trin (Nguyn c
Lng, 2002).
c) Nui cy nh sinh trng
Chi ngn c ra sch v kh trng bng cn, hypoclorite calcium. Sau
dng dao m tch ri nh sinh trng (gm vng m phn sinh v c phn di
ngn) ra khi ngn v cy trn mi trng ti sinh cy hon chnh. Vi phng
php ny, chng ta to c cy sch bnh, sch virus.
d) Nui cy m so
To m so c tin hnh ln u tin vo nm 1920, m so l mt khi t
bo khng c t chc, hnh thnh t cc m hoc c quan phn ho di cc
iu kin c bit (vt thng, x l ho cht, tia phng x...).Cc t bo ca m
so phi chu s phn phn ho trc ln phn chia u tin (Halperin, 1969).
M so tng trng nhanh trn mi trng c cht auxin v trong mi trng
khng c cht kch thch th m so c th ti sinh thnh cy hon chnh. Cm m
so c th ti sinh cng mt lc nhiu chi hn l nui cy nh sinh trng, nhng
kh nng b bin d t bo soma li cao hn.
e) Nui cy huyn ph t bo
Huyn ph t bo c to t cc mnh m so nui cy trong mi trng lng
lc lin tc. Trong mi trng lng, m so phng thch ra nhng t bo ring l
hay nhng cm t bo dnh nhau hp th dinh dng. Sau t bo c chuyn
sang mi trng c ti sinh thnh cy.
11
f) Nui cy th n bi
Cy th n bi mang b nhim sc th gim i mt na, s dng nhng phn
sau cho vic nui cy in vitro:
Ti phn: Thu ti phn t chi hoa th ch kh trng chi hoa. Cn thu ti phn
ca hoa n th phi kh trng ti phn.
Ht phn: c nui cy trn mi trng to m so.
Cm hoa: Thng nui cy trong mi trng lng.
g) Nui cy protoplast( t bo trn)
T bo thc vt c x l bng ho l tch lp v cenlulose, nhng vn cn
gi chc nng ca t bo. Trong mi trng thch hp, protoplast c th phn chia
t bo hay ti sinh thnh cy (Nguyn c Lng, 2002)
Vi cch nui cy ny, ta c th p dng chuyn gene vo t bo trn hay to
cy a bi bng cch dung hp hai t bo trn vi nhau.
2.2.3 Cht dinh dng trong mi trng nui cy in vitro
V nguyn tc, m thc vt nui cy trong mi trng in vitro cng cn cc
cht dinh dng ging nh cy trng trn t.
a) Cc cht v c
Cht v c nng ln hn 0,5 mM/l c xp vo nhm a lng, cn
nng thp hn 0,05 mM/l c xp vo nhm vi lng.
Ngun m trong mi trng in vitro c cung cp hai dng ion NH4+ v
NO3-. Lng m tng s cn trong khong t 12-60 mM/l, trong c NH4+ bin
thin t 6-20 mM/l v NO3- t 6-40 mM/l ( Bi B Bng, 1995). Phn ln cy a
NO3- hn NH4+ nhng cng c nhng trng hp ngc li. Lng K+ cn trong
mi trng bin thin t 2-25 mM/l. Cn cc ion khc Ca2+, H2PO4-, SO42-, Mg2+,
thng thp hn khong 1-5 mM/l
Khong vi lng thng c dng nng 0,1-100 M/l
b) Ngun carbon
M cy trong mi trng t dng khng c kh nng t dng (autotrophic),
do khng tin hnh c quang hp y trong iu kin khng thun li nh
12
thiu s trao i kh bn trong chai v bn ngoi chai. Do ngun carbon phi

c cung cp cho mi trng nui cy nh cc dng ng, ch yu l ng
sucrose, thng chim khong 2-3%.
c) Vitamin v amino acid
Cc vitamin thng dng l thiamine (B1), nicotinic acid (B3), pyridoxine
(B6), calcium pentothenate (B5) v myo- inositol. Nng t 0,1 n 10 mg/l.
Cc amino acid thng dng l L- asparagine (100 mM/l), L-glycine (2mM/l),
L-arginine (10 mM/l), L-cysteine (10 mM/l). Khc vi m v c, cc amino acid
thng c t bo hp th nhanh hn. Tuy nhin, c th dng cc cht hu c
trong nui cy nh nc da (5-20%), cc cht trch t khoai ty, cam, c chua,
chui.
d) Cc cht iu ho sinh trng
Auxin: Cc dng auxin thng dng trong nui cy in vitro l 2,4-D,NAA,
IBA v IAA nng 0,1-5 mg/l. NAA v IBA thng dng cho ra r v dng
phi hp vi cytokinin trong s nhn chi. Cn 2,4-D rt hu hiu trong to ra m
so.c
Cytokinin: C tc dng trong s phn chia t bo, s phn ho chi, s nhn
chi. Cc cytokinin thng dng l kinetin, BAP, nng khong 1-10 mg/l, kch
thch s ra chi bn, lm gim nh hng ngn v c ch s thnh lp r.
Gibberellin: C khong 20 loi nhng GA3 l loi thng dng nht, c tc dng
lm tng s vn lng v s pht trin ca nh sinh trng hoc chi. Tuy nhin
gibberellin khng c dng ph bin trong nui cy m
e) Agar (thch)
c bo ch t rong bin, dng lm cht nn cho mi trng nui cy, c
ngun gc thc vt t nhin, khng gy c i vi cy. Agar ho tan thnh mt th
keo c th bm nc v hp th cc hp cht. Lng agar thng dng t 0,60,8%, nu nng thp (0,4%) mi trng mm long. Nng cao (1%) mi
trng b cng, kh cy v m kh tip xc vi mi trng.
13
f) Nc
Nn s dng nc ct bo m cht lng nc, chim khong 95% mi
trng nui cy. Ngoi nc ct c th dng nc kh ion nhng khng dng nc
my.
g) Yu cu pH
pH mi trng cy m t 5,0-6,5 v thng c iu chnh t khong
6,0. Mi trng c pH thp (thp hn 4,5) hoc cao (cao hn 7,0) c ch s pht
trin ca m. Nu pH bt u khong 5,0-7,0, sau khi hp s gim i 0,3-0,5 n v.
2.2.4 Cc bc nhn ging in vitro
Bc 1: Nui cy nh sinh trng.
Mu cy thng l chi, mt lng, nh sinh trng, l, r... em ra sch v
kh trng bng cn, natri hypochlorit, calci hypochlorit. Sau , mu c a vo
mi trng to iu kin phn chia v phn ho mnh. i vi mu d b ho nu,
mi trng thng b sung than hot tnh hoc ngm mu vi ascorbic acid v
citric acid (25-150 mg/l mi loi) trc khi cy (Bi B Bng, 1995).
Bc 2: To th nhn ging in vitro
Th chi (multiple shoot) v th ct t (cutting) cy vo mi trng c cht
kch thch sinh trng (cytokinin, GA3...) to m so, to cm chi.
Bc 3: Nhn ging in vitro
y l giai on quan trng nht nhm mc ch tng sinh khi th nhn
ging. S nhn ging ny tu thuc vo kh nng to chi nch hoc pht khi chi
bt nh t nhng khi ging nh callus di gc chi, ri ra r.
Bc 4: Ti sinh cy hon chnh in vitro
Cc chi, callus hay m so s c cy vo mi trng c auxin to r, to
thn, l y . Cy phi c nui cy trong iu kin ging nh bn ngoi vn
m thun ho cho n khi cy con khe mnh.
14
Bc 5: Chuyn cy in vitro ra vn m
Cy con pht trin tt trong mi trng in vitro c ly ra v ra sch mi
trng nhn to bm gc r, ri em trng ra vn m. Do thay i v iu kin
sng, cy con d b tress, mt nc v mau ho nn vn phi mt, m cao.
2.2.5 Cc yu t nh hng nhn ging in vitro
a) La chn mu cy
M non (chi nh), chi nch hay chi bt nh ti sinh tt hn so vi m gi
ca cng mt cy.
b) Mi trng nui cy
Mi trng MS (Murashige v Skoog) cho kt qu tt trong nhn ging cy
ban u v nhn chi tip theo. to chi nch nng auxin v cytokinin tng
i thp, cn to chi bt nh cn nng cytokinin cao v auxin thp; Cn to m
so th ngc li.
Mi trng rn hay lng cng nh hng n kh nng pht trin r. Agar
gip cy ng vng v tng kh nng thong kh to iu kin hp thu dng cht.
i vi cy tit cc c t nh phenol th cn thm than hot tnh hay PVP
(polyvinyl pyrrolidone) hp th cc cht ny gip cy tng trng tt hn.
Mi trng c th b sung cc cht ngn cn s oxyt ho phenols nh citric
acid, ascorbic acid, thiourea hoc L- cystine. Hoc b sung thm amino acid nh
glutamine, arginine v asparagine.
Thng xuyn cy chuyn vo mi trng mi nhm to iu kin dinh dng
y cho cy nui cy in vitro sinh dng tt.
c) nh sng v nhit
nh sng: Cng nh sng ph hp trong khong 1000-5000 lux. Thi
gian chiu sng thng 16 gi sng/8 gi ti.
Nhit : nh hng rt ln n s sinh trng v pht trin ca cy in
vitro. Nhit ti u khong 20-250C.
15
2.3 Cng ngh to ht nhn to

2.3.1 Khi nim
Nm 1978, Murashige pht hin u tin v k thut ht nhn to. Nm 1987,
Redenbaugh v cng s pht trin thnh cng k thut to ht nhn to. V c
bn, ht nhn to ging ht t nhin, c cu to l phi v tnh c bc bi lp o
bn ngoi bi lp gel v c kh nng ny mm nh ht t nhin, nhng khc bit l
ht nhn to khng c phi nh.
Hnh 2-2: Cu to ht nhn to

Nm 1988, McKersie v cng s to thnh cng ht nhn to cy c linh
lng, 65% ht bin i sau bc lm kh. Nm 1985, Kitto v Janick bc ht nhn
to c rt bng polyxyethylene t t l sng 3%. Janick (1988), to ht nhn to
thnh cng trn cn ty. Gray (1987) thnh cng trn nho v cy c nn. Nm 1988,
la m c to ht nhn to do Carman v cng s.
2.3.2 Mc ch
S pht trin ca cng ngh nui cy in vitro, ti sinh thnh cy hon chnh t
phi v tnh, m rng phm vi ng dng mi cho vic to ht nhn to nhm
mc ch vn chuyn, bo qun, lu tr ging d dng hn. Trong iu kin ti u
th ht s ny mm khi lp v nhn to cn khng s chuyn sang trng thi ngng
sinh trng cho n khi gp iu kin thun li. Ngoi ra, cn c nhiu li ch cho
vic nhn ging nhanh nhng thc vt nh:
16
- Khng to c ht
- Ht to thnh ny mm vi mt s lng thp
- Kh nng ht sng st rt thp sau khi bo qun
2.3.3 Cc nhn t to ht nhn to
2.3.3.1 To phi v tnh
Phi soma c hnh thnh khng thng qua qu trnh to m so c gi l
phi v tnh, phi v tnh c th ti sinh cy hon chnh hoc nhn sinh khi trc
tip bng h thng nui cy ph hp.
Phi soma khc vi phi hp t, phi hp t thnh lp do s phi hp gia
giao t c v giao t ci, khi u c thnh lp c cu trc n cc ni lin vi
cc m mch ca m so. Con phi soma thnh lp t m t bo , c cu trc lng
cc v khng c mch ni lin vi m so.
Nm 1958, khi nhng t bo phi thc vt u tin c thu nhn t m dinh
dng ca cy c rt (Daucus carota). Sau , hng lot cc m thc vt khc s
dng nui cy to phi (Triticum aestivum v Glycine max)
Giai on trc khi hnh thnh t bo soma gi l tin phi (PEDCpreembryogenic determined cell) (Sharp v csv, 1980). V t bo tin phi c th
phn chia hnh thnh t bo phi trc tip, gi l pht sinh phi trc tip.
Bng 2.1: S pht trin ca m trn mi trng nui cy
17
Tuy c nhiu t bo pht sinh t bo phi khng cn cc cht kch thch sinh
trng nh t bo cy c rt, u vn nhng cng c nhiu t bo li cn auxin
phn bo trc khi pht sinh t bo phi (IEDC- induced embryonic determined
cell) (Sharp v scv, 1980). V c nhiu t bo hnh thnh phi t m so, s pht
sinh phi tin hnh gin tip. Ngoi ra, cn s dng t bo phi hp t to phi,
ngoi tr ht c to trong qu trnh th phn m kiu di truyn khng xc nh.
2.3.3.2 C ch pht sinh phi soma
Nhng quan st chi tit v qu trnh pht sinh phi pht hin c 4 pha : 0, 1, 2
v 3, c nhn thy trong giai on u ca tin trnh pht sinh phi trong h
thng nui cy ni trn (Fujimura v Komamine, 1979).
pha 0, nhng t bo n (giai on 0) hnh thnh nhng cm t bo c kh
nng pht sinh phi (giai on 1) trn mi trng c auxin. Trong sut giai on
ny, nhng cm t bo hnh thnh t nhng t bo n c kh nng to phi khi
mi trng nui cy khng c auxin, hnh thnh nhng cm t bo giai on 1.
Sau , pha 1 xut hin khi cy chuyn nhng cm t bo giai on 1 qua mi
trng khng c auxin. Trong sut pha 1, nhng cm t bo tng sinh chm v
dng nh khng bit ha.
Sau pha 1 s phn bo xut hin nhanh trn mt phn ca nhng cm t bo,
dn n vic hnh thnh nhng t bo phi hnh cu. Pha ny c gi l pha 2.
Pha tip theo sau, pha 3, cy con in vitro pht trin t nhng phi hnh cu qua
phi hnh tim v phi hnh thy li.
18
Hnh 2-3: Cc giai on pht sinh phi soma

2.3.3.3 To v bc bng sodium alginate
C rt nhiu tc nhn to gel c s dng lm v bc cho phi. c th l
agar, alginate, polyco 2133 (Bordon Co.), carboxyl methyl cellulose, carrageenan,
gelrite (Kelko. Co.), guargum, sodium pectate, tragacanth gum, dextran, xanthan
gum nhng hp cht ny s ng li thnh gel khi c cho vo mi trng c
cht in phn thch hp nh ng sulphate, calcium chloride hoc ammonium
chloride nh vo nhng lin kt ion. Phng php c s dng rng ri nht
to v bc cho phi l s dng sodium alginate (Bajaj, 1995; Jeon v csv, 1986;
Redenbaugh v csv, 1993). Alginate c s dng nhiu do n c nhng c tnh
rt thun li nh tnh dnh va phi, khng gy c cho phi sinh dng, c cc c
tnh tng hp sinh hc, kh nng to thnh gel nhanh, r tin, c lu, cng
ca gel va phi c th va to thun li cho s h hp ca phi v va bo v
c phi khi nhng tn thng bn ngoi.
19
Hnh 2-4: Cng thc ho hc ca alginate

Alginate, cht hu c mch thng, k nc, mui ca acid alginic, l mt
polyuronic bao gm -D-mannuronate (M) v mnh bn C5
-L-guluronate (G)
(Aoyagi v csv, 1998).

Nguyn tc chnh trong qu trnh to v bc alginate l sodium alginate
cha phi s to thnh tng ht nh, trn v cng khi nh vo trong hn hp
NaCl2.2H2O nh vo s trao i ion gia ion Na+ c trong hn hp sodium alginate
vi Ca2+ c trong hn hp CaCl2.2H2O. V bc cng nhiu hay t l ph thuc vo
s lng ion Na+ trao i vi ion Ca2+. Do nng ca hai tc nhn to gel,
sodium alginate v CaCl2.2H2O, v thi gian cho vic to lin kt ion phi tht ti
u c th to thnh v bc mt cng thch hp nht.
Khng nh phi hp t, phi v tnh thiu lp ni nh cha cht dinh dng
bn ngoi nui phi, do bng vic thm vo cht nn gel nhng cht dinh dng,
cht iu ha tng trng, carbohydrate s to mt ni nh nhn to thch hp ti a
cho tng trng v sng st ca phi (Gray, 1990; Gray v Purohit, 1991;
Redenbaugh v Walker, 1990). Ngoi ra nhm trnh cho phi b mt nc, hay cc
tn thng c hc, tng sc khng cho phi ngi ta c th thm vo cht nn
gel cht khng sinh, thuc tr nm, tr su, vi sinh vt.
Mc d vic to ht nhn to t cch bc phi v tnh bng sodium alginate
cho thy c mt s thnh cng nht nh trong vic ti sinh cy con, vn cn nhiu
vn kh khn khi s dng v bao alginate ny, cht dinh dng c th b mt i
khi v bao (Redenbaugh v csv, 1987), s trao i kh km (Redenbaugh v csv,
1993) c mt s ngh cho rng, vic thm vo than hot tnh s gip nng
20
cao kh nng sng st, pht trin ca phi v tnh hn. Nguyn nhn l do than hot
tnh tng cng kh nng h hp ca phi, v n gi li nhng cht dinh dng
nhiu hn trong v bc, phng thch chng rt chm dng cho s pht trin ca
phi.
2.3.3.4 Quy trnh to ht nhn to
Hnh cu
Hnh tim
Hnh thu li
Hnh l mm
Hnh 2-5: Cc dng phi soma

Bc 1: Dng dao ct chi (shootbud) c kch thc 2-3 mm t cy nui cy in
vitro. Sau , dng dao tch ri cc phi hnh tim.
Bc 2: Dng pipette hay ng tim ht dung dch sodium alginate ri cho nh
tng git ri xung.
Bc 3: Dng pince gp chi hay phi a vo trong git alginate c nh
xung dung dch CaCl2.2H2O 100 mM trong 15 pht.
Bc 4: Dng pince gp ht cho vo nc ct v trng trong 5 pht lm
sch lng CaCl2.2H2O cn st li.
21
Dung dch sodium alginate

a phi/chi vo git sodium alginate
Ngm trong dung dch CaCl2.7H2O 100
mM trong 15 pht
Ra ht trong nc ct 5 pht
sch CaCl2.2H2O
Hnh 2-6: Quy trnh to ht nhn to
2.4 Nm r cng sinh- mycorrhiza

2.4.1 Khi nim
Cc loi thc vt khai thc t t trng t s gip ch ca vi sinh vt c li
c gi l mycorrhiza (nm vng r). Cc si nm len li gia cc ht t, pht
trin bn trong cht hu c, thm ch chng cn xm nhp vo v cn trng cht, t
s dng cc dng cht hu c v photphorus, sau tr li cho cy trng. Nh
vy, c s kt hp cao gia nm t v r cy trng trong cc nm to nn
mycorrhiza thng l mt s nm Basidiomycetes, Ascomycetes, v Zygomycetes
(theo Harley & Smith 1983, Kendrrick 1992, Brundrett 1992)
Nm r l mt thut ng c Frank s dng ln u vo nm 1885 khi pht
hin mi lin h gia si nm v r trn cy thng v mt s cy l rng, c ly
t ch Hy Lp Mykes v Rhiza. Sau ghp t ting Anh Myco vo thnh
t Mycorrhiza
Ngi ta d on, trn th gii ngy nay c 1000 chi 200 h thc vt c nm
r. Theo thng k ca Trappe nm 1962 c 535 loi nm thuc 81 chi 30 h 10 b
nm c th cng sinh vi trn 1500 loi cy g khc nhau.
22
2.4.2 Cc dng mycorrhiza

2.4.2.1 Ectomycorrhiza
c hnh thnh ch yu trn cy rng do nhiu loi nm m v mt vi loi
nm ti. Hu ht bo t ca Ectomycorrhiza c sinh ra trn mt t v c pht
tn nh gi. Si nm ca Ectomycorrhiza thng sinh ra mt lp nm ph chung
quanh pha ngoi ca cc r dinh dng. Cc nm ny cng i vo bn trong r
nhng ch pht trin xung quanh cc t bo v r thay th mt phn phin mng v
hnh thnh ci gi l mng li Hartig (Hartig net). Tu thuc vo mu sc ca nm
pht trin trn r m Ectomycorrhiza c mu vng, nu, trng, en.
2.4.2.2 Endomycorrhiza
c hnh thnh trn hu nh tt c cy trng v trn mt s cy rng bi cc
nm tip hp, ch yu l ging Glomus v mt s nm khc nh Acaulospora.
Endomycorrhiza cng c hnh thnh bi mt vi nm m, r cy c
Endomycorrhiza bn ngoi ging vi r khng c mycorrhiza v hnh dng v mu
sc (Agrios, 1997)
a)Vesicular arrbuscular mycorrhiza (=arrgbusular mycorrhiza, VAM hoc AM)
y l nhm ph bin, hin din trn c cy thn g, l nhm mycorrhiza
quan trng nht trong Endomycorrhiza. Cc si nm mc bn trong t bo v ca r
dinh dng hnh thnh vi ht gi l bi (arbuscules) hoc hnh thnh cc u sng
ln d tr thc n gi l ti nang (vesicules). Bo t c hnh thnh trong t.
b)Erioid mycorrhiza
Si nm xon cun bn trong t bo lng r ca cy thuc b Ericales. Si nm
mc tha khng lan rng vo mi trng t, chng c th mc t bo t nhng t
c kh nng ti sinh.
c)Orchid mycorrhiza
Si nm xon cun bn trong r hoc thn ca cc cy thuc h lan. Nhng
cy lan nh v mt s cy trng thnh khng c dip lc t (chlorophyll) u cn
cho s sinh tn ca chng.
23
2.4.3 Tc ng c ch ca mycorrhiza
Mi quan h gia lan v nm l mi quan h mt thit, trn 80% loi lan u
c mi quan h vi nm. Theo Currah et al (1997), nhm nm chnh r lan l
Basidiomycetes mc d cng c Ascomycetes, tuy nhin Ba sidiomycetes cng l
tc nhn gy bnh trn nhng cy khc nh nm Rhizoctonia solani (theo Harley,
1982). Thm ch, nm cng sinh trn loi lan ny l tc nhn gy bnh cho loi lan
khc, v lan c kh nng iu tit s xm nhim v pht trin ca nm cng sinh.
2.4.3.1 M rng din tch hp thu ca r cy
Si nm cng sinh l c quan hp thu ch yu ca r nm, si nm c th ko
di ra xung quanh r lm tng tc ht P ln gp 6 ln. Tui th ca th si nm
cng vt xa lng ht.
2.4.3.2 S trao i dinh dng
Do ht lan khng d tr cht dinh dng nn s ny mm ca ht ph thuc
vo mycorrhiza. Mycorrhiza lan khc vi loi mycorrhiza khc v bn cht trao
i dinh dng. Sau khi hnh thnh mycorrhiza, cacbon hu c v nhng cht dinh
dng i vo ht. Bi v s ny mm ca ht lan khi khng c s cng sinh vi nm
i hi phi c cc cht ng, acid amin v vitamin nn ngi ta gi nh rng ht
lan nhn c cc cht ny t nm ny mm. Sau , nm tip tc cung cp
ngun nng lng cho cy lan.
Cn i vi cc cy trong t, do lng P c nh, thng cc gc phosphat
kt hp vi Fe, Na, Al kh di ng. Do , nm r ngoi cng sinh lm nhim v
sn sinh enzyme phosphorase chuyn P khng tan thnh P ha tan, tng kh nng
hp th ca r, cung cp cho cy trng.
2.4.3.3 S hnh thnh cht kch thch sinh trng mycorrhiza
Trong qu trnh cng sinh vi r cy, nm hnh thnh nhiu cht kch thch
sinh trng nh auxin, cytokinin, gibberellin, vitamin B1, indole-3acetic acid (IAA).
24
2.4.3.4 Nng cao sc chng chu ca cy

Nhiu nghin cu u chng minh, sau khi nhim nm cho cy, cy ch c kh
nng chng kh hn, chng chu mn, nhit , m v pH cc oan, kim loi
nng.
2.4.3.5 Tng kh nng khng bnh ca cy trng
Nm 1942, Davis pht hin nm r cng sinh c th lm gim bt bnh hi r.
Nm 1968, 1982, 1994 nhiu tc gi u cp n nm cng sinh c th gim
bnh thi r thng xung 25%, cn lm gim bnh do tuyn trng, bnh mc sng,
bnh bu r (Trn Vn Mo, 1999)
2.4.4 S xm nhp ca mycorrhiza
Nm c th xm nhp qua ht, r, hoc thn. i vi ht, nm mc xuyn qua
ht vo bn trong lng biu b. Cn r, nm thng mc xuyn qua v r ni gn
u r li vo l mt t bo ca ca ngoi b. Nm nhanh chng lan rng khp v.
Trong mi t bo v r, si nm xon cun dy c, c gi l peleton (Harley,
1982)
25
CHNG 3
VT LIU V PHNG PHP NGHIN CU
3.1 i tng th nghim
C hai ni dung u lm th nghim trn cy lan
3.2 Thi gian thc hin

Thi gian thc hin ti t thng 3/2007 n thng 8/2007 ti phng nui
cy m_ B Mn Cng Ngh Sinh Hc v phng thc tp bnh cy _ khoa Nng
Hc_ trng i Hc Nng Lm Tp H Ch Minh.
3.3 Ni dung nghin cu

ht nhn to
Phi soma em bc v ht cc nng alginate khc nhau xc nh nh
hng ln tnh cht v ht nhn to.
S dng mi trng MS c nng khong a lng khc nhau xc nh
t l sng ca ht nhn to.
Th nghim 3: Kho st mi trng bo qun ht nhn to.
Ht nhn to sau khi to xong em lu tr cc mi trng khc nhau Xc
nh iu kin mi trng bo qun ti u.
Ht nhn to cho ny mm trn cc gi th khc nhau xc nh t l ny
mm ca ht nhn to.
26
Ni dung 2: Kho st s hin din ca mycorrhiza trn mt s ging lan

Ly mu r lan Dendrobium lai, Dendrobium rng, Catleya, Ngc im em
nhum quan st hnh dng v s c mt ca mycorrhiza trn cc ging lan.
3.4 Vt liu nghin cu

3.4.1 Trang thit b v dng c nghin cu
Thit b: T cy v trng, ni hp (autolave), my o pH, cn in t, n
neon....
Dng c: Pince, ko, dao cy, chai nc bin, a petri, n cn, ng ong,
pipet...
3.4.2 Mu s dng v iu kin nui cy

Mu: Dng phi v tnh hay chi cy lan.
iu kin phng nui cy:
Cng nh sng 2000 lux
Nhit 26 20C
m 60-80%
Thi gian chiu sng:16 gi/ngy
3.4.3 Mi trng s dng

Mi trng s dng trong th nghim: Mi trng MS (Murashige v Skoog
1962)
Thnh phn
Khong a lng
(Macro MS)
Khong vi lng
Dang s dng
nng (mg/l)
NH4NO3
1650
KNO3
1900
KH2PO4
170
MgSO4.7H2O
370
CaCl2.2H2O
440
H3BO3
6,2
MnSO4.4H2O
22,3
27
(Micro MS)
Fe_EDTA
Vitamine
CoCl2.6H2O
0,025
CuSO4.5H2O
0,025
ZnSO4.4H2O
8,6
Na2MoO4.2H2O
0,25
KI
0,83
FeSO4.7H2O
27,8
Na2EDTA.2H2O
37,3
Myo-Inositol
100
Nicotinic acid
0,5
Thiamine-HCl (B1)
0,1
Glycine
Pyridoxine-HCl (B6)
0,5
Cc cht khc:
Nc ct
ng: 30 g/l
Agar: 8 g/l
pH mi trng trc khi hp: 5,8
Ngoi ra cn s dng thm mi trng MS c b sung thm sodium alginate
v CaCl2.2H2O 7mM bc v nhn to
Nhum r: Nc ct, Alkaline H2O2, HCl 1%, Acid Glycerol, Trypan blue.
3.5 Phng php nghin cu

3.5.1 Chun b mi trng
Tt c mi trng u phi c kh trng bng ni hp autoclave 1210C
trong 25 pht, k c CaCl2.2H2O
28
3.5.2 B tr th nghim
Cc th nghim c b tr theo kiu hon ton ngu nhin, lp li 3 ln. Ht
nhn to sau khi bc v xong cho ny mm trn mi trng MS khng c cht kch
thch sinh trng.
ht nhn to
Nng alginate
Nghim thc Mi trng

1
MS
10 g/l
MS
20 g/l
MS
30 g/l
MS
40 g/l
MS
50 g/l
Ghi ch:
S nghim thc: 5
S bnh mi nghim thc: 1
S ht mi bnh: 10
Tng s bnh: 15
Tng s ht: 150 ht
Ch tiu theo di: Theo di tnh cht v ht, t l sng ca phi sau khi bc v
ht
Nghim thc Mi trng
Nng alginate
MS
30 g/l
MS
30 g/l
MS
30 g/l
1/8MS
30 g/l
29
Ghi ch
S nghim thc: 4
S bnh mi nghim thc:1
S ht mi bnh: 10
Tng s bnh: 12
Tng s ht: 120
Ch tiu theo di: Theo di t l sng ca ht nhn to trn mi trng MS
Th nghim 3: Kho st mi trng bo qun ht nhn to
Nghim thc Mi trng tao ht
Nng alginate
Mi trng bo qun
MS
30 g/l
Khng c dinh dng
MS
30 g/l
Nc ct
MS
30 g/l
Nc ng
MS
30 g/l
Agar
MS
30 g/l
Agar c ng
MS
30 g/l
MS
Ghi ch:
S nghim thc:6
S ht mi bnh: 10
Tng s bnh: 18
Tng s ht: 180
Cc dng mi trng bo qun ht nhn to:
Mi trng khng dinh dng: Bnh nc bin khng, em hp kh trng sau
cho ht nhn to vo.
Mi trng nc ct: Nc ct cho vo mi bnh 150 ml em hp kh trng
Mi trng nc ng: Nc ct b sung 30 g/l ng sucrose, cho vo mi
bnh 150 ml em hp kh trng cho ht nhn to vo.
Mi trng agar: Nc ct cho vo 8,6 g/l agar, cho vo mi bnh 70- 80ml
em hp kh trng.
30
Mi trng agar ng: Nc ct c 8,6 g/l agar v b sung thm 30 g/l ng

cho vo mi bnh 70-80 ml em hp kh trng.
Ch tiu theo di: Theo di mu sc ca phi khi bo qun, t l sng ca ht
nhn to khi bo qun v khi cho ny mm trn mi trng MS
Nghim thc
Mi trng to ht
Nng alginate
iu kin ny mm
MS
30 g/l
MS
MS
30 g/l
Cu giy
MS
30 g/l
Bng gn
Ghi ch:
S nghim thc: 3
S ht mi bnh:10
Tng s bnh: 9
Tng s ht: 90
Phng php to gi th cu giy, bng gn
Cu giy: Giy thm xp thnh 4 lp cho vo bnh em hp kh trng, sau
cho vo thm 40 ml nc ct hp tit trng, cho ht nhn to nm trn b mt
giy.
Bng gn: Bng gn ct lp dy 2 cm cho vo bnh em hp kh trng, sau
cho 40 ml nc ct tit trng , cho bng gn thm t ri b ht nhn to ln b
mt bng gn
Ch tiu theo di: T l ht nhn to ny mm trn cc gi th
Ni dung 2: Kho st s hin din ca Mycorrhiza trn mt s ging lan
Ly mu r lan Dendrobium lai, Dendrobium rng, Cattleya, Ngc im bo
qun trong ti nylon. Mi ging lan thu 15 mu r. R sau khi ly xong phi ct
mng ri em nhum theo quy trnh sau:
Ra sch r
thm kh
hp trong ni autoclave 1210 C-3
Ra r lm nhiu ln trong nc ct
31
Ngm r trong dung dch Alkaline H2O2 t 10 30 (tu theo mu

nu ca r )
Ra r nhiu ln
Ngm r trong dung dch HCl 1% trong 1-2 gi. Khi ngm trong ho
cht cn phi lm ngp mu r v lng ho cht gp 20 th tch r.
Nhum r trong Acid Glycerol/Trypan Blue 30 ti 900C trong t sy.
Tn tr mu trong Acidic Glycerol
Quan st hnh thi v nh gi t l hin din ca mycorrhiza trn cc ging lan.
3.6 X l s liu
S liu c x l thng k bng phn mm STATGRAPHIS 7.0 v phn mm
Excel
32
CHNG 4
KT QU V THO LUN
4.1 Ni dung 1: Nghin cu k thut to ht nhn to t phi cy lan
A: Phi hnh tim
B: Phi hnh cu
C: Phi hnh l
D: Phi hnh thu li
Hnh 4-2: Cc dng phi v tnh

Phi soma l cu trc lng cc vi c nh ngn v vng m phn sinh c
bn, m kh nng ti sinh hnh thnh cnh non v r. T bo c kh nng sinh phi
l nhng t bo ng knh, nhn to, t bo cht m c v nhiu ht tinh bt
(Nguyn c Lng, 2002). Nhng t bo ny c hm lng protein v RNA cao
v tri qua cc bc:
-S bit ho cc t bo c kh nng sinh phi thnh t bo phi.
-S pht trin ca nhng t bo phi mi hnh thnh thng qua cc giai on
sau
Phi hnh cu
Phi hnh thu li
Phi hnh tim
Phi hnh l
Sau 4 tun nui cy, bt u xut hin nhng t bo c cu trc ging phi,
phi hnh cu c cu to non hn, c mu xanh l t hn so vi nhng dng phi
khc.
33
A: Phi hnh cu
B: Phi hnh tim
C: Phi hnh thu li
D: Phi hnh l
Hnh 4.2: Cu trc ca phi v tnh
A: Nc ct
B: CaCl2.2H2O
C: Sodium alginate
Hnh 4.3: Mi trng to ht nhn to
34
A: Ht sodium alginate
B: Ly phi
C: a phi vo git alginate
D: Nh git phi xung CaCl2
Hnh 4.4: Cc bc to ht nhn to
A: Ht nhn to ngm trong CaCl2.H2O 100 mM trong 20 pht

B: Ht nhn to ra trong nc ct 5 pht
Hnh 4.5: S hnh thnh ht nhn to
35
Lp alginate
Phi v
tnh
Hnh 4.6: Cu to vi th ca ht nhn to
4.1.1 Th nghim 1: Kho st nh hng ca nng sodium alginate n s

hnh thnh ht nhn to
* nh hng ln hnh thi v ht nhn to
Ht nhn to sau khi to cc nng sodium alginate 10 g/l, 20 g/l, 30 g/l, 40
g/l, 50 g/l cho ra cc ht nhn to c hnh thi v khc nhau. Ht nhn to nng
10 g/l khng nh hnh c r rng, v alginate khng bc c phi.
Bng 4.1: Nng alginate nh hng ln hnh thi v ht nhn to
NT
Mi trng
Sodium alginate
Hnh thi v ht
MS
10 g/l
Rt mm, rt d v
MS
20 g/l
Mm, khng bc ton phi
MS
30 g/l
Va, bc ht trn p
MS
40 g/l
Dy, ht trn u
MS
50 g/l
Rt dy, ht trn khng u
36
A: Nng alginate 10 g/l
B: Nng alginate 20 g/l
C: Nng alginate 30 g/l
D: Nng alginate 40 g/l
E: Nng alginate 50 g/l

Hnh 4-7: Ht nhn to cc nng alginate khc nhau
*nh hng ln kh nng bo qun
Ht nhn to nng sodium alginate 20 g/l,30 g/l, 40 g/l, 50g/l to hnh
dng ht r rng, a nhng ht nhn to ny ln mi trng MS khng c cht
kch thch sinh trng, theo di t l sng ca ht nhn to trong khong thi gian
dn cch 1 tun.
37
Bng 4.2: T l sng ca ht nhn to cc nng alginate

NT
Nng alginate
T l sng (%)
1tun
1tun
3tun
10 g/l
20 g/l
80,43B
78,02B
71,63B
30 g/l
75,41AB
70,25AB
69,82B
40 g/l
62,68A
58,02 A
55,28A
50 g/l
60,66 A
58,98 A
55,59 A
18,82
23,25
17,87
CV
Cc gi tr theo sau bi ch ci khng cng k t c s khc bit rt c ngha v mt

thng k hc
Tun
Biu 4-1: T l sng ca ht nhn to cc nng alginate khc nhau

*nh hng ln t l ny mm
Ngoi theo di t l sng ca ht nhn to, t l ny mm ca ht nhn to trn
mi trng MS cng quan trng
38
Bng 4.3: T l ny mm ca ht nhn to cc nng alginate khc nhau

NT Nng alginate T l ny mm sau 3 tun T l ny mm sau 4 tun
1
10 g/l
20 g/l
61%B
66,41B
30 g/l
56 %B
71,60B
40 g/l
31%A
39,68A
50 g/l
26% A
42,62A
14,66%
20,8%
CV

thng k hc
Kt lun s b: nng alginate 10 g/l, 20 g/l v ht nhn to rt d v,

khng hnh thnh v ht r rng. Vi nng alginate 30 g/l, 40 g/l, 50 g/l v ht
nhn to cng v bc ton b phi/chi. Trong , nng 30 g/l lm cho ht nhn
to trn p, c cng va phi. nng 20 g/l v 30 g/l, t l ht nhn to
sng st v ny mm cao hn so vi 40 g/l v 50 g/l c s khc bit v mt thng
k. Do , c th s dng nng alginate 30 g/l to ht nhn to, va c hnh
thi p v t l ny mm cao.
Hnh 4-8: Ht nhn to ny mm
4.1.2 Th nghim 2: Kho st nh hng ca mi trng to ht nhn to

* Kho st kh nng bo qun ht trn mi trng MS
V ht nhn to bn cnh b nh hng bi nng sodium alginate cn b nh
hng bi mi trng dinh dng b sung trong v ht. V nh mi trng ny m
cung cp dinh dng cho phi sng v ny mm khi gp iu kin thun li
39
Mi trng s dng b sung cho v ht nhn to l mi trng MS v mi

trng MS gim dn khong a lng nh mi trng MS, MS, 1/8 MS. Ht
nhn to sau c em gieo trn mi trng MS, theo di t l sng ca phi ht
nhn to sau khong thi gian 1 tun.
Bng 4.4: T l sng ca ht nhn to cc mi trng dinh dng khc nhau
NT
T l sng (%)
Mi trng
1 tun
2 tun
3 tun
MS
91,00NS
73,33NS
49,67B
MS
86,00NS
70,67NS
55,67B
MS
87,33NS
81,00NS
34,67A
1/8 MS
77,33NS
70,00NS
36,67A
CV
9,42%

thng k hc
Biu 4-2: T l sng st ca ht nhn to mi trng khc nhau
40
*Kho st t l ht ny mm
Khong a lng c trong v ht nhn to cng nh hng n t l ny
mm ca ht nhn to trn mi trng MS
Bng 4.5: T l ny mm ca ht nhn to cc mi trng dinh dng khc
nhau
NT Mi trng
T l ny mm sau
T l ny mm sau
3 tun (%)
4 tun (%)
MS
52,33B
53,80A
MS
71,67A
71,42B
MS
40,00B
50,40A
1/8 MS
39,00B
47,78A
18,05
14,65
CV

thng k hc
Kt lun s b: th nghim 2, chng ti nhn thy ht nhn to ca phi cy

lan c v bc mi trng nhn to MS,1/2MS c t l sng st cao hn cc mi
trng to ht khc (1/4 MS, 1/8MS) c s khc bit v mt thng k. Cn mi
trng MS t l ny mm ca ht nhn to cao nht (72%), thch hp cho vic
to ht nhn to phi cy lan Vanda.
4.1.3 Th nghim 3: Kho st mi trng bo qun ht nhn to

* nh hng n v ht
Ht nhn to c v bc ti u (1/2 MS, 30 g/l alginate) c th nghim bo
qun cc mi trng khc nhau.
41
Bng 4.6:Kho st nh hng ca mi trng bo qun ht nhn to

Mi
Nng
Mi trng bo
trng
alginate
qun
MS
30 g/l
Khng dinh dng
V ht kh cng, mu xanh
MS
30 g/l
Nc ct
V ht mm v ra, mu xanh
MS
30 g/l
Nc ng
V ht mm n ra, mu xanh
MS
30 g/l
Agar
Ht mu vng, nu en
MS
30 g/l
Agar c ng
Ht xanh vng,
MS
30 g/l
MS
Ht vn gi c hnh dng c
NT
Hnh thi v ht
A: Nc ct
B: Nc ng
C: Khng c cht d tr
D: Agar
E: Agar ng
F: Mi trng MS
Hnh 4-9: Ht nhn to trn cc mi trng bo qun

Kt lun s b:Trong mi trng lng (nc ct v nc ng), v ht b
nc thm thu mm v v ra, khng bao c phi, ht nhn to vn gi c
mu xanh m. Trong mi trng khng c cht dinh dng, v ht nhn to kh
bm cht vo phi. Cn mi trng agar v agar c ng, ht b vng hoc nu
en. Mi trng MS, ht nhn to vn gi c nguyn hnh thi ban u, mu sc
v hnh thi u khng thay i.
42
*Kho st t l ny mm sau khi bo qun

Ht nhn to sau khi bo qun trong 3 tun, em tt c ht nhn to mi loi
cho ny mm trn mi trng MS, theo di t l ny mm ca ht nhn to.
Bng 4.7: T l ny mm ca ht nhn to sau khi bo qun
Mi
Nng
Mi trng bo
T l ny mm
trng
alginate
qun
(%)
MS
30 g/l
Khng dinh dng
18,67A
MS
30 g/l
Nc ct
15,33A
MS
30 g/l
Nc ng
20,00A
MS
30 g/l
Agar
10,00A
MS
30 g/l
Agar c ng
24,67A
MS
30 g/l
MS
71,67B
NT
CV
2,6

thng k hc
Kt lun s b: Qua bng trn, chng ti nhn thy ht nhn to t phi cy

lan bo qun trn mi trng MS c t l ny mm cao hn trn cc mi trng bo
qun khc v mt thng k hc.
4.1.4 Th nghim 4: Kho st s ny mm ca ht nhn to trn cc gi th

khc nhau
Ht sau khi to xong cho ny mm 3 mi trng khc nhau cu giy, bng
gn, MS
43
Bng 4.8: T l ht nhn to ny mm trn cc gi th

Mi trng
Nng
iu kin
T l ny mm
to ht
alginate
ny mm
(%)
MS
30 g/l
MS
65,00A
MS
30 g/l
Cu giy
86,33AB
MS
30 g/l
Bng gn
94,33B
NT
CV
17,08 %

thng k hc
A: Bng gn
B: Cu giy
C: Mi trng MS
Hnh 4-10: Ht nhn to ny mm trn cc gi th

Kt lun s b: Qua th nghim 4, chng ti nhn thy gi th bng gn c ht
nhn to ny mm vi t l cao (88,63) so vi cu giy (68,64) v mi trng MS
c (53,65), c s khc bit v mt thng k.
44
4.2 Ni dung 2: Kho st s hin din ca mycorrhiza cc ging lan

Sau khi thu thp, nhum r quan st di knh hin vi, chng ti quan st
mycorrhiza bt mu xanh m, c cc dng sau:
-Si nm n mu xanh, phn nhiu nhnh n xuyn qua nhiu lp t bo
-Si nm cun trn trong 1 t bo
-Si nm l cc cun trn nm cc t bo, ni vi nhau bng si nm
Bng 4.9: S hin din ca mycorrhiza trn cc ging lan
Ging
S cy ly
S cy c
% cy c mycorrhiza
mu
mycorrhiza
DENDROBIUM RNG
15
13
87
DENDROBIUM LAI
15
10
67
CATTLEYA 1 L
15
12
80
CATTLEYA 2 L
15
12
80
NGC IM
15
47
ONCIDIUM
50
32
64
VANDA
50
12
MOKARA
50
15
30
(Ring ging Oncidium, Vanda, Mokara tham kho s liu ca Ng Hunh Thanh Vn,
2005.)
45
Biu 4.3: S hin din ca mycorrhiza trn cc ging lan

Qua bng s liu v biu , chng ti nhn thy hu ht cc ging lan u c
mycorrhiza cng sinh, gip cy lan pht trin tt hn. Tuy nhin s hin din cc
ging lan c s khc bit r rt. Mycorrhiza hin din nhiu nht Dendrobium
(77%), t nht Vanda (12%). nhng ging lan c tr nh Mokara, Ngc im,
Vanda th s c mt ca mycorrhiza km hn so vi cy lan a thn. Cy
Dendrobium rng (87%) c t l mycorrhiza nhiu hn so vi cy Dendrobium lai
(67%). Cn gia loi Cattleya 1 l v Cattleya 2 l th t l hin din ca
mycorrhiza l ngang bng nhau, khng c s khc bit.
Nguyn nhn:
Do hnh thc pht trin ca r
Do ngun gc cy lan trng
Do cch chm sc nh phun thuc tr su lm gim lng nm.
46
Khng c
mycorrhiza
C mycorrhiza
Hnh 4-11: Vng m c mycorrhiza
A, B: Si nm a bo, xuyn qua cc t bo

C: Nm c nhng u to trn cui si nm
D: Nm cun trn trong t bo.
Hnh 4-12: Cc dng mycorrhiza
47
A: Si nm cun li c 2 u a ra bn ngoi
B: Cc cun nm ni vi nhau bng cc si nm, si nm c vch ngn
Hnh 4-13: Dng mycorrhiza cun trn
48
CHNG 5
KT LUN V NGH
5.1 Kt lun
* Nghin cu k thut to ht nhn to t phi cy lan Vanda
Ht nhn to t phi cy lan Vanda c hnh dng p, t l ny mm cao khi
nng sodium alginate (30 g/l) v mi trng dinh dng (1/2 MS) b sung vo
v ht nhn to thch hp cho phi cy lan Vanda.
Mi trng bo qun (MS) thch hp cho vic lu tr ht nhn to, bo m
cht lng v t l ny mm cao cho ht nhn to.
Ht nhn to ny mm cho t l cao (bng gn thm 40 ml nc ct), v si
gn xp to mi trng thng thong, kh ro, gi m lu nn thch hp cho ht
nhn to ny mm
* Kho st s hin din ca mycorrhiza trn mt s ging lan

Mycorrhiza xut hin ging lan a thn cao hn ( 64-87%) cn lan n
thn th t l ny thp hn ( 12-47%) v vy cy a thn ht dinh dng tt hn, t
b hin tng thiu ht khong hn so vi cy n thn.
Mycorrhiza cy cy m (Dendrobium lai 67%) c t l thp hn so vi cy
lan c ngun gc t nhin (Dendrobium rng 87%)
5.2 ngh
* Nghin cu k thut to ht nhn to t phi cy lan Vanda
Tip tc kho st cc iu kin v mi trng bo qun ht nhn to lu hn
vn cho t l ny mm cao hn khi gieo ht nhn to iu kin thch hp.
Tm gi th khc gieo ht nhn to c gi r nhng c t l ny mm cao.
* Kho st s hin din ca mycorrhiza trn mt s ging lan
Tip tc nghin cu phn lp v nh danh tn cc dng mycorrhiza trn cy
lan.
49
Tip tc thu thp nhiu mu r lan v thc hin trn nhiu loi lan a ra
kt lun chnh xc, chi tit hn.
Nghin cu nh hng ca mycorrhiza trn s sinh trng ca cy lan trong
nui cy in vitro.
50
TI LIU THAM KHO

* Ti liu ting Vit
1. Bi B Bng, 1995. Nhn ging c y bng nui cy m. Nh Xut Bn S Khoa
Hc Cng Ngh V Mi Trng An Giang.
2. Lee S. Cooke, 1995. Su bnh hi cy hoa lan. (Nguyn Minh Trc dch). Nh
Xut Bn Nng Nghip.
3. Dng Cng Kin, 2002. Nui cy m thc vt (tp 1). Nh Xut Bn i Hc
Quc Gia Thnh Ph H Ch Minh
4. Nguyn c Lng, L Th Thu Tin, 2002. Cng ngh t bo. Nh Xut Bn
5. Trn Vn Mo, 1999. S dng vi sinh vt c ch (tp 2). Nh Xut Bn Nng
Nghip
6. Nguyn Cng Nghip, 1999. Trng hoa lan. Nh Xut Bn Tr
7. Liu Hng Ph, 2005. Kho lun tt nghip (i Hc Nng Lm)
8. Trnh Pari, 2004. Lun vn thc s (i Hc Nng Lm
9. Nguyn Thin Tch, on Th Hoa, Trn S Dng, Hunh Th Ngc Nhn, 1996.
K thut nui trng hoa lan. Nh Xut Bn Nng Nghip.
10. Trn Ngc Thu Tin, 2004. Kho lun tt nghip (i Hc Khoa Hc T
Nhin)
11. Ng Hunh Thanh Vn, 2005. Kho lun tt nghip (i Hc Nng Lm)
12. Cu lc b hc vin hoa lan, 1999. Tm hiu hoa lan. Nh Xut Bn Nng
Nghip
*Ti liu nc ngoi:
1. Supriyanto, Jahja T. K., 1995. Biology and biotechnology of mycorrhiza.
Southeast Asian Regional Centre for Tropical Biology Bogor, Indonesia
2. Mukerji K.G., Manoharachary C., Chamola B.P., 2001. Techniques in
mycorrhiza studies. Kluwer Academic Publishers.
51
3. Sharp W.R., Sondah M.R., Caldas L.S and Maraffo S.B.,1980. The phisiology of
in vitro asexual embryogenesis. Hortie
4. Fujimura T. and Komamine A., 1979. Synchronization of somatic embryogenesis
in carrot cell suspension culture. Plant physiol
5. Redenbaugh K. et al., 1986. Somatic seed: encapsulation of asexual plant
embryo. Biotechnology
*Ti liu t internet
http://www.academicjournals.org/AJB
http://www.jstor.org/jstor/gifcvtdir/di001521/0028646x/di011905/01p03843_1.1.gif
?config
http://helios.bto.ed.ac.uk/bto/microbes/mycorrh.htm
http://botit.botany.wisc.edu:16080/courses/mpp/OrchidMyco116.htm
http://www.esf.edu/resorg/rooseveltwildlife/Research/Orchid/Fam/Family.htm
52
PH LC
Th nghim 1:T l sng st ca ht nhn to sau 1 tun
One-Way Analysis of Variance
-------------------------------------------------------------------------------Data: tn1.ss
Level codes: tn1.t
Labels:
Means plot: LSD
Confidence level: 95
Range test: LSD
Analysis of variance
-----------------------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level
-----------------------------------------------------------------------------------------Between groups
1672.8675
3 557.62249
3.230 .0442
Within groups
3452.8019 20 172.64009
------------------------------------------------------------------------------------------Total (corrected)
5125.6693 23
0 missing value(s) have been excluded.
Tests for Homogeneity of Variances
---------------------------------- ---------------------------------------------------------Cochran's C test: 0.528615 P = 0.123637
Bartlett's test: B = 1.67499 P(9.52264) = 0.0230917
Hartley's test: 21.2296
Multiple range analysis for tn1.ss by tn1.t
-----------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
Average Homogeneous Groups
-----------------------------------------------------------------------------------------4
6 60.656667 X
3
6 62.676667 X
2
6 75.410000 X X
1
6 80.430000
X
----------------------------------------------------------------------------------------contrast
difference +/- limits
2-3
12.7333
15.8279
2-4
14.7533
15.8279
2-1
-5.02000
15.8279
3-4
2.02000
15.8279
3-1
-17.7533
15.8279 *
4-1
-19.7733
15.8279 *
----------------------------------------------------------------------------------------* denotes a statistically significant difference.
53

-------------------------------------------------------------------------------Data: Tn1.ss
Level codes: tn1.t
Labels:
Means plot: LSD
Range test: LSD
----------------------------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level
----------------------------------------------------------------------------------------------Between groups
1649.8414
3 549.94713
2.312
.1069
Within groups
4756.3131 20 237.81565
----------------------------------------------------------------------------------------------Total (corrected)
6406.1544 23
---------------------------------------------------------------------------------------------Cochran's C test: 0.566161 P = 0.0720209
-------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
-------------------------------------------------------------------------------3
6 58.020000 X
4
6 58.983333 X
2
6 70.250000 X X
1
6 78.016667
X
-------------------------------------------------------------------------------contrast
2-3
12.2300
18.5768
2-4
11.2667
18.5768
2-1
-7.76667
18.5768
3-4
-0.96333
18.5768
3-1
-19.9967
18.5768 *
4-1
-19.0333
18.5768 *
-------------------------------------------------------------------------------* denotes a statistically significant difference.
54

------------------------------------------------------------------------------------Data: Tn1.ss
Level codes: Tn1.t
Labels:
Means plot: LSD
Range test: LSD
---------------------------------------------------------------------------------------------Between groups
1490.3449
3 496.78165
3.882
.0245
Within groups
2559.2726 20 127.96363
---------------------------------------------------------------------------------------------Total (corrected)
4049.6176 23
---------------------------------Cochran's C test: 0.644031 P = 0.0191185
----------------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
----------------------------------------------------------------------------------------------3
6 55.280000 X
4
6 55.586667 X
2
6 70.725000
X
1
6 71.633333
X
----------------------------------------------------------------------------------------------contrast
2-3
15.4450
13.6268 *
2-4
15.1383
13.6268 *
2-1
-0.90833
13.6268
3-4
-0.30667
13.6268
3-1
-16.3533
13.6268 *
4-1
-16.0467
13.6268 *
----------------------------------------------------------------------------------------------* denotes a statistically significant difference.
55
Th nghim 1: T l ny mm sau 3 tun

---------------------------------------------------------------------------------------Data: Tn1.nm
Level codes: Tn1.t
Labels:
Means plot: LSD
Range test: LSD
----------------------------------------------------------------------------------------------Between groups
.2740917
3
.0913639 22.559
.0003
Within groups
.0324000
8
.0040500
----------------------------------------------------------------------------------------------Total (corrected)
.3064917 11
---------------------------------Cochran's C test: 0.598765 P = 0.258378
Multiple range analysis for Tn1.nm by tn1.t
Level Count
---------------------------------------------------------------------------------------------4
3
.2633333 X
3
3
.3066667 X
2
3
.5566667
X
1
3
.6100000
X
----------------------------------------------------------------------------------------------contrast
2-3
0.25000
0.11986 *
2-4
0.29333
0.11986 *
2-1
-0.05333
0.11986
3-4
0.04333
0.11986
3-1
-0.30333
0.11986 *
4-1
-0.34667
0.11986 *
56

-------------------------------------------------------------------------------Data: TN12.nm
Level codes: TN12.t
Labels:
Means plot: LSD
Range test: LSD
-------------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level
-------------------------------------------------------------------------------Between groups
.2380639
3
.0793546
5.890 .0201
Within groups
.1077792
8
.0134724
-------------------------------------------------------------------------------Total (corrected)
.3458431 11
---------------------------------Cochran's C test: 0.423889 P = 0.764855
Multiple range analysis for TN12.nm by TN12.t
Level Count
-------------------------------------------------------------------------------3
3
.3968254 X
4
3
.4262372 X
1
3
.6641026
X
2
3
.7160062
X
-------------------------------------------------------------------------------contrast
1-2
-0.05190
0.21860
1-3
0.26728
0.21860 *
1-4
0.23787
0.21860 *
2-3
0.31918
0.21860 *
2-4
0.28977
0.21860 *
3-4
-0.02941
0.21860
57

----------------------------------------------------------------------------------------Data: TN2.ss
Level codes: tn2.t
Labels:
Means plot: LSD
Range test: LSD
----------------------------------------------------------------------------------------------Between groups
144.6845
3
48.22816
.100 .9578
Within groups
3863.6698
8 482.95872
---------------------------------------------------------------------------------------------Total (corrected)
4008.3543 11
----------------------------------------------------------------------------------------------Cochran's C test: 0.361364 P = 1
Multiple range analysis for TN2.ss by tn2.t
---------------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
---------------------------------------------------------------------------------------------4
3 69.486667 X
2
3 75.213333 X
3
3 76.106667 X
1
3 79.070000 X
---------------------------------------------------------------------------------------------contrast
2-3
-0.89333
41.3896
2-4
5.72667
41.3896
2-1
-3.85667
41.3896
3-4
6.62000
41.3896
3-1
-2.96333
41.3896
4-1
-9.58333
41.3896
---------------------------------------------------------------------------------------------* denotes a statistically significant difference.
58

-------------------------------------------------------------------------------------------Data: tn2.ss
Level codes: tn2.t
Labels:
Means plot: LSD
Range test: LSD
---------------------------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level
---------------------------------------------------------------------------------------------Between groups
68.09663
3 22.698875
.317
.8131
Within groups
573.17567
8 71.646958
--------------------------------------------------------------------------------------------Total (corrected)
641.27229 11
---------------------------------Cochran's C test: 0.503208 P = 0.490437
-------------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
--------------------------------------------------------------------------------------------4
3 57.703333 X
2
3 59.106667 X
1
3 59.696667 X
3
3 64.076667 X
------------------------------------------------------------------------------------------contrast
2-3
-4.97000
15.9417
2-4
1.40333
15.9417
21
-0.59000
15.9417
3-4
6.37333
15.9417
3-1
4.38000
15.9417
4-1
-1.99333
15.9417
------------------------------------------------------------------------------------------* denotes a statistically significant difference.
59

--------------------------------------------------------------------------------------------Data: tn2.ss
Level codes: tn2.t
Labels:
Means plot: LSD
Range test: LSD
--------------------------------------------------------------------------------------------Source of variation Sum of Squares d.f. Mean square F-ratio Sig. level
--------------------------------------------------------------------------------------------Between groups
308.28820
3 102.76273
6.689
.0143
Within groups
122.90087
8
15.36261
----------------------------------------------------------------------------------------------Total (corrected)
431.18907 11
---------------------------------- ----------------------------------------------------------Cochran's C test: 0.834815 P = 0.0180291
-------------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
--------------------------------------------------------------------------------------------3
3 36.123333 X
4
3 37.266667 X
1
3 44.843333
X
2
3 48.193333
X
---------------------------------------------------------------------------------------------contrast
2-3
12.0700
7.38191 *
2-4
10.9267
7.38191 *
2-1
3.35000
7.38191
3-4
-1.14333
7.38191
3-1
-8.72000
7.38191 *
4-1
-7.57667
7.38191 *
60

--------------------------------------------------------------------------------------------Data: tn2.nm
Level codes: tn2.t
Labels:
Means plot: LSD
Range test: LSD
---------------------------------------------------------------------------------------------Between groups
.2080917
3
.0693639
8.266
.0078
Within groups
.0671333
8
.0083917
-------------------------------------------------------------------------------------------Total (corrected)
.2752250 11
-------------------------------------------------------------------------------------------Cochran's C test: 0.480636 P = 0.560372
Multiple range analysis for tn2.nm by tn2.t
--------------------------------------------------------------------------------------------Method: 95 Percent LSD
Level Count
--------------------------------------------------------------------------------------------4
3
.3900000 X
3
3
.4000000 X
1
3
.5233333 X
2
3
.7166667
X
---------------------------------------------------------------------------------------------contrast
2-3
0.31667
0.17253 *
2-4
0.32667
0.17253 *
2-1
0.19333
0.17253 *
3-4
0.01000
0.17253
3-1
-0.12333
0.17253
4-1
-0.13333
0.17253
61

-------------------------------------------------------------------------------Data: TN22.nm
Level codes: TN22.t
Labels:
Means plot: LSD
Range test: LSD
-------------------------------------------------------------------------------Between groups
.1024765
3
.0341588
5.100 .0291
Within groups
.0535860
8
.0066982
-------------------------------------------------------------------------------Total (corrected)
.1560625 11
---------------------------------Cochran's C test: 0.335218 P = 1
Level Count
-------------------------------------------------------------------------------4
3
.4777778 X
3
3
.5039683 X
1
3
.5379915 X
2
3
.7142191 X
-------------------------------------------------------------------------------contrast
1-2
-0.17623
0.15414 *
1-3
0.03402
0.15414
1-4
0.06021
0.15414
2-3
0.21025
0.15414 *
2-4
0.23644
0.15414 *
3-4
0.02619
0.15414
62
Th nghim 3: T l ny mm ca ht nhn to sau khi bo qun

-------------------------------------------------------------------------------Data: TN32.nm
Level codes: TN32.t
Labels:
Means plot: LSD
Range test: LSD
-------------------------------------------------------------------------------Between groups
.7630944
5
.1526189 22.154 .0000
Within groups
.0826667 12
.0068889
-------------------------------------------------------------------------------Total (corrected)
.8457611 17
---------------------------------Cochran's C test: 0.51371 P = 0.163166
Bartlett's test: B = 260862 P(125.298) = 0
Hartley's test: 7.34998E31
Level Count
-------------------------------------------------------------------------------4
3
.1000000 X
2
3
.1533333 X
1
3
.1866667 X
3
3
.2000000 X
5
3
.2466667 X
6
3
.7166667 X
-------------------------------------------------------------------------------contrast
2-3
-0.04667
0.14769
2-4
0.05333
0.14769
2-5
-0.09333
0.14769
2-6
-0.56333
0.14769 *
2-1
-0.03333
0.14769
3-4
0.10000
0.14769
3-5
-0.04667
0.14769
* denotes a statistically significant difference.
63
Th nghim 4: T l ny mm ca ht nhn to tr n cc gi th
---------------------------------------------------------------------------------------------Data: tn4.nm
Level codes: tn4.t
Labels:
Means plot: LSD
Range test: LSD
---------------------------------------------------------------------------------------------Between groups
1847.6009
2 923.80043
6.408 .0324
Within groups
865.0445
6 144.17409
----------------------------------------------------------------------------------------------Total (corrected)
2712.6454
8
----------------------------------------------------------------------------------------------Cochran's C test: 0.895096 P = 0.0330148
Multiple range analysis for tn4.nm by tn4.t
Level Count
---------------------------------------------------------------------------------------------1
3 53.653333 X
2
3 68.636667 XX
3
3 88.630000
X
--------------------------------------------------------------------------------------------contrast
difference limits
2-3
-19.9933
23.9965
2-1
14.9833
23.9965
3-1
34.9767
23.9965 *
--------------------------------------------------------------------------------------------* denotes a statistically significant difference.
64
Bng s liu
Th nghim 1:T l ht nhn to sng st cha bin i arcsin
alginate
20 g/l
30g/l
40 g/l
50g/l
1 tun
90
82
75
89
1 tun
100
100
75
76
1 tun
88
77
86
60
2 tun
89
73
73
89
2 tun
100
100
62
68
2 tun
80
63
80
60
3 tun
85
73
64
80
3 tun
99
100
57
65
3 tun
80
60
80
58
Th nghim 1:T l ny mm cc nng alginate sau 3 tun

20 g/l
30 g/l
40 g/l
50 g/l
ln 1
0,69
0,56
0,27
0,20
ln 2
0,64
0,58
0,36
0,27
ln 3
0,50
0,53
0,29
0,32
Th nghim 1:T l ny mm cc nng alginate sau 4 tun

20 g/l
30 g/l
40 g/l
50 g/l
Ln 1
0,70
0,77
0,33
0,33
Ln 2
0,69
0,83
0,50
0,59
Ln 3
0,60
0,55
0,36
0,36
65
Th nghim 2: T l sng st cc mi trng dinh dng cha bin i

arsin
MS
1/2 MS
1/4 MS
1/8 MS
1 tun
0,83
1,00
1,00
0,65
1 tun
1,00
0,83
0,80
0,67
1 tun
0,90
0,75
0,82
1,00
2 tun
0,91
0,70
0,79
0,90
2 tun
0,65
0,69
0,78
0,60
2 tun
0,64
0,73
0,86
0,60
3 tun
0,45
0,60
0,33
0,38
3 tun
0,40
0,57
0,37
0,36
3 tun
0,64
0,50
0,34
0,36
Th nghim 2: T l ht nhn to ny mm sau 3 tun

MS
1/2 MS 1/4 MS
1/8 MS
ln 1
0,45
0,8
0,33
0,4
ln 2
0,67
0,73
0,5
0,35
ln 3
0,45
0,62
0,37
0,42
Th nghim 2: T l ht nhn to ny mm sau 4 tun

MS
1/2 MS
1/4 MS
1/8 MS
Ln 1
0,48
0,73
0,43
0,45
Ln 2
0,54
0,80
0,58
0,58
Ln 3
0,59
0,62
0,50
0,40
66
Th nghim 3: T l ht nhn to ny mm sau khi bo qun

Khng dinh
Nc
Nc
dng
ct
ng
Ln 1
0,24
0,18
0,3
0,1
0,11
0,7
Ln 2
0,22
0,08
0,2
0,1
0,4
0,73
Ln 3
0,1
0,2
0,1
0,1
0,23
0,72
Agar
Agar ng
MS
Th nghim 4: T l ht nhn to ny mm cha bin i arcsin

MS
Cu giy
Bng gn
0,60
0,87
1,00
0,73
0,80
0,83
0,62
0,92
1,00
67

Nội Dung 1: Nghiên Cứu Kỹ Thuật Tạo Hạt Nhân Tạo Từ Phôi Cây Lan Vanda Nội Dung 2: Khảo Sát Sự Hiện Diện Của Mycorrhiza Trên Một Số Giống Lan

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Nội Dung 1: Nghiên Cứu Kỹ Thuật Tạo Hạt Nhân Tạo Từ Phôi Cây Lan Vanda Nội Dung 2: Khảo Sát Sự Hiện Diện Của Mycorrhiza Trên Một Số Giống Lan

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B GIO DC V O TO

TRNG I HC NNG LM THNH PH H CH MINH

NI DUNG 1: NGHIN CU K THUT TO HT NHN

MYCORRHIZA TRN MT S GING LAN

Ngnh hc: CNG NGH SINH HC

NI DUNG 1: NGHIN CU K THUT TO HT NHN

MYCORRHIZA TRN MT S GING LAN

Gio vin hng dn

Sinh vin thc hin

Nguyn Hong Qun

Th nghim 3: Kho st mi trng bo qun ht nhn to

Supervisor: TRAN THI DUNG, ph.D

Resulting in MS nutritional environment gave ratio of life and germination

The hyphae scrolled in one cell.

2.2 Phng php nui cy m lan ...10

3.4.1 Trang thit b v dng c nghin cu.

3.4.2 Mu s dng v iu kin nui cy.. 27

3.5 Phng php nghin cu ............................................................................. 28

4 KT QU V THO LUN ....................................................................... 33

4.1.4 Th nghim 4: Kho st kh nng ny mm ca ht.

4.2 Ni dung 2: Kho st s hin din ca mycorrhiza cc ging lan........... 45

DANH SCH CC BNG

DANH SCH CC BIU

DANH SCH CC HNH

thi, y mnh lai to ging chn ra nhng ging mi l, kh nng chu ng vi

Vit Nam, nm 1789 Joanis Loureiro, nh truyn gio ngi B o Nha ln

-Th trng tiu th cn nhiu kh khn, cha n nh kinh t lu di cho

b) S pht trin cy n thn

Hnh 2-1: Cu to hoa lan

- S t th phn: Khi phn hoa hoa ny c ri vo num ca chnh hoa

Rhizoctonia repens gip ht lan Cattleya, Laelia ny mm.

Rhizoctonia mucoroides gip ht lan Vanda, Phalaenopsis ny mm.

Rhizoctonia lanugiosa gip ht lan Oncidium, Miltonia ny mm.

thch c b sung dinh dng (thm 2% ng )

2.1.6.2 Tch chit

Phalaenopsis c lc cy con mc trn cng pht hoa n khi c r tt th tch

2.2 Phng php nui cy m lan

thiu s trao i kh bn trong chai v bn ngoi chai. Do ngun carbon phi

2.3 Cng ngh to ht nhn to

Hnh 2-2: Cu to ht nhn to

Hnh 2-3: Cc giai on pht sinh phi soma

Hnh 2-4: Cng thc ho hc ca alginate

(Aoyagi v csv, 1998).

Hnh 2-5: Cc dng phi soma

Dung dch sodium alginate

2.4 Nm r cng sinh- mycorrhiza

2.4.2 Cc dng mycorrhiza

2.4.3.4 Nng cao sc chng chu ca cy

3.2 Thi gian thc hin

3.3 Ni dung nghin cu

Ni dung 2: Kho st s hin din ca mycorrhiza trn mt s ging lan

3.4 Vt liu nghin cu

3.4.2 Mu s dng v iu kin nui cy

3.4.3 Mi trng s dng

3.5 Phng php nghin cu

Nghim thc Mi trng

Khng c dinh dng

Mi trng agar ng: Nc ct c 8,6 g/l agar v b sung thm 30 g/l ng

hp trong ni autoclave 1210 C-3

Ngm r trong dung dch Alkaline H2O2 t 10 30 (tu theo mu

A: Phi hnh tim

D: Phi hnh thu li

Hnh 4-2: Cc dng phi v tnh