Ảnh Hưởng Của Môi Trường Và Giá Thể Mô Rễ Đến Khả Năng Nhân Sinh Khối Cộng Sinh Nấm Rễ AM (Arbuscular Mycorhiza) in Vitro

VIN KHOA HC V CNG NGH VIT NAM
VIN SINH THI V TI NGUYN SINH VT
NGUYN TH GIANG
NGHIN CU
NH HNG CA MI TRNG V GI TH M R N
KH NNG NHN SINH KHI CNG SINH NM R AM
(ARBUSCULAR MYCORRHIZA)IN VITRO
Chuyn ngnh: VI SINH VT HC

M s: 60 42 40
LUN VN THC S SINH HC
Ngi hng dn khoa hc: TS.L QUC HUY
H Ni Nm 2012
S ha bi Trung tm Hc liu i hc Thi Nguyn
http://www.lrc-tnu.edu.vn
LI CM N
Trong sut qu trnh hc tp, nghin cu v hon thnh lun vn, ti nhn
c nhiu s gip ca cc thy c, cc anh ch v gia nh.
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ngh sinh hc Lm nghip, Vin Khoa hc Lm nghip Vit Nam, ngi tn
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Ti xin by t lng bit n su sc n tp th cn b, gio vin b mn Vi
sinh vt, Vin Sinh thi v Ti nguyn Sinh vt, nhng ngi Thy gip ,
ng vin ti trong sut qu trnh hc tp, to mi thun li cho ti trong qu trnh
thc hin v hon thnh lun vn ny.
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to iu kin thun li, hng dn,gip ti thc hin lun vn ny.
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tm Cng ngh sinh hc Lm nghip - Vin Khoa hc Lm nghip Vit Nam
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H Ni, ngy 15 thng 11 nm 2012
Tc gi lun vn
Nguyn Th Giang
LI CAM OAN
Ti xin cam oan y l cng trnh nghin cu khoa hc ca ti.
Cc s liu v kt qu trong lun vn l trung thc v cha tng c ai
cng b trong bt k cng trnh nghin cu no khc./.
Tc gi lun vn
Nguyn Th Giang
MC LC
M U.
1.1. t vn .. 2
1.2. Mc tiu ti... 2
1.2.1. Mc tiu chung....
1.2.2. Mc tiu c th........
1.3. ngha khoa hc v thc tin ca ti..
1.3.1. ngha khoa hc.
1.3.1. ngha thc tin.
1.4. Phm vi nghin cu... 2

Chng 1. TNG QUAN VN NGHIN CU... 3
1.1. Tng quan v nm r ni cng sinh AM... 3
1.1.1. Khi nim.
1.1.2. c im ca Nm r ni cng sinh AM(Arbuscular mycorrhiza).
1.1.3. Vai tr ca nm r ni cng sinh vi cy ch..
1.2. Tng quan v vi khun Agrobacterium rhizogense
12
1.3. Nghin cu nm r ni cng sinh trn Th gii v Vit Nam..
13
1.3.1. Trn th gii ...
13
1.3.2. Trong nc
19
Chng 2. VT LIU - NI DUNG - PHNG PHP NGHIN CU 22

2.1. Vt liu nghin cu..
22
2.2. Ni dung nghin cu.... 23

2.2.1. Nghin cu to vt liu gi th m r in vitro..
23
2.2.2. nh gi nh hng ca mi trng nui cy n kh nng nhn sinh

khi cng sinh nm r AM in vitro...
23
2.2.3. nh gi nh hng ca pH mi trng nui cy n kh nng nhn

sinh khi cng sinh nm r AM in vitro
23
2.2.4. nh gi nh hng ca gi th m r n kh nng nhn sinh khi

cng sinh nm r AM in vitro..
2.3. Phng php nghin cu....
23
24
2.3.1. Phng php b tr th nghim...
24
2.3.2. Phng php to vt liu m r in vitro 24

2.3.3. Phng php cy chuyn v nhn sinh khi m r
28
2.3.4. Phng php to cng sinh AM in vitro
28
2.3.5. Phng php nhn sinh khi cng sinh AM in vitro ..
29
2.3.6. Phng php thu thp, phn tch v x l thng k s liu th

nghim.
29
Chng 3. KT QU V BN LUN 31
3.1. Kt qu to vt liu gi th m rin vitro ..
31
3.2.nh gi nh hng ca mi trng nui cy n nhn sinh khi cng

sinh AM in vitro..
32
3.3. nh gi nh hng ca pH mi trng nui cy n nhn sinh khi cng

sinh AM in vitro..
38
3.4. nh gi nh hng ca gi th m r n nhn sinh khi cng sinh AM

in vitro.
44
Chng 4. KT LUN - TN TI - KIN NGH..... 50

4.1. Kt lun 50
4.2. Tn ti v kin ngh.......... 51
TI LIU THAM KHO.............................................
52
PH LC........................................................................ 58
BNG NHNG T VIT TT TRONG LUN VN
STT
Vit tt
Vit y
AM
Arbuscular mycorrhiza
EM
Ectomycorrhiza
IBA
Indole butylic acid
IP
Infective propagules
Minimal medium
MS
Murashige and Skoog medium
MSR
Strullu and Romand medium
PCR
Polymerase chain reaction
Ri-tDNA
Root inducing transfer Deoxyribonucleic acid
10
rRNA
Ribosomal Ribonucleic acid
11
TY
trypton-yeast extract medium
12
VAM
Vesicular arbuscular mycorrhiza
13
VM
Vesicular mycorrhiza
DANH MC BNG
Bng 3.1: nh hng ca mi trng nui cy n nhn sinh khi cng sinh
AM in vitrogia chng 41833 vi gi th m r C rt chuyn gen RitDNA
32
AM in vitrogia chng M7 vi gi th m r C rt chuyn gen RitDNA.
34
AM in vitro gia chng 41833 vi gi th m r Medicago chuyn gen RitDNA........
35
AM in vitrogia chng M7 vi gi th m r Medicago chuyn gen RitDNA.
36
Bng 3.5: nh hng ca pH mi trng nui cy n nhn sinh khi cng

sinh AM in vitrogia chng 41833 vi gi th m r C rt chuyn gen RitDNA
38

sinh AM in vitrogia chng M7 vi gi th m r C rt chuyn gen RitDNA
40
Bng 3.7:nh hng ca pH mi trng nui cy n nhn sinh khi cng

sinh AM in vitro gia chng 41833 vi gi th m r Medicago chuyn gen RitDNA..
41

sinh AM in vitrogia chng M7 vi gi th m r Medicago chuyn gen RitDNA.........
42
Bng 3.9: nh hng ca cc loi gi th m r khc nhau n nhn sinh

khi cng sinh AM in vitro trn chng 41833..
45
Bng 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh

khi cng sinh AM in vitro trn chng M7..
48
DANH MC BIU
Biu 3.1: nh hng ca mi trng nui cy n nhn sinh khi cng
sinh AM in vitrogia chng 41833 vi gi th m r C rt chuyn gen RitDNA.
33

sinh AM in vitrogia chng M7 vi gi th m r C rt chuyn gen RitDNA.
35
Biu 3.3: nh hng ca mi trng nui cy n nhn sinh khi cng sinh AM
in vitro gia chng 41833 vi gi th m r Medicago chuyn gen RitDNA.
36

sinh AM in vitrogia chng M7 vi gi th m r Medicago chuyn gen RitDNA.
37
Biu 3.5: nh hng ca pH mi trng nui cy n nhn sinh khi

cng sinh AM in vitro gia chng 41833 vi gi th m r C rt chuyn gen
Ri-tDNA
39
Biu 3.6: nh hng ca pH mi trng nui cy n nhn sinh khi cng

sinh AM in vitrogia chng M7 vi gi th m r C rt chuyn gen RitDNA
41
Biu 3.7:nh hng ca pH mi trng nui cy n nhn sinh khi cng sinh
AM in vitro gia chng 41833 vi gi th m r Medicago chuyn gen Ri-tDNA. 42
Biu 3.8: nh hng ca pH mi trng nui cy n nhn sinh khi cng
sinh AM in vitrogia chng M7 vi gi th m r Medicago chuyn gen RitDNA.
43
Biu 3.9: Kt qu nhn sinh khi AM in vitro ca 41833-C rt Ri-tDNA,

M7-C rt Ri-tDNA, 41833-Medicago Ri-tDNA, M7-Medicago Ri-tDNA
trn mi trng MSR 0,5% agar, pH 5,5..
44
Biu 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh

khi cng sinh AM in vitro trn chng 41833..
Biu 3.11: Kt qu nhn sinh khi AM in vitro ca 4 loi gi th m r
46
cng sinh vi chng 41833 trn mi trng MSR 0,5% agar, pH 5,5
47
Biu 3.12: nh hng ca cc loi gi th m r khc nhau n nhn sinh

khi cng sinh AM in vitro trn chng M7..
49
Biu 3.13: Kt qu nhn sinh khi AM in vitro ca 4 loi gi th m r

cng sinh vi chng M7 trn mi trng MSR 0,5% agar, pH 5,5.
49
DANH MC HNH
Hnh 1.1: Cy phn loi nm r ni cng sinh AM..........5
Hnh 1.2.a: Bi si nm (Arbuscules).....6
Hnh 1.2.b: Ti si nm (Vesicules)........................6
Hnh 1.3.a : Si nm ngoi bo(extraradical hyphae) .....7
Hnh 1.3.b : Bo t (spores) .........7
Hnh 1.4: S cu trc AM in hnh8
Hnh 1.5.a: Cy Medicago truncatula pht trin bnh thng............11
Hnh 1.5.b: Cy Medicago truncatula c cng sinh nm r.......11
Hnh 1.6: Cu trc vng Ri-plasmids ca vi khun A. rhizogenes(Veena and
Taylor 2007)....13
Hnh 2.1.a: Gieo ht Medicago...........24
Hnh 2.1.b: R Medicago pht trin sau 5 ngy.........24
Hnh 2.2.a: Ht C rt ny mm sau 4 ngy gieo ht.............................................25
Hnh 2.2.b: R C rt khng chuyn gen Ri-tDNA pht trin sau 30 ngy...........25
Hnh 3.1.a : R C rt khng c gen Ri-tDNA......................................................31
Hnh 3.1.b :R C rt c gen Ri-tDNA.................................................................31
Hnh 3.2. Phn tch PCR cho m r C rt chuyn gen Ri-tDNA v khng chuyn
gen Ri-tDNA. Bng 1: c gen rolB; bng 2: c gen rolC (cho mu chuyn gen);
bng 3 v 4: khng c gen rolB v rolC (cho mu khng chuyn gen); M: DNA
thang chun 100 bp (Fermentas).......................................................................32
Hnh 3.3.a: R Medicago khng c gen Ri-tDNA.................................................32
Hnh 3.3.b: R Medicago c gen Ri-tDNA...........................................................32
Hnh 3.4.a: R cng sinh pht trin trn mi trng MSR 0,5% agar...37
Hnh 3.4.b: R cng sinh pht trin trn mi trng MSR lng........37
Hnh 3.4.c: R cng sinh pht trin trn mi trng MS 0,5% agar......37
Hnh 3.5.a: AM cng sinh vo r C rt v sinh trng si nm mi.......45
Hnh 3.5.b: AM cng sinh vo r Medicago v sinh trng si nm mi.....45
Hnh 3.6.a: Sinh sn bo t AM trn gi th C rt c Ri-tDNA sau 1 thng.......50
Hnh 3.6.b: Sinh sn bo t AM trn gi th C rt c Ri-tDNA sau 4 thng.......50
M U
1.1. t vn
Ngy nay, nhn loi ang rt n lc trong vic gii quyt 3 vn ln, l
(i) Tng sinh trng v nng sut cy trng, m bo an ninh lng thc v nng
lng, (ii) Gim thiu thin tai, nhim mi trng v thch ng vi bin i kh
hu, (iii) Pht trin bn vng v nng cao cht lng cuc sng(AFCconference
2012).
Cc gii php sinh hc theo hng tip cn xanh (Green approach) c
nghin cu v hng ng p dng mnh m nhm lm tng nng sut cy trng,
vt nui, gim thiu thin tai, nhim mi trng v thch ng tt nht vi bin
i kh hu. Nghin cu pht trin ng dng cc ch phm sinh hc, vi sinh, dn
thay th cc loi sn phm ha hc cho tng nng sut cy trng v bo v mi
trng ang ngy cng c quan tm v u t pht trin.
Nm r ni cng sinh AM (Arbuscular mycorrhiza) c nghin cu s dng
nh mt loi phn bn sinh hc, mt mt c tc dng lm tng cng hp th dinh
dng ca cy trng, c bit l hp th Ln v gi nc trn nhng lp a thoi
ha, do lm tng sinh trng v nng sut, mt khc n cng c tc dng lm
n nh cu trc, c tnh sinh hc ca t v l yu t ch th cho mc suy
thoi ca mi trng t.
Tuy nhin, cc nghin cu ng dng nm r ni cng sinh AM mi ch tp
trung nhiu cho cc cy trng ngn ngy, cng ngh ch phm AM vn ph bin
p dng dng th s truyn thng l cht nhim t (soil innoculum), by thc
vt (AM trap plant), cha p ng c cc nhu cu i hi ca xn xut c v mt
s lng, cht lng sn phm, cng nh quy m v hiu qu ca vic p dng vo
sn xut. Do vy, hng i t ph mi trong nghin cu AM l cng ngh nhn
sinh khi AMinvitroc kh nng gp phn gii quyt c cc vn tn ti nu
trn ca cc loi ch phm AM truyn thng, trong mi trng nui cy v gi
th r thc vt ch l nhng yu t rt quan trng trong nghin cu v cng ngh
nhn sinh khi AM invitro.
Nhm gp phn gii quyt cc vn tn ti nu trn ca nghin cu ng

dng cng ngh AM, c bit trong lnh vc Lm nghip, ti nghin cu Thc
s Nghin cu nh hng ca mi trng v gi th m r n kh nng
nhn sinh khi cng sinh nm r AM (Arbuscular mycorhiza) in vitro c
xut thc hin. ti Thc s ny ca ti c thc hin trong khun kh ti
cp Nh nc v Nghin cu sn xut nm r ni cng sinh AM (Arbuscular
mycorrhiza) cho cy Lm nghipthuc Chng trnh trng im pht trin v
ng dng cng ngh sinh hc trong lnh vc nng nghip v pht trin nng thn
n nm 2020.
1.2. Mc tiu ca ti
1.2.1. Mc tiu chung
Nhm nghin cu mt s c s khoa hc cho cng ngh nhn sinh khi cng
sinh AM invitro v sn xut ch phm ng dng cho cy trng v bo v mi
trng.
1.2.2. Mc tiu c th
- Nhm nghin cu la chn c mi trng ph hp cho hnh thnh cng
sinh v nhn sinh khi cng sinh nm r AM in vitro.
- Nhm nghin cu la chn c gi th m r ph hp cho nhn sinh khi
cng sinh nm r AM in vitro.
1.3. ngha khoa hc v thc tin ca ti
1.3.1. ngha khoa hc
Kt qu nghin cu ca ti s cung cp cc c s khoa hc quan trng
cho cng ngh nhn sinh khi cng sinh AM invitro v sn xut ch phm ng
dng cho cy trng v bo v mi trng.
1.3.2. ngha thc tin
Kt qu nghin cu ca ti s xut c loi mi trng v gi th m
r ph hp nht cho cng ngh nhn sinh khi cng sinh AM in vitro, lm nguyn
liu sn xut ch phm AM phc v gy trng cy lm nghip.
1.4. Phm vi nghin cu
ti c tin hnh nghin cu trong phm vi phng th nghim.
Chng 1: TNG QUAN VN NGHIN CU

1.1. Tng quan v nm r ni cng sinh AM
1.1.1.Khi nim
Mycorrhiza l th cng sinh gia h si nm trong t vi r ca thc vt bc
cao.Frank l ngi u tin pht hin ra c im kt hp c bit ny r ca cy
Cupulifereae vo nm 1885 v gi l mycorrhiza. T mycorrhiza c ngha l
nm- r, tc gi dng t ny nhn mnh mi quan h gia nm v r cy
(Roger et al. 2004a).
Nm r ni cng sinh AM c xc nh l mi quan h khng th thiu
hu ht cc loi thc vt (hn 90% cc loi thc vt c kh nng hnh thnh cng
sinh AM). S kt hp mang li li ch cho c thc vt v vi sinh vt, qua ,
nm c c cc hp cht ng ha t thc vt sng, ng thi nm li gip r
cy tng cng kh nng hp th nc, cc cht hu c ha tan trong t c bit
l phospho, chng chu cc yu t bnh hi cng nh cc cht c kim loi nng.
Do , c tc dng ci to v n nh cu trc t, cn bng h sinh thi.Quan h
cng sinh ny c bit th hin vai tr trn nhng vng t kh cn, h sinh thi b
xo trn nghim trng, ngho dinh dng hay c tim nng c hi cao. V vy
cng ngh AM c kh nng p dng rng cho nhiu loi cy lm nghip, khng ch
gip to ra c nguyn liu cy trng rng c cht lng cao, kh nng thch nghi
v nng sut tt trn nhng lp a cn ci m cn p ng tt nht cho nhu cu s
dng hiu qu ngun ti nguyn t ai theo mc tiu m rng din tch cy trng
rng nhng khng cnh tranh vi t trng cy nng nghip, tng cng hiu qu
s dng cc vng t hoang ha theo cch bn vng v thn thin vi mi trng.
Mycorrhiza c phn b hu khp cc ni, thy cy c, ru, dng x, mt
s cy l kim, v hu ht cc cy l rng. S ph bin cng vi nhng vai tr tch
cc ca nm r kch thch vic nghin cu v mycorrhiza ngy cng m rng v
su sc hn. Trong khong 20 nm tr li y, nhng nghin cu c bn c thc
hin bi hng trm cc nh nghin cu t cc nc khc nhau trn th gii em
li nhiu kt qu ht sc ngha cho ng dng mycorrhiza trong h sinh thi nng
nghip,lm nghip v mi trng.
Da trn c im xm nhim ca nm vo r cy ch, mycorrhiza c phn

thnh 2 nhm chnh l ngoi cng sinh (Ectomycorrhiza, EM), v ni cng sinh
(Endomycorrhiza, AM).
Ectomycorrhiza:Ectomycorrhiza c nhng cy g ln, in hnh l thng,
si, cng l, nhng cy c gi tr kinh t cao, tuy nhin ectomycorrhiza c tnh c
trng loi. c im ca ectomycorrhiza l si nm ni bo ch xm nhp vo
khong gian bo ca cc t bo vng v r v si nm ngoi bo phn nhnh mnh
to thnh lp v bao quanh r nn lm bin i hnh thi bn ngoi ca r. Hu ht
ectomycorrhiza
thuc
LpBasidiomycetes
nh
Agaricales,
thuc
LpAscomycetes.
Endomycorrhiza:Hnh thnh khong 80% thc vt bc cao. c im ca
endomycorrhyza l si nm ca chng xm nhp vo bn trong t bo v r ca
thc vt bc cao v khng gy nn nhng bin i hnh thi bn ngoi ca r,
thng c mt phn ca si nm cn nm pha ngoi nhng chng khng to lp
v bao ngoi r. Cu trc in hnh ca endomycorrhiza l s hnh thnh nhng
cu trc c bit vesicules v arbuscules. mt s nhm endomycorrhiza ngi ta
quan st thy c vesicules(Vesicular mycorrhiza, VM) hoc arbuscules
(Arbuscular mycorrhiza, AM) hoc ng thi c hai cu trc ny trong t bo v r
(Vesicular arbuscular mycorrhiza, VAM).
Vy AMl th cng sinh gia nm vi r cy thc vt bc cao m hnh thnh
nn cu trc c bit vesicules, arbuscules trong t bo v r v khng gy bin i
hnh thi ngoi ca r.
Do tnh ph bin, c li v khng c hu cho 1 loi nn nhm
vesiculesarbuscular mycorrhiza rt c quan tm nghin cu ng dng trong
nng nghip cng nh trong lm nghip.
1.1.2. c im ca Nm r ni cng sinh AM (Arbuscular mycorrhiza)
a. Phn loi
Trong mt thi gian di AM c xp vo ngnh ph nm tip hp
(Zygomycota) do cu trc si nm khng c vch ngn, lp nm tip hp
(Zygomycetes). Hin nay, bng nhng nghin cu mc phn t h thng pht
sinh loi cho thy Zygomycota l ngnh a h (poli-phyletic), do nm AM

c tch ra khi ngnh Zygomycota hnh thnh ln ngnh mi l Glomeromycota.
Phn loi n cp h cho AM c da trn 4 tiu ch c bn:
- Cu trc mycorrhiza cng sinh trong r.
- Phng thc hnh thnh bo t khi c phn lp trong t.
- Cu trc ni bo t.
- Phng thc ny mm bo t.
H thng phn loi AM hin nay ( da trn trnh t ca rRNA ) c tm tt
trong hnh sau:
Hnh 1.1: Cy phn loi nm r ni cng sinh AM

(Ngun:http://www.google.com.vn/url?source=imglanding&ct=img&q=http://invam.caf.
wvu.edu/fungi/taxonomy)
N
b. Cu trc
Nm r ni cng sinh (AM) c cu to in hnh bao gm cu trc ni bo
(arbuscules, vesicules, si nm ni bo) v cu trc ngoi bo (si nm ngoi bo,
bo t).
Nhm cu trc ni bo:
Hinh 1.2: Bui si nm (Arbuscules) (a)

Tui si nm (Vesicules) (b)
Ngun:(H 2011)
- Arbuscules: l th gic mt, lng phn, dng nh lng bn chi, l phn

trao i dinh dng chnh gia thc vt ch v nm (Gianinazzi et al. 2002).Chng
c hnh thnh bn trong tbo v r(Mosse and Hepper 1975b) v l du hiu
cho bit c hot ng ca mycorrhiza. Ty vo tng loi khc nhau m arbuscules
cng c nhng c trng ring v hnh dng v s phn nhnh.
- Vesicules: c dng hnh cu hoc trng, c thnh t bo dy, l c quan d
tr dinh dng cho nm, c cha lipit v glycolipit (Mosse and Thompson 1981a).
N c to thnh bi on gia hay u li tn cng ca si nm ni bo, phn
b trong khong gian bo hoc bn trong t bo v r.
- Si nm ni bo: si nm ni bo khng c vch ngn,dng thng hoc phn
nhnh hnh ch H hoc Y,chng cng hnh thnh dng cun, tn s xut hin ca
chng ph thuc vo v tr trong r v c im ca tng loi nm (Morton2000).
Si nm va l phn cha cht d tr va l mt phn ca con ng vn chuyn
cc cht hp th bi cc si nm bn ngoi t t ti arbuscules hoc trc tip ti
t bo r ca cy ch (Bieleski 1973).
Nhm cu trc ngoi bo:
b
Hinh 1.3: a: Si nm ngoai bao (extraradical hyphae)
b: Bao t (spores)
Ngun: (H 2011)
- Si nm ngoi bo:Si nm ngoi bo khng c vch ngn, vai tr lm tng

r rt din tch hp th ca r cy (Bieleski 1973), cu si nm hnh thnh con
ng vn chuyn cht dinh dng gia thc vt cng sinh v khi t bm quanh
r (Koske and Gemma 1989). Si nm ngoi bo to ra ch c ng quan trng ca
h nm r (Jasper et al. 1989, 1991).
- Bo t:Bo t c th dng n hoc a bo, ch yu hnh thnh u ca
si sinh bo t ni tip vi si nm ngoi bo, i khi bo t cng xut hin bn
trong r (Koske et al. 1985), trn b mt t (BeCard and Fortin 1988), trn b mt
thc vt hay cc mnh phn gii (Blaszkowski et al. 1998). S lng bo t hnh
thnh ph thuc vo tng loi nm (Blaszkowski, 1993), loi cy ch v tnh a
dng ca n (Blaszkowski1993; Hetrick and Bloom1986), mu m ca t v
ch phn bn (Koske et al. 1989), c im vt hu ca cy ch (Giovannetti
and Avio 2002), cng nh sng (Daft and El Giahmi 1978), v kh nng cnh
tranh ca tng loi nm (Koske et al. 1989). Bo t c kch thc tng i ln
(50 500 m), ln hn nhiu so vi bo t ca nhng loi nm khc. Vai tr ca
bo t l pht tn n ni sng mi, v khi u qu trnh sinh trng khi c
tch ra t c th m. Do cim cu trc cc thnh phn cu to nn bo t n
nh trong nhng iu kin sinh thi khc nhau nn chng c coi l tiu ch quan
trng trong phn loi AM.
Hnh 1.4: S cu truc AM in hnh

(Bao gm arbuscules, vesicles, si nm ngoi bo v bo t)
(Ngun: http://mycorrhizas.info/vam/vamsoil2.gif)
c. Sinh trng
AM l th cng sinh bt buc. AM c th tn ti mt thi gian di trong t,
thm ch khi t b hn hay bng gi di dng cc mnh si nm trong cc r cht
hoc t do trong t. Tuy nhin, sinh trng c trong mt thi gian di AM
cn c cy ch thu nhn cacbon v nng lng cn thit. Do c im ny m
khng th tin hnh nui cy AM trc tip trn mi trng nhn to m cn phi
c gi th l r ca thc vt bc cao.
d. Chu trnh sinh sn v vng i
Khng c chng liu v s sinh sn hu tnh ca AM. Nghin cu bng ch
th phn t xc nh khng c s ti t hp hoc mc rt thp (Kuhn et al.
2001). V th thng gi nh rng bo t c hnh thnh bng sinh sn v tnh

bng bo t hoc si nm.
Phng thc nay mm cua bo t: Trong nhng iu kin nhit thi ch hp ,
bo t AM s ny mm trong t , trn r cy chu , hnh thnh si nm to mng
li h si phn b trong t va n su vao r cy chu . Bo t c th to ra h si
nm bn trong cung nh bn ngoai r.
Sinh san bng si nm: Nhiu loai AM co th nhn ln t manh si nm trong
t hoc trc tip t th cng sinh trn r cy
. c im ca nm r ni cng sinh AM
l th cng sinh bt buc vi r cy ch, do o nu khng co r cy chu cho cac si
nm nay mm va cng sinh thi sinh trng cua nm se bi ngng tr lai sau mt thi
gian va t bao cht co th co lai bn trong bao t
.
Phng thc nui cy: nui cy nm r ni cng sinh AM , c th s dng
hai phng thc nui cy , o la : nui cy in vitro v in vivo. i vi nui cy in
vivo, c th nui cy trong chu bng t hin trng c cha bo t hay si nm
(Bianciotto va cng s , 2000; Hijri va cng s , 2002). Cn i vi nui cy in
vitro, c th to ra mt s lng ln nm r thng qua nui cy m r trn mi
trng nui cy nhn tao (Fortin et al. 2002). c bit l trong nui cy m r, sinh
khi nm r tao ra thng khng cha tap cht va cac vi sinh vt khac nn phng
php ny c s dng nhiu.
1.1.3. Vai tr ca nm r ni cng sinh vi cy ch
Nm r l th sng cng sinh bt buc. Nhng hot ng ca nm cng nh
ca thc vt c vai tr h tr cho nhau. Nm s dng ngun cacbon t thc vt
di dng ng hexoses v cc vitamin. S vn chuyn cacbon t thc vt sang
nm c thc hin nh arbuscules hoc cc si nm ni bo. Ti cc si nm ni
bo din ra qu trnh bin i dinh dng th cp cung cp glycogen, pentose,
lipit cho hot ng ca nm (Turmel 2004). Gn 20% cacbon do thc vt tng
hp c chuyn sang nm v khong 25% cacbon ngun gc t thc vt c
nm bin i v d tr nhng si nm ngoi bo, vic ny gp phn lm tng
thm ngun hu c trong t.
Li ch ca AM i vi thc vt ch yu l tng cng ci thin hp th cc

cht dinh dng v nc, trong quan trng nht l tng cng hp th dinh
dng ln (P), hp th nc, chng chu vi cc yu t bt li ca mi trng, c
bit trn cc hin trng t ai cn ci, kh hn. Hot ng ny c tng cng
l do nm r ni cng sinh hnh thnh nhiu h si nm phn nhnh mnh to
thnh v s cu ni gia mi trng t vi cc t bo r, tng din tch tip xc
vi t, bin i mi trng quanh r lm cho cc cht tr nn linh ng v thc
vt c th hp th c (n 80% nhu cu v P v 25% nhu cu v N ca cy c
cung cp nh nm) (Turmel 2004). Si nm c th lan rng n 8cm quanh r v
hp th cht dinh dng vn chuyn li vo r, tng kh nng hp th cc cht dinh
dng cao hn so vi lng r 10 ln. Tc thm nhp ca P qua si nm cao gp
6 ln so vi qua cc lng r.
Khng nhng th, thc vt cn c hng li nh c tng cng kh
nng hp th nc v bo v r khi ngun bnh. AM c vai tr kch thch sinh
trng thc vt bng cch tit ra rt nhiu cc cht kch thch sinh trng nh
auxins, cytokinins, gibberellic acids v mt s cht khng sinh bo v cy ch
chng li cc mm bnh t trong t.
S cng sinh AM c th lm tng cng sinh trng ca cy con ln n
400%, gip cy c kh nng chng chu vi iu kin kh hn v ngho dinh
dng (Pope 2007). Thc t nhng loi g ln c cng sinh AM cho thy cy
vn tn ti v sinh trng trong iu kin m rt thp trong khi nhng cy
khng c cng sinh th khng sng c hoc sinh trng yu. Kt qu nghin cu
cng cho thy, sinh trng ca cy con nhng loi g ln c s cng sinh AM t
ngang vi cy khng c s cng sinh nhng chng ch s dng lng dinh dng
bng mt na. Nhng loi sng ph thuc vo AM ch yu l nhng ni t
cht, kh cn, c h r t phn nhnh v lc lng cht dinh dng tn ti nhiu
dng kh tiu, r cy khng th hp th c m phi nh n s lan rng ca h
si nm.
Hnh 1.5. Cy Medicago truncatula pht trin bnh thng (a)

Cy Medicago truncatula c cng sinh nm r (b)
Ngun:(Lan 2011)
Nm AM khng phi l mt thnh phn chnh trong t nhng c vai tr

quan trng trong s iu chnh hot tnh sinh hc ca t nh s phn b rng
khp ca h si nm AM trong lp t tng mt. Nm AM hp th trc tip hp
cht carbon do cy c nh v cu thnh u vo chnh ca carbon v nng lng
trong t, chng phn phi carbon ny khp c khu vc t quanh r cy cho vi
sinh vt t s dng. Lng cacbon ng k c vn chuyn bng h si nm t
cy ny sang cy khc c xc nh (Simard et al. 1997). iu ny gip gim
s cnh tranh gia cc loi khc nhau v gp phn vo tnh n nh v a dng ca
h sinh thi.
AM tham gia vo bin i mi quan h t- cy- nc, tng cng s thch
nghi ca thc vt vi cc iu kin bt li (kh hn, nhim kim loi). nhng ni
c hm lng kim loi nng cao, AM c vai tr kh c mi trng gip cy sinh
trng tt (Songul and Sevinc 2002).Bt u t ci thin hp th nc v cc cht
dinh dng quan trng cho s sinh trng ca cy s dn ti tng hiu sut quang
hp, s vn chuyn cht dinh dng v s trao i cht ca cy. Cy sinh trng
nhanh, gim bt vic s dng phn bn ha hc (i khi ln ti mt na lng d
kin) dn ti tng thu nhp cho nng dn. Kt qu th nghim phn bn sinh hc
mycorrhiza trn cy Asparagus chng minh c iu , khi nhng nng dn
s dng lng phn bn ha hc cng vi phn bn sinh hc mycorrhiza th nng
sut cy trng c ci thin hn 50% v thu nhp ca nng dn tng 61% so vi

khi s dng ch mi phn bn ha hc.
1.2. Tng quan v vi khun Agrobacterium rhizogenses
Vi khun Agrobacterium rhizogenses ln u tin c pht hin cch y
hn 70 nm (Riker et al. 1930, Hildebrand 1934, White 1972) nh mt yu t gy
nn bnh lng r (hairy-root disease) mt s thc vt. A. rhizogenses thuc
Chi Agrobacterium, l vi khun t, gram m, c hnh que. A. rhizogenses c quan
h gn gi vi vi khun A. tumefaciens, mt loi vi khun c bit nhiu n v
gy nn bnh u biu thc vt (crown gall disease) rt ph bin (Conn 1942).
C ch nhim thng qua qu trnh tip xc, vi khun A. rhizogenses chuyn
mt on T-DNA, trong c mang cc gen lin quan n to lng r(rol-genes),
gen lin quan ti qu trnh sinh tng hp v c cc gen cha xc nh c chc
nng trong vng Ri- Plasmid ca chng vo b gen ca t bo thc vt ch; cc
gen (rol-genes) trong T-DNA tng tc vi gen ca thc vt ch to ra lng r.
Cc biu hin gen c cha cc thng tin c m ha trong T-DNA ny thc
y qu trnh pht trin v sn xut ra lng r vi hu ht cc thc vt hai l mm
(Veena and Taylor 2007) (Hnh 1.6). Qu trnh chuyn cc gen (rol-genes) trong TDNA t vi khun A. rhizogenses sang thc vt ch c th c xy ra ngu nhin,
cng c th c thc hin bng cc k thut ng nui cy n gin vi cc
chng A. rhizogenses hoang di t nhin (Abdoulaye 2003, Chabaud et al. 2006),
v cng c th thc hin bng k thut bin np gen hin i (Pak et al. 2009, Park
et al. 2011).
Vi khun A. rhizogenses tip cn chuyn gen T-DNA sang thc vt ch
thng qua cc vt thng, cc hp cht phenolic tit ra t cc vt thng ca t
bo thc vt hp dn A. rhizogenses, sau qu trnh nhim l qu trnh hp nht
cc nguyn liu di truyn T-DNA ca A. rhizogenses vi h gen thc vt ch, v
kt qu to nn bnh lng r (Veena and Taylor 2007). V hnh thi hc, lng r
to ra do vi khun A. rhizogenes c hnh dng v cu trc rt ging vi cc loi r
thng thng, tuy nhin n c nhng tnh trng rt c trng, d dng nhn bit l:
tc r di, sinh khi r ln, phn nhnh nhiu, tc r pht trin nghing v khng
c tnh hng t (negative geotropism)
Bnh lng r t vi khun A. rhizogenses c nghin cu p dng rng ri
nh mt cng c hu hiu cho cc nghin cu trao i cht th cp, chc nng
gen, cng sinh vi khun c nh m rhizobium, nm r ni cng sinh AM
(Abuscular mycorhizae), v cc c tnh sinh hc r thc vt khc (Tsuro et al.
2005, Chabaud et al. 2006, Veena and Taylor 2007, Sidwa-Gorycka et al. 2009,
Park et al. 2011).
Hnh 1.6:Cu trc vng Ri-plasmids ca vi khun A. rhizogenes

(Veena and Taylor 2007)
1.3. Nghin cu nm r ni cng sinh trn Th gii v Vit Nam
1.3.1. Trn Th gii
Giai on u, cc nghin cu v AM trn th gii tp trung cc mt: a
dng sinh hc ca cc chng bn a, phn loi, chn lc cc chng hiu lc vi
tng i tng cy trng v t trng khc nhau, ng thi pht trin nhng
phng php nh lng v nhn nhanh chng. Phng php nhn sn xut cht
nhim AM truyn thng l s dng cc thc vt ch (trap plant) nhn AM trong

nh knh, vn m t h r ca chng trn cc gi th gy trng khc nhau (in
vivo). Bng phng php ny, cht nhim AM thu c cn c gi l soil
innoculum dng th, thng c sch AM khng cao, s lng, cht lng
AM (IP- infective propagules) v hiu qu p dng cn b hn ch. Vn t ra l
cn c mt cng ngh cao hn, hiu qu hn c th nhn sinh khi ln, sn xut
ch phm cht nhim AM thun khit, cht lng v c hiu qu p dng cao cho
tng sinh trng, cht lng sn phm cy trng v bo v mi trng t. Trong
cng ngh ny, chng ta cn phi ch ti mt c tnh quan trng ca nm r ni
cng sinh AM, l, trong bt k iu kin nui cy no, bn ngoi mi trng
t (in vivo) hay trong iu kin nui cy v trng (invitro), sinh khi ca chng
ch c th c nhn ln khi hnh thnh c cng sinh vi t bo v r ca cc
thc vt ch. Do , vic s dng mi trng dinh dng nhn to khng c gi
th m r khng p ng c nhu cu sinh trng, pht trin ca AM (Roger et
al. 2004b).
Nghin cu v AM thng xuyn b cn tr bi c tnh cng sinh bt buc
ca chng (Bago and Bcard 2002). Mt hng nghin cu mi l, nui cy in
vitro pht huy c rt nhiu li th, n cho php nghin cu c c bn
nhng c tnh sinh l ca s cng sinh nm-r. Nhng nghin cu th nghim
u tin c tin hnh, c th k n nh (Mosse and Hepper 1975a). Sau
c b sung bi (Fortin et al. 2002)(Declerck et al. 2005).
Trong nhng nm 1980, s pht trin cng ngh nui cy m r m ra
mt hng i mi, cho php nui cy thnh cng mt s chng AM trong iu
kin in vitro. S dng phng php ny, nm AM v r cy c nhn ln ng
thi v nhanh chng qua mt s ln cy chuyn, thi gian c rt ngn rt nhiu
v m bo c thun khit cao (Fortin et al. 1996b).
Hai loi mi trng c s dng thng xuyn nht trong nui cy AM in
vitro l mi trng M(BeCard and Fortin 1988) v mi trng MSR (Strullu and
Romand 1986, Declerck et al. 1998). C hai loi mi trng ny u c cha cc
nguyn t a lng, vi lng, vitamin v ng (Cranenbrouck et al. 2005). ng
thi, chng c th c lm rn bi hp cht Agargel hoc Phytagel. Nm 2006,

nhm nghin cu ca Gadkar v cng s (Gadkar et al. 2006) tin hnh nui cy
AM trn mi trng M lng bng phng php chia ngn. Theo , mt ngn ca a
petri nui cy nm c cung cp thm ng, ngn cn li khng c ng cng nh
vitamin. Ging nh trong phng php nui cy in vivo, vic cung cp y xi cho
mi trng lng l vic lm cn thit trong nui cy in vitro(Jolicoeur et al. 1999).
Tuy nhin, mt vi nghin cu ch ra rng, mi trng MSR (Strullu and
Romand 1986, Declerck et al. 1998) l mi trng c bin i thnh phn vi
mc ch ti u ha cho s pht trin ca nm r trong iu kin in vitro. Trong ,
thnh phn a lng ca MSR l tng t trong mi trng M (Minimal medium).
S khc nhau gia hai loi mi trng ny l thnh phn vitamin: MSR va thiu
it, myo-inositol v glycin cn trong thnh phn vitamin ca mi trng M thiu
panthotenate, biotin v cyanocobalamine. Tuy nhin, nghin cu cng ch ra rng,
s vng mt ca nhng thnh phn ny khng c hiu qu tiu cc r rt n s
cng sinh AM.
Trong nhng nm 2000, rt nhiu nghin cu v cng ngh nui cy AM in
vitro c tin hnh trn mi trng MSR (Strullu and Romand 1986, Declerck et
al. 1998) khng b sung ng v vitamin (Voets et al. 2005a)(De Boulois et al.
2006)v c lm rn bi Phytagel hoc Agargel. Nghin cu ch ra rng, vic
b sung ng v vitamin l khng cn thit trong h thng nui in vitro v thc
vt c kh nng t quang hp to ng v chuyn ha vitamin cn thit cho s
pht trin ca chng. Nghin cu ca (Declerck et al. 2009) cng s dng MSR m
khng c ng v vitamin.
Nm 1996, St-Arnaud v cng s (St-Arnaud et al. 1996) ln u tin s
dng phng php chia ngn. Trong phng php ny, a petri c chia thnh
hai ngn ring bit, mt ngn c cha gi th r, ngn cn li ch c AM pht trin.
S dng phng php phn chia a, (Douds 2002a) chng minh c rng, bo
t AM tip tc c hnh thnh sau khi c s b sung mi trng c ng t
ngn cha gi th r.
Vic nghin cu v AM c thc hin bi rt nhiu nh khoa hc, c th

k n nh (Chabot et al. 1992)(Declerck et al. 2001)(Douds 2002b)(St-Arnaud et
al. 1996). Gi th u tin c s dng cho nui cy ny l r C rt. Tuy nhin,
trong nhng nm gn y, nhiu loi gi th khc nhau c a vo s dng
thnh cng nh Chicory (Cichorium intybus L.) v Medicago(Medicago truncatula
Gaertn)(Boisson-Dernier et al. 2001)(Fontaine et al. 2004). Tuy nhin, vic s
dng cc loi gi th khc nhau c nh hng khc nhau n vic sn sinh bo t
AM (Tiwari and Adholeya 2003). Voets v cng s, 2005 (Voets et al. 2005b)
nghin cu s dng gi th l r cy khoai ty (Solanum tuberosum L.) v thu c
gn 12.000 bo t/ a petri sau 12 tun nui cy. Gn y, mt kt qu nghin cu
khc thu c gn 50.000 bo t/a petri khi cng sinh gia chng G.
intraradices (MUCL 41.833) vi r cy Medicago sau 14 tun nui cy. Mt s gi
th khc nh chui (Koffi et al. 2009) hay nho cng c kh nng cng sinh vi AM
nhng khng p ng c nhu cu p dng sn xut bo t trn quy m ln.
Nm 1996, Fortin v cng s (Fortin et al. 1996a) trnh by phng php
nui cy AM in vitro s dng 2 ngun nguyn liu l r c chuyn gen RitDNA hoc cha c chuyn gen. Phng php chn cy ch, k thut cng sinh
r AM, thnh phn mi trng nui cy. Nghin cu cng ng thi trnh by kh
nng nui cy v to bo t lin tc trong mi trng in vitro, phng php bo
qun bo t trong thi gian di. Nhng li ch khi nui cy AM in vitro, cho php
pht trin nghin cu su v rng hn nh hnh thi AM, phn loi v tm hiu c
ch cng sinh vi r.
Trong nghin cu nhn sinh khi r c mang Ri-plasmid ca vi khun
Agrobacterium rhizogenes, nhm tc gi (Mugnier and Mosse 1987) quan st
thy c s hnh thnh nhiu lng r trong mi trng nhn to v thc y c s
cho nhn sinh khi AM in vitro. Tc gi quan st thy s cng sinh hnh thnh
gia AM v r chuyn gen khi nui cy cng vi bo t Glomus mosseae. Tuy
nhin c s sinh trng gii hn v chu trnh sinh trng pht trin hon chnh
ca AM trong mi trng in vitro cha c hon thin. Trong nhiu nghin cu
sau , cc nhm tc gi nghin cu AM in vitro theo nhiu hng khc nhau
nhm tm ra iu kin thu hi c sinh khi AM cht lng nhiu nht trong
thi gian ngn nht. Cc nghin cu bao gm thnh phn mi trng nui cy,
phng thc cng sinh, phng thc nui cy, phng thc thu hi AM Mt
trong nhng kt qu ni bt c th k n l (Fortin et al. 1996b) thu hi c
30.000 bo t sau 6 tun nui cy AM in vitro qua r C rt c chuyn gen Ri trn
a petri v thu hi c 120.000 bo t AM sau 2 thng nui cy bng bioreator.
y l kt qu ht sc c ngha pht trin v hon thin cng ngh nhn
nhanh AM in vitro.
(Jolicoeur et al. 1999) cng b sn sut ch phm t loi Glomus
intraradices bng bioreactor. Bng phng php nui cy Glomus intraradices vi
r cy C rt c chuyn gen Ri-tDNA ca vi khun Agrobacterium rhizogenes
trn a petri v bioreactor cho kt qu tng sinh khi AM cc i t ~ 0,6g
sinh khi kh trong 1lt mi trng, khi pht trin qu gi tr sinh trng ca r
s b suy gim. Trong iu kin ti thch nui cy trn a petri, tc sinh trng
r ti a v bo t G. intraradices ln lt l 0,021 v 0,035/ngy. Nghin cu
bc u nhn sinh khi AM trong bioreactor lun chuyn kh cho kt qu thp
hn so vi nui cy trn a petri 10 ln v cng thy s hnh thnh cng sinh t
hn (0,13g sinh khi kh trong 1 lt mi trng). Kt qu nghin cu m ra kh
nng nhn nhanh bo t AM bng bioreactor.
Gn y, thng 1 nm 2009, nhm tc gi Venter, Marianne, Wilma bo
co kt qu nghin cu trnh by thng tin kh y v ton din v nhn sinh
khi AMin vitro,bao gm: phng php to cng sinh AM, nhn sinh khi bo t
AM trong mi trng in vitro, cc bc phn lp bo t t mi trng nui cy.
Nghin cu cng to ra ht bo t c nhiu c im u tri nh mt bo t
cao, c tnh n nh v phn tn tt trong mi trng t. Ht bo t c th p
dng trong sn xut nng lm nghip nh mt loi ch phm c th dng trc tip.
Nghin cu cng cp n mt s thnh phn c th b sung vo ht bo t lm
tng tc dng hin trng nh cht kch thch sinh trng, cc cht trao i th
cp.
Cng ngh nhn sinh khi AMinvitro v ch phm AM tinh khit ca Vin
Nng lng v Ti nguyn (TERI), n l mt bc t ph cng ngh quan

trng trong lch s nghin cu AM trn th gii, mang li c hi p dng ln
hn v hiu qu hn. Cng ngh AM in vitro c TERI chuyn giao p dng
ti 5 nc khc trn th gii, hiu qu cho gy trng nhiu loi cy nng lm
nghip, trong c Cc ro, c bit hiu qu cho cc vng sinh thi suy thoi
nghim trng nh vng ct kh cn, bi thi khai thc v vng nhim cht
thi(Adholeya and Singh 2006).
Trong nghin cu nm r ni cng sinh AM in vitro, m r C rt (Daucus
carrota L.) chuyn gen Ri-tDNA c s dng kh ph bin v hiu qu cho cc
nghin cu cng sinh AM in vitro, sn xut sinh khi AM invitro v sn xut ch
phm AM (Declerck et al. 1996, Douds Jr 1997, Abdoulaye 2003, Ijdo et al.
2011).
Mc d, hin nay, phng php nui cy AM in vitro vn cn mt vi hn
ch nh vic cn u t kh tn km v nhn lc cng nh trang thit b phc v
cho nghin cu nhng vi nhng u th ca n, c bit l vic c th kim sot
cht lng ch phm AM cng nh gim nguy c nhim th trong tng lai, y
s l mt hng i mi c kh nng p ng c tiu chun cht lng cho sn
xut thng mi hng lot.
Trong nhng thp k qua, c rt nhiu cng trnh nghin cu v AM theo
c chiu rng ln chiu su. Vi hn 40 bng sng ch lin quan n AM, trong
c nghin cu v nhng c tnh c li ca AM. Mt s nghin cu li tp trung
vo k thut nui cy (Sylvia and Jarstfer 1992), mt s khc nghin cu cc ng
dng ca n (Cano and Bago 2007)(Fernandez et al. 2006), v phng php nui
cy trn cc loi gi th khc nhau (Mosse and Thompson 1981b) hoc nui cy
trong iu kin in vitro(Declerck et al. 1996)(Cranenbrouck et al. 2005)(Declerck
et al. 2009)(Fortin et al. 1996b)(Mugnier et al. 1986)(Wang 2003).
Hin nay trn th gii, nghin cu pht trin cng ngh AM cho mc tiu
phc v cy trng nng lm nghip, lm tng sinh trng, nng sut v cht lng
sn phm cng nh phc v cc mc tiu lm sch mi trng, gp phn lm cn
bng n nh cc h sinh thi ang ngy cng c quan tm u t nhiu hn v
mi kha cnh, nghin cu chuyn su v m rng lnh vc p dng cng ngh v

sn phm. Nhiu trung tm nghin cu ln v AM trn th gii c thnh lp
tip tc pht trin v ng dng khng ch nhng sn phm AM m cn c nhng
gii php cng ngh cho thc tin sn xut, cc h sinh thi v mi trng sng.
Mt s trung tm ln c th k n nh INVAM (M), GINCO (Canada),
CESAMM (B), Mycorrhiza Association (c), TERI (n ), ACT (i Loan)
v.v.
1.3.2. Trong nc
ng dng nm r ngoi cng sinh Ectomycorrhiza cho sn xut cy con thng
nha c nghin cu bng vic s dng t mn rng thng nh mt loi cht
nhim t nhin (natural soil innoculum) v tuyn chn sn xut cht nhim nhn
to (Giao and Nhm 1988). Tuy nhin, quy m & kt qu p dng cn rt hn ch.
L Quc Huy vi nghin cu k thut mycorrhiza cho lm nghip (Vin
nghin cu nng lng TATA, New Delhi - n , 1999)(Huy 1999) a ra
mt hng i mi cho nghin cu mycorrhiza Vit Nam
Nghin cu Ectomycorrhiza cho Bch n v Phi lao (Phm Quang Thu, Vin
Khoa hc Lm nghip Vit Nam, (Nguyn Vn Mo, Trng i hc Lm nghip)
c nhng kt qu bc u ng dng th nghim vn m v rng trng .Sn
xut ch phm nm cng sinh a chng chc nng cho cy Lm nghip (Phm
Quang Thu, 2001- 2004). ti nghin cu sn xut ch phm nm cng sinh
cho Thng v Bch n di dng bt cha bo t hu tnh ca nm Pisolithus
tinctorius v mt s vi sinh vt chc nng. Hiu qu ca ch phm khi nhim cho
bch n PN2 trng trn cc lp a thoi ho cho thy c s khc bit r rt so vi
i chng.Nghin cu cng ngh sn xut ch phm vi sinh hn hp dng vin nn
cho bch n v thng trn cc lp a thoi ho, ngho cht dinh dng(Thu
2006-2010).
Ch phm nm r ngoi cng sinh (ECM) dng vin nang (gel) c
nghin cu v p dng th nghim cho i tng cy Sao en (Hopea odorata),
nhm tng sinh trng, sn xut cy con Sao en cht lng cao cung cp cho
trng rng. Kt qu p dng lm tng sinh trng cy con sao en ti vn m
ln 50-80% so vi i chng. Tuy nhin, vn tn ti cn c tip tc nghin

cu gii quyt i vi ch phm ECM dng ny l t l sng st ca cc si
nm ECM trong vin Gel gim nhanh chng sau mt thi gian bo qun, v lm
sao bo qun cho s dng dng ch phm ny trong mt thi gian di, ti thiu
l 6 thng cng c t ra (Chu and Huy 2007).
Cc nghin cu trn ch yu tp trung nm r ngoi cng sinh
Ectomycorrhiza. Dotnh cht c bit ca Nm r ni cng sinh Abuscular
mycorrhiza (AM), vic nghin cu AM cn c phng php tip cn khc; cc
chng AM phn lp tuyn chn t mt i tng cy ch ny c kh nng ng
dng hiu qu cho nhiu loi cy trng khc, nghin cu v ng dng AM trong
lm nghip hin nay ti Vit Nam l mt i hi ht sc cn thit.
ti Nghin cu, p dng k thut pht trin cng sinh mycorhiza cho mt
s cy trng chnh ti mt s vng sinh thi phc v sn xut nng nghip bn
vng Vit Nam (Vin Th Nhng Nng Ho 2004 -2006) cho thy vai tr
ca cng sinh nm r cy khng ch thc y sinh trng m cn nh hng tt
n nng sut cy trng nng v lm nghip.
Nghin cu s a dng nm cng sinh Arbuscular mycorrhiza c Vetiver t
t nhim ch(Chnh and Cng 2007).
Nghin cu phng php nui cy dung dch lng (MS ci tin) in vitro Keo
lai v Keo tai tng (hay gi l phng php nui cy trn cu giy) nhm lm
ny mm bo t AM v xc tin qu trnh hnh thnh cng sinh AM trong r Keo
c tin hnh thnh cng(Huy and Chu 2004). Kt qu phng php l mt
c s quan trng cho th nghim pht trin phng php nui cy nhn nhanh sinh
khi AM in vitro v Bioreactor ci tin cho sn sut ch phm AM.
Nhng kt qu bc u v phn lp, tuyn chon cc chng AM cho sn xut
th ch phm in vivo v nghin cu nh gi nh hng ca bn nhim mt s loi
ch phm AM ti sinh trng v nng sut qu ht ca cy Cc ro ti vn m
v rng trng rt kh quan (thc hin ti Trung tm CNSH Lm nghip, Vin
Khoa hc Lm nghip Vit Nam). Ti vn m, khi p dng bn nhim ch
phm AM 25mg/cy (200IP) t c phn ng sinh trng cao nht ca cy con
Cc ro v ng knh (D), chiu cao (H) v sinh khi (P) sau 24 tun th nghim,
tng hn so vi i chng tng ng cho cc ch tiu l 18,8%, 27,4% v 63% (V

Qu ng, 2009).
Tng t, kt qu nh gi mc cng sinh AM trong t bo r cy Cc ro
vn m vi cc loi ch phm AM v cng thc nhim khc nhau c
nghin cu tin hnh ti Trung tm CNSH Lm nghip: cng thc FM1 25mg/cy
(200IP), loi ch phm AM in vitro ca TERI cho thy mc cng sinh AM - r
cao nht (F: 96,67%, v A: 39,79%, cao hn so vi i chng (F: 83,33% v A:
16,62%) (V Qu ng, 2009).
Kt qu p dng bn nhim ch phm AM cho Cc ro trng rng ti vng ct
kh hn Ninh Phc, Ninh Thun sau 01 nm trng lm tng nng sut ht
trung bnh t 25-35% so vi i chng khng bn nhim(Huy et al. 2009).
Tuy nhin nhng nghin cu v nm r ni cng sinh AM mi ch khi u,
cha c nhng nghin cu chuyn su v h thng v cng ngh ch phm AM in
vitro v do kt qu v s ng dng cn rt hn ch.Pht trin nghin cu ng
dng cng ngh AM trong nng lm nghip, phc v gy trng rng, qun l phc
hi sinh thi l mt hng i quan trng, ph hp vi nhng n lc pht trin trn
th gii.
Chng 2: VT LIU -NI DUNG - PHNG PHP NGHIN CU

2.1. Vt liu nghin cu

- Ht C rt (Daucus carrota L.), ht Medicago (Medicago truncatula) c
s dng to vt liu m r in vitro khng c gen Ri-tDNA ti Trung tm Cng
ngh sinh hc Lm nghip, Vin Khoa hc Lm nghip Vit Nam.
- M r C rt (Daucus carrota L.)in vitro c nghin cu to ra ti Phng
th nghimCng ngh vi sinh v Sinh hc mi trng, Trung tm Cng ngh sinh
hc Lm nghip, Vin Khoa hc Lm nghip Vit Nam; trong khi m r
Medicago ( Medicago truncatula) in vitro c Ri-tDNA c cung cp t Phng th
nghim AM ca Trung tm CESSAM (Trung tm nghin cu Nm r ni cng
sinh in vitro, i hc Catholique, Louvain-la-Neuve, Vng quc B).
- Chng AM s dng cho nghin cu bao gm (i) chng 41833 (Glomus
intraradices)c cung cp t trung tm CESAMM (Trung tm nghin cu Nm
r ni cng sinh in vitro, i hc Catholique, Louvain-la-Neuve, Vng quc B),
trong bo co ny c gi tt l chng AM 41 (ii) chng M7 (Glomus sp.) l
chng bn a c phn lp, tuyn chn ti Phng th nghim Cng ngh vi sinh
v Sinh hc mi trng, Trung tm Cng ngh sinh hc Lm nghip, Vin Khoa
hc Lm nghip Vit Nam, trong bo co ny c gi tt l chng AM7.
- Chng AM s dng cho nghin cu bao gm (i) chng 41833 (Glomus
intraradices) c cung cp t trung tm CESAMM (Trung tm nghin cu Nm
r ni cng sinh in vitro, i hc Catholique, Louvain-la-Neuve, Vng quc B),
(ii) chng M7 (Glomus sp.) l chng bn a c phn lp, tuyn chn ti Phng
th nghim Cng ngh vi sinh v Sinh hc mi trng, Trung tm Cng ngh sinh
hc Lm nghip, Vin Khoa hc Lm nghip Vit Nam.
- Vi khun Agrobacterium rhizogenses: Chng A4 t Phng th nghim
CESAMM (Trung tm nghin cu Nm r ni cng sinh in vitro, i hc
Catholique, Louvain-la-Neuve, BELGIUM).
2.2. Ni dung nghin cu

2.2.1. Nghin cu to vt liu gi th m r in vitro

- Nghin cu to vt liu gi th m r C rt (Daucus carrota L.) v
Medicago (Medicago truncatula) in vitro khng c gen Ri-tDNA.
- Nghin cu to vt liu m r C rt (Daucus carrota L.) in vitro c gen
Ri-tDNA.
2.2.2. nh gi nh hng ca mi trng nui cy n kh nng nhn sinh
khi cng sinh nm r AM in vitro
CTTN
CT1
CT2
CT3
Mi trng
MSR 0,5% agar
MSR lng (khng agar)
MS 0,5% agar
2.2.3. nh gi nh hng ca pH mi trng nui cy n kh nng nhn

sinh khi cng sinh nm r AM in vitro
CTTN
CT1
CT2
CT3
pH
5,0
5,5
6,0
2.2.4. nh gi nh hng ca gi th m r n kh nng nhn sinh khi cng

sinh nm r AM in vitro
CTTN
CT1.1
CT1.2
CT2.1
CT2.2
Loi gi th m r
M r C rt c gen Ri-tDNA
M r C rt khng gen
M r Medicago c gen Ri-tDNA
M r Medicago khng gen
2.3. Phng php nghin cu

2.3.1.Phng php b tr th nghim

- Trong th nghim v nh gi nh hng ca mi trng nui cy n kh
nng nhn sinh khi cng sinh AM in vitro, s dng gi th l m r C rt v
Medicago c chuyn gen Ri-tDNA cng sinh vi chng 41833 v chng M7.
- Trong th nghim v nh gi nh hng ca pH mi trng n kh nng
nhn sinh khi cng sinh AM in vitro, s dng mi trng MSR 0,5% agar (l loi
mi trng c la chn vi mc ch ti u ha cho kh nng nhn sinh khi
cng sinh AM in vitro trong th nghim mc 2.2.2).
- Trong th nghim v nh gi nh hng ca gi th m r n kh nng
nhn sinh khi cng sinh AM in vitro, s dng mi trng nui cy l mi trng
MSR 0,5% agar, pH 5,5.
2.3.2. Phng php to vt liu m r in vitro
a. To vt liu m r khng chuyn gen Ri-tDNA(Abdoulaye 2003, Chabaud et al.
2006)
Hnh 2.1: Gieo ht Medicago (a)

R Medicago pht trin sau 5 ngy (b)
- Ht C rt v Medicago u c kh trng b mt bng cch ngm 15
pht trong cn etanol 700, sau ngm trong dung dch Sodium hypochlorite 5%
trong 30 pht, tip theo l ra k nhiu ln bng nc ct v trng.
- Ht v trng b mt c cy vo a thch 0,5% agar (5 g/lt; thc hin
trong bc cy v trng), 5-7 ht/a, y kn parafin v p ngc trong t m
270C v theo di ny mm trong thi gian 10 ngy.
- Sau 7-10 ngy, ht ny mm vi on r di khong 3 - 5 cm, dng panh

v dao v trng ct on r (khong 3 - 4 t phn nhnh) v chuyn sang mi
trng MS (Murashige and Skook 1962)c b sung IBA (Indole butylic acid) vi
nng 1,5 ppm, cy chuyn v la chn c cc dng m r C rt v
Medicago in vitro thun, c sinh trng tt nht s dng cho cc th nghim
tip theo.
b. To vt liu m r chuyn gen Ri - tDNA
To m r invitro(Abdoulaye 2003, Chabaud et al. 2006):
Hnh 2.2: Ht C rt ny mm sau 7 ngy gieo ht (a)

Nhn r C rt (b)
- Ht C rt (Daucus carrota L.) c kh trng b mt bng cch ngm 15
pht trong cn etanol 700, sau ngm 30 pht trong dung dch Sodium
hypochlorite 5%; tip theo l ra k nhiu ln bng nc ct v trng.
- Ht C rt v trng b mt c cy vo a thch 0,5% agar (5 g/lt;
thc hin trong bc cy v trng), y kn, parafin v p ngc trong t m 270C
v ch ny mm trong 10 ngy.
- Sau khong 10 ngy, ht ny mm vi r tr khong 1 cm, dng panh v
dao v trng ct b u r (khong 2 mm) v chuyn sang cy nhim vi khun
Agrobacterium rhizogenseschng A4 bng cch ngm nhng v ph ton b b
mt r tr v vt ct vo dung dch vi khun A. rhizogense vi nng t bo 8
triu cfu/ml.
Chun b dung dch vi khun Agrobacterium rhizogensesChng A4
(Abdoulaye 2003, Chabaud et al. 2006):
Vi khun Agrobacterium rhizogenses Chng A4c duy tr nui cy trn

mi trngTY/calcium [5g/l Bacto-tryptone, 3g/l Yeast extract, 6mM CaCl2 (c
b sung sau khi hp v trng); pH = 7,2; Agar 15g/l] . Sau 48 gi nui cy trn
mi trng TY/calcium, sinh khi khun lc A. rhizogenses c s dng cho cy
nhim m r invitro vi nng t bo l 8 triu cfu/ml.
Cy nhim, ng nui cy A. rhizogenses vi m r invitro v chuyn gen:
- S dng mi trng nui cy m r cy nhim A. Rhizogenses l mi
trng MS (Murashige and Skoog 1962) (a lng, vi lng, vitamin, khng cht
kch thch sinh trng thc vt) 0,5% agar (Park et al. 2011).
- Ht C rt ny mm vi r tr ct b tp r c ngm nhng trong sinh
khi khun lc A. rhizogenses (sau 48 gi nui cy) trong 5 - 7 pht, v c ph 1
lp vi khun, sau c cy chuyn vo mi trng trn nui cy trong t m
250C trong 48 gi (ti).
- Cy chuyn sang mi trng MS mi (0,5% agar), b sung khng sinh
carbenicillin (500 mg/lt) loi b vi khun A. rhizogenes v nui cy phng sinh
trng 200C, quang chu k l 16 gi trong 5 - 7 ngy.
- Theo di v chn la gi th m r c s hnh thnh pht trin lng r tt
sau khong 7 - 21 ngy.
Nui cy trn mi trng chn lc c khng sinh v to dng thun m r
invitro c Ri-tDNA:
- Sau 48 gi ng nui cy vi vi khun A. rhizogenses, chng A4 trong t
m 250C (ti), Cc m r C rt c cy chuyn sang mi trng MS mi (0,5%
agar), b sung khng sinh carbenicillin (500 mg/lt) loi b vi khun A.
rhizogenes cn li v nui cy phng sinh trng 20 - 250C, quang chu k l 16
gi: nh gi hiu qu loi b vi khun A. rhizogensesca khng sinh carbenicillin
bng phn tch PCR (polymerase chain reaction) (Sidwa-Gorycka et al. 2009).
- Cc m r hnh thnh v pht trin lng r tt nht, phn nhnh v nhiu
nht (du hiu hnh thi l c chuyn gen Ri-tDNA t vi khun A.
rhizogenses) c chn lc, ct v cy chuyn sang mi trng MS mi (0,5%
agar), b sung khng sinh carbenicillin (500 mg/lt) tip tc loi b vi khun A.
rhizogenes v nui cy phng sinh trng 20 - 250C (ti) to cc dng thun

m r C rt invitro c Ri-tDNA (cy 1 m r 3cm/a petri) (Abdoulaye 2003,
Sidwa-Gorycka et al. 2009).
- Cc dng thun m r invitro c Ri-tDNA (khng cn vi khun A.
rhizogenes) sau c cy chuyn trn mi trng MSR (0,5% agar), nui duy
tr trong phng sinh trng hoc t m 270C v s dng lm gi th cho nghin
cu cng sinh AM (Abuscular mycorrhizae) invitro(Declerck et al. 1998, Ijdo et al.
2011).
Kim tra m r chuyn gen Ri-tDNA v hiu qu loi b vi khun A.
rhizogensesca khng sinh carbenicillin bng phn tch PCR (polymerase
chain reaction):
- Tch DNA t m r C rt chuyn gen Ri-tDNA v r C rt khng
chuyn gen Ri-tDNA (i chng) bng kit tch chit GeneJET Plant Genomic
DNA Purification Mini Kit (Fermentas), cc bc thc hin theo hng dn ca
nh sn xut v c b sung 1% PVP vo Lysis Buffer.. DNA sau c in di
trn gen agarose 1% kim tra cht lng v c o trn my quang ph
NanoDrop ND1000 kim tra tinh sch ca mu. Phn ng PCR c thc
hin trn my lun nhit C1000 Thermal Cycler (BioRad). Thnh phn v chu
trnh nhit ca phn ng PCR c thc hin theo Krolicka et al. (2001). Tng th
tch cho mi phn ng l 20 l vi thnh phn bao gm 10 l PCR Master Mix
(Fermentas), 2 l mi mi (10 pmol), 2 l ca DNA (10 ng/l) v 6 l nc kh
ion v trng. Chu trnh nhit gm: 940C - 3 pht, sau lp li 35 chu k ca 940C
- 1 pht, 540C 1 pht. 720C 1 pht, cui cng l 720C 10 pht v lu 120C.
S
dng
cc
mi
5-GCTCTTGCAGTGCTAGATTT-3
5-
GAAGGTGCAAGCTACCTCTC-3 pht hin cc gen rolB v 5CTCCTGACATCAAACTCGTC-3v
5-TGCTTCGAGTTATGGGTACA-3
trong phn ng PCR pht hin cc gen rolC ca vi khun A. rhizogenses (c

trn on T-DNA) trong b gen ca m r C rt.
- Nhm khng nh cc m r C rt chuyn gen Ri-tDNA hon ton sch vi
khun A. rhizogenes, phn ng PCR c tin hnh vi cc mi virG5S ha bi Trung tm Hc liu i hc Thi Nguyn
ACTGAATATCAGGCAACGCC-3v5-CGTCAAAGAAATAGCCAGC
(Aoyama et al. 1989) xc nh c hay khng cc gen virG (gen nm trn vng
Ri plasmid, ngoi tr on T-DNA) c chuyn sang m r: Chu trnh nhit gm:
940C - 3 pht, sau lp li 35 chu k ca 940C - 1 pht, 540C 1 pht. 720C 1
pht, cui cng l 720C 10 pht v lu 120C
2.3.3. Phng phpcy chuyn v nhn sinh khi m r
- R pht trin sau khong 15 ngy trn mi trng ph hp, tin hnh cy
chuyn nhn nhanh sinh khi r, cc bc tin hnh nh sau:
- t dng c trn ngn la n cn.
- Quan st bng knh hin vi chn la nhng a petri cha mu tt,
khng b nhim tp khun.
- nh du nhng on r cy chuyn ( mu trng hoc vng nht, c
phn nhnh cp 1 hoc 2, ng knh tri hn).
- Dng dao ct on r xc nh di khong 3-5 cm.
- Dng kp gp mnh r cy chuyn sang a petri c sn mi trng,
mi a petri cy 2 on r vi chiu ngc nhau.
- Bc kn bng parafilm.
- Ghi y thng tin trn a petri (loi mu, ngy/thng/nm cy chuyn).
- a vo t nui cy trong iu kin ti 270C.
- Thc hin cy chuyn nhiu ln trong bc cy v trng nhn sinh khi
m r to ngun vt liu a vo cng sinh vi AM.
2.3.4.Phng php to cng sinh AM in vitro
- Trn a thch cha mi trng, ct b mnh thch phn chnh gia a.
- Trn a thch cha vt liu AM v trng ny mm, ct ly AM sao cho
mnh nguyn liu nm trn bn trong phn thch loi b a mi trng.
Dng dao cn thn chuyn vt liu AM t va vo phn thch va loi b trn
mi trng MSR (the modified Strullu Romand) (Strullu and Romand 1986,
Declerck et al. 1998). Cy chuyn on r in vitro ln a thch cha AM sao cho
chiu hng pht trin ca r mi s chm si nm.
- Nui cy trong iu kin thch hp, trong ti, 270C. Theo di tin trin
ca r, si nm, cng sinh v sinh bo t mi trn a cy.
2.3.5. Phng php nhn sinh khi cng sinh AM in vitro
Sau thi gian 4-5 thng, s dng ngun vt liu AM cng sinh vi r lm
nguyn liu AM v trng to cng sinh vi r in vitro trong bc tip theo vi mc
tiu nhn sinh khi cng sinh AM in vitro. Cc bc tin hnh nh sau:
- Trn a thch cha mi trng, ct b mnh thch phn chnh gia a.
- Trn a thch cha vt liu AM v trng ny mm, ct ly AM sao cho
mnh nguyn liu nm trn bn trong phn thch loi b a mi trng.
Dng dao cn thn chuyn vt liu AM t va vo phn thch va loi b trn
mi trng MSR (the modified Strullu Romand) (Strullu and Romand 1986,
Declerck, Stullu et al. 1998). Cy chuyn on r in vitro ln a thch cha AM
sao cho chiu hng pht trin ca r mi s chm si nm.
- Nui cy trong iu kin thch hp, trong ti, 270C. Theo di tin trin
ca r, si nm, cng sinh v sinh bo t mi trn a cy.
2.3.6. Phng php thu thp, phn tch v x l thng k s liu th nghim
a. Phng php thu thp s liu
S lng bo t (bt/a petri): m trc tip bng quan st trn knh hin vi
soi ni trn cc m c thit k sn, tng cc quan st bng 2/3 din tch a
petri, thu s lng bo t cc khong thi gian 1 thng, 2 thng, 3 thng, 4 thng
v 5 thng sau khi tin hnh cng sinh gi th r vi chng AM tng ng, trong
bo co ny cc khong thi gian trn c gi tt tng ng l T1, T2, T3, T4 v
T5.
b. Phng php x l s liu
Cc s liu thu c s c x l thng k bng phn mm SPSS 20. So snh
ngha khc bit cc gi tr trung bnh ca cc cng thc th nghim bng phn
tch ANOVA Post Hoc Multiple Comparison Test theo tiu chun Bonferroni v
Duncan nu phng sai bng nhau v Tamhanes T2 nu phng sai khng bng
nhau, p<0,05 c xem l c ngha.
Chng 3: KT QU V BN LUN
3.1. Kt qu to vt liu gi th m r in vitro
b
Hnh 3.1: R Ca rt khng c gen Ri-tDNA (a)
R Ca rt c gen Ri-tDNA (b)
Bng cc phng php th nghim v k thut kh trng b mt ht, k thut

nui cy trn mi trng thch agar, chng ti to c vt liu gi th r C
rt (Daucus carrota L.) v Medicago (Medicago truncatula) in vitro khng c gen
Ri-tDNA s dng cho cc nghin cu ca ti.
Bng k thut ng nui cy vi A. rhizogenes trn mi trng MS 0,5 % agar,
v sau mi trng MS 0,5 % b sung 500 mg Canbenicilin/lt chn lc v
loi b vi khun A. rhizogenes, chng ti nghin cu to c vt liu gi th r
C rt (Daucus carrota L.) in vitro chuyn gen Ri-tDNA hon ton sch khun.
Vt liu gi th r C rt (Daucus carrota L.) in vitro chuyn gen Ri-tDNA c
khng nh v mt di truyn bng phn ng PCR. Kt qu PCR xc nh trong b
gen ca cc gi th m r C rt invitro ny u pht hin thy c cc gen rolB v
rolC ca vi khun A. rhizogenses (c trn on T-DNA) cc v tr khong 380bp
(cho gen rolB) v 580bp (cho gen rolC) (Hnh 3.2), y l v tr biu hin s c
mt hay khng c mt ca 2 gen rolB v rolC trong cy chuyn gen Ri-tDNA
(Veena and Taylor 2007) cn trn cy i chng (khng chuyn gen) th khng
thy c s xut hin ca 2 gen ny. Nh vy v mt di truyn khng nh l cc
gi th m r C rt invitro ny c chuyn gen Ri-tDNA t vi khun A.
rhizogenes thng qua ng nui cy.
580 bp
380 bp
Hnh 3.2: Phn tch PCR cho m r C rt chuyn gen Ri-tDNA v khng chuyn
gen Ri-tDNA. Bng 1: c gen rolB; bng 2: c gen rolC (cho mu chuyn gen);
bng 3 va 4: khng c gen rolB va rolC (cho mu khng chuyn gen); M: DNA
thang chun 100 bp (Fermentas)
Hnh 3.3: R Medicago khng c gen Ri-tDNA (a)

R Medicago c gen Ri-tDNA (b)
3.2. nh gi nh hng ca mi trng nui cy n nhn sinh khi cng
sinh AM in vitro
Bng 3.1: nh hng ca mi trng nui cy n nhn sinh khi cng sinh AM
in vitro gia chng 41833 vi gi th r C rt chuyn gen Ri-tDNA
Cng thc
S lng bo t
T1
T2
T3
T4
T5
CT1
1809a 47
3453a 62
5057a 43
8938a 64
10363a 77
CT2
147b 8
263b 6
330b 10
402b 3
636b 8
CT3
644c 9
1226c 14
1465c 24
1805c 8
2021c 17
Post Hoc Test: Gi tr trong cng ct thi gian c ch ci ging nhau l khc bit khng ngha, =0,05
CT1: MSR 0,5% agar
CT2: MSR lngCT3: MS 0,5% agar
T1: thng th nht, T2: thng th 2, T3: thng th 3; T4: thng th 4, T5: thng th 5
Sau thi gian 5 thng tin hnh b tr v theo di th nghim, kt qu Bng

3.1 cho thy, t thng th nht n thng th nm s lng bo t trung bnh cao
nht thu c l CT1. S lng bo t sn sinh mc bnh thng trong cc

thng th nht, thng th hai v thng th ba, sn sinh nhiu nht thng th t,
v sau gim dn thng th nm. Trong thng th 5 (thng cui), s lng bo
t t cao nht CT1 (trung bnh t 10363 bo t/petri), tip theo l CT3 (2021
bo t/petri) v thp nht CT2 (ch t trung bnh 636 bo t/petri).
Kt qu phn tch Post Hoc Test v ngha khc bit ca s lng bo t
trong cc cng thc th nghim cho thy rng, s lng bo t gia cc cng thc
th nghim CT1, CT2 v CT3 khc bit c ngha (=0,05) trong tt c cc thng
th nghim t T1 n T5. iu ny c ngha rng, yu t th nghim mi trng
nui cy nh hng quan trng ti s sn sinh bo t in vitro ca chng 41833
S lng bo t/dia petri
cng sinh vi gi th r C rt c gen Ri-tDNA.

10363
12000
10000
8000
6000
2021
4000
636
2000
0
CT1
CT2
CT3
C rt - 41833
Biu 3.1:nh hng ca mi trng nui cy n nhn sinh khi cng sinh AM
in vitro gia chng 41833 vi gi th r C rt chuyn gen Ri-tDNA
Bng 3.2:nh hng ca mi trng nui cy n nhn sinh khi cng sinh AM in
vitro gia chng M7 vi gi th r C rt chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
2061a 45
4031a 8
7055a 12
9238a 31
9948a 31
CT2
120c 3
252c 3
308c 6
493c 13
575c 14
CT3
722b 5
1245b 32
1455b 18
1804b 8
2049b 40
Post Hoc Test: Gi tr trong cng ct thi gianc ch ci ging nhau l khc bit khng ngha, =0,05
CT1: MSR 0,5% agar
CT2: MSR lng
CT3: MS 0,5% agar

nui cy nh hng quan trng ti s sn sinh bo t in vitro ca chng M7
S lng bo t/dia petri
9948
10000
8000
6000
4000
2049
575
2000
0
CT1
CT2
CT3
C rt - M7
in vitro gia chng M7 vi gi th r C rt chuyn gen Ri-tDNA
vitro gia chng 41833 vi gi th r Medicago chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
1632a 11
3354a 25
4025a 52
7110a 16
8271a 12
CT2
81c 5
169c 5
217c 5
288c 6
407c 6
CT3
408b 6
1056b 36
1250b 12
1411b 8
1740b 6
CT1: MSR 0,5% agar
CT2: MSR lng
CT3: MS 0,5% agar
S lng bo t/petri

cng sinh vi gi th r Medicago c gen Ri-tDNA.
8271
10000
8000
6000
4000
1740
407
2000
0
CT1
CT2
CT3
Medicago - 41833
in vitro gia chng 41833 vi gi th r Medicago chuyn gen Ri-tDNA
vitro gia chng M7 vi gi th r Medicago chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
1949a 23
3656a 39
5873a 74
8266a 75
9229a 29
CT2
104c 3
218c 6
254c 6
406c 7
457c 6
CT3
548b 8
1123b 4
1356b 13
1704b 12
1827b 10
CT1: MSR 0,5% agar
CT2: MSR lng
CT3: MS 0,5% agar

10000
9229
S lng bo t/ petri
9000
8000
7000
6000
5000
4000
1827
3000
2000
457
1000
0
CT1
CT2
CT3
Medicago - M7
in vitro gia chng M7 vi gi th r Medicago chuyn gen Ri-tDNA
Hnh3.4: R cng sinh pht trin trn mi trng MSR c (a), trn mi trng
MSR lng (b), trn mi trng MS c (c)
Kt qu s lng bo t thu c trong th nghim mi trng sau 5 thng
tin hnh th nghim trn 4 loi gi th C rt-41833, C rt-M7, Medicago-41833,
Medicago-M7 tng ng vi nhng kt qu nghin cu trc y (Declerck,
DOr et al. 2001) v mi trng nui cy AM in vitro. Kt qu CT1 cho thy,
mi trng MSR c b sung 0,5% agar ph hp hn cho nui cy AM so vi mi
trng cng loi MSR lng bi AM cn c s c nh tt pht trin, so vi mi
trng MS, thnh phn dinh dng trong MSR c bin i vi mc ch ti
u ha cho s pht trin ca AM, hn ch mt s cht gy c ch s cng sinh.
Mi trng MS tt cho s sinh trng ca r, do gim s ph thuc ca r vo
quan h cng sinh vi AM, t lm gim mc cng sinh nm - r. Trong
nghin cu ny, chng ti cn la chn c loi mi trng ti u cho kh nng
nhn sinh khi cng sinh AM. V vy, mi trng MSR c b sung 0,5% agar l
loi mi trng c la chn v s dng cho nhng th nghim nghin cu tip
theo.
Mt s nghin cu ch ra rng, s dng mi trng MSR khng c ng
cho kt qu cng sinh AM tt hn khi s dng mi trng MSR c b sung ng
(Ijdo, Cranenbrouck et al. 2011). Tuy nhin, trong phng php ny, Ijdo v cng
s s dng a petri c cu trc 2 ngn, trong , mt ngn c b sung ng
cung cp dinh dng cho gi th r pht trin, ngn cn li l mi trng khng
ng cho s pht trin ca nm, ng thi c s b sung mi trng theo tng
t. Vic p dng phng php ny cn hn ch Vit Nam do vn v vic cn
u t trang thit b rt tn km v yu cu cao v cng ngh. Do vy, kt qu
nghin cu ca chng ti l ph hp vi vic tip cn nghin cu theo hng k

thut n gin.
3.3. nh gi nh hng ca pH mi trng nui cy n nhn sinh khi cng
sinh AM in vitro
Bng 3.5:nh hng ca pH mi trng nui cy n nhn sinh khi cng sinh
AM in vitro gia chng 41833 vi gi th rC rt chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
1108b 25
2127c 20
3147c 14
4216c 17
5412c 7
CT2
1809a 47
3453a 60
5057a 43
8938a 64
10363a 78
CT3
1025b 8
2523b 18
4357b 31
6164b 71
7138b 14
CT1: pH = 5,0 CT2: pH = 5,5CT3: pH = 6,0

bo t/petri) v thp nht CT1(ch t trung bnh 5412 bo t/petri).
th nghim CT1v CT2 , gia CT2 v CT3 khc bit c ngha (=0,05) trong tt
c cc thng th nghim t T1 n T5. S khc bit c ngha gia CT1 v CT3
xy ra t T2 n T5. iu ny c ngha rng, yu t th nghim pH mi trng
S lng bo t/petri
10363
12000
10000
7138
8000
5412
6000
4000
2000
0
CT1
CT2
CT3
C rt - 41833
AM in vitro gia chng 41833 vi gi th r C rt chuyn gen Ri-tDNA
AM in vitro gia chng M7 vi gi th r C rt chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
1072b 14
2060c 32
2953c 18
4041c 22
5207c 4
CT2
2061a 45
4031a 8
7055a 12
9238a 32
9948a 31
CT3
949c 30
2352b 40
4153b 34
6046b 27
7043b 13
CT1: pH = 5,0 CT2: pH = 5,5CT3: pH = 6,0

th nghim t T1 n T5. iu ny c ngha rng, yu t th nghim pH mi trng

S lng bo t/ petri
9948
10000
9000
8000
7000
6000
5000
4000
3000
2000
1000
0
7043
5207
CT1
CT2
CT3
C rt - M7
AM in vitro gia chng M7 vi gi th r C rt chuyn gen Ri-tDNA
AM in vitro gia chng 41833 vi gi th r Medicago chuyn gen Ri-tDNA
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1
533c 23
1362c 13
2469c 22
3655c 45
4161c 22
CT2
1632a 11
3354a 25
4052a 52
7110a 16
8271a 12
CT3
749b 5
1816b 12
3377b 24
4998b 32
5738b 32
CT1: pH = 5,0 CT2: pH = 5,5CT3: pH = 6,0

8271
S lng bo t/ petri
9000
8000
5738
7000
6000
5000
4161
4000
3000
2000
1000
0
CT1
CT2
CT3
Medicago - 41833
AM in vitro gia chng 41833 vi gi th r Medicago chuyn gen Ri-tDNA
AM in vitro gia chng M7 vi gi th r Medicago chuyn gen Ri-tDNA
Cng thc
S lng bo t
T1
T2
T3
T4
T5
CT1
663c 34
1537c 25
2589c 56
3934c 70
4342c 15
CT2
1949a 23
3656a 39
5873a 74
8266a 75
9229a 28
CT3
822b 9
2051b 38
3560b 17
4947b 35
5531b 19
CT1: pH = 5,0 CT2: pH = 5,5CT3: pH = 6,0

9229
10000
S lng bo t/petri
9000
8000
7000
6000
5531
4342
5000
4000
3000
2000
1000
0
CT1
CT2
CT3
Medicago - M7
AM in vitro gia chng M7 vi gi th r Medicago chuyn gen Ri-tDNA
Kt qu v ni dung nh gi nh hng ca pH mi trng n kh nng
nhn sinh khi cng sinh AM in vitro, vi vic s dng gi th r c gen Ri-DNA
trn mi trng MSR 0,5% agar cho thy, s lng bo t thu c cao nht
mc pH 5,5 trn c 4 loi gi th C rt-41833, C rt-M7, Medicago-41833,
Medicago-M7. Nh vy, mc pH ti u cho s pht trin ca AM l 5,5. Vi mc

pH 6,0, s lng bo t thu c cao hn so vi mc pH 5,0. iu ny c gii
thch l do AM chu baz tt hn axit (Hawkins and George 1997).
12000
41833-C rt
Ri-tDNA
M7-C rt
Ri-tDNA
M7-Medicago
Ri-tDNA
S lng bo t/petri
10000
41833-Medicago
Ri-tDNA
8000
6000
4000
2000
T1
T2
T3
T4
T5
Biu 3.9: Kt qu nhn sinh khi AM in vitro ca 41833-C rt Ri-tDNA, M7C rt Ri-tDNA, 41833-Medicago Ri-tDNA, M7-Medicago Ri-tDNA trn mi
trng MSR 0,5% agar, pH 5,5
Kt qu tng hp trong biu 3.9 cho thy, sau 5 thng tin hnh th
nghim trn mi trng MSR 0,5% agar, pH 5,5 nng sut nhn sinh khi bo t
AM in vitro ca chng 41833 cao hn chng M7 khi cng sinh vi gi th r C
rt c Ri-tDNA. Kt qu thu c ngc li khi tin hnh cng sinh 2 chng AM
trn vi gi th r Medicago c Ri-tDNA (kt qu trong bng 3.1, 3.2, 3.3, 3.4, 3.5,
3.6, 3.7 v 3.8). iu c ngha rng, cc chng AM c nh hng khc nhau
n nng sut nhn sinh khi bo t AM in vitro khi cng sinh vi cc loi gi th
m r khc nhau.
Kt qu tng hp trong biu 3.9 cng ch ra rng, nng sut nhn sinh
khi bo t AM in vitro ca gi th r C rt c Ri-tDNA cao hn gi th r
Medicago c Ri-tDNA khi cng sinh trn cng chng 41833. Kt qu thu c
tng t khi tin hnh cng sinh 2 loi gi th trn chng M7 (kt qu trong bng
3.1, 3.2, 3.3, 3.4, 3.5,3.6, 3.7 v 3.8). iu c ngha rng, gi th m r C rt
c Ri-tDNA ph hp hn cho nhn sinh khi AM in vitro trong nghin cu ny.
Hnh 3.5: AM cng sinh vo r va sinh trng si nm mi trn r C rt (a),

trn r Medicago (b)
3.4.nh gi nh hng ca gi th m r n kh nng nhn sinh khi cng
sinh AM in vitro
Bng 3.9: nh hng ca cc loi gi th m r khc nhau n nhn sinh khi
cng sinh AM in vitro trn chng 41833
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1.1
1809a 47
3454a 61
5057a 43
8938a 64
10363a 78
CT 1.2
282c 23
646c 30
1066c 34
1527c 13
2119c 22
CT 2.1
1456b 38
2756b 47
4053b 28
7110b 16
8271b 12
CT 2.2
225c 8
527c 20
847d 5
1226d 11
1720d 20
CT 1.1: C rt c Ri-tDNACT 1.2: C rt khng Ri-tDNA

CT 2.1: Medicago c Ri-tDNACT 2.2: Medicago khng Ri-tDNA
Sau thi gian 5 thng tin hnh b tr v theo di th nghim, kt qu Bng 3.9
cho thy, t thng th nht n thng th nm s lng bo t trung bnh cao nht
thu c l CT1.1. S lng bo t sn sinh mc bnh thng trong cc

t t cao nht CT1.1 (trung bnh t 10363 bo t/petri), tip theo l CT2.1
(8271 bo t/petri), sau l CT 1.2 (2119 bo t/petri), thp nht CT2.2(ch
t trung bnh 1720 bo t/petri).
Kt qu phn tch Post Hoc Test v ngha khc bit ca s lng bo t trong cc
cng thc th nghim cho thy rng, s lng bo t gia cc cng thc th nghim
CT1.1 v CT1.2, gia CT2.1 v CT2.2, gia CT1.1 v CT2.1 khc bit c ngha
(=0,05) trong tt c cc thng th nghim t T1 n T5. S khc bit c ngha
(=0,05) gia CT1.2 v CT2.2 xy ra t T3 n T5. iu ny c ngha rng, yu t
th nghim pH mi trng nui cy nh hng quan trng ti s sn sinh bo t
in vitro ca chng 41833 cng sinh vi gi th r C rt v Medicago c gen RitDNA v khng gen Ri-tDNA.
12000
10363
S lng bo t/petri
10000
8328
8000
6000
4000
2119
1720
2000
0
CT1.1
CT1.2
CT2.1
CT2.2
Chng 41833
Biu 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh khi

cng sinh AM in vitro trn chng 41833
12000
41833-C rt
Ri-tDNA
S lng bo t/petri
10000
41833-Medicago
Ri-tDNA
8000
6000
4000
41833-C rt
khng
Ri-tDNA
2000
41833-Medicago
khng
Ri-tDNA
0
T1
T2
T3
T4
T5
Biu 3.11: Kt qu nhn sinh khi AM in vitro ca 4 loi gi th m r cng

sinh vi chng 41833 trn mi trng MSR 0,5% agar, pH 5,5
Kt qu tng hp trong biu 3.11 cho thy, nng sut nhn sinh khi bo
t AM in vitro trn gi th m r C rt c Ri-tDNA cao hn rt nhiu trn gi th
m r C rt khng Ri-tDNA (cao hn khong 5 ln T5). iu ny ng cho c 2
loi gi th m r Medicago c Ri-tDNA v khng c Ri-tDNA (kt qu trong
bng 3.9). iu ny c ngha rng, s dng gi th r m r c chuyn gen RitDNA cho hiu qu nhn sinh khi bo t AM in vitro tt hn gi th m r khng
chuyn gen Ri-tDNA.
khi bo t AM in vitro trn gi th r C rt c Ri-tDNA cao hn trn gi th m
r Medicago c Ri-tDNA. iu ny ng cho c hai loi gi th m r C rt
khng c Ri-tDNA v Medicago khng c Ri-tDNA (kt qu trong bng 3.9). iu
ny c ngha rng, trong nghin cu ny, gi th m r C rt ph hp hn gi th
m r Medicago cho nhn sinh khi cng sinh AM in vitro.
Bng 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh khi

cng sinh AM in vitro trn chng M7
S lng bo t
Cng thc
T1
T2
T3
T4
T5
CT1.1
2060a 47
4031a 8
7055a 12
9238a 32
9948a 31
CT 1.2
170c 26
534c 6
926c 21
1341c 24
1918c 19
CT 2.1
944b 39
2059b 19
3208b 158
8266b 75
9229b 29
CT 2.2
135c 6
430d 17
743c 11
1075d 9
1534d 6
CT 1.1: Carot c Ri-tDNA

CT 1.2: Carot khng Ri-tDNA
CT 2.1: Medicago c Ri-tDNA

CT 2.2: Medicago khng Ri-tDNA

nht thu c l CT1.1. S lng bo t sn sinh mc bnh thng trong
cc thng th nht, thng th hai v thng th ba, sn sinh nhiu nht thng th
t, v sau gim dn thng th nm. Trong thng th 5 (thng cui), s lng
bo t t cao nht CT1.1 (trung bnh t 9948 bo t/petri), tip theo l CT2.1
(9229 bo t/petri), sau l CT 1.2 (1918 bo t/petri), thp nht CT2.2(ch
t trung bnh 1534 bo t/petri).
th nghim CT1.1 v CT1.2, gia CT2.1 v CT2.2, gia CT1.1 v CT2.1 khc bit
c ngha (=0,05) trong tt c cc thng th nghim t T1 n T5. S khc bit
c ngha (=0,05) gia CT1.2 v CT2.2 xy ra t T2 n T5. iu ny c ngha
rng, yu t th nghim pH mi trng nui cy nh hng quan trng ti s
sn sinh bo t in vitro ca chng M7 cng sinh vi gi th r C rt v Medicago
c gen Ri-tDNA v khng gen Ri-tDNA.
9948
10000
S lng bo t/petri
9000
7119
8000
7000
6000
5000
4000
1918
3000
1534
2000
1000
0
CT1.1
CT1.2
CT2.1
CT2.2
Chng M7
Biu 3.12: nh hng ca cc loi gi th m r khc nhau n nhn sinh khi

cng sinh AM in vitro trn chng M7
12000
M7-C rt
Ri-tDNA
10000
M7-Medicago
Ri-tDNA
S lng bo t/petri
8000
6000
4000
M7-C rt khng
Ri-tDNA
2000
M7-Medicago
khng
Ri-tDNA
0
T1
T2
T3
T4
T5
Biu 3.13: Kt qu nhn sinh khi AM in vitro ca 4 loi gi th m r cng

sinh vi chng M7 trn mi trng MSR 0,5% agar, pH 5,5
Kt qu tng hp trong biu 3.13 cho thy, nng sut nhn sinh khi bo
t AM in vitro trn gi th m r C rt c Ri-tDNA cao hn rt nhiu trn gi th
m r C rt khng Ri-tDNA (cao hn khong 5 ln T5). iu ny ng cho c 2
loi gi th m r Medicago c Ri-tDNA v khng c Ri-tDNA (kt qu trong

bng 3.10). iu ny c ngha rng, s dng gi th r m r c chuyn gen RitDNA cho hiu qu nhn sinh khi bo t AM in vitro tt hn gi th m r khng
chuyn gen Ri-tDNA.
khi bo t AM in vitro trn gi th r C rt c Ri-tDNA cao hn trn gi th m
r Medicago c Ri-tDNA. iu ny ng cho c hai loi gi th m r C rt
khng c gen Ri-tDNA v Medicago khng c gen Ri-tDNA (kt qu trong bng
3.10). iu ny c ngha rng, trong nghin cu ny, gi th m r C rt ph hp
hn Medicago cho nhn sinh khi cng sinh AM in vitro.
Hnh 3.6: Sinh sn bo t AM sau 1 thng (a), sau 4 thng (b) trn gi th C
rt c Ri-tDNA
Mt kt qu theo di quan trng t c trong nghin cu ny ca chng
ti, l, trong tt c cc mi trng nui cy th nghim, c bit trong mi
trng MSR 0,5% agar vi 10 gam ng/lt mi trng: trong cc thng u th
nghim (T1, T2), khi hm lng ng trong mi trng cn cao, qu trnh hnh
thnh cng sinh v nhn sinh khi AM in vitro din ra chm v thp; tuy nhin qu
trnh ny bt u tng mnh trong thng th 3 v c bit tng rt t bin trong
thng th 4, v gim dn trong thng cui th nghim, thng th 5. iu ny c
gii thch l trong cc thng sau (T3, T4) hm lng ng trong mi trng
MSR gim mnh do gi th r tiu th trc tip, v do khng cn c ch qu
trnh nhn sinh khi AM in vitro.
Chng 4: KT LUN - TN TI - KIN NGH

4.1. Kt lun
- ti nghin cu to c vt liu gi th r C rt (Daucus carrota L.)
in vitrochuyn gen Ri-tDNA hon ton sch khun bng k thut ng nui cy
vi A. rhizogenes trn mi trng MS 0,5% agar, khng cht kch thch sinh
trng v chn lc, loi b vi khun A. rhizogenes trn mi trng MS 0,5% agar
b sung 500 mg Cacbenicilin/lt. Bng phn ng PCR, vt liu gi th r C rt
ny c xc nh c cc gen rolB v rolC ca vi khun A. rhizogenses(c trn
on T-DNA) cc v tr khong 380bp (cho gen rolB) v 580bp (cho gen rolC).
- Sau 5 thng nui cy, nhn sinh khi cng sinh AM in vitrotrn cc loi
mi trng th nghim khc nhau, sinh khi bo t cao nht t c l vi mi
trng MSR 0,5% agar cho c 2 chng AM th nghim 41833 & M7 khi cng sinh
vi 2 loi gi th r chuyn gen Ri-tDNA l C rt (Daucus carrota L.) v
Medicago (Medicago truncatula) vi s liu s lng bo t/a petri tng ng l
10363, 9948, 8271 v 9229; v sinh khi bo t thp nht l vi mi trng MSR
lng, khng agar.
- Sau 5 thng nui cy, nhn sinh khi cng sinh AM in vitrovi cc loi gi
th r in vitro khc nhau, s lng bo t cao nht t c l vi gi th r C rt
chuyn gen Ri-tDNA, tip theo l gi th r Medicago chuyn gen Ri-tDNA v s
lng bo t thp nht l vi 2 loi gi th r C rt v Medicago khng c gen RitDNA, kt qu ny ng cho c 2 chng AM th nghim l 41833 v M7 (kt qu
trong bng 4.9 v bng 4.10).Kt qu tng hp cho thy mt iu r rng rng, s
dng gi th r chuyn gen Ri-tDNA cho mc tiu nhn sinh khi cng sinh AM in
vitro mang li hiu qu cao hn rt nhiu so vi gi th r khng chuyn gen RitDNA; iu ny ng cho c 2 loi gi th r l C rt v Medicago, v c 2 chng
AM th nghim l 41833 v M7.
- Trong s cc th nghim v nh hng ca pH mi trng MSR 0,5% agar
n nhn sinh khi cng sinh AM in vitro,s lng bo t AM t c cao nht
trong mi trng c pH l 5,5 v thp nht trn mi trng c pH 5,0. Kt qu ny
ng cho c 2 chng AM th nghim l 41833 & M7 khi cng sinh tng ng vi

2 loi gi th r in vitro l C rt chuyn gen Ri-tDNAv Medicago chuyn gen
Ri-tDNA (kt qu trong bng 4.5; 4.6; 4.7 v 4.8).
4.2. Tn ti v kin ngh
Do nhng hn ch v thi gian v dng c thit b cng ngh, ti khng
tin hnh c th nghim nghin cu hnh thnh v nhn sinh khi cng sinh AM
in vitro trn a petri 2 ngn vi 2 loi mi trng MSR khc nhau l MSR agar c
ng v MSR agar khng c ng, nhm tng cng hn kh nng nhn sinh
khi cng sinh AM trn mi trng MSR khng ng. Tc gi lun vn xut
tip tc th nghim nghin cu b sung v iu chnh gim hm lng ng trong
mi trng MSR 0,5% agar gim c ch v kch thch qu trnh hnh thnh
cng sinh v nhn sinh khi AM in vitro ngay t nhng thng u ca th nghim;
v do c th thay th vic phi s dng loi dng c a petri 2 ngn (gi t v
quy trnh phc tp hn).
TI LIU THAM KHO

Ti liu Ting Vit
1) Nguyn Minh Chu, L Quc Huy, 2007. Kt qu nghin cu p dng th
nghim ch phm nm r ECM dng vin nang (Alginate beads) cho cy con
Sao en (Hopea odorata). Tp ch KH & CN, B NN&PTNT 18:81-86.
2) Tng Th Chnh, Bi Vn Cng, 2007. Nghin cu s a dng nm cng
sinh Arbuscular mycorrhiza c Vetiver t t nhim ch. Bo co khoa
hc v Sinh thi v Ti nguyn sinh vt, Hi ngh khoa hc ton quc ln
th hai, Vin sinh thi v Ti nguyn sinh vt 1:216-221.
3) Nguyn S Giao, Nguyn Th Nhm, 1988. Nghin cu s dng nm r cng
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PH LC
Ph lc 1: Kt qu phn tch cc i lng c trng ca s lng bo t
trong th nghim mi trng trn gi th r C rt cng sinh vi chng 41833
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
1809.00
47.149
27.221
1691.88
1926.12
1776
1863
147.00
7.937
4.583
127.28
166.72
138
153
644.00
9.165
5.292
621.23
666.77
636
654
Total
866.67
739.190
246.397
298.47
1434.86
138
1863
3453.00
61.828
35.697
3299.41
3606.59
3383
3498
263.00
6.245
3.606
247.49
278.51
258
270
1226.00
13.937
8.047
1191.38
1260.62
1217
1242
Total
1647.33
1417.353
472.451
557.86
2736.81
258
3498
5057.00
43.405
25.060
4949.18
5164.82
5007
5085
330.00
7.937
4.583
310.28
349.72
324
339
1465.00
24.109
13.919
1405.11
1524.89
1448
1493
Total
2284.00
2137.179
712.393
641.22
3926.78
324
5085
8938.33
63.721
36.789
8780.04
9096.62
8892
9011
402.00
3.000
1.732
394.55
409.45
399
405
1805.33
7.506
4.333
1786.69
1823.98
1798
1813
Total
3715.22
3964.314
1321.438
667.98
6762.46
399
9011
10362.50
77.397
44.685
10170.24
10554.76
10305
10451
636.00
7.937
4.583
616.28
655.72
627
642
2021.50
17.385
10.037
1978.31
2064.69
2006
2040
Total
4340.00
4556.718
1518.906
837.40
7842.60
627
10451
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5
Ph lc 2: Kt qu phn tch Post Hoc Test v s lng bo t ca cc cng

thc th nghim mi trng trn gi th r C rt-41833
Dependent Variable
(I)
(J)
CT
CT
Bonferroni
Bonferroni
S lng bo t 3
Bonferroni
S lng bo t 4
Bonferroni
Upper Bound
1586.56
1737.44
1165.000*
22.949
.000
1089.56
1240.44
-1662.000
22.949
.000
-1737.44
-1586.56
-497.000*
22.949
.000
-572.44
-421.56
22.949
.000
-1240.44
-1089.56
-1165.000
497.000*
2
3
22.949
.000
421.56
572.44
3190.000
30.022
.000
3091.30
3288.70
2227.000
30.022
.000
2128.30
2325.70
30.022
.000
-3288.70
-3091.30
30.022
.000
-1061.70
-864.30
30.022
.000
-2325.70
-2128.30
-3190.000
-963.000
-2227.000
30.022
.000
864.30
1061.70
4727.000
23.703
.000
4649.08
4804.92
3592.000
23.703
.000
3514.08
3669.92
-4727.000
23.703
.000
-4804.92
-4649.08
-1135.000
23.703
.000
-1212.92
-1057.08
-3592.000
23.703
.000
-3669.92
-3514.08
1135.000
23.703
.000
1057.08
1212.92
8536.333
30.279
.000
8436.79
8635.87
7133.000
30.279
.000
7033.46
7232.54
-8536.333
30.279
.000
-8635.87
-8436.79
-1403.333
30.279
.000
-1502.87
-1303.79
-7133.000
30.279
.000
-7232.54
-7033.46
1403.333
30.279
.000
1303.79
1502.87
9726.500
37.581
.000
9602.95
9850.05
8341.000
37.581
.000
8217.45
8464.55
-9726.500*
37.581
.000
-9850.05
-9602.95
-1385.500
37.581
.000
-1509.05
-1261.95
-8341.000*
37.581
.000
-8464.55
-8217.45
37.581
.000
1261.95
1509.05
2
3
1
3
2
3
1
3
963.000
Bonferroni
Lower Bound
.000
S lng bo t 5
Sig.
22.949
S lng bo t 2
(I-J)
Error
95% Confidence Interval
1662.000*
Difference
Std.
2
1
S lng bo t 1
Mean
3
1385.500
*. The mean difference is significant at the 0,05 level.

trong th nghim mi trng trn gi th r C rt cng sinh vi chng M7
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
2061.00
45.398
26.211
1948.22
2173.78
2025
2112
120.00
3.000
1.732
112.55
127.45
117
123
722.00
4.583
2.646
710.62
733.38
717
726
Total
967.67
860.740
286.913
306.04
1629.29
117
2112
4030.50
8.352
4.822
4009.75
4051.25
4023
4040
252.00
3.000
1.732
244.55
259.45
249
255
1244.50
32.392
18.702
1164.03
1324.97
1211
1275
Total
1842.33
1696.547
565.516
538.25
3146.41
249
4040
7055.00
12.031
6.946
7025.11
7084.89
7043
7067
308.00
6.245
3.606
292.49
323.51
303
315
1454.50
17.514
10.112
1410.99
1498.01
1436
1470
Total
2939.17
3126.561
1042.187
535.88
5342.45
303
7067
9238.00
31.261
18.049
9160.34
9315.66
9204
9266
492.67
12.741
7.356
461.02
524.32
478
501
1804.00
7.937
4.583
1784.28
1823.72
1798
1813
Total
3844.89
4084.532
1361.511
705.24
6984.54
478
9266
9947.50
30.790
17.776
9871.01
10023.99
9914
9974
575.00
13.528
7.810
541.40
608.60
561
588
2049.00
39.686
22.913
1950.41
2147.59
2004
2079
Total
4190.50
4364.747
1454.916
835.46
7545.54
561
9974
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim mi trng trn gi th r C rt-M7
Dependent Variable
(I)
(J)
CT
CT
Bonferroni
Bonferroni
Bonferroni
Bonferroni
2011.86
1339.000*
21.556
.000
1268.14
1409.86
-1941.000
21.556
.000
-2011.86
-1870.14
-602.000*
21.556
.000
-672.86
-531.14
21.556
.000
-1409.86
-1268.14
-1339.000
602.000*
2
3
Bonferroni
21.556
.000
531.14
672.86
3778.500
15.832
.000
3726.45
3830.55
2786.000
15.832
.000
2733.95
2838.05
15.832
.000
-3830.55
-3726.45
15.832
.000
-1044.55
-940.45
15.832
.000
-2838.05
-2733.95
-3778.500
-992.500
-2786.000
15.832
.000
940.45
1044.55
6747.000
10.440
.000
6712.68
6781.32
5600.500
10.440
.000
5566.18
5634.82
-6747.000
10.440
.000
-6781.32
-6712.68
-1146.500
10.440
.000
-1180.82
-1112.18
-5600.500
10.440
.000
-5634.82
-5566.18
1146.500
10.440
.000
1112.18
1180.82
8745.333
16.347
.000
8691.59
8799.08
7434.000
16.347
.000
7380.26
7487.74
-8745.333
16.347
.000
-8799.08
-8691.59
-1311.333
16.347
.000
-1365.08
-1257.59
-7434.000
16.347
.000
-7487.74
-7380.26
1311.333
16.347
.000
1257.59
1365.08
9372.500
24.522
.000
9291.88
9453.12
7898.500*
24.522
.000
7817.88
7979.12
-9372.500
24.522
.000
-9453.12
-9291.88
-1474.000*
24.522
.000
-1554.62
-1393.38
-7898.500
24.522
.000
-7979.12
-7817.88
1474.000*
24.522
.000
1393.38
1554.62
1
3
1
3
992.500
S lng bo t 5
Upper Bound
1870.14
3
S lng bo t 4
Lower Bound
.000
3
So luong bao tu 3
Sig.
21.556
S lng bo t 2
(I-J)
Error
1941.000*
Difference
Std.
2
1
S lng bo t 1
Mean

trong th nghim mi trng trn gi th r Medicago cng sinh vi chng 41833
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
1632.00
10.817
6.245
1605.13
1658.87
1623
1644
81.00
5.196
3.000
68.09
93.91
78
87
408.00
6.000
3.464
393.10
422.90
402
414
Total
707.00
708.084
236.028
162.72
1251.28
78
1644
3353.50
25.159
14.526
3291.00
3416.00
3338
3383
169.00
4.583
2.646
157.62
180.38
165
174
1056.00
36.000
20.785
966.57
1145.43
1020
1092
Total
1526.17
1423.474
474.491
431.99
2620.35
165
3383
4052.00
51.752
29.879
3923.44
4180.56
4001
4104
217.00
4.583
2.646
205.62
228.38
213
222
1249.67
11.590
6.692
1220.87
1278.46
1239
1262
Total
1839.56
1718.734
572.911
518.42
3160.69
213
4104
7110.33
16.258
9.387
7069.95
7150.72
7092
7123
288.00
6.000
3.464
273.10
302.90
282
294
1411.33
7.638
4.410
1392.36
1430.31
1403
1418
Total
2936.56
3167.914
1055.971
501.48
5371.63
282
7123
8271.00
11.715
6.764
8241.90
8300.10
8258
8279
407.00
6.245
3.606
391.49
422.51
402
414
1740.00
6.245
3.606
1724.49
1755.51
1733
1745
Total
3472.67
3644.753
1214.918
671.06
6274.27
402
8279
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim mi trng trn gi th r Medicago-41833
Dependent Variable
(I)
(J)
CT
CT
Mean
Difference (I-
Bonferroni
6.325
.000
1530.21
1571.79
1224.000
6.325
.000
1203.21
1244.79
-1551.000*
6.325
.000
-1571.79
-1530.21
6.325
.000
-347.79
-306.21
6.325
.000
-1244.79
-1203.21
-327.000
-1224.000*
2
3
Bonferroni
Bonferroni
Bonferroni
.000
306.21
347.79
3184.500
20.817
.000
3116.07
3252.93
2297.500
20.817
.000
2229.07
2365.93
20.817
.000
-3252.93
-3116.07
20.817
.000
-955.43
-818.57
20.817
.000
-2365.93
-2229.07
-887.000
-2297.500
20.817
.000
818.57
955.43
3835.000
25.094
.000
3752.51
3917.49
2802.333
25.094
.000
2719.84
2884.83
-3835.000
25.094
.000
-3917.49
-3752.51
-1032.667
25.094
.000
-1115.16
-950.17
-2802.333
25.094
.000
-2884.83
-2719.84
1032.667
25.094
.000
950.17
1115.16
6822.333
8.928
.000
6792.98
6851.68
5699.000
8.928
.000
5669.65
5728.35
-6822.333
8.928
.000
-6851.68
-6792.98
-1123.333
8.928
.000
-1152.68
-1093.98
-5699.000
8.928
.000
-5728.35
-5669.65
1123.333
8.928
.000
1093.98
1152.68
7864.000
6.916
.000
7841.26
7886.74
6531.000*
6.916
.000
6508.26
6553.74
-7864.000
6.916
.000
-7886.74
-7841.26
-1333.000*
6.916
.000
-1355.74
-1310.26
-6531.000
6.916
.000
-6553.74
-6508.26
1333.000*
6.916
.000
1310.26
1355.74
1
3
1
3
887.000
S lng bo t 5
6.325
-3184.500
3
S lng bo t 4
327.000
3
S lng bo t 3
Bonferroni
Upper Bound
1551.000*
S lng bo t 2
Lower Bound
Sig.
J)
S lng bo t 1
Std. Error

trong th nghim mi trng trn gi th r Medicago cng sinh vi chng M7
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
1949.00
22.913
13.229
1892.08
2005.92
1929
1974
104.00
3.464
2.000
95.39
112.61
102
108
548.00
7.550
4.359
529.25
566.75
540
555
Total
867.00
834.053
278.018
225.89
1508.11
102
1974
3656.00
39.125
22.589
3558.81
3753.19
3623
3699
218.00
6.245
3.606
202.49
233.51
213
225
1123.00
3.775
2.179
1113.62
1132.38
1119
1127
Total
1665.67
1543.459
514.486
479.26
2852.07
213
3699
5873.00
73.856
42.641
5689.53
6056.47
5792
5936
254.00
6.245
3.606
238.49
269.51
249
261
1355.50
12.933
7.467
1323.37
1387.63
1344
1370
Total
2494.17
2578.895
859.632
511.85
4476.48
249
5936
8265.67
74.969
43.283
8079.43
8451.90
8196
8345
405.67
6.658
3.844
389.13
422.21
398
410
1703.67
11.930
6.888
1674.03
1733.30
1690
1712
Total
3458.33
3649.244
1216.415
653.28
6263.39
398
8345
9229.00
28.710
16.576
9157.68
9300.32
9206
9261
457.00
6.245
3.606
441.49
472.51
450
462
1827.50
9.644
5.568
1803.54
1851.46
1817
1835
Total
3837.83
4086.722
1362.241
696.50
6979.17
450
9261
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim mi trng trn gi th r Medicago-M7
Dependent Variable
(I)
(J)
CT
CT
Bonferroni
Difference (IJ)
S lng bo t 3
Bonferroni
1882.77
1401.000*
11.489
.000
1363.23
1438.77
-1845.000*
11.489
.000
-1882.77
-1807.23
11.489
.000
-481.77
-406.23
11.489
.000
-1438.77
-1363.23
Bonferroni
-444.000
-1401.000*
Bonferroni
444.000
11.489
.000
406.23
481.77
3438.000
18.762
.000
3376.32
3499.68
2533.000
18.762
.000
2471.32
2594.68
18.762
.000
-3499.68
-3376.32
18.762
.000
-966.68
-843.32
18.762
.000
-2594.68
-2471.32
18.762
.000
843.32
966.68
-3438.000
-905.000
-2533.000
905.000
5619.000*
35.468
.000
5502.40
5735.60
35.468
.000
4400.90
4634.10
-5619.000
35.468
.000
-5735.60
-5502.40
-1101.500
35.468
.000
-1218.10
-984.90
-4517.500
35.468
.000
-4634.10
-4400.90
1101.500
35.468
.000
984.90
1218.10
7860.000
35.923
.000
7741.91
7978.09
6562.000
35.923
.000
6443.91
6680.09
-7860.000
35.923
.000
-7978.09
-7741.91
-1298.000
35.923
.000
-1416.09
-1179.91
-6562.000
35.923
.000
-6680.09
-6443.91
1298.000
35.923
.000
1179.91
1416.09
8772.000
14.577
.000
8724.08
8819.92
7401.500*
14.577
.000
7353.58
7449.42
-8772.000
14.577
.000
-8819.92
-8724.08
-1370.500*
14.577
.000
-1418.42
-1322.58
-7401.500
14.577
.000
-7449.42
-7353.58
1370.500*
14.577
.000
1322.58
1418.42
1
3
1
3
S lng bo t 5
3
S lng bo t 4
Upper Bound
1807.23
3
2
Lower Bound
.000
Bonferroni
Sig.
11.489
S lng bo t 2
Error
1845.000*
Std.
2
1
S lng bo t 1
Mean
4517.500

trong th nghim pH mi trng trn gi th r C rt cng sinh vi chng 41833
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
1108.33
25.423
14.678
1045.18
1171.49
1079
1124
1809.00
47.149
27.221
1691.88
1926.12
1776
1863
1025.33
8.327
4.807
1004.65
1046.02
1016
1032
Total
1314.22
373.804
124.601
1026.89
1601.55
1016
1863
2127.33
19.655
11.348
2078.51
2176.16
2115
2150
3453.33
61.647
35.592
3300.19
3606.47
3383
3498
2523.33
17.898
10.333
2478.87
2567.79
2508
2543
Total
2701.33
590.445
196.815
2247.48
3155.19
2115
3498
3147.00
13.748
7.937
3112.85
3181.15
3135
3162
5057.00
43.405
25.060
4949.18
5164.82
5007
5085
4357.33
30.665
17.704
4281.16
4433.51
4322
4377
Total
4187.11
837.300
279.100
3543.51
4830.72
3135
5085
4216.33
16.921
9.770
4174.30
4258.37
4202
4235
8938.33
63.721
36.789
8780.04
9096.62
8892
9011
6164.33
70.571
40.744
5989.02
6339.64
6114
6245
Total
6439.67
2055.654
685.218
4859.55
8019.78
4202
9011
5412.33
6.658
3.844
5395.79
5428.87
5408
5420
10362.67
77.681
44.849
10169.70
10555.64
10305
10451
7138.00
13.528
7.810
7104.40
7171.60
7125
7152
Total
7637.67
2176.428
725.476
5964.72
9310.62
5408
10451
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim pH mi trng trn gi th r C rt-41833
Dependent Variable
(I)
(J)
Mean
Std.
CT
CT
Difference (I-J)
Error
-700.667*
25.555
83.000
1
3
783.667
-83.000

Sig.
Lower Bound
Upper Bound
.000
-784.68
-616.66
25.555
.053
-1.01
167.01
700.667*
25.555
.000
616.66
784.68
25.555
.000
699.66
867.68
25.555
.053
-167.01
1.01
25.555
.000
-867.68
-699.66
S lng bo t 1
Bonferroni
S lng bo t 2
Bonferroni
S lng bo t 3
Bonferroni
S lng bo t 4
Bonferroni
-783.667
-1326.000*
31.647
.000
-1430.04
-1221.96
-396.000*
31.647
.000
-500.04
-291.96
1326.000
31.647
.000
1221.96
1430.04
930.000
31.647
.000
825.96
1034.04
396.000
31.647
.000
291.96
500.04
1
2
31.647
.000
-1034.04
-825.96
-1910.000
25.877
.000
-1995.07
-1824.93
-1210.333
25.877
.000
-1295.40
-1125.26
1910.000
25.877
.000
1824.93
1995.07
1
2
-930.000
699.667
25.877
.000
614.60
784.74
25.877
.000
1125.26
1295.40
1210.333
25.877
.000
-784.74
-614.60
-4722.000
45.527
.000
-4871.67
-4572.33
-1948.000
45.527
.000
-2097.67
-1798.33
4722.000
45.527
.000
4572.33
4871.67
2774.000
45.527
.000
2624.33
2923.67
1948.000
45.527
.000
1798.33
2097.67
-2774.000
45.527
.000
-2923.67
-2624.33
-4950.333
37.303
.000
-5072.96
-4827.70
-1725.667
37.303
.000
-1848.30
-1603.04
4950.333*
37.303
.000
4827.70
5072.96
3224.667
37.303
.000
3102.04
3347.30
1725.667*
37.303
.000
1603.04
1848.30
-3224.667*
37.303
.000
-3347.30
-3102.04
3
1
2
2
-699.667
S lng bo t 5
Bonferroni

trong th nghim pH mi trng trn gi th r C rt cng sinh vi chng M7
95% Confidence Interval for

N
Mean
Std. Deviation
Mean
Std. Error
Lower Bound
Upper Bound
Min
Max
1071.67
14.012
8.090
1036.86
1106.47
1058
1086
2061.00
45.398
26.211
1948.22
2173.78
2025
2112
948.67
30.072
17.362
873.96
1023.37
927
983
Total
1360.44
528.857
176.286
953.93
1766.96
927
2112
2060.33
32.130
18.550
1980.52
2140.15
2024
2085
4030.67
8.622
4.978
4009.25
4052.08
4023
4040
2352.33
40.017
23.104
2252.93
2451.74
2313
2393
Total
2814.44
921.256
307.085
2106.30
3522.58
2024
4040
2953.33
17.616
10.171
2909.57
2997.09
2939
2973
7055.33
12.014
6.936
7025.49
7085.18
7043
7067
4153.00
33.719
19.468
4069.24
4236.76
4124
4190
Total
4720.56
1826.620
608.873
3316.49
6124.62
2939
7067
4041.33
22.301
12.875
3985.93
4096.73
4016
4058
9238.33
31.533
18.206
9160.00
9316.67
9204
9266
6045.67
26.633
15.377
5979.51
6111.83
6023
6075
Total
6441.78
2270.013
756.671
4696.89
8186.66
4016
9266
5206.67
4.163
2.404
5196.32
5217.01
5202
5210
9948.00
30.790
17.776
9871.51
10024.49
9914
9974
7043.33
12.503
7.219
7012.27
7074.39
7031
7056
Total
7399.33
2070.414
690.138
5807.87
8990.79
5202
9974
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim pH mi trng trn gi th r C rt-M7
Dependent Variable
(I)
(J)
Mean Difference
CT
CT
(I-J)
Bonferroni
Bonferroni
S lng bo t 3
Bonferroni
S lng bo t 4
Bonferroni
Bonferroni
.000
-1076.47
-902.19
123.000*
26.506
.011
35.86
210.14
989.333
26.506
.000
902.19
1076.47
1112.333*
26.506
.000
1025.19
1199.47
-123.000
26.506
.011
-210.14
-35.86
-1112.333*
26.506
.000
-1199.47
-1025.19
-1970.333
24.531
.000
-2050.98
-1889.69
24.531
.000
-372.65
-211.35
1970.333
24.531
.000
1889.69
2050.98
1678.333
24.531
.000
1597.69
1758.98
-292.000
24.531
.000
211.35
372.65
-1678.333
24.531
.000
-1758.98
-1597.69
-4102.000
18.807
.000
-4163.83
-4040.17
-1199.667
18.807
.000
-1261.49
-1137.84
4102.000
18.807
.000
4040.17
4163.83
2902.333
18.807
.000
2840.51
2964.16
1199.667
18.807
.000
1137.84
1261.49
-2902.333
18.807
.000
-2964.16
-2840.51
-5197.000
22.116
.000
-5269.70
-5124.30
-2004.333
22.116
.000
-2077.04
-1931.63
5197.000
22.116
.000
5124.30
5269.70
3192.667
22.116
.000
3119.96
3265.37
2004.333
22.116
.000
1931.63
2077.04
-3192.667
22.116
.000
-3265.37
-3119.96
-4741.333*
15.788
.000
-4793.24
-4689.43
-1836.667
15.788
.000
-1888.57
-1784.76
4741.333*
15.788
.000
4689.43
4793.24
2904.667
15.788
.000
2852.76
2956.57
1836.667*
15.788
.000
1784.76
1888.57
2
2
3
1
2
2
3
1
292.000
2
3
Bound
26.506
1
S lng bo t 5
Upper
-989.333
S lng bo t 2
Lower Bound
Sig.
S lng bo t 1
Std. Error
-2904.667*
15.788
.000
-2956.57
-2852.76
Ph lc 13: Kt qu phn tch cc i lng c trng ca s lng bo t trong

th nghim pH mi trng trn gi th r Medicago cng sinh vi chng 41833

N
Mean
Std.
Std.
Deviation
Error
for Mean
Lower
Upper
Bound
Bound
Minimum
Maximum
533.00
22.517
13.000
477.07
588.93
510
555
1632.00
10.817
6.245
1605.13
1658.87
1623
1644
749.00
4.583
2.646
737.62
760.38
744
753
Total
971.33
504.410
168.137
583.61
1359.06
510
1644
1362.33
12.583
7.265
1331.08
1393.59
1349
1374
3354.00
25.159
14.526
3291.50
3416.50
3338
3383
1816.33
11.930
6.888
1786.70
1845.97
1803
1826
Total
2177.56
904.097
301.366
1482.60
2872.51
1349
3383
2469.33
22.030
12.719
2414.61
2524.06
2448
2492
4052.33
51.501
29.734
3924.40
4180.27
4001
4104
3377.33
23.756
13.715
3318.32
3436.35
3350
3393
Total
3299.67
688.602
229.534
2770.36
3828.97
2448
4104
3654.67
45.347
26.181
3542.02
3767.31
3627
3707
7211.33
77.391
44.682
7019.08
7403.58
7122
7258
4997.67
32.332
18.667
4917.35
5077.98
4963
5027
Total
5287.89
1556.118
518.706
4091.75
6484.03
3627
7258
4161.33
22.234
12.837
4106.10
4216.56
4148
4187
8271.33
11.590
6.692
8242.54
8300.13
8258
8279
5735.33
31.565
18.224
5656.92
5813.74
5705
5768
Total
6056.00
1795.972
598.657
4675.49
7436.51
4148
8279
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim pH mi trng trn gi th r Medicago-41833
Dependent Variable
(I)
(J)
CT
CT
Mean
Difference (IJ)
Bonferroni
.000
1530.21
1571.79
1224.000
6.325
.000
1203.21
1244.79
-1551.000*
6.325
.000
-1571.79
-1530.21
6.325
.000
-347.79
-306.21
6.325
.000
-1244.79
-1203.21
Bonferroni
S lng bo t 4
Bonferroni
-327.000
-1224.000*
327.000
6.325
.000
306.21
347.79
3184.500
20.817
.000
3116.07
3252.93
2297.500*
20.817
.000
2229.07
2365.93
-3184.500
20.817
.000
-3252.93
-3116.07
20.817
.000
-955.43
-818.57
20.817
.000
-2365.93
-2229.07
-887.000
-2297.500
20.817
.000
818.57
955.43
3835.000
25.094
.000
3752.51
3917.49
2802.333
25.094
.000
2719.84
2884.83
-3835.000
25.094
.000
-3917.49
-3752.51
-1032.667
25.094
.000
-1115.16
-950.17
-2802.333
25.094
.000
-2884.83
-2719.84
1032.667
25.094
.000
950.17
1115.16
6822.333*
8.928
.000
6792.98
6851.68
8.928
.000
5669.65
5728.35
-6822.333
8.928
.000
-6851.68
-6792.98
-1123.333
8.928
.000
-1152.68
-1093.98
-5699.000
8.928
.000
-5728.35
-5669.65
1123.333
8.928
.000
1093.98
1152.68
7864.000
6.916
.000
7841.26
7886.74
6531.000*
6.916
.000
6508.26
6553.74
-7864.000
6.916
.000
-7886.74
-7841.26
-1333.000*
6.916
.000
-1355.74
-1310.26
6.916
.000
-6553.74
-6508.26
1
3
1
3
887.000
3
S lng bo t 3
Upper Bound
6.325
Bonferroni
Lower Bound
S lng bo t 2
Sig.
1551.000*
Error
2
1
S lng bo t 1
Std.
5699.000
1
S lng bo t 5
Bonferroni
2
3
-6531.000
1333.000*
6.916
.000
1310.26
1355.74
Ph lc 15: Kt qu phn tch cc i lng c trng ca s lng bo t trong

th nghim pH mi trng trn gi th r Medicago cng sinh vi chng M7
95% Confidence
N
Mean
Std.
Std.
Deviation
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
663.00
33.808
19.519
579.02
746.98
624
684
1949.00
22.913
13.229
1892.08
2005.92
1929
1974
822.00
9.000
5.196
799.64
844.36
813
831
Total
1144.67
607.526
202.509
677.68
1611.65
624
1974
1537.33
24.583
14.193
1476.27
1598.40
1509
1553
3656.33
38.850
22.430
3559.82
3752.84
3623
3699
2050.67
37.501
21.651
1957.51
2143.82
2013
2088
Total
2414.78
957.789
319.263
1678.56
3151.00
1509
3699
2589.33
56.146
32.416
2449.86
2728.81
2553
2654
5873.33
73.793
42.604
5690.02
6056.64
5792
5936
3560.33
17.243
9.955
3517.50
3603.17
3545
3579
Total
4007.67
1461.816
487.272
2884.02
5131.32
2553
5936
3934.00
70.235
40.550
3759.53
4108.47
3853
3978
8315.33
82.008
47.347
8111.61
8519.05
8234
8398
4947.33
35.445
20.464
4859.28
5035.38
4923
4988
Total
5732.22
1987.215
662.405
4204.71
7259.73
3853
8398
4342.33
14.640
8.452
4305.97
4378.70
4329
4358
9229.33
28.431
16.415
9158.71
9299.96
9206
9261
5531.33
19.296
11.141
5483.40
5579.27
5516
5553
Total
6367.67
2207.218
735.739
4671.05
8064.28
4329
9261
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5

thc th nghim pH mi trng trn gi th r Medicago-M7
Dependent Variable
(I)
(J)
Mean
Std.
CT
CT
Difference (I-J)
Error
-1286.000
3
Bonferroni
Bonferroni
So luong bo t 3
Bonferroni
S lng bo t 4
Bonferroni
-1221.19
-159.000*
19.715
.001
-223.81
-94.19
1286.000
19.715
.000
1221.19
1350.81
1127.000*
19.715
.000
1062.19
1191.81
19.715
.001
94.19
223.81
Bonferroni
159.000
-1127.000*
19.715
.000
-1191.81
-1062.19
-2119.000
27.968
.000
-2210.94
-2027.06
27.968
.000
-605.28
-421.39
2119.000
27.968
.000
2027.06
2210.94
1605.667
27.968
.000
1513.72
1697.61
-513.333
27.968
.000
421.39
605.28
-1605.667
27.968
.000
-1697.61
-1513.72
-3284.000
44.460
.000
-3430.16
-3137.84
-971.000*
44.460
.000
-1117.16
-824.84
3284.000*
44.460
.000
3137.84
3430.16
2313.000
44.460
.000
2166.84
2459.16
971.000*
44.460
.000
824.84
1117.16
-2313.000*
44.460
.000
-2459.16
-2166.84
-4381.333
53.572
.000
-4557.45
-4205.22
-1013.333
53.572
.000
-1189.45
-837.22
4381.333
53.572
.000
4205.22
4557.45
3368.000
53.572
.000
3191.89
3544.11
1013.333
53.572
.000
837.22
1189.45
-3368.000
53.572
.000
-3544.11
-3191.89
-4887.000
17.607
.000
-4944.88
-4829.12
-1189.000*
17.607
.000
-1246.88
-1131.12
4887.000
17.607
.000
4829.12
4944.88
3698.000*
17.607
.000
3640.12
3755.88
1189.000
17.607
.000
1131.12
1246.88
-3698.000*
17.607
.000
-3755.88
-3640.12
3
1
2
S lng bo t 5
Upper Bound
-1350.81
S lng bao tu 2
Lower Bound
.000
19.715
S lng bo t 1
Sig.
513.333
Ph lc 17: Bng kt qu cc i lng c trng v s lng bo t ca cc

loi gi th r cng sinh vi chng 41833
N
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5
Mean
Std.
Deviation
for Mean
Std. Error
Lower
Upper
Bound
Bound
Min
Max
1.1
1809.00
47.149
27.221
1691.88
1926.12
1776
1863
1.2
282.00
23.431
13.528
223.79
340.21
255
297
2.1
1456.33
38.004
21.942
1361.93
1550.74
1413
1484
2.2
225.00
7.937
4.583
205.28
244.72
219
234
Total
12
943.08
732.759
211.529
477.51
1408.66
219
1863
1.1
3454.00
61.286
35.384
3301.76
3606.24
3384
3498
1.2
646.00
30.050
17.349
571.35
720.65
612
669
2.1
2755.67
46.758
26.996
2639.51
2871.82
2712
2805
2.2
527.00
19.975
11.533
477.38
576.62
510
549
Total
12
1845.67
1341.412
387.232
993.37
2697.96
510
3498
1.1
5057.00
43.405
25.060
4949.18
5164.82
5007
5085
1.2
1066.00
33.719
19.468
982.24
1149.76
1029
1095
2.1
4053.33
28.449
16.425
3982.66
4124.00
4034
4086
2.2
847.00
4.583
2.646
835.62
858.38
843
852
Total
12
2755.83
1917.429
553.514
1537.56
3974.11
843
5085
1.1
8938.33
63.721
36.789
8780.04
9096.62
8892
9011
1.2
1526.67
12.741
7.356
1495.02
1558.32
1512
1535
2.1
7110.33
16.258
9.387
7069.95
7150.72
7092
7123
2.2
1226.33
10.693
6.173
1199.77
1252.90
1217
1238
Total
12
4700.42
3538.592
1021.503
2452.10
6948.73
1217
9011
1.1
10362.67
77.681
44.849
10169.70
10555.64
10305
10451
1.2
2119.33
21.939
12.667
2064.83
2173.83
2102
2144
2.1
8327.67
13.317
7.688
8294.59
8360.75
8313
8339
2.2
1720.33
19.858
11.465
1671.00
1769.66
1706
1743
Total
12
5632.50
3952.807
1141.077
3121.01
8143.99
1706
10451
Ph lc 18: Kt qu phn tch Post Hoc Test v s lng bo t ca cc loi

gi th m r cng sinh vi chng 41833
Dependent Variable
(I)
(J)
CT CT
Upper Bound
1434.09
1619.91
1.1 2.1
352.667*
26.706
.000
259.76
445.57
2.2
1584.000
26.706
.000
1491.09
1676.91
1.1
-1527.000*
26.706
.000
-1619.91
-1434.09
1.2 2.1
26.706
.000
-1267.24
-1081.43
-1174.333
26.706
.392
-35.91
149.91
26.706
.000
-445.57
-259.76
2.1 1.2
1174.333*
26.706
.000
1081.43
1267.24
2.2
1231.333
26.706
.000
1138.43
1324.24
1.1
-1584.000*
26.706
.000
-1676.91
-1491.09
-57.000
26.706
.392
-149.91
35.91
57.000
-352.667
2.1
-1231.333*
26.706
.000
-1324.24
-1138.43
1.2
2808.000
34.748
.000
2687.12
2928.88
1.1 2.1
34.748
.000
577.45
819.22
34.748
.000
2806.12
3047.88
-2808.000
34.748
.000
-2928.88
-2687.12
1.2 2.1
-2109.667
34.748
.000
-2230.55
-1988.78
2.2
119.000
2.2
1.1
1.1
2.1 1.2
2.2
698.333
2927.000
34.748
.054
-1.88
239.88
34.748
.000
-819.22
-577.45
2109.667
34.748
.000
1988.78
2230.55
2228.667
34.748
.000
2107.78
2349.55
34.748
.000
-3047.88
-2806.12
-698.333
1.1
-2927.000
2.2 1.2
-119.000
34.748
.054
-239.88
1.88
2.1
-2228.667
34.748
.000
-2349.55
-2107.78
1.2
3991.000
25.336
.000
3902.86
4079.14
1003.667
25.336
.000
915.53
1091.81
4210.000
25.336
.000
4121.86
4298.14
-3991.000
25.336
.000
-4079.14
-3902.86
1.2 2.1
-2987.333
25.336
.000
-3075.47
-2899.19
2.2
25.336
.000
130.86
307.14
1.1 2.1
Bonferroni
Lower Bound
.000
Bonferroni
Bonferroni
Sig.
26.706
2.2 1.2
S lng bo t 3
(I-J)
Error
1527.000*
1.1
S lng bo t 2
Difference
Std.
1.2
2.2
S lng bo t 1
Mean
2.2
1.1
219.000
1.1
-1003.667*
25.336
.000
-1091.81
-915.53
2.1 1.2
2987.333
25.336
.000
2899.19
3075.47
3206.333
25.336
.000
3118.19
3294.47
25.336
.000
-4298.14
-4121.86
25.336
.000
-307.14
-130.86
25.336
.000
-3294.47
-3118.19
2.2
1.1
-4210.000
2.2 1.2
2.1
1.1
7411.667*
27.693
.000
7315.33
7508.01
2.1
1828.000
27.693
.000
1731.66
1924.34
7712.000
27.693
.000
7615.66
7808.34
-7411.667
27.693
.000
-7508.01
-7315.33
-5583.667
27.693
.000
-5680.01
-5487.33
300.333
27.693
.000
203.99
396.67
-1828.000
27.693
.000
-1924.34
-1731.66
5583.667
27.693
.000
5487.33
5680.01
5884.000
27.693
.000
5787.66
5980.34
-7712.000
27.693
.000
-7808.34
-7615.66
-300.333
27.693
.000
-396.67
-203.99
-5884.000
27.693
.000
-5980.34
-5787.66
1.2
8243.333
34.369
.000
8123.77
8362.90
2.1
2035.000*
34.369
.000
1915.43
2154.57
2.2
8642.333
34.369
.000
8522.77
8761.90
1.1
-8243.333*
34.369
.000
-8362.90
-8123.77
2.1
34.369
.000
-6327.90
-6088.77
2.2
399.000*
34.369
.000
279.43
518.57
1.1
34.369
.000
-2154.57
-1915.43
1.2
6208.333*
34.369
.000
6088.77
6327.90
2.2
6607.333
34.369
.000
6487.77
6726.90
1.1
-8642.333*
34.369
.000
-8761.90
-8522.77
1.2
-399.000
34.369
.000
-518.57
-279.43
2.1
-6607.333*
34.369
.000
-6726.90
-6487.77
1.1
S lng bo t 4
2.1
2.2
Bonferroni
1.1
2.1
1.2
2.2
1.1
2.2
1.2
2.1
1.1
1.2
S lng bo t 5
-3206.333
1.2
2.2
1.2
-219.000
-6208.333
Bonferroni
2.1
2.2
-2035.000
Ph lc 19: Bng kt qu cc i lng c trng v s lng bo t ca cc

loi gi th r cng sinh vi chng M7
95% Confidence
N
S lng bo t 1
S lng bo t 2
S lng bo t 3
S lng bo t 4
S lng bo t 5
Mean
Std.
Deviation
Std.
Error
Interval for Mean

Lower
Upper
Bound
Bound
Minimum
Maximum
1.1
2059.67
47.014
27.144
1942.88
2176.46
2021
2112
1.2
170.00
26.211
15.133
104.89
235.11
141
192
2.1
944.00
39.038
22.539
847.02
1040.98
906
984
2.2
135.00
6.000
3.464
120.10
149.90
129
141
Total
12
827.17
816.863
235.808
308.16
1346.18
129
2112
1.1
4030.67
8.622
4.978
4009.25
4052.08
4023
4040
1.2
534.00
6.000
3.464
519.10
548.90
528
540
2.1
2059.33
18.610
10.745
2013.10
2105.56
2042
2079
2.2
430.00
17.059
9.849
387.62
472.38
411
444
Total
12
1763.50
1524.165
439.988
795.09
2731.91
411
4040
1.1
7055.33
12.014
6.936
7025.49
7085.18
7043
7067
1.2
926.00
21.284
12.288
873.13
978.87
903
945
2.1
3208.00
158.329
91.411
2814.69
3601.31
3102
3390
2.2
743.00
10.536
6.083
716.83
769.17
732
753
Total
12
2983.08
2657.779
767.235
1294.41
4671.76
732
7067
1.1
9238.33
31.533
18.206
9160.00
9316.67
9204
9266
1.2
1341.00
24.062
13.892
1281.23
1400.77
1316
1364
2.1
8265.67
74.969
43.283
8079.43
8451.90
8196
8345
2.2
1075.67
9.292
5.364
1052.59
1098.75
1068
1086
Total
12
4980.17
3957.272
1142.366
2465.84
7494.50
1068
9266
1.1
9948.00
30.790
17.776
9871.51
10024.49
9914
9974
1.2
1918.33
18.610
10.745
1872.10
1964.56
1901
1938
2.1
9229.00
28.710
16.576
9157.68
9300.32
9206
9261
2.2
1534.00
6.245
3.606
1518.49
1549.51
1529
1541
Total
12
5657.33
4117.040
1188.487
3041.49
8273.18
1529
9974
Ph lc 20: Kt qu phn tch Post Hoc Test v s lng bo t ca cc loi

gi th m r cng sinh vi chng M7
Dependent Variable
(J)
Mean
CT
CT
Difference (I-J)
1.2
1889.667*
27.256
2.1
1115.667
2.2
1.1
-1889.667
2.1
1.1
1.2
S lng bo t 1
2.2
.000
1794.85
1984.49
27.256
.000
1020.85
1210.49
1924.667*
27.256
.000
1829.85
2019.49
27.256
.000
-1984.49
-1794.85
-774.000*
27.256
.000
-868.82
-679.18
2.2
35.000
27.256
1.000
-59.82
129.82
1.1
-1115.667*
27.256
.000
-1210.49
-1020.85
27.256
.000
679.18
868.82
27.256
.000
714.18
903.82
27.256
.000
-2019.49
-1829.85
774.000
2.2
809.000*
1.1
-1924.667
1.2
-35.000
27.256
1.000
-129.82
59.82
27.256
.000
-903.82
-714.18
3496.667
11.163
.000
3457.83
3535.50
1971.333
11.163
.000
1932.50
2010.17
3600.667
11.163
.000
3561.83
3639.50
-3496.667
11.163
.000
-3535.50
-3457.83
2.1
-1525.333
11.163
.000
-1564.17
-1486.50
2.2
2.1
1.1
2.1
1.1
11.163
.000
65.17
142.83
1.1
-1971.333
11.163
.000
-2010.17
-1932.50
1.2
1525.333
11.163
.000
1486.50
1564.17
1629.333
11.163
.000
1590.50
1668.17
Bonferroni
11.163
.000
-3639.50
-3561.83
-3600.667
1.2
11.163
.000
-142.83
-65.17
2.1
-1629.333
11.163
.000
-1668.17
-1590.50
1.2
6129.333
65.544
.000
5901.31
6357.35
3847.333
65.544
.000
3619.31
4075.35
6312.333
65.544
.000
6084.31
6540.35
-6129.333
65.544
.000
-6357.35
-5901.31
2.1
-2282.000
65.544
.000
-2510.02
-2053.98
2.2
183.000
2.1
1.1
1.2
2.1
104.000
1.1
2.2
S lng bo t 3
-809.000
2.2
2.2
2.2
Bonferroni
1.2
1.2
1.2
Sig.
Upper Bound
2.1
1.1
Std. Error
Lower Bound
Bonferroni
2.1
S lng bo t 2
(I)
-104.000
65.544
.141
-45.02
411.02
1.1
-3847.333
65.544
.000
-4075.35
-3619.31
1.2
65.544
.000
2053.98
2510.02
2282.000
2.2
2465.000*
65.544
.000
2236.98
2693.02
1.1
-6312.333*
65.544
.000
-6540.35
-6084.31
2.2
1.2
-183.000
65.544
.141
-411.02
45.02
2.1
-2465.000
65.544
.000
-2693.02
-2236.98
7897.333
34.833
.000
7776.15
8018.51
972.667
34.833
.000
851.49
1093.85
8162.667
34.833
.000
8041.49
8283.85
-7897.333
34.833
.000
-8018.51
-7776.15
-6924.667
34.833
.000
-7045.85
-6803.49
265.333
34.833
.000
144.15
386.51
-972.667
34.833
.000
-1093.85
-851.49
6924.667
34.833
.000
6803.49
7045.85
7190.000
34.833
.000
7068.82
7311.18
-8162.667
34.833
.000
-8283.85
-8041.49
-265.333
34.833
.000
-386.51
-144.15
-7190.000
34.833
.000
-7311.18
-7068.82
8029.667
18.963
.000
7963.70
8095.64
2.1
719.000*
18.963
.000
653.03
784.97
2.2
8414.000
18.963
.000
8348.03
8479.97
1.1
-8029.667*
18.963
.000
-8095.64
-7963.70
2.1
18.963
.000
-7376.64
-7244.70
2.2
384.333*
18.963
.000
318.36
450.30
1.1
-719.000
18.963
.000
-784.97
-653.03
1.2
7310.667*
18.963
.000
7244.70
7376.64
2.2
7695.000
18.963
.000
7629.03
7760.97
1.1
-8414.000*
18.963
.000
-8479.97
-8348.03
1.2
-384.333
18.963
.000
-450.30
-318.36
2.1
-7695.000*
18.963
.000
-7760.97
-7629.03
1.2
1.1
2.1
2.2
1.1
1.2
Slng bo t 4
2.1
2.2
Bonferroni
1.1
2.1
1.2
2.2
1.1
2.2
1.2
2.1
1.2
1.1
1.2
S lng bo t 5
-7310.667
Bonferroni
2.1
2.2

Ảnh Hưởng Của Môi Trường Và Giá Thể Mô Rễ Đến Khả Năng Nhân Sinh Khối Cộng Sinh Nấm Rễ AM (Arbuscular Mycorhiza) in Vitro

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Ảnh Hưởng Của Môi Trường Và Giá Thể Mô Rễ Đến Khả Năng Nhân Sinh Khối Cộng Sinh Nấm Rễ AM (Arbuscular Mycorhiza) in Vitro

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VIN KHOA HC V CNG NGH VIT NAM

VIN SINH THI V TI NGUYN SINH VT

Chuyn ngnh: VI SINH VT HC

LUN VN THC S SINH HC

Ngi hng dn khoa hc: TS.L QUC HUY

S ha bi Trung tm Hc liu i hc Thi Nguyn

S ha bi Trung tm Hc liu i hc Thi Nguyn

S ha bi Trung tm Hc liu i hc Thi Nguyn

1.2.2. Mc tiu c th........

1.3. ngha khoa hc v thc tin ca ti..

1.3.1. ngha khoa hc.

1.3.1. ngha thc tin.

1.4. Phm vi nghin cu... 2

1.1.2. c im ca Nm r ni cng sinh AM(Arbuscular mycorrhiza).

1.1.3. Vai tr ca nm r ni cng sinh vi cy ch..

1.2. Tng quan v vi khun Agrobacterium rhizogense

1.3. Nghin cu nm r ni cng sinh trn Th gii v Vit Nam..

1.3.1. Trn th gii ...

Chng 2. VT LIU - NI DUNG - PHNG PHP NGHIN CU 22

2.2. Ni dung nghin cu.... 23

2.2.2. nh gi nh hng ca mi trng nui cy n kh nng nhn sinh

2.2.3. nh gi nh hng ca pH mi trng nui cy n kh nng nhn

2.2.4. nh gi nh hng ca gi th m r n kh nng nhn sinh khi

2.3.1. Phng php b tr th nghim...

2.3.2. Phng php to vt liu m r in vitro 24

2.3.4. Phng php to cng sinh AM in vitro

2.3.5. Phng php nhn sinh khi cng sinh AM in vitro ..

2.3.6. Phng php thu thp, phn tch v x l thng k s liu th

3.2.nh gi nh hng ca mi trng nui cy n nhn sinh khi cng

3.3. nh gi nh hng ca pH mi trng nui cy n nhn sinh khi cng

3.4. nh gi nh hng ca gi th m r n nhn sinh khi cng sinh AM

Chng 4. KT LUN - TN TI - KIN NGH..... 50

S ha bi Trung tm Hc liu i hc Thi Nguyn

BNG NHNG T VIT TT TRONG LUN VN

Indole butylic acid

Murashige and Skoog medium

Strullu and Romand medium

Polymerase chain reaction

Root inducing transfer Deoxyribonucleic acid

Ribosomal Ribonucleic acid

trypton-yeast extract medium

Vesicular arbuscular mycorrhiza

S ha bi Trung tm Hc liu i hc Thi Nguyn

Bng 3.5: nh hng ca pH mi trng nui cy n nhn sinh khi cng

Bng 3.6: nh hng ca pH mi trng nui cy n nhn sinh khi cng

Bng 3.7:nh hng ca pH mi trng nui cy n nhn sinh khi cng

Bng 3.8: nh hng ca pH mi trng nui cy n nhn sinh khi cng

Bng 3.9: nh hng ca cc loi gi th m r khc nhau n nhn sinh

Bng 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh

S ha bi Trung tm Hc liu i hc Thi Nguyn

Biu 3.2: nh hng ca mi trng nui cy n nhn sinh khi cng

Biu 3.4: nh hng ca mi trng nui cy n nhn sinh khi cng

Biu 3.5: nh hng ca pH mi trng nui cy n nhn sinh khi

Biu 3.6: nh hng ca pH mi trng nui cy n nhn sinh khi cng

Biu 3.9: Kt qu nhn sinh khi AM in vitro ca 41833-C rt Ri-tDNA,

Biu 3.10: nh hng ca cc loi gi th m r khc nhau n nhn sinh

Biu 3.12: nh hng ca cc loi gi th m r khc nhau n nhn sinh

Biu 3.13: Kt qu nhn sinh khi AM in vitro ca 4 loi gi th m r

S ha bi Trung tm Hc liu i hc Thi Nguyn

S ha bi Trung tm Hc liu i hc Thi Nguyn

S ha bi Trung tm Hc liu i hc Thi Nguyn

Nhm gp phn gii quyt cc vn tn ti nu trn ca nghin cu ng

Chng 1: TNG QUAN VN NGHIN CU

Da trn c im xm nhim ca nm vo r cy ch, mycorrhiza c phn