NGHIEN
CQU
KHA NANG
SINH
TONG HOP
lAA
CUA CHUNG
RH
NGUYEN
THI
KIM QUY,
NGUYEN
THI HOAI
HA, PHAM
VAN
TY
Trung
tarn
Cong nghe Sinh hge, DHQG, Ha Ngi
1.
Md
dau
Vice
sir
dung phan bdn vi sinh se lam tang giau dit mot cich ed
hieu qua va do ton kem ban vice su dung
cac loai
phan hda hge, nhiing
loai
cd the gay hai cho mdi trudng vi
lim

vu cho vice
thiic
diy sinh trudng ciia cay,
xir
ly
miii hdi. Cic vi sinh vat cd trong phan bdn sinh hge con tiet mot sd
chit
kich thieh
sinh trudng, chit hoi hge ehdng nam, ciing nhu lam
tang su nay mim eua hat vi su sinh trudng ciia
xh.
Vice
sir
dung phan
bdn vi sinh se lam tang giau dit mot cich ed hieu qui va dd ton kem
ban
viee sir
dung cic loai phan hoi hge,
nhflng
loai cd the
gay
hai cho
mdi trudng va lim can kiet cic ngudn nang lugng khdng the tii tag.
Vi vay viec nghien
ciiu,
tuyen cbgn vi sinh vat cd boat tinh cao,
vdi cic
ehiie
nang
khae

"Nghien eifu
khd ndng sinh tong hgp
lAA
eua ehung Rh".
2. Nguyen lieu va
phi/dng
phap
2.1. VI sinh vat
Chiing vi khuan phan lap tir dit dugc
giiJ
vi bio quin tai Bao ting
Gidng
chuan
Vi sinh vat, Trung tam Cdng nghe Sinh
hge
- Dai
hge
Qudc gia Hi Ngi.
2.2. Trinh
ti/cac
doan mdi
dCing
trong nghien cuu
27f 5' - agagtttgatcctggctcag - 3'
780f
5'
- GATTAGATACCCTGGTAG -
3'
920r
5'-GTCaaTrCCTTTGAGTTT-3'

phuong phap sac ky ban mong
Thieu hut boocmon sinh trudng
li
van de ma
cay
trdng
thudng
gap
phii.
Dieu niy cd the khic phue dugc neu bd sung vao dit
viing
rd
cic chiing vi khuan cd boat
tinh
sinh cac chit kich
thich
sinh
trudng cao.
169
c
•a,
<
<
B
X
26
2
il
I
16

viing
tiep
xiic
vdi be mat
Idng
hiit,
do dd, cay
trdng cd the su dung mot cich cd hieu qui.
Trude
day ngudi ta cho
ring khi nang kich thich sinh trudng ciia nhdm vi khuin niy la do
qui trinh cd dinh nita.
Nbung sau dd bing thue nghiem ngudi ta da cbiing minb ring ngay
trong nhiing dieu kien mi qui trinh cd dinh nita khdng
di6n
ra vin xiy
ra su
kich
thich sinh trudng thue vat. Ket qui cho thiy ea 10 ehung
(Hinh 1) deu cd kha nang sinh
lAA
rd ret. Tuy nhien, him lugng
lAA
ciia cic chiing vi khuan sinh ra la khdng gidng nhau
13-27mg/l.
Dae biet cd cbiing ky hieu Rh ed khi nang tdng hgp
lAA
him
lugng dat
tir

nudi ciia chiing Rh vi (Hinh 2) cbiing
bieu hien khi rd bang boat tinh khi
nang sinh
lAA
tuang
ling
vdi chim
lAA
chuin. Tien hinh cao vet d vi tri
sd 3 li ciia chiing Rh.
Cbiet miu va tinh sach miu bing
^*''*
lOOfil
metanol trong dng nghiem cd
niit 1234
xoay. Lge qua ban
Igc
0,45|am.
Ly tam
Hinh
2.
Xac dinh
lAA ciia
chiing
lanh
4"C
d 10
OOOvdng/phiit
trong Rh bang sac ky ban mong
20phut.

1:1
ml
lAA,
dng II (dd'i
chiing):
1
ml
nude
cat. Them vao mdi dng 2ml thud'c
thit
Van Urk Salkowski (0.5M
FeCl^
1ml, 35%
HCIO4
50ml) phan irng
dugc
lie
deu. Dung dich cd miu
hong nhat sau 15-30
pbiit.
Thdi gian
(phiil)
Hinh 3. Hinh hap phu
lAA ciia
chiing Rh tren bieu d6 HPLC.
171
Chi sd OD dugc dd'i
chi6'u
vdi phuong trinh dd thi chuin de
tinh

nhau ve hinh thii ciing cd the khic nhau rit rd ret ve cic dac diem
sinh ly, sinh boa hge. Vi vay, nhieu vi sinh vat dugc xep vao cac nhdm
sinh ly khic nhau khdng phu
'thude
vio vi tri phan loai ma phu
thude
vao
tac
dgng ciia chiing
len
mot
-cbit
nao dd ciia ea chit hoae vao ban
chit eua cic sin pham trao ddi chit dugc tao ra. Do vay, 2 ehung HA4
va Rh cung khdng nim ra khdi qui
luat
ehung dd. Hai chiing niy dugc
nudi tren cic mdi trudng dac trung cho tirng loii nhu da md ti d phin
phuong phip.
- Chiing Rh cd hinh que kich thude
0,5-0,9x1,20-2,0|j,m,
pbd bien
la da hinh trong nhirng dieu kien bit
Igi.
Thudng
ehiia
nbung hat
poly-|3-hydroxybutyrat,
nhiing hat nay
khtic

(X7.500)
Can
cii
vio cic dac diem hinh thii (Hinh
),
sinh ly, sinh hoi, ddng
dua vao
khoi
phan loai Bergeys [66], ehung tdi xic dinh cd kha nang
cbiing Rh thude chi Sinorhizobium.
De khing dinh
cbinb
xac den loai ehung Rh
thude
loii nao trong
chi Sinorhizobium chiing tdi tien hinb tich cbiet ADN
ciia
cbiing Rh
ddng thdi tien hinb phin
ling
PCR.
1
M
"2"
1500 -2000 bp
Ket
time
qud trinh phdn
ling
sau

16S. Chiing tdi
173
dung cac cap mdi thdng dung va pbd bi6'n ma cac phdng
thi
nghiem
trfin
the gidi vin dang dung, bao gdm cic doan 27F, 780F, 980R va
1525R.
Diing
ehuang trinh Blast search ket qui phan tich trinh tu cho
thiy doan gen dugc tdng hgp eua Rh ed do dii li 493bp day la
nhtrng
doan gen bao tdn va cbgn lge trong qui trinh
diing
4 mdi cho phan
tich trinh tu song song. Khi so sinh Rh vdi trinh tu gen ma hoi cho
16S da dugc dang ky trong ngan hing dir lieu gen Nhat Ban. Do tuang
ddng rit cao dat 97% vdi trinh tu
ciia
ehung thude loai Sinorhizobium
xinjiangensis vdi ky hieu AJ420774, day la nhirng chi so tuang ddng
so sanh dugc phep chip nhan. Tit cac thdng sd thu dugc eua
Sinorhizobium xinjiangensis Rh se lam co sd va cin tbiet cho viec lap
so dd
cay
phi he (Hinh 7).
Mdi quan he gin giira chiing
xinjiangensis
Sinorhizobium xinjiangensis Rh
vdi 14 trat tu nucleotit thude

Rh
S.xlnjlangenslsAF250354
Rh.melilotiD12783
Rh.fredilD12792
S.fredliD14516
- S.kummerowlaeAF364067
S.morelenseA Y559079
S.morelenseA Y024335
H
Hinh 7. Cay pha he dua tren phan tich
trinh
tu rADN 168 ciia chiing Rh va cac
loai
CO
quan he
gdn
174
Vdi muc dich nghien
eiiu
phdi trdn bdn hgp cic cbimg vi khuan
trong che pham phan
hiJu
ca da
chiie
nang, thi khdng the khdng nghien
ciiu
ky dac tinh
ciia
tirng cbiing dugc.
Do vay,

Tai lieu tham khao
1.
Hoben H. J., Somasegaran P., Handbook for
Rliizobia,
Springer-
Verlag, New York, pp. 1-6.
2.
Holt J. G., N. R. Krieg, P. H. A. Sneath, J. T. Staley, and S. T.
Williams (1994), Bergey's Manual of Systematic Bacteriology,
Williams and Wilkins. Baltimore, Maryland, pp. 235-242.
3.
Lynch, J. M. (1990), "Some consequences of microbial
175
Rhizosphere competence for plant and soil". In The
Rhizosphere, ed. J. M. Lynch, pp. 1-10.
4.
Michalczuk L., Ribnicky D. M., Cooke T. J. and Cohen J. D
(1992),
"Regulation of indole-3-aeetic acid biosynthesis in
carrot cell
cultures".
Plant Physiol, (100), pp. 1346-1353.
5.
Ribnicky D. M., Cohen J. D., Hu W. S. and Cooke T. J. (2002),
"An auxin surge following fertilization in carrot: Its
significance for plant totipotency", Planta, (214), pp. 505-509.
6. Ridge R. W. and B. G.
Rolfe
(1985),
''Rhizobium