B GIO DC V O TO

TRNG I HC PHNG NG

n tt nghip
Khoa: Cng Ngh Sinh Hc v Mi Trng
ti:
S dng ch th phn t trong vic xc nh ging mang gen khng
bnh mc sng phc v cho cng tc chn to ging khoai ty
GVHD

: TS. L Hng Lnh

SV

: Th L Hng

MSSV

: 510301006

NI DUNG N
CHNG 1
TNG QUAN V CY KHOAI TY
CHNG 2
VT LIU V PHNG PHP
CHNG 3
KT QU V THO LUN

Mc tiu v ni dung nghin cu

Mc tiu:
To c ging khoai ty mang gen khng bnh
mc sng, phc v cho cng tc chn to ging.

Ni dung:
nh gi vt liu b m v s dng ch th phn
t trong vic xc nh ging mang gen khng bnh
mc sng phc v cho cng tc chn to ging
cy khoai ty.

Tn khoa hc Solanum tuberosum L.

Gii : Plantae.
B : Solanales.
H : Solanaceae.
Chi : Solanum.
Loi : S.tuberosum.

L loi cy lng thc ngn ngy v cha nhiu cht dinh

dng
C ngun gc t Nam M v sau c trng kh ph bin
trn Th Gii.

1.1. B phn ca cy khoai ty

Hoa mu trng
pht tm, qu
mng hnh cu
.

Thn cy
khoai ty c 2
loi l thn b
v loi thn
ng.

R khoai ty phn b ch
yu tng t su 30cm,
phnh to ln thnh c.

1.2. Tnh hnh sn xut khoai ty Vit

Nam
Khoai ty c nhp vo Vit Nam nm 1890.

T nm 1980, khoai ty c quan

tm v c ti nghin cu cp
Nh nc m Vin Khoa hc v k
thut Nng nghip Vit Nam l c
quan ch tr. Nh vy, nng sut
khoai ty c nng cao, trc
thng l 8 tn/ha, cao nht l 18-20
tn/ha.
T nm 1981 n nay, nng sut
bnh qun t gn 12 tn/ha, cao nht
t 35-40 tn/ha.

1.3. Bnh mc sng hi khoai ty

Nguyn nhn: Bnh do nm phytophthora thuc b sng mai,
lp nm to khun gy ra.

Bnh ph hoi tt c cc b
phn l, thn, cnh, c v k
c lc ang tn tr.

1.4. Nghin cu v chn ging khoai ty

khng bnh mc sng Vit Nam
Theo nh gi mt vng ngoi thnh H Ni nhng nm
1965, thit hi do bnh mc sng gy ra cho rung trng khoai
ty trung bnh t 30 70% gy mt nng sut hon ton cho v
khoai ty.
Nhiu bin php phng tr v cha bnh cho cy khoai ty
nhng u khng mang li hiu qu nh mong mun.
Vi s tin b ca cng ngh sinh hc, nh vic kt hp ch
th phn t vi phng php chn ging truyn thng, cc nh
khoa hc d dng chuyn np cc gen khng bnh vo cc
ging mi cho nng sut, cht lng tt, gip tit kim thi gian
v chi ph nghin cu

1.5. ng dng ch th phn t trong nghin cu di

truyn v chn to ging khoai ty khng bnh mc
sng.
Gene khng bnh mc sng u tin c lp bn v
gii trnh t khoai ty l R1 trn nhim sc th s 5.
Vit Nam, vic ng dng ch th phn t trong chn to
ging khoai ty khng bnh mc sng c rt t cng trnh
nghin cu. Vic chn ging khng trn khoai ty trong
nc ch yu c tin hnh theo phng php truyn
thng.

Chng 2:Vt liu v phng php

2.1. Vt liu nghin cu
Vt liu thc vt gm: 9 ging khoai ty :
Ging 11-106; Ging 11-113; Ging 11-115;
Ging 11-116; Ging 11-119; Ging 11-129;
Ging 11-140; Ging Solara v Ging KT3.
Cc cp mi : ch th phn t LP2, LP3

Bng 1
Tn ch th

Loi ch th

phn t

phn t

Trnh t

Chiu di

Nhit

(bp)

ngn mi
(0C)

LP2

SNP

F: CAA TTG ATT CAT TTT ATG TAG CGA G

R: TCT TGA CGC AAA CCT CTG CGA G

510

56

LP3

SNP

F: GTA GTA CAT CAA CAT ACA TTT TGC GG

R: CTC AGA ATT CAG AGC TTC AAC TGA TG

510

58

Bn phn t locus gen R1 v v tr ca cc ch th phn t

Hnh 1
LP4
LP6
STI 0006

STI 0032
STI 0038
STI 0049

STM 1056

LP2

STM 5146

LP3

STM 5149
STPoAc58
STPoAc85
Pota1
Pota3
Pota6

Pota8
Pota6
Pota
11
Pota 12
Pota 14
Pota 16
Pota 20

R1

2.2. phng php nghin cu

Phng php nghin cu trong phng th nghim

- Tch DNA tng s

- Kim tra nguyn vn ca DNA thu c
- Xc nh nng DNA v kim tra cht lng
DNA(kim tra nguyn vn)
- Tin hnh phn ng PCR
- Phn tch kt qu PCR bng k
thut in di trn gel polyacrylamide

Phng php nghin cu ngoi ng rung

- Chun b t trng v lm t
- Chun b khoai ging
- Gieo trng
- Bn phn
- Bn thc
- Ti nc

My soi gel

My in di

My nghin mu

My PCR

Chng 3: Kt qu v tho lun

3.1. nh gi mc nhim bnh mc sng ca cc ging
khoai ty trng ti Sa Pa- Lo Cai (ngy trng 3/4/2013, ngy
thu hoch 16/7/2013)
Bng 2

TT

1
2
3
4
5
6
7
8
9

Vit Nam
code
11-106

11-113
11-115
11-116
11-119
11-129
11-140
solara
KT3

Ngy sau trng

35

42

49

56

63

70

77

84

10

15

10

15

20

30

10

25

10

10

20

10

10

15

20

40

10

15

23

50

3.2. Nng sut v cc yu t cu thnh nng sut ca

cc ging khoai ty c trng ti Sa Pa Lo Cai
Bng 2
S c
Vit Nam code

/khm

T l c c (%)
Nng sut c
(tn/ha)

>50mm

30-50mm

<30mm

11-106

2,7

6,50

11

85

11-113

3,9

10,25

15

78

11-115

4,7

13,50

17

78

11-116

4,5

9,50

34

50

16

11-119

2,9

5,50

18

64

18

11-129

3,9

14,00

86

11-140

3,1

8,25

36

55

Solara

6,5

16,00

16

78

KT3

4,8

18,00

56

38

3.3. Kt qu nh gi mc nhim bnh ca khoai ty ngoi ng rung

ti Trung tm Nghin cu v Pht trin cy c C - Vin Cy Lng
thc v Cy Thc phm, Thanh Tr- H Ni (ngy trng 30/10/2013,
ngy thu hoch 20/1/2014)
Bng 3
Ging

30 ngy sau mc

TT

Mc sng
Ho
Virus (%)
xanh (%)

11-129

33,3

11-113

3,3

11-116

26,7

11-106

1,7

11-140

35,0

11-115

45,0

Solara

5,7

KT3

5,0

60 ngy sau mc

45 ngy sau mc

m l
(1-9)

(1-9)

Ho
Virus (%)
xanh (%)

Mc sng
(1-9)
m l
(1-9)

xanh (%)

33,3
0

1,7
5,0

35,0
1

45,0
0

1,7

36,0
0

26,7

1,7
0

3,3

26,7
0

m l
(1-9)

33,3

3,3
0

Ho
Virus (%)

Mc
sng
(1-9)

45,0
0

1,7
5,0

3.4. Nng sut v cc yu t cu thnh nng sut ca cc ging

khoai ty ngoi ng rung ti Thanh Tr- H Ni, nm 2013 .
Bng 4

TT

Ging

S c/
khm

T l c c ( %)

Nng
sut
(tn/ha)

>5 cm

3-5 cm

<3 cm

11-129

7,2

15,83

11,0

54,8

34,2

11-113

4,9

9,00

8,4

47,4

44,2

11-116

5,0

19,42

30,3

52,6

17,1

11-106

6,4

15,71

13,5

58,4

28,1

11-140

12,6

26,25

13,1

63,4

23,5

11-115

9,4

14,67

9,8

60,7

29,5

Solara (/c)

11,5

21,08

10,1

57,6

32,3

3.5. Kt qu nghin cu xc nh ch th phn t lin

kt gen khng bnh mc sng ca cc ging khoai ty
Kt qu tch chit v tinh sch DNA tng s
Hnh 2

Bc tip theo, chng ti tin hnh xc nh nng v

tinh sch ca cc mu DNA thu c sau khi tch bng
phng php quang ph k.
Bng 5
STT

Tn mu

Nng (ng/l)

Ch s tinh sch A260/A280

11-106

908,7

1,90

11-113

1096,1

1,92

11-115

1269,6

1,92

11-116

849,5

1,91

11-119

731,7

1,88

11-129

1114,8

1,92

11-140

782,0

1,80

Solara

699,1

1,85

KT3

977,1

1,90

3.6. Nghin cu xc nh ch th phn t lin kt locus

gen khng bnh mc sng
xc nh ch th phn t lin kt locus gen R1 khng bnh
mc sng trong th nghim ny, chng ti chn lc nhng cp
mi SNP t kt qu nghin cu cng b trn tp chi quc t v cc
bn lin kt gen cy khoai ty cng b xc nh ging
mang gen khng bnh mc sng R1.
Kt qu chng ti xc nh v s dng ch th phn t SNP,
gm c ch th LP2 v ch th LP3 y l nhng ch th phn t thit
k c hiu da trn trn trnh t cc Nucleotic ca gen khng bnh
mc sng.

3.7. Kt qu xc nh gen khng bnh mc sng R1 trn 9

ging khoai ty bng ch th phn t LP2

K hiu
1: Ging 11-47; 2: Ging 11-106; 3: Ging 11-113; 4: Ging 11-115;
5: Ging 11-116; 6: Ging 11-119; 7: Ging 11-124; 8: Ging 11-129; 9:
Ging 11-140; 10: Ging Solara; 11: Ging KT3.

Kt qu nh gi phn tch di truyn tnh khng bnh mc

sng
Bng 6
Ch th phn t

Ch th phn t

LP2

LP3

11-106

11-113

11-115

11-116

11-119

11-129

11-140

Solara

KT3

Dng/ging

KT LUN
Sau khi kt thc nghin cu ti nh gi
c cc c im nng sinh hc, kh nng khng
bnh mc sng ca ging khoai ty tham gia th
nghim c trng trong nh li ti Trung tm
nghin cu cy c c ti Sa Pa Lo Cai v khi c
trng vi iu kin ngoi ng rung ti Thanh Tr
H Ni. Chng ti thu c kt qu:
C 4 ging khoai ty chu bnh mc sng tt c
th s dng lm vt liu khi u cho chn to ging
l : ging 11-113, ging 11-116, ging 11-106 v ging
11-129.

KT LUN
V nng sut : cc ging khoai ty trng ngoi ng
rung ti Thanh Tr H Ni ( dao ng trong khong
9,00 tn/ha- 26,25 tn/ha )cho nng sut cao hn hn so vi
cc ging khoai ty c trng trong nh li ti Sa Pa
Lo Cai ( t 5,00 tn/ha- 18,00 tn/ha ).
V kich thc c : cc ging khoai ty trng ngoi ng
rung ti Thanh Tr H Ni c kich thc c >5cm ( 8,4 30,3% ) t hn so vi cc ging khoai ty trng trong nh
li ti Sa Pa Lo Cai ( 9-56% ).
V s lng c/khm : cc ging khoai ty trng trong
nh li ti Sa Pa Lo Cai ( 2,7 6,5 c/khm) t hn so
vi cc ging khoai ty trng ngoi ng rung ti Thanh
Tr H Ni ( 4,9 12,6 c/khm).

KT LUN
ti xc nh c ch ch th phn t LP2 lin kt gen
R1 khng bnh mc sng v xc nh c cc ging Ging
11-106, Ging 11-115, Ging 11-116, Ging 11-129, Ging 11140 l nhng ging mang gen R1 khng bnh mc sng .
i vi ch th phn t LP3 xc nh c 3 ging: Ging 11106, Ging 11-116, Ging 11-119 l nhng ging mang gen R1
khng bnh mc sng.

 NGH
Cn c tip tc nhiu cng trnh nghin cu v ti ny hn
na.
Trong th nghim ny ti ch mi s dng c 2 ch th phn
t vo vic xc nh lin kt gen khng bnh mc sng ca mt
s ging. Trong thi gian ti ti s tin hnh th nghim thm
nhiu ch th phn t hn na phc v cho cng tc chn to
ging khoai ty khng bnh mc sng.