Ti liu chia s ti: wWw.SinhHoc.edu.

vn I HC THI NGUYN
Khoa khoa hc t nhin v x hi ------------ ------------

Nguyn Th Kiu Lin

Nghin cu h thng ti sinh

cy c chua (Lycopersicum esculentum

L.) phc v chuyn gen

Bo co ti khoa hc Chuyn ngnh: Di truyn hc Gio vin hng dn: ThS. Nguyn Th Hi Yn

Thi nguyn, Thng 6 - 2008

Ti liu chia s ti: wWw.SinhHoc.edu.vn

NI DUNG bo co

M u Vt liu v phng php nghin cu Kt qu nghin cu v tho lun Kt lun v ngh

Ti liu chia s ti: wWw.SinhHoc.edu.vn M U

L do chn ti

C chua l ngun thc phm quan trng trong i sng thng ngy ca con ngi y l loi rau n qu c gi tr dinh dng v gi tr kinh t cao, l loi thc phm d ch bin v s dng. Nng sut v cht lng c chua thng b gim ng k do c chua l loi cy trng d b su bnh gy hi. Vic to ra nhng ging c chua c kh nng khng su bnh, nng cao nng sut, cht lng l rt cn thit. K thut chuyn gen l mt trong nhng bin php hu hiu to ra ging mi. Xut pht t nhng l do trn chng ti tin hnh ti: "Nghin cu h thng ti sinh cy c chua (Lycopersicum esculentum L.) phc v chuyn gen"

Ni dung nghin cu

Tm hiu nh hng ca mt s cht kch thch sinh trng ln kh nng ny mm ca ht. Tm hiu kh nng ti sinh t thn mm.

Ti liu chia s ti: wWw.SinhHoc.edu.vn

Vt liu v phng php

2.1. Vt liu
Ht ging ca ging c chua bi (cn gi l c chua ta hay c chua kiu - Lycopersicum esculentum Mill var. Cerasiforme Alef).

Hnh cy c chua dng trong th nghim.

Ti php s ti: wWw.SinhHoc.edu.vn 2.2. Phngliu chianghin cu

2.2.1. X l mu
- Chn nhng qu c chua chn , khng b su bnh. Lc ly ht, loi b nhng ht ni, phi kh. - Kh trng ht bng cn 700 trong mt pht v javen 60 % trong 20 pht.

2.2.2. Mi trng nui cy

Mi trng MS (Murashige and Skoog 1962) c b sung ng sucrose 15g/l i vi mi trng ny mm v 30g/l i vi mi trng ti sinh: agar 9g/l v cc cht kch thch sinh trng nng thay i tu mc ch th nghim Qu trnh nui cy c tin hnh trong iu kin c chiu sng 10h/ngy nhit khong 25-300C

Ti liu chia s ti: wWw.SinhHoc.edu.vn

2.3.2. Phng php b tr th nghim

* Th nghim 1: nh hng ca BAP ln kh nng ny mm ca ht. * Th nghim 2: nh hng phi hp ca BAP v kinetin ln kh nng ny mm ca ht. * Th nghim 3: nh hng phi hp ca BAP v -NAA ln kh nng ny mm ca ht. * Th nghim 4: nh hng ca BAP ln kh nng ti sinh t thn mm.

2.3.3. Cc ch tiu nh gi

Ti liu chia s ti: wWw.SinhHoc.edu.vn

Tng s ht ny - T l ny mm (%) = mm Tng s ht cy vo mi tr100 ng

Tng s mu to chi
T l to Tng s mu cy vo mi chi (%) trng =

*100

Ti liu chia s ti: wWw.SinhHoc.edu.vn

Kt qu nghin cu v tho lun

3.1. nh hng ca mt s cht kch thch sinh trng ln kh nng ny mm ca ht
3.1.1. nh hng ca BAP Bng 3.1. nh hng ca BAP ln kh nng ny mm ca ht
K hiu C1-0 Nng ) S ht cy S ht ny T l (%) BAP(mg/l) vo mm 0 45 27 60

C1-1
C1-2 C1-3 C1-4 C1-5

1
2 3 4 5

45
45 45 45 45

26
23 35 24 26

57.8
51.1 77.8 53.3 57.8

Ti liu chia s ti: wWw.SinhHoc.edu.vn

90 80 70 60 60 50 40 30 20 10 0 C1-0 C1-1 C1-2 C1-3 C1-4 C1-5 tl(%) 57.8 51.1 53.3 57.8 77.8

Biu 3.1. nh hng ca BAP ln kh nng ny mm ca ht

Hnh 3.1. Ht ny mm mi trng C1-0

Ti liu chia s ti: wWw.SinhHoc.edu.vn

3.1.2. nh hng phi hp ca BAP v kinetin

Bng 3.2. nh hng phi hp ca BAP v kinetin ln kh nng ny mm ca ht
K hu Nng BAP(mg/l) Nng Kinetin(mg/l) S ht cy vo S ht ny mm T l (%)

C2-0 C2-1 C2-2

0.2 0.2 0.2

0 0.5 1

45 45 45

25 26 29

55.6 57.8 64.4

C2-3
C2-4 C2-5

0.2
0.2 0.2

1.5
2 2.5

45
45 45

23
24 24

51.1
53.3 53.3

C2-6

0.2

45

17

37.8

Ti liu chia s ti: wWw.SinhHoc.edu.vn

70 60 50 40 30 20 10 0 C2-0 C2-1 C2-2 C2-3 C2-4 C2-5 C2-6 t l(%) 37.8 55.6 57.8 51.1 53.3 53.3 64.4

Biu 3.2. nh hng phi hp ca BAP v kinetin ln kh nng ny mm ca ht

Hnh3.2. Ht ny mm cc mi trng C2

3.1.3. nh hng phi hp ca BAP v -NAA

Bng 3.3. nh hng phi hp ca BAP v -NAA ln kh nng ny mm ca ht
Khiu Nng - NAA (mg/l) Nng BAP(mg/l) S ht cy vo S ht ny mm T l (%)

Ti liu chia s ti: wWw.SinhHoc.edu.vn

C3-0 C3-1 C3-2 C3-3 C3-4

0.2 0.2 0.2 0.2 0.2

0 1 2 3 4

45 45 45 45 45

17 23 26 24 21

37.8 51.1 57.8 53.3 46.7

C3-5

0.2

45

17

37.8

Ti liu chia s ti: wWw.SinhHoc.edu.vn

70 60 51.1 50 40 30 20 10 0 C3-0 C3-1 C3-2 C3-3 C3-4 C3-5 37.8 57.8 53.3 46.7 37.8

t l(%)

Biu 3.3. nh hng phi hp ca BAP v -NAA ln kh nng ny mm ca ht

Hnh3.3. Ht ny mm cc mi trng C3

Ti liu chia s ti: wWw.SinhHoc.edu.vn

3.2. nh hng ca kch thch sinh trng ln kh nng ti sinh cy

Bng3.4. nh hng ca BAP ln kh nng ti sinh cy K hiu R1 R2 R3 R4 R5 Nng BAP (mg/l) 0 0,25 0,5 0,75 1 T l mu to chi (%) 80,0 55,5 86,6 80,0 50,0 S chi c to thnh 19 15 18 27 15

Ti liu chia s ti: wWw.SinhHoc.edu.vn

Biu 3.4. nh hng ca BAP ln kh nng ti sinh cy Hnh 3.4. Cc mu ti sinh t mi trng R3

1.

Kt lun Cc nng BAP khc nhau trong mi trng nui cy c

nh hng khc nhau ln t l ny mm ca ht c chua. Trong nng 3mg/l BAP trong mi trng MS cho t l ny mm cao nht (77.8%).

Ti KT LUNwWw.SinhHoc.edu.vn liu chia s ti: V NGH

2.

Nng phi hp 0.2 mg/l BAP v 1mg/l kinetin trong mi trng MS cho t l ny mm cao nht (64.4%).

3.

S phi hp ca BAP v -NAA nng 0.2 mg/l -NAA v 2 mg/l BAP trong mi trng MS cho t l ny mm cao nht (57.8%). Mi trng R3 ( MS + 0.5mg/l BAP) l mi trng cho t l mu to chi cao nht

4.

Mi trng R4 (MS+ 0.75mg/l BAP) l mi trng thch hp cho vic ti sinh t thn mm c chua

Ti liu chia s ti: wWw.SinhHoc.edu.vn

ngh
1. Kho st kh nng ti sinh t l mm. 2. Tm hiu nh hng ca tui thn mm v l mm ln kh nng ti sinh. 3. Tip tc nghin cu v hon thin quy trnh ti sinh thnh cy hon chnh phc v cho vic chuyn gen thng qua vi khun Agrobacterium tumfaciens sau ny.

Ti liu chia s ti: wWw.SinhHoc.edu.vn

XIN CHN THNH CM N !