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vn I HC THI NGUYN
KHOA KHOA HC T NHIN V X HI B MN SINH HC
TI SINH VIN NGHIN CU KHOA HC

NGHIN CU A DNG CC GING CH TRNG THI NGUYN BNG K THUT SINH HC PHN T

Gio vin hng dn: ThS. Hong Th Thu Yn Sinh vin thc hin: Nguyn Th Thu Phng

Thi Nguyn 2008

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Ni dung bo co

M u Tng quan ti liu i tng v phng php nghin cu Kt qu nghin cu v tho lun Kt lun v kin ngh

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Cy ch Vit Nam ng gp mt phn khng


nh vo s pht trin t nc theo hng cng nghip ha - hin i ha Cy ch cn gp phn gi gn bn sc vn ho dn tc. Ung ch tr thnh loi hnh ngh thut trong ngh thut m thc Vit Nam Ch l mt dc liu qu trong dn gian. ch cha mt s bnh ng rut, li tiu, bnh tim, bo ph, ung thv nhiu loi vitamin v axitamin cn thit cho c th

M U

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Vic tm ra nhng bin php y mnh nng


sut v nng cao nng lc cnh tranh l vn cn thit vi ngi trng ch, cn c nhiu nghin cu su rng v cy trng ny Sinh hc phn t l mt cng c h tr chnh xc trong vic phn tch di truyn chn ging Hin nay, vic nh gi h gen cy ch vn cn l vn kh mi m. lm r hn vn ny chng ti tin hnh thc hin ti: Nghin cu a dng cc ging ch trng Thi Nguyn bng k thut sinh hc phn t .

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Ni dung nghin cu ti bao gm

Thu mu l ch. Tch chit DNA tng s. o quang ph DNA Nghin cu tnh a hnh ca genome ch bng k thut RAPD. Phn tch sn phm RAPD bng phn mm sinh hc phn t.

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Tng quan ti liu

1. Khi qut v cy ch 1.1. Ngun gc ca cy ch Vn Nam-Trung Quc. Vng Atxam thuc n Vit Nam.

1.2. S phn loi cy ch


Ngnh
Lp

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Hn kn
Song t dip

Angiospermae
Dicotylleonae

Ch

Theales

Ch

Theaceae

Chi

Ch

Camellia

Loi

Camellia

sinensis

1.1.2. c im sinh hc ca cy ch

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H r bao gm: r tr, r bn, r hp thu. R tr c


th di n 2m nhng thng ch di n 1m.

Cy ch c thn thng v trn. Thn ch lm 3 loi:


thn bi, thn g, thn g nh .

Cnh ch mc t chi dinh dng. Trn cnh chia


thnh nhiu t cnh, bin i t 1-10cm

Ch c hai loi mm chnh l mm dinh dng v


mm sinh thc

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L ch l loi l n, nguyn, c h gn l rt r, c
nhiu hnh dng khc nhau: thun, mi mc, van,
trng g v gn trn.

Qu ch thuc loi qu nang, c t 1- 4 ht; hnh


trn, tam gic, vung

Ht c ng knh 10 - 15 mm, trng lng trung


bnh l 1,0 - 1,6g, hnh cu, tam gic, bn cu

Hoa ch l loi lng tnh, c 5 - 9 cnh mu trng

PHNG PHP V NGUYN LIU

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2.1. Nguyn liu

Mu l ch ly t nng trng ch ca cng ty ch


Sng Cu, huyn ng H, tnh Thi Nguyn

Cc ging ch c cung cp: PH1, keo Tam tch v


trung du l nh - Trung Quc, Iakatamidori Nht Bn , LDP2 Vit Nam, PT95- , Tri2024 v Tri777 Srilanca

2.2. Cc ho cht v thit b

Cc ho cht: ethidium bromid, phenol, Cc primer


dNTP, Buffer Taq DNApolymerase

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Cc thit b
My li tm T m

Sn xut
Hettich Germany Germany

T lnh My in di
My o quang ph My chy PCR My soi DNA My chp nh Gel - Doc B n nhit T - 200C L vi sng

Janpan USA
Phermo Electriction Eppendorf - USA Wealtea USA Applied BioSciences Jeiotech Italy Shorp

Ti liu chia s cu 2.3. Quy trnh nghin ti: wWw.SinhHoc.edu.vn Nguyn liu l ch Tch chit v thu nhn genome tng s in di kim tra kt qu Kim tra tinh sch ca DNA bng my quang ph

Tin hnh k thut RAPD


Thit lp mi di truyn gia cc ging

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2.3. Cc phng php nghin cu

Phng php tch chit DNA tng s Phng php o quang ph trn my quang ph t ghi Phng php in di Phng php RAPD Phng php x l s liu bng phn mm NTSYS 2.0

KT QU V THO LUN
3.1. Kt qu tch chit DNA tng s
1 2 3 4 5 6 7 8
1. LDP2 2. PH1 3. TDLN 4. Iakatamidori 5. Tri777 6. Keo Tam Tch 7. tri 2024 8. PT 95

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Ti liu chia s ti: wWw.SinhHoc.edu.vn 3.2. Kim tra tinh sch ca DNA bng my quang ph

Hnh 1: Ph hp th DNA t ging Tri 2024

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Hnh 2: Ph hp th DNA t ging Iakatamidori

Ti liu chia s + Kt qu o tinhti: wWw.SinhHoc.edu.vn sch DNA

A 260 nm= OD260nm v A 260nm= OD280nm ; tnh t l

A = OD260nm/OD280nm
Ging LDP2
A 1.589

PH1
1.981

TDLN Iakatamidori Tri777


1.941 1.734 1.223

Tri2024
1.752

PT95
1.239

KTT
1.788

+ Kt qu nng DNA ca cc mu thu c nh sau


Bng 1: Nng ca DNA tch chit c
Ging PH1 LDP2 TDLN Iakatamidori Tri777 Keo tam tch Tri2024 PT95 pha long 400 400 400 400 400 200 400 400 Gi tr OD260nm 0.46 0.336 0.179 0.370 0.031 0.254 0.308 0.022 Nng DNA (g/ml) 9200 6720 3580 7400 620 5080 6160 440

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Ti liu chia s ti: wWw.SinhHoc.edu.vn 3.3. Kt qu phn tch a hnh bng k thut RAPD

+ Mi RA32
1. LDP2

2. PH1
3. TDLN 4. Iakatamidori 5. Tri777 6. Keo Tam tch 7. tri 2024 8. PT 95

Hnh 3. Hnh nh in di sn phm RAPD vi on mi RA32 ca 8 ging

Ti liu + Mi RA40 chia s ti: wWw.SinhHoc.edu.vn

1.LDP2 2. PH1

3. TDLN
4. Iakatamidori 5. Tri777

6.Keo Tam tch


7. tri 2024 8. PT 95

Hnh 4. Hnh nh in di sn phm RAPD Vi on mi RA40 ca 8 ging ch

Bng 3: BngTi liu chia son wWw.SinhHoc.edu.vn thng k phn ti: DNA c nhn bn ngu nhin trong phn ng RAPD vi 2 mi ngu nhin ca 8 ging ch M G RA32 1,5 (kb) 1,25 1,0 8,5 8,0 0,7 0,64 0,6 0,5 0 1 1 1 1 0 1 1 1 0 1 1 1 1 0 1 1 1 0 0 0 1 1 1 1 0 1 0 0 0 1 0 0 0 1 1 0 0 0 0 0 0 0 0 1 0 0 0 1 1 0 0 1 1 0 1 1 1 1 0 1 1 1 1 0 1 0 0 1 0 0 0 1 2 3 4 5 6 7 8

0,2
Tng

0
7

0
7

0
5

0
3

0
1

0
4

0
7

1
4

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M G

RA40 1,5 (kb) 1,0 0,9 0,8 0,7 6,6 1 1 0 1 1 1 0 1 0 1 1 1 0 1 0 1 1 1 1 1 0 0 1 0 1 1 1 0 1 1 0 1 0 1 1 1 1 1 0 0 1 0 0 1 1 1 1 1

0,5
0,3

0
0

0
0

0
0

1
0

0
1

0
0

1
1

1
1

0,2
0,18 Tng

0
1 6

0
0 3

1
0 5

0
0 4

0
0 6

0
0 4

1
0 6

1
1 9

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Bng 4: S phn on DNA xut hin v s phn on DNA a hnh i vi mi mi


Mi Phn on DNA nhn bn Phn on DNA a hnh T l % phn on a hnh

RA32 RA40

7 9

7 7

100% 78%

Tng s phn on DNA ngu nhin l 16 v c kch thc t 0,18 1.5 kb. Trong c 14 phn on DNA a hnh chim 87,5%, 2 phn on DNA khng a hnh chim 12,5%.

3.4. Mi quan h di truyn gia cc ging ch trng Thi Ti liu chia s ti: wWw.SinhHoc.edu.vn Nguyn da trn phn tch RAPD * Kt qu h s tng ng di truyn gia cc ging ch
1 1.000 0.900 0.650 0.600 0.500 0.750 0.700 0.300 1.000 0.750 0.600 0.500 0.850 0.700 0.300 1.000 0.550 0.550 0.800 0.550 0.450 1.000 0.700 0.750 0.700 0.300 1.000 0.650 0.500 0.500 1.000 0.550 0.350 1.000 0.300 1.000 2 3 4 5 6 7 8 1.LDP2 2. PH1 3. TDLN 4. Iakatamidori 5. Tri777 6. Keo Tam tch 7. Tri 2024 8. PT 95

Bng 5. H s tng ng gia cc ging ch

S ph h ca 8 ging ch trng Thi Nguyn.


1.LDP2 2. PH1 3. TDLN 4. Iakatamidori 5. Tri777 6.Keo Tam tch

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7. tri 2024
8. PT 95

Hnh 5. S quan h di truyn gia cc ging ch

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KT LUN V KIN NGH

Kt lun
1. tch chit c DNA ca 8 ging ch vi hm lng DNA kh cao. 2. Kt qu nhn c sau khi nghin cu tnh a hnh DNA c nhn bn ngu nhin chng t s sai khc nhau trong cu trc h gen ca cc ging ch. 3. Khi s dng 2 mi ngu nhin cho thy c 2 mi u biu hin tnh a hnh vi t l cc phn on dao ng t 14,3% - 90%. 4. H s di truyn gia cc ging dao ng t 0,3 0,9

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Kin ngh
1. S lng mi ngu nhin s dng trong nghin cu a hnh cc ging ch cn c nhiu hn. 2. Cn kt hp nhiu phng php sinh hc phn t khc: SSR (Simple sequence region), RFLP, xc nh trnh t DNA cng vi k thut RAPD c kt qu chnh xc trong nghin cu a dng ging ch.

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Xin CHN THNH CM N !

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