Nghiên cứu ứng dụng nanochitosan trong phòng trừ bệnh thán thư hại ớt sau thu hoạch

I H C QU

H
IH

NGUY

U

NG

NG D NG NANOCHITOSAN TRONG

B

I

th c ph
: 605402

LU

TP. H

CS

T SAU THU HO CH

u ng

ng d n khoa h c: TS. Tr

cv
ch m nh

..............................................

cv
ch m nh
cv

Lu

cb ov t

ih

nH

m:
cv c aH

ng ch m b o v lu

1.

............................................................

2.

............................................................

3.

............................................................

4.

............................................................

5.

............................................................
n c a Ch t ch H

ng Khoa qu
c s a ch a (n


p-

-

NHI M V LU
H

Nguy

ng

MSHV: 12110193

23/02/1985
Ngh An
th c ph

u ng

: 605402

u ng d ng nanoch

b

t sau thu ho
II. NHI M V

I DUNG:

- T o ch ph m nanochitosan b
Sodium tripolyphosphat (STTP).
-

nh ng

tri n c a n m
tri n c a n m
-

nh c a n

nh h

o gel ion gi

t.

ng c a ch ph

n s sinh tr

u ki n in vitro.
nh h ng c a ch ph
u ki n in vivo.

n s sinh tr

hi u qu b o qu n t b
a t sau thu ho ch

: 20/06/2014
V.

i


TS. Tr


i

L IC

c r t nhi u s

c

L
TS. Tr

nh s c g ng c a b
y

il ic
, tr c ti

su

c hi n lu

l i nh
h

n
ng nghi p, b n

ng d n cho em trong
o nh

u ki n thu n

t lu

nh kh e,

p nhi u may m

Em xin
ngh ,

m vui trong cu c s ng.

n Th
ng d

ch b o

h c t

th c hi

i

.
Con xin c

, anh ch

nh con h c t p t

tr

ih

trong su
H

K

n t p th

ngh th c ph m, Khoa K thu

p th
y
n d t, ch d y cho

em ki n th

i c n thi
c t p, th c hi
cs
c kh
c nhi u ni m vui trong s nghi p tr ng

ic
il ic
ngh th c ph

tr

c hi

h

th c ph
c t p, sinh ho t t

Th y
nh c
tr

ng.

TP. H
c hi n
Nguy

ng


ii

T LU

S d

i thu

c trong b o qu n rau qu
ng s ng. Vi

ngu n g c t

ch

n s c kh e

d ng m t ch ph m sinh h
gi m thi u t n th t sau thu ho

o gel ion v

n

cs d

Colletotrichum gleoesporioides
nanochitosan 0,03%

ng x

t.

c ch

c ch 50% (EC50) s

m

n ym mc

n khu n l

u ki n in vitro, n ng
n m; n

ng PDA t i 0,04%.

u

ki n in vivo, m
0

. Kh

c

p ch t
-1,3-

u.


iii

ABSTRACT

Using chemical pesticides in vegetable reservation made adversely affect to human
health and the environment. Finding and using a natural biological product, which is
non-toxic to minimize the post-harvest lost, are being concerned. In this study,

nanochitosan was formed by gel ion method with sodium tripolyphosphate, used as
an antifungal agent against Colletotrichum gleoesporioides causing anthracnose on
red chilli. In vitro, nanochitosan concentration of 0,03% inhibited over 50%
germination of fungal spore; the concentration inhibiting growth by 50% of colony
on PDA was 0,04%. In vivo, nanochitosan coating at concentration of 0,4% had
well effective in the preservation of red chilli. Results in this study show that
physiological and biochemical quality of red chilli were insignificant changed after
12 days stored at 25oC compared with red chilli before storage. The antifungal
stimulation ability of nanochitosan by increased such compounds as polyphenol,
chitinase and -1,3-glucanase was studied.


iv

L
uc ab
k t qu
ngu

t lu n trong lu
b t k m t ngu
ib tk
c th c hi
u.

o. Vi c tham kh
u tham kh

lu

Nguy

ng


v

M CL C
Trang
M

U .................................................................................................................... 1
p thi t c

.......................................................................................... 1

2. M

u ............................................................................................... 2

3. N

n c u ............................................................................................... 2
c ti n................................................................................. 3
c ............................................................................................. 3

4.2.

c ti n.............................................................................................. 3
U ...................................................................... 4

1.1. Gi i thi u v

t ................................................................................................ 4
m sinh v t h c c

t ........................................................... 4
c a t ..................................................... 5

1.1.3. M t s lo i t ph bi n .......................................................................... 6
n xu
nh sau thu ho
1.2. T ng quan v

gi i ................ 7
t .............................................................. 8

m c a Colletotrichum gloeosporioides

i t sau thu ho ch ............................................................................................ 10
1.2.1.
1.2.2.
1.2.3. S
1.3. Bi

c

c c a ch ng n m C. Gloeosporioides .. 11
c ........................................................................................... 12
n

........................................ 12
nh sau thu ho

t ............ 15

1.3.1. Bi

a ........................................................................ 15

1.3.2. Bi

............................................................................ 15
........................................ 18
.......................................................................... 18
nochitosan.................................................................. 19
..... 25


vi

.............................................. 28
1.5. Nh

c li

........................ 30

1.5.1.

........................................................ 30

1.5.2.

c ........................................................ 31
U ....................... 33
u .................................................................................... 33

2.1.1.

u

...................................................................................... 33

2.1.2. N m Colletotrichum gloeosporioides .................................................. 33
2.1.3. Ch ph m chitosan

t o nanochitosan ..................................... 33

u ............................................................................... 34
t

m............................................................. 34
a t ................ 40
................ 43
.................................................................... 45
.............................. 46

3.1. K t qu
ionic ........................................................................................................................... 46

3.2. Kh

i t c a nanochitosan

u ki n in

vitro ........................................................................................................................... 48
3.2.1.

ng c

gloeosporioides
3.2.2.

n

n n m Colletotrichum

ng PDA ..................................................................... 48
ng c

n sinh kh i s i n m Colletotrichum

gloeosporioides ......................................................................................................... 51
3.2.3. Kh

c ch c

ns n ym mc a

n m

Colletotrichum gloeosporioides ................................................................................ 54
3.3. Kh

h i t c a nanochitosan

u ki n in

vivo ............................................................................................................................ 57
3.3.1. K t qu

nh c a n m Colletotrichum

gloeosporioides ........................................................................................................ 57


vii

3.3.2. Kh

i t c a nanochitosan

u ki n in

vivo ............................................................................................................................ 59
u qu b o qu n t sau thu ho ch b ng nanochitosan ................ 64
............................. 65
.......................... 68
............................... 70

a nanochitosan

t .............................. 72

3.5.1. S

ng polyphenol t ng s ......................................... 72

3.5.2. S

................ 75

K T LU

N NGH ............................................................................ 81

1. K t lu n ............................................................................................................. 81
2. Ki n ngh ........................................................................................................... 82
3

u tham kh o ............................................................................................ 83


viii

DANH M C B NG BI U
Trang
B ng 1.1.
B ng 1.2. S
B ng 1.3.

B ng 1.4.

n

cc a

ng

gi i g m
bi

c ...................................... 5
................................. 8

............................................................................. 9

c ch E. coli, S. choleraesuis
........................................................................................ 27

B ng 2.1.

a chitosan .............................................................. 33
Colletotrichum

gloeosporioides. ........................................................................................................ 48

C. gloeosporiodes ...................................................................................................... 54
.......................... 57
............. 58

.......................................................................................................... 60
.............. 61


ix

DANH M
Trang
M t c t ngang qu
.B

t ................................................................................. 5
t ................................................................................ 10

i th

c a ch ng n m C.gloeosporioides ................. 12
...................... 13
............................ 14
p tr c ti p c

t

n m qua b m

n b ng

................................................................................................................... 14
C

chitosan, chitin ............................................................ 18
....................................................... 21

1.9.

t o gi t b

1.10.

o gi t/k t t a ................................... 21
p nh t gi

......................................... 22

t o nanochitosan b

t

......................................................................................................................... 23
t o nanochitosan b
3.

y phun ........................... 23

t o h t nanochitosan b

o gel ionic ................ 24
........................................ 27

nh ch

nt

FE

SEM c

ph m

nanochitosan ............................................................................................................. 46
ng c

n n m C. gloeosporioides sau

y ............................................................................................................ 49
ng c

n sinh kh i s i C.gloeosporioides sau

y ............................................................................................................ 51

........................................................................ 52
.5.

ng c a n

n m C.
............................................................................... 55

x

Vt b

o v

n m

............................................ 59

............................................................................ 62
ng c

n kh

o qu n

... 64
nhi

H

nhi
S

cc a

S
nhi

o qu n

ng ch t r

a

nhi

25oC ..... 65
25oC ... 66
25oC ...... 68
o qu n

25oC ............................................................................................................. 69

nh 3.13. S

a

o qu n

nhi t

25oC ...................................................................................................................... 69
S bi

ng ethylen s n sinh c a t sau

o qu n

nhi

25oC ............................................................................ 71

ng c
o qu n
6.
o qu n

nhi
ng c
nhi
ng c

o qu n

nhi

t sau 10
25oC ................................................................................. 73
n ho

t sau 8

25oC ................................................................................. 76
n ho

t sau

25oC .............................................................................. 76


xi

DANH M

VI T T T

C. gloeosporioides: Colletotrichum gloeosporioides
CS:

Chitosan

CT:

c

DD:

(Degree of Deacetylation)
i ch ng

h:

Gi

PDA:

Potato Dextrose Agar

PDB:

Potato Dextrose Borth

STPP:

Sodium Tripolyphosphat

PIRG:

T l ph

c ch t

nl c

(Percentage Inhibition of Radial Growth)
AUDPC:

ng cong ti n tri n b nh
(Area Under Disease Progress Curve)

EC50:

N

c ch 50% s

n

(The concentration inhibiting growth by 50%)
MIC:

N

c ch t i thi u 90% s

ns in m

(The minimum concentration showing over 90% inhibition of
mycelial growth)
MBC:

N

di t khu n t i thi u

(Minimum Bactericidal Concentration)
SEM:

nt

GLU:

Glucanase

CHI:

Chitinase

(Scanning Electron Microscope)

DNS:
FAOSTAT:

D li u Th

a T ch

(Food and Agriculture Organization Corporate Statistical
Database)
DNA:

Deoxyribo Nucleic Acid

mRNA:

messenger Ribo Nicleic Acid

LMW:

Kh

th p (Low Molecular weight)

HMW:

Kh

cao (High Molecular weight)

TLB:

T l b nh


xii

CFU:

S

khu n l c (Cell Forming Unit)

MAP:

B

POD:

Peroxidase

PPO:

Polyphenol oxidase

u bi n (Modified Atmosphere Packaging)

1

M

U

p thi t c
N m trong khu v

u nhi

t trong nh ng

n ph m rau qu
ti t
ng
lo i rau qu

gi

u ki n th i
ng
i [9].

r t thu n l i cho vi c tr
ng
r ng cho s n ph

ph c v th
rau gia v

t kh u.
c tr ng ch y u

kinh t

B . Nh
t v i di

c ch bi n

ts t
n nh m cung c

t lo i
nh mi

ng b

c ph

u tr ng
n xu t

t kh

i l i nhu n cao [10].

c tr ng t mang l i hi u qu kinh t
n th t sau thu ho

nh t. B
B

t ph bi

c bi
i v i t, b
ch. V t b

ph n ranh gi i gi
Nh ng v t b

i

u nhi
nhi u lo i qu

th

qu

tr ng i l n nh t
ng n ng n
c ta.
i,

h

y
ng,

qu

y theo v t b nh.

tv

th
m Colletotrichum capsici,

Colletotrichum nigrum, Colletotrichum
[1].
gloeosporioides
t
thu ho ch n ng n nh
t h i l n v kinh t [62].
h n ch b nh
i thu
c s d ng [10], [45].
th

n s c kh
t ch ph
u qu

i rau qu sau thu ho

c h n ch t n
e (TBZ)

cs d

i thu
nv

ng,

ng ct
t sau thu ho

t th c.

Colletotrichum

u c n thi t


2

Chitosan, s n ph m deacetyl c
h i v i nhi
sinh

t polymer t

c quan tr
d
t s lo i qu sau thu ho

n

t

ng m i trong vi
i
m t l n v i nhi u

u qu

Xu

nh

c nhi

k t qu r t kh quan [6], [39], [58]. Hi n nay, m
hi u qu ng d ng ch ph
c t 20m di
2

c

t tr

nh ng lu n c
i mong mu
n m b nh h i t sau thu ho
u ng d

u so v i

xu t m t gi
c hi
b

i t sau

i

t sau thu ho ch c a

thu ho
2. M

u
-

nh kh

nanochitosan.
-

ng d

b

3. N

i t.

u

- T o ch ph m nanochitosan b
Sodium tripolyphosphat (STTP).
-

nh ng

tri n c a n m
tri n c a n m
ch

nh c a n

nh h ng c a ch ph
u ki n in vitro.
nh h

ng c a ch ph

u ki n in vivo.
u qu b o qu n t b ng
a t sau thu ho ch

o gel ion gi

t.
n s sinh tr

n s sinh tr

3

c ti n
c
+ Cung c p d li u khoa h c v kh
ph m nanochitosan.
+ Cung c p d
nanochitosan.
4.2.

li u khoa h c v

i t c a ch

kh

a ch

ph m

c ti n
xu

m b o an t

o qu n t sau thu ho ch s d ng ch ph m nanochitosan
c ph

nv

ng


4

U

1.1. Gi i thi u v

t

m sinh v t h c c
t

t

s

thu n l i
c
25-280

n
n
u

i. Nhi
ng 180C.

th i k

ng trong vi
mc
tri

ng l
n, n

ng

n ch

u th p
r

[18].

ng d
su

t tr ng, d

t ch u
t ch m tr

thu ho
8-

u ki n che r
u qu , r ng n
tt
-

-7. V
tr ng t trong v

-

tr

p nhi
c gieo tr ng
-

-

-7, tr
-3,

-8 [1], [14].

R :

c

ts r

tri

r c a t kh

c nhi

r c c
ng cho

n.
ng m c th
ng

, d c theo chi
ng cao 125-135 cm, chi

u ki

ng
-5 c nh.

ng.
c xo
n b u d c, phi

ng m
n

xanh nh t ho c

m.

Hoa:
hoa h
ti t m

-3 hoa. Hoa nh
t th ph n hay th ph

ng ho


5

Qu

h d ng r

m t qu

th ph ng ho c g
ho
[2].

Khoang qu

Th t qu
H t

M t c t ngang qu

t

tr c a t
T

t

t ph n trong m th c c

t lo i gia v t o

c bi
d ng r
vitamin c n thi t, nhi

gi

c tr

nh vi c cung c p m
ng v y h c c a
cc a
cc a

h
B ng 1.1.

i theo gi
u ki
c th hi n trong b ng 1.1.
cc a

n

c ng d ng ph bi n

c [1]

ng

n

ng

m

85,7g

P

80mg

Protein

2,9g

Fe

1,2mg

0,6g

Na

6,5mg

Ch

1,0g

K

2,7mg

Cacbonhydrat

3,0g

S

34mg

6,8g

Cu

1,55mg

Thiamin

0,19mg

Ca

30mg

Riboflavin

0,39mg

Vitamin A

292mg

Axit oxalic

67mg

Vitamin C

111mg

Ch

Ch


6

. Trong qu
c bao g m ch t d u d

protein, ch t s
ng quan tr

ch a nhi u ch t

t. Nhi

n trong qu
c. Qu

gi m nhi m x
vitamin E [77].
Trong th t qu

a m

ng capsaicin (C18H27NO3

t lo i

cay, t o c
ch

t. S

capsaicin

t ng gi

ch

gi

cay n

i c a
y h c, ho t

cs
u ch

c a

[1].

1.1.3. M t s lo i t ph bi n
t (Capsicum sp.) thu c h
t ph bi
Trong s

(Solanaceae), chi Capsicum.

t cay (Capsicum frutescens L.)
ng thu c h
(Solanaceae)

ch

t ng t (Capsicum annuum L.).
m quan tr ng th hai

Qu
8], [13].

tT

Chi Capsicum bao g m kho ng 20 -

c thu
c

tr ng

nhi

m t trong nh
[78].
t ng

n th gi i. Trong s
c tr ng nhi u nh

Capsicum

c tr ng, C. annuum

gi i k ti

C. frutescens

c tr ng nhi u

cs d

c
t lo i rau xanh ho

ch bi n

[70].
c tr ng ph bi n
cs d
t ng t [70].

.

c nhi
kinh t

i

ng ph bi n


7

ng 50 gi ng
d
[10].

ts

ir
tc

t ch

t hi m, t ng t...

n xu

gi i

n xu

c

c ta hi n nay
p trung ch y u

c tr ng
nh mi

kh

c

Trung

Nam. Tuy
ng Tr , Th a

t s t nh Mi n B
n
ng b
c tr ng ph bi

ng

cm r
[14].

ts

t ch

i t

t hi m,

Theo s li u th
a T ng c c H i quan trong n
2007, kim ng ch xu t kh u t c a c
Hi n nay,
Trung Qu
ng xu t kh u t cao nh t v i 40% t ng kim ng ch xu t kh u,
i Loan v i kim ng ch xu t kh u l
c

t chi
ng xu t kh u c

ng t. M t s
Qu nh Ph

i di

ho ch

ng

ch b

c bi
kinh t c

n xu
Xu
tr ng ph bi
quan tr ng th 2 ch

canh

ha
i L c (Qu ng Nam) v i
nh) v i 253 ha, B Tr ch (Qu ng
c, Ch M
i

i 350 ha, m t s huy
2.146 ha v
ts t

ki
c

nv

nh h i t sau thu
t.

gi i
ng, hi u qu kinh t
gi i. Trong h
(Solonaceae),
c tr ng r

c bi

ts

c
m
gi i,
c,

8].
c s n xu
ut n

gi
ng s

ng 25,9 tri u t n cho
ng t c a c th gi

,


8

26.056.900 t n.

t (Capsicum sp.)

th c v t quan tr

ng th

gi i [77].

Hi n Trung Qu c v

cd

s
ng t c a Trung Qu
ti
th hi n qua b ng 1.2.
B ng 1.2. S

cx p h

u trong vi c s n xu

t,

ng th nh
gi i trong nhi
S
ng t c a m t s

ng

n,
c

gi

gi i g m
:t n)

Qu c gia

2004

Trung Qu c

12.031.031

12.530.180

13.031.000

14.033.000

Mexico

1.431.258

1.617.264

1.681.277

1.690.000

Indonesia

1.100.514

1.058.023

1.100.000

1.100.000

1.700.000

1.829.000

1.842.175

1.090.921

1.077.025

1.063.501

1.074.100

1.065.000

978.890

959.070

998.210

855.870

720.000

721.000

721.500

723.000

467.433

460.000

470.000

475.000

24.587.124

25.261.259

26.252.907

26.056.900

Nigeria

2005

nh sau thu ho
Theo c m nang chu
ph bi
b

2006

2007

t
Vi
c xu t b n b i
p Qu c t Australia (ACIAR) th
nh

t g m: b nh th i r Phytophthora, th i g
tuy

- B nh th i r Phytophthora do n m Phytophthora capsici, tri u ch
ir

[4].

Tri u ch ng c a b
b r ng. Ti
Trong nhi
[13].

ng xu t hi n mu
mn

ng h p, ch

ch t.
c m t ph n c

u ch


9

- B nh th i g c hay b
ch
thu ho ch. N

cu

ab

c m c tr ng (Sclerotium rolfsii): Tri u

c th hi
tt
n th
sau v t b nh lan r

c, lan r ng xu ng t n c r
ig
nt

- B
Colletotrichum sp. B

t, v t b

im

u nh

t b nh d n d n b

t do n m
h

i ch y

uv tb
qu sau 2-

m nh

m tv

cv t b

t b nh
i gi

ch y d c theo v t b
nh ng n

m tv tb
tb
n. N

th i ng n t. Ch i h
d n ho

nn n
t ng ph

nh do n m
b
c ab
tuy

nh ng ch m nh
tv
t s ch

n do vi khu
t hi n d ch khu n
cv tb
Meloidogyne sp.

b th i, v
ng
ch t
[10], [19].

b nh do vi khu
Ralstonia solanacearum

u ch ng

bi
n. B

[4].

B ng 1.3.

a

bi

[10].