BO NONG NGHIEP VA PHAT TRIEN VIEN KHOA HQC VA CONG NGHE
NONG
THON VIET
NAIM
CHUONG TRINH UD CNSH TRONG NONG NGHIEP
BAO CAO TONG
HOP
KET QUA KHOA HQC CONG NGHE
DU
AN
Hoan thien cong nghe siin xuat che pham vi sinh vat doi khang
phdng benh thoi re cho cay ca phe, bong vai viing Mien Trung,
Tay Nguyen.
Co'
quan chii tri Du
an:
Lien hiep KHSX Cong nghe Sinh hoc va Mdi trudng,
Vien Cdng nghe sinh hoc, Vien Khoa hpc va Cdng nghe Viet Nam
Chii
nhiem Du
an:
TS. Pham Viet Cudng
Dien thoai:
04.37568261
Fax:
04.37568261
Co"
quan chii quan : Vien Cdng nghe Sinh hoc
Ha Noi
-12/2009

THANH
Ha Noi - 12/2009
Lien hiep KHSX CNSH & MT
Du an san xuat thu nghiem
DANH SACH CAC CAN BO THAM GIA THUC HIEN
DU
AN
1
2
3
4
5
6
7
8
9
Ho va ten
Nguyen Thi Kim
Ctic
Pham Viet Cudng
Pham
Due
Thuan
Truang Ba Hiing
Le Thi Hdng Minh
Le Quang Quyen
Le Hieu Phung
Nguyen Thanh San
Le Quang Dinh
Do'n vj cong tac

Htru ca vi sinh Ddi khang
Budn Ma Thuat
Mdi trudng san xuat
Co phan
Lien hiep KHSX CNSH & MT
Dir
an san xudt
thir
nghiem
DANH MUC CAC BANG
Bang
I:
Bd tri thi nghiem thu dpc tinh cap
mSu
Bang 2: Anh hudng
ciia
thanh phan mdi trudng len sinh khoi vi khuan
Bang 3: Anh
hirdng ciia
nhiet dp len mat dp
ciia
TH
10
va DC29
Bang 4: Mat dp cua cac chiing vi khuan nghien ciru dudi cac pH mdi trudng ban
dau khac nhau
Bang 5: Mat dp cua cac chung vi khuan tuyen chpn theo luu luang khi
Bang 6: Tdng hap cac thdng sd cdng nghe cho qua trinh len men chim thu hdi
sinh khdi 2 chiing vi khuan sir dung de san xuat che pham
Bang 7: Anh hudng ctia nhiet dp len sinh khdi

Anh hudng cua thanh
phSn
co chat len sinh khdi
ciia
2 chiing VSV
Bang 16: Anh hudng
cila
nhiet dp len mat dp cua vi sinh vat
Lien hiep KHSX CNSH & MT Du an san xudt thi nghiem
Bang 17: Mat dp cua cac chimg vi sinh vat theo
luu
lupng khi
Bang 18: Anh hudng ciia dp am len mat dp vi sinh vat
Bang 19: Mat dp cua vi khuan trong mdi trudng sau ly tam
Bang 20: Mat do vi sinh vat tdng sd va hoat tinh ddi khang
ciia
che pham
Bang
21:
Danh gia thanh phan ca chat len mat dp cac chung vi sinh vat
Bang 22: Anh hudng
ciia
nhiet dp len mat dp vi sinh vat
Bang 23: Danh gia mat dp vi sinh vat cua che pham ddi khang (CFU/g) theo thdi
gian
Bang 24: Ket qua theo doi trpng lupng
ciia
chudt sau
10
r^ay

Bang 34: Cac
ySu
td
ciu
thanh nang suat va nang suat ly thuyet
Bang
35:
Hieu qua kinh te
Bang 36: Nang
suit
tai cac dia diem khao nghiem
Bang 37:
K6t
qua tap hop thu nghiem dien rdng cua ba vimg khao nghiem
Bang 38: Tinh hinh benh hai cua cac cdng thiic
Bang 39: Nang suat ca phe hat
Lien
hiep KHSX CNSH
&
M
T
Du an san xudt
thu-
nyhiem
Bang 40: Luang toan hieu qua kinh te d cac cdng thdc
sii'
dting
phan huu ca vi sinh
Bang
41:

14
1.3.
Sukhac
nhau giua cdng nghe da nghien cuu va Duan
17
Chuong II: Noi dung cac
kit
qua thuc hien da dat dugc 19
2.1.
Nguyen lieu va phuang phap nghien cuu
19
2.1.1.
Nguyen lieu
19
2.1.2.
Phuong phap 20
2.1.3.
Dia dilm va ca sd vat chat de thuc hien
Duan
22
2.2.
Kit
qua dat dugc 23
A.
Hoin thien cong nghe
2i
2.2.1.
Hoan thien cdng nghe len men chim 23
2.2.1.1.
Lira

2.2.2.4. Xac dinh dp am cho qua trinh len men xdp
hai (da) chung vi sinh vat 36
2.2.2.5.
Xac djnh thdi gian bao hp chat lupng
ciia
che pham ddi
Idiang
37
2.2.3.
Thiet lap cdng nghe len men xdp thu nhan sinh khdi da
chimg cac vi sinh vat ddi khang vdi budng len men 5
m'
38
2.2.4. Danh gia dp an loan
ciia
cac chirng vi sinh vat
len chupt thi nghiem 42
2.2.5.
Hoan thien cdng nghe len men
ii
cac loai ban thanh pham
hiru
CO
va che pham vi sinh ddi khang goc
45
So dd quy trinh san xuat che pham vi sinh vat ddi khang
51
Thuyet minh quy trinh san xuat che pham vi sinh ddi khang
gde
52

2.2.7.1.
Danh gia hieu luc
ciia
san pham phan bdn HCVSDK
len ddi tupng cay ca phe tren dien hep 66
2.2.7.2. Danh gia hieu luc
ciia
san pham len ddi tugng cay
ca phe tren dien rdng qui md
10
ha 70
C. To chiic sdn xudt 75
D.
U'ng
dung mo hinh ton phdn HCVSDK Polyfa 75
E.
Bdo tao 76
Chuffng
III. Ket luan 82
Phu luc 84
Lien hiep KHSX CNSH & MT
Dir an
san xudt
thir
nghiem
CHU'ONG I: TONG QUAN
Trong nhung nam gan day phan hoa
hge,
thudc trir cd dugc su dung vdi
sd lugng ldn trong san xuat ndng nghiep d Viet Nam, la nguyen nhan gay 6

Mien trimg, Tay Nguyen nam trong khu vuc cd khi hau nhiet ddi gid mua,
vdi dac trung khi hau ndng am, lupng mua hang nam Idn, la dieu kien thuan Ipi
cho cae vi sinh vat, ma dien hinh la nam biing phat, gay hai cho cay trdng, dan
den tdn that ldn cho san xuat ndng nghiep. Mdi nam chi phi trong viec phdng,
trir benh cay la rat cao, ma phan ldn la sii dung thudc hoa hpc. Tren thuc te,
phuang phap nay cd hieu qua tuc thdi nhung neu sii dung lau dai se dan den tinh
trang thoai hoa dat va khang thudc cda cac ngudn benh. Vi vay, bien phap sir
dung vi sinh vat ddi khang se la mpt trong nhirng giai phap thiet thuc de c6 mpt
nen san xuat ndng nghiep ben virng.
Lien hiep
KHSX CNSH & MT
Duan
san xudt thu nghiem
1.1. Mot so benh cay
third'ng
gap do vi sinh vat
Hang nam tren the gidi va d Viet Nam, benh cay trdng da gay ra nhung
tdn that to Idn cho nen san xuat ndng nghiep, chiing pha huy tdi 537,3 trieu tan
cac loai ndng san, chiem
11,6%
tdng san lugng ndng nghiep. Hang loat cac loai
benh d cay trdng deu bat ngudn tir vi sinh vat, dien hinh nhu cac loai benh heo
vang va heo xanh d khoai tay, ca chua do vi khuan, nam gay ra, vi du nhu benh
dao dn {Pyricularia orisea), khd van
(Rhizoclonia
solani) va thdi than, thdi re
(Fitsariuin
oxysporum) vv.
Ddi vdi cay ca phe, thudng gap mdt sd benh do vi sinh vat gay ra nhu sau:
Benh Id co re tren cdv ed phe:

thii'
nghiem
ngang, chdp re, phan re duoi chudt xuit hien nhung
vit
thdi
mim
cd mau tham
den. Cay bi benh sinh trirdng can cdi, la vang, heo, it canh
thii
cip va chdi vugt,
la chuyen mau vang, re to va cd re bj thdi. Ca phe kiln thilt ca ban thudng bi
nghiem trpng trong mua mua, re cpc bi thdi, cay bj nang se chet.
DI
phdng benh
nay hien nay mdi
chii
y tdi bien phap tham canh, tang cudng bdn phan huu co,
cai thien dac diem ly va hda tinh
cira
dat, dac biet la giam do chua
ciia
dit. Chua
cd nhQng loai thudc hda hpc dac tri nao de phdng tru benh thdi re cd hieu qua.
Ngudi ta chi cd gang thuc hien cac bien phap tdng hgp nhu: thudng xuyen kiem
tra vtrdn cay, phat hien kip thdi cay bj benh nang dao cay ddt, tudi thudc Benlate
C hay Bendazol ndng do 0,5%, 5 lit dung dich/hd, tudi 2 lan
each
nhau
15
ngay

sail
xudt thu nghiem
kien sdng bang
each
thay ddi dac diem hinh thai sinh ly trong qua trinh phat
triiin
va ket qua dan den
sy
gia tang sd loai trong nhung dieu kien dia ly khac nhau.
Chi Fusarium bao gdm tren 20 loai, trong dd loai Fusariuni
oxysporum
la
phd bien va eung la loai gay benh nguy hiem d thuc vat. F. oxysporum gay ra hien
tu'png
tic maeh dan
ciia
cay va
cOng
la nguyen nhan tryc
liep ciia
benh thdi than,
thdi rl d mdt sd cay rau qua, cay luang thyc va cay cdng nghiep nhu: ca chua,
klioai tay, bdng, hd tieu, ca phe Benh it khi xuat hien sdm ma thudng gay hai
manh nhit vao giiia giai doan sinh trudng
ciia
cay. Do dac diem va tinh chat gay
benli
nguy
hiim ciia
F. oxysporum nen rat khd kiem soat va tieu diet bang phuang

Du an dirac
hinh
thdnh tren kit qud cua cdc de tdi
du-
dn sau:
1-
" Nghien eiru cdng nghe san xuit phan bdn da chiing chiic nang cho cac viing
sinh thai" Ma sd KC 04-04, thudc Chuang trinh cdng nghe sinh hpc giai doan
Lien hiep KHSX CNSH & MT Du an sdn xudt
thil'
nghiem
2001-2005
da dugc danh gia nghiem thu nam 2005 do PGS.TS. Pham Van Toan
lam
chii
nhiem.
2-
"Nghien
eii'u
cdng nghe san xuat phan bdn cho cay bdng vai". De tai cap bd
Vien Khoa hpc va Cdng nghe Viet nam nam 2004-2005, da nghiem thu do PGS.
TS Pham Viet Cudng lam
chii
nhiem.
3-
" Ung dung ky thuat sinh hpc phan tir phan lap va nghien
ciiu
mdt sd vi sinh
vat ddi khang gay benh viing re cay trdng" 2003-2005 da nghiem
thti.

dung cac chung nay van cdn han che bdi
cdng nghe va hoat tinh cua cac chung chua dupe dn djnh. Hon
nOa,
hien nay cay
bdng vai dugc trdng rdng rai d viing mien Trung va Tay Nguyen dang bj benh
thdi re do nam gay ra, nhung chua cd che pham vi sinh vat ddi khang dua ra thi
thudng de phuc vu loai cay trdng nay. Ddi vdi cay ea phe d Tay Nguyen benh
vang la thdi ed re do Fusarium oxysporum da gay tdn that rat Idn, dac biet la d
Die
Lie,
cd noi hon 50% vudn cay bi benh. Mdt sd viing ddng che pham
Triehoderma eho ca phe va mdt so cay trdng can khac nhung hieu qua chua cao.
DI
dua dugc cdng nghe san xuat che pham vi sinh vat ddi khang phdng chdng
Lien hiep
KHSX CNSH & MT Du an san xuat
thii
nghiem
benh thdi cd re, vang la tren ddi tupng cay ca phe va cay bdng virng mien Trung,
Tay Nguyen, gdp phan nang cao hieu qua kinh te, dam bao nen ndng nghiep
sach, ben virng, chiing tdi da xay dyng va thyc hien dy an
"Hocxn
thien cong nghe
sdn xudt che pham vi sinh
vgi
dot khdng phuc vu cho
cdy
bong vdi, ca phe vung
Mien Trung, Tay Nguyen " Dy an su dung mpt sd chiing vi sinh vat dugc phan
lap va tuyen chpn tai cae viing sinh thai mien Trung Tay Nguyen, nhu vay

nguyen lieu trong virng, gia nhan cdng re giam dupe mdt phan chi phi dau vao.
16
Lien hiep KHSX CNSH & MT Du an san
xuJt
thu nghiem
1.3.
Sir
khac nhau giua cdng nghe da nghien cuu va Du' an
Cdng nghe san xuat che pham ddi khang hien tai dugc irng dung trong
Du an la mdt sd ket qua cua cac de tai nghien eiru khoa hpc cap nha
nude,
Chuang trinh Cdng nghe sinh hpc
KC04
- 04 va De tai cap bg (Vien Khoa hpc
va Cdng nghe Viet Nam). Viec nghien ciru cdng nghe chi thyc hien tren mirc do
nghien ciru ca ban va phdng thi nghiem, quy md nhd vdi he thdng len men thi
nghiem dung tich 5 lit. Cac thanh phan mdi trudng dugc su dung trong nghien
ciru cd gia thanh cao nen hieu qua kinh te thap.
Ddi vdi cdng nghe len men chim san xuat che pham vi sinh vat
gde
trong
cac ngi dung cua cac de tai trude day chua danh gia hieu qua
ciia
viec len men
don le vdi len men tap hgp cac ehiing vi sinh vat. Vi vay, Dy an can nghien cuu
khang djnh hieu qua cua len men don le va len men tap hop cac chimg vi sinh vat
vdi cac dieu kien mdi trudng dinh dudng va cac thdng sd ky thuat khac, tren he
thdng len men Idn han la
1001.
Cung nhu ddi vdi cdng nghe len men chim, du an can xae djnh thanh

Lien hiep KHSX CNSH & MT Du an san xuat thir nghiem
CHUONG
H:
NOI
DUNG CAC KET QUA
THyC
HIEN DA DAT DUQC
2.1.
Nguyen lieu va phu'o'ng phap nghien cii'u
2.1.1.
Nguyen lieu
VI sinh vat:
- Cac chiing nam va vi khuan kiem djnh nhan tir Vien Bao ve Thuc vat, Ha Ndi.
- Cac chiing vi sinh vat dugc cung cap tir ngan hang gidng la ket qua cua cac de
tai va du an da thyc hien. Cac chiing nay da dugc djnh danh den loai bang ky
thuat sinh hge phan tir, bao gdm:
Vi khuan: (1) Serratia marcescens
THIO,
(2) P. chlororaphis DC29
Vi nam: Chlomocleista cinnaba
CC510
Xa khuan: 5. nigrisceus VCM
1172
Cac moi trirdng nuoi cay:
- Mdi trudng MPB, MPA, LB, Czapek-Dox, 1SP4, KingB, mdi trudng
MTI(cho 1
Ih:
Urea 1,0 %;
K2O
1,0 %;

-
0,1
g;
MnSOi
va
FeSOi
- 0,001 g; glucose -
0,5 g; pepton - 0,5 g.
Doi tirgng cay trong khao nghiem
- Cay bdng:
sii
dung gidng bdng ludi
Lien hiep KHSX CNSH & MT Du an san xuat thu nghiem
- Cay ca phe: khao nghiem tren loai ca phe kinh doanh dn djnh 6 nam tudi
Robusta (
cci
phe voi)
2.1.2.
Phuong phap
Phuong phap nghien
ciiu
vi sinh
vpt:
- Phuang phap giu' gidng vi sinh vat
- Phuang phap nudi cay vi sinh vat
- Phuang phap xac djnh hoat tinh ddi khang: Phuang phap
due
lo hoac phuang
phap thdi thach
HTDK = D - d (mm); (D: dudng kinh vdng khang vi sinh vat, d: dudng

Lien hiep KHSX CNSH & MT
Du'
an san xuat
Ihu'
nghiein
Tien hanh thu nghiem chinh thuc moi chung tren 40 con chupt, chia thanh
4 nhom gom 1 nhom doi chung diing nuoc cat va 3 nhom thu theo muc lieu da
dy tinh. Cac nhom thiv duac dimg cac dung dich thu 6 cac muc lieu va so lan dua
mau thu nhu sau:
Bang
I:
Bo tri thi nghiem
thti-
doc
tinh
cdp
mau
Nhdi-n
chudt
Nhdm ddi chirng
Muc lieu 1
Muc lieu 2
Muc lieu
Lieu diing (ml hdn
djch nudi/20g chudt)
0,4 ml
nude
x 3 lan
0,4inl
hd

cac cdng thire nhu: tudi nude, lam cd, phun thudc phdng trir sau benh, thu
hoach
+ Kilm tra chit lugng
ciia
phan bdn.
+ Quan
trie
cac ehi tieu ddng rupng: sinh trudng phat trien cua cay, cac
chi tieu ciu thanh nang suat, ehi tieu nang suat.
-I-
Phan tich hieu qua kinh te.
Cic thi nghiem trong Du in
dieac
lap lai 3 lan vi xu ly thong ke
Lien hiep KHSX CNSH & MT
Dir
an san xuat thir nghiem
2.1.3.
Dia diem va co'
so'
vat chat de thuc hien du an
Du'
an du'o'c thirc hien tai:
A. Lien hiep Khoa hpc san xuat Cdng nghe Sinh hpc va Mdi trudng,
Nghia Dd, Cau Giay, Ha Npi vdi eac ndi dung:
- Hoan thien cdng nghe san xuat che pham goc cac chimg VSV ddi khang
- San xuat che pham vi sinh vat ddi khang gde va mdt phan phan bdn ddi
khang tren nen hOu co khdng thanh trting
- Dao tao can bd cdng nhan ky thuat
- Tham gia tieu thu san pham

He thdng
gdm 5-8 him u
thi
tich 70-100
m^;
He thdng bang chuyen; He thdng nen khi
Lien hiep KHSX CNSH & MT
Dir
an san xuat thu nghiem
cudng hire; He thdng phun mua; Cac loai may dap, may nghiin, may sang, may
trdn, may ddng
bao,can
djnh lugng
2.2.
Ket qua dat duoc
A. Hoan thien cong nghe
2.2.1.
Ngi dung
15.3.1.1:
Hodn thien cong nghe ten men chim, thu nhan sinh
khoi cdc chdng vi khuan doi khang (Serratia marcenscen THIO, P.
chlororaphis), thu nhan sinh khoi cdc chung vi nam gdi khdng, thu nhan sinh
khoi chdng xa khudn doi khdng (Streptomyces nigrisceus) v&i he thdng len
men 100 lit vd tgo che phdm doi khdng goc
2.2.1.1.
Lua chgn moi trudng nudi cay thich hop cho len men chim 2 chiing
vi khuan doi khang
Ba loai mdi trudng MPB, LB va
MTl
da dugc su dyng de nudi cay chung

3,2.10'
5,8.10'
5,1.10'
MTl
5,6.10'
4,5.10'
3,4.10'
2,3.10*
2,1.10'
1,5.10'
P.
chlororaphis DC29
KingB
1,3.10'
1,2.10*
3,9.10'
4,5.10'
7,8.10'
5,6.10'
MTl
3,5.10'
3,2.lO'
2,4.10*
3,6.10*
2,9.10'
1,3.10'
23
Lien hiep KHSX CNSH & MT
Du an san xuat thu
nghiem

miu
va xac dinh mat do vi sinh vat sau cae
khoang thdi gian len men nhat dinh. Ket qua dugc the hien trong bang 3:
Bdng
3: Anh huang
ciia
nhiet dp len mat dp
ciia THIO
va DC29 (CFU/ml)
Thdi g
THIO
DC29
ian (h)
25°C
30"C
37"C
45°C
25°C
30°C
37°C
45°C
24h
2,6.10"
2,2.10'
2,7.10'
1,7.10"
1,1x10"
1,7x10'
2,5x10"
3,6x10'

2,8.10'
1,3.10'
5,3x10*
3,5x10*
1,9x10*
3,0x10"
24

Trích đoạn CHUONG III: KET LUAN

---