Hoàn thiện công nghệ sản xuát chế phẩm vi sinh vật đối kháng phòng bệnh thối rễ cho cây cà phê, bông vải miền trung, tây nguyên

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

Lien hiep KHSX CNSH & MT
Dtr
ail
san xuat thu nghiem
BO NONG NGHIEP VA PHAT TRIEN
NONG THON
VIEN KHOA HOC VA CONG NGHE
VlfT
NAM
CHUONG TRINH UD CNSH TRONG NONG NGHIEP
BAO CAO
TONG
HOP
KET QUA KHOA HQC CONG NGHE
DU
AN
Hoan thien cong nghe san xuat che pham vi sinh vat doi khang
phong benh thoi re cho cay ca phe, bong vai
viing
Mien Trung,
Tay Nguyen.
Chii
nhiem Du an
(ky ten)
Co quan chd tri Du an
ddu)
PGS.TS.
Pham Viet Cudng
~TRi^GPH6NQ0LTH
MAI 1H(

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

Vien Cdng nghe Sinh
hpc
Lien hiep KHSX Cdng nghe Sinh hoc & Moi trudng
Lien hiep KHSX Cdng nghe Sinh hpc & Mdi trudng
Lien hiep KHSX Cdng nghe Sinh hpc & Mdi trudng
Lien hiep KHSX Cdng nghe Sinh hpc & Moi trudng
Vien NC cay bdng va phat trien ndng nghiep Nha Hd
Cdng ty Phan bdn Hoa sinh Sdng Kon Binh Dinh
Xi nghiep phan bdn
POLYFA
cdng ty ca phe Buon
Ma Thudt
Cdng ty Lam nghiep Gia NghTa
Lien hiep KHSX CNSH & MT
Du an san xuat thu nghiem
DANH MUC CAC KY HIEU, CAC
CHU
VIET TAT
UD CNSH
KHSX
MT
NC
HCVSDK
BMT
MTSX
CP
U'ng dung Cdng nghe Sinh hpc
Khoa hpc san xuat
Mdi trudng
Nghien cuu

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

ciia
Chromocleista cirmaba
CC510
Bang 8: Mat dp cua chung vi nam Chlomocleista cinnaba
CCS 10
vdi cac tri sd
pH ban dau khac nhau cda qua trinh len men
Bang 9: Mat dp cua chiing vi nam Chlomocleista cinnaba CC510 theo luu
lugng
khi
Bang 10: Cdng thiic mdi trudng thir nghiem
Bang
11:
Mat dp vi sinh vat sau cac khoang thdi gian len men
Bang 12: Tdng hop cac thdng sd cdng nghe tdi uu cho qua trinh len men
ciia
chung Chromocleista cinnaba
CCS 10
sir dung cho qua trinh san xuat
che pham
Bang 13:
KSt
qua sir dung mdi trudng trong qua trinh len men chung xa khuan
Bang 14: Cac
diSu
kien cdng nghe tdi uu cho qua trinh len men cua chung xa
khuan Steptomyces nigrisceus VCM
1172
Bang
15:

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

udng dich nudi
DC29vaTH10
Bang
2S:
Kit
qua theo doi trpng lupng
ciia
chupt sau
10
ngay udng dich nudi
Bang 26: Mat dp vi sinh vat tdng sd theo loai ca chat va thdi gian
ii
Bang 27: Dpng hpc
ciia
nhiet dp trong qua trinh len men co chat hiiu ca
Bang 28: Thanh phin trong phan bdn hiiu ca vi sinh ddi khang
Bang 29: Danh gia mat dp vi sinh vat cua che pham ddi khang
Bang 30: Tinh hinh benh
Id
cd re
Bang
31:
Anh hudng
ciia
che pham len su phat trien cua bp re va chieu cao cua
cay sau 20 ngay nay mam
Bang 32: Danh gia anh hudng cua che pham len cac yeu td cau thanh nang suat
Bang 33: Tinh hinh benh hai va thdi gian phat
due ciia
cay

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:

Nang suat ca phe nhan lai Dak Genh
Bang 42: Nang suat ca phe nhan tai Krdng Bdng
Bang 43: Nang suat ca phe nhan tai Budn Ddn
Bang 44: Bang tdng
hgp
kit qua su dung phan bon HCVSDK tren ddi lugng
cii
phe vdi cdng tuc bon CT3
Bang 45: Cdng thuc bdn phan cho cay bdng ludi
Bang 46: Cdng
tliiic
bdn phan cho cay ca phe vdi kinh doanh nam thu 5
Bang 47:
Kk
qua md hinh sir dung phan bdn ddi khang ddi vdi cay Bdng vai
Bang 48:
Kit
qua md hinh sir dung phan bdn ddi khang ddi vdi cay ca phe
Lien hiep KHSX CNSH & MT Du an san xudt
thii-
nghiem
MUC LUC
Chuong I: Tong quan
11
1.1.
Mgt sd benh cay thudng gap do vi sinh vat
12
1.2. Xuit
xii ciia
Du an

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

chgn mdi trudng nudi cay thich hgp cho len men chim
2 chung vi khuan ddi khang 23
2.2.1.2.
Dilu kien len men toi uu cho chiing vi khuan
Serratia marcenscen
TH 10
va
P.
chlororaphis DC29 24
2.2.1.3.
Hoan thien cdng nghe len men chim, thu nhan sinh khdi
chung vi nam ddi khang 27
2.2.1.4. Hoan thien cdng nghe len men chim, thu nhan sinh khdi
chung xa khuan ddi khang 32
2.2.2. Thilt lap cdng nghe len men xop thu nhan sinh khdi dan
chiing cac vi sinh vat ddi khang vdi budng len men Sm 33
2.2.2.1.
Lira
chpn
CO chit
nudi cay phu
hgp
vdi cac chung 33
Lien hiep KHSX CNSH
&.
MT Du an sdn xuat thu nghiem
2.2.2.2. Xac dinh nhiet dp len men 34
2.2.2.3.
Xac dinh dieu kien cung cap khdng khi
cho len men xdp 35

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

Sa dd quy trinh san xuat phan bdn ddi khang tren nen co chat khdng
thanh trimg 55
Thuyet minh quy trinh san xuat phan bdn HCVS ddi khang 56
B.
Banh gid hieu
lure
cda
sdn
phdm len idi tuang cdy trong 57
2.2.6. Danh gia hieu luc
ciia
san pham len ddi tuong cay bdng vai 57
2.2.6.1.
Danh gia hieu
lire ciia
phan bdn ddi khang POLYFA
len cay bdng vai tren dien hep quy md 3.000 m" 57
2.2.6.2. Danh gia khao nghiem hieu luc
cila
Phan bdn Vi sinh
ddi khang vai quy md
lOha
60
2.2.7.
Danh gia hieu luc
ciia
san pham len ddi tugng cay ca phe 64
Lien hiep KHSX CNSH & MT
Du
an san xudt thu nghiem

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

nhiem mdi trudng nghiem trpng, dat dai bac mau, lam
chit
cac sinh vat c6 ich,
lam tang kha nang khang thudc
ciia
sinh vat gay hai, ket qua la viec san xuat
ndng nghiep roi vao tinh trang khd khan. Dac biet la khu
virc
Mien Trung, Tay
Nguyen, cay bdng vai va cay ca phe thudng bj thdi cd re va heo xanh do
Fusariuni
oxysporum.
R.
solani,
P. solanacearium va mpt sd tac nhan khac.
De giam tdi mirc thap nhat hau qua xau do phuang phap canh tac hoa hpc
mang lai, ddng thdi lam tang hieu qua kiem soat, dan tdi tieu diet ngudn benh vi
sinh vat gay hai cho cay trdng, cac nha khoa hpc dang
chil
y den ngudn tai
nguyen ldn thir ba
ciia
gidi tu nhien - Tai nguyen vi sinh vat cd ich de bao ve
mdi trudng sinh thai. Phuang phap sir dung vi sinh vat trong bao ve mdi trudng
khdng nhung mang lai hieu quii cao, an toan, san pham thu hoach khdng anh
hudng tdi ngudi su dung, cd Ipi cho can bang sinh thai, ma cdn giam phan ldn
lugng thudc hoa
hge
sii'
dung.

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:

Benh nay phd bien tren cay ea phe con trong vudn uom va cay d giai doan
kiln thilt ca ban(chua cho san pham), trdng tren dat cd thanh phan co gidi nang.
Cay bj thdi toan phin hoac mdt phin cd re khien eay sinh trudng cham, vang la
va dan
din
chit.
Benh do nim Rhizoclonia solani gay nen. Benh gay hai
chii
yeu
len eay con trong thdi ky vudn uom va thdi ky kien thiet ca ban. Vj tri benh xam
nhap gay tac hai la d phin cd rl (cd re bi teo, khd that lai, ngan can hay lam dinh
tre qua trinh van chuyen chat dinh duang de nudi cay).
Din
nay, nhCrng nai da bi benh nang thi ngudi ta bd di khdng lam lai dugc
vudn uom d vi tri cu. Nhiing cay da bi benh nhd dem ddt, cay cdn bj nhe hoac d
vung cd cay bi benh thi diing cac loai thudc hoa hpc de tudi hay phun
xii
ly:
TMTD (Thiuram) 0,5%, Maneb, Zineb tir 0,2 - 0,3%, Benlate 0,5%, Validacin
3%.
TUy nhien mim benh khdng diet
trir
duac hoan toan, kha nang tai phat ldn.
Benh vine li thdi re tren cdv cd phe:
La loai berth nguy hiem cd the gay
chit
hang loat. Benh do su phdi hgp
tin cdng cda tuyin triing (Pratylenchus coffeae) va cac nam (Fusarium
oxysporum), gay tac hai vao bd re cua cay ca phe. Trieu chiing: Tren cac rl
Lien hiep KHSX CNSH & MT Du an san xudt

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

cho cac cay xung quanh viing benh.
Benh do vi sinh vat chu yeu hai bong:
Benh xanh
liin:
Tac nhan gay benh do virus va dugc lan truyen trong ttr
nhien nhd cdn triing mdi gidi la rep bdng.
Benh moc trdng: Tac nhan gay benh do nam Ramulariopsis gossypii gay
ra. Benh gay hai tren la, tan cdng tir la gia den la banh te. Cay bi benh nang la
vang va lam nu, hoa, qua non bj rung, qua chin ep, giam nang suat.
Benh
dom
- chdy
Id:
Benh xuat hien trong
miia
mua, lam thdi qua d ting
dudi, lam giam nang suat. Tac nhan gay benh do nam Rhizoclonia solani gay ra.
Benh la co re: Tac nhan gay benh do nam va vi khuan nhu: Rhizoclonia
solani, Pythium spp., Thielaviopsis basicola, Pythium haryanum va mdt sd loai
vi khuan. Benh gay hai tir khi eay bdng vira nay mam den giai doan 3 - 4 la that
trong dieu kien nhiet dp thap va am dp dat cao.
Nam Fusarium oxysporum gay thiet hai nang ne cho cay trdng, chiem tdi
83%
(theo
OU,1972).
Fusarium oxysporum cd mat hau het d cac vung trdng trpt
tren the gidi (Rebell,
1981).
Chiing thich ling rat manh vdi nhiing thay ddi dieu
Lien hiep KHSX CNSH & MT Du an

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

phap don le ma chi cd the ket hpp mdt sd bien phap de kiem soat dugc benh.
R.
solani tdn tai tren tan du cay trdng va trong dat, dudi dang sgi va hach
nim. Hach nim cd
thi
sdng trong dat hang nam, gap dieu kien thich hgp, hach
cd
thi
mge ra cac sgi nim xam nhap vao gde cay chd giap mat dat.
Nhiing nghien
ciiu
vl phan bdn cho cay bdng tai Viet Nam da dugc thuc
hien trong nhiing nam gan day, nhung chi tap trung vao cac yeu td da va trung
lugng, chua cd chl phim phan bdn sinh hge nao ket hgp giila vi sinh vat va che
phim kich thich phat triln trong mdi trudng ty nhien dugc nghien
ciiu
va san
xuit cho tirng loai cay trdng ndi ehung ciing nhu cho cay bdng vai ndi rieng.
Chua cd san phim phan bdn nhap ngoai thudc dang vi sinh sir dung ngudn hiru
CO
til'
phu lieu cua nganh bdng, cung nhu nghien
ciiu
bd sung vi sinh vat cd ich
vao ngudn phan hilu co sinh hpc, phuc vu chuyen cho cho cay bdng vai.
1.2. Xuat
xir
cua
Dir
an

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.

Thudc
chuong trinh nghien eiru co ban do PGS.TS Pham Viet Cudng lam
chii
nhiem.
Trong giai doan
2001-2005,
Dl tai KC04-04 do Vien KTNNVN
chil
tri,
De tai Khoa hge Co ban Ma sd 82- 03.04, De tai cap Bd Vien Khoa hpc & Cdng
nghe Viet Nam do Vien Cdng nghe sinh hpc chu tri da phan lap tuyen chpn dugc
mdt sd chung vi sinh vat ddi khang Serratia marcenscen TH
10,
P chlororaphis
DC29,
Steptomyces nigrisceus VCM
1172,
S. hygrosropicus VCM
1158
cd
hoat tinh khang cac vi sinh gay benh cho mdt sd cay trdng nhu: ca phe, tieu,
cac chiing nay da dugc dinh danh bang ky thuat sinh
hge
phan ni va danh gia
hoat tinh ddi khang vdi
P.oxysporrium,
R. solani ed vdng khang mam tre 20 mm.
Cac chung nay da duge img dung lam che pham chiic nang budc dau cho ket
qua. Tuy nhien viec san xuat va
sir

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

phii
hgp vdi dieu kien ty nhien d noi diiy.
Viec trien
kliai
cdng nghe san xuat che pham phan bdn ddi khang cho cay
trdng dugc thuc hien tai LHKHSX CNSH va MT ciing vdi cac cdng ty cd kinh
nghiem va trang thilt bi chuyen san xuat phan bdn vi sinh d vung mien Trung
Tay Nguyen tham gia. Cdng nghe
chii
yeu la len men chim va len men xdp cac
chiing vi sinh vat ddi khang, cd bd sung cac chung vi sinh vat hilu ich khac va
cac nguyen td da vi lupng, hoat chat sinh hge vao chat nen. Mien Trung va Tay
Nguyen la vung san xuit hang hoa ndng nghiep tap trung
eiia
cay bdng, ca phe
va cac loai cay trdng khac,
rJiu
cau phan bdn ddi khang rat ldn.
- Khi ap dung san phim do Dy an tao ra se han ehe dugc viec sir dung hoa
chit bao ve thuc vat, giam mdt phan phan bdn vd
eg.
Ngoai viec giam chi phi
trong san xuit ndng nghiep, cdn gdp phan bao ve mdi trudng dat, tao nen su can
bang sinh thai.
- Viec su dung phan bdn ddi khang lam giam du lugng thudc bao ve thyc
vat, dam bao duge an toan va nang cao chat lugng san pham ca phe, nhu vay
lam tang tinh canh tranh trong xuat khau.
- Viec to chirc xay dung dy an san xuat phan bdn ehire nang ddi khang eho
virng miln Trung, Tay Nguyen phye vu tai chd, nen trong mdt khu vuc tap trung
cac loai eay cdng nghiep se giam chi phi van chuyen, sir dung dugc ngudn

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

phan dinh dudng, thanh phan co chat va cac thdng sd cdng nghe len men xdp khi
len men rieng va len men tap hgp chiing vdi budng len men 5
m'.
Ddi vdi cdng doan san xuat phan bdn hiru co vi sinh ddi khang, van de
can hoan thien la: Xay dyng cdng nghe len men u cac loai ban thanh pham huu
CO khac nhau vdi che pham vi sinh ddi khang
gde
cd quy md the tich khdi u tir
90 -
100m'
tai mien Trung va Tay Nguyen.
Vi vay de di vao thuc te san xuat cdng nghiep can phai hoan thien, lam
ehii
quy trinh va cac thdng sd ky thuat; dn djnh chat lupng san pham, chung loai
san pham; nang cao khdi lupng san pham can san xuat thu nghiem de khang dinh
cdng nghe, lam tiln dl cho viec thye hien cac du an san xuat tiep theo.
Lien hiep KHSX CNSH & MT Du an san xuit thir nghiem
Ddi vdi tat ca cac san pham phan bdn viec danh gia hieu lyc ddi vdi cay
trdng la dilu bat budc. Vdi dy an nay, ddi tugng danh gia la cay bdng va cay ca
phe vdi hai dang md hinh dien hep va dien rdng theo quy djnh
ciia
Bd NN va
PTNT.
Dy an cin san xuit dugc 5 tin che pham ddi khang gde va
5.000
tan che
phim vi sinh vat ddi khang tren nen ca chat khdng thanh triing phuc vu
dii
cho
cay trdng.

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 %;

P2O5
0,5%; NaCl 0,2g;
MgSO,
0,2g;
CaCl,
0,lg;
MnS04
va
FeSO^
0,001g; Ri dudng 10 ml; Nude thit bd 5%).
- Hoat tinh khang nam dugc xac djnh tren mdi trudng thach Waksman
(WA):
pepton - 5 g; glucose -
10
g; cao thjt - 3 g; NaCl - 5 g; thach - 20 g; nude
cit -
1
lit.
Thanh phin mdi trudng san xuit (MTSX)
(g/1):
K2O
-
1,0
g;
P2O5
- 0,5 g;
NaCl - 0,2 g;
MgSOi
- 0,2 g;
CaCh

-
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

kinh thdi thach hoac khuan
lac),
tren cac mdi trudng dac hieu cho vi khuan, vi
nam va xa khuan.
- Phuang phap len men: Phuang phap len men chim: diing cho nhan gidng cap I
cip 2. Phuang phap len men xop (len men be mat): diing de nhan sinh khdi cac
chung ua khi lam che pham.
Phuong phap danh gia do an toan cua cac chiing vi sinh vat
irng
dung trong
che pham:
Thd dp an toan cua cac chung nghien cim len chudt nhat trang
Bdn chung nghien ciru duge tien hanh kiem tra dp an toan len ddng vat
nghien ciru la chudt nhit trang gidng Swits cung cap tir Vien ve sinh djch te.
- Chudt dugc
nhjn
an
15
gid trudc khi thi nghiem,
nude
udng theo nhu
ciu. Kilm tra can nang trudc khi thir nghiem. Chudt dat cac yeu can vl can nang
dugc dua vao thir nghiem
- Dua mau thir dudi dang hon djch chiia lugng vi khuin 2,1
10*te bao/ml
theo dudng udng. Liy
thi tich miu
thir theo quy djnh dua thing vao da day
chudt bang kim cong dau tu.
2D

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

X 1
lan
0,4ml hd
X
2 lan
0,4inl
hd x 3 lan
Lieu diing (ml mau
thu/kg chudt)
I5inl/kg
chudt
30ml/kg chupt
45ml/kg chupt
sd chudt thi
nghiem
^To^
10
10
10
Chung tdi theo doi bilu hien
ciia
chudt sau khi udng trong 24 gid dau va
theo ddi hoat ddng cua ddng vat thi nghiem trong thdi gian 10 ngay sau khi
udng.
Phirong
phap khao nghiem ngoai thuc dia
+ Thi nghiem duge bd tri theo khdi day dd
ngiu
nhien.
+ Cac cdng doan ky thuat khae dugc tien hanh thuc hien nhu nhau d tat ca

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

B.
Xi nghiep phan bdn POLYFA thudc Cdng ty ca phe Budn Me Thudt,
Die
Lie,
Cdng ty TNHH hda sinh hiru co Polyfa, Cdng ty Lam nghiep Gia
NghTa, Dac Ndng, Cdng ty cd phan phan bdn sinh hda Sdng Kon, Binh Djnh
Tilp nhan cdng nghe va
siin
xuat phan bdn ddi khang tren nen co chat
huu
CO
Polyfa khdng thanh triing.
Thuc hien tieu thy
C. Vien Nghien ctru Bdng va phat trien ndng nghiep Nha Hd, Ninh Thuan
Thuc hien khao nghiem danh gia hieu qua che pham len cay bdng vai
Thilt bj va co sd vat chat phuc vu cho Du an:
- Lien hiep KHSX CNSH va MT cd : Budng ciy vi sinh BAS
11
vd triing;
He thdng len men 100 lit; Budng len men xdp gia nhiet cd dieu khien 5 m'; May
ly tam lanh; Phdng thi nhiem vdi cac thiet bj, dung cu phuc vu nghien ciru phan
tich vi sinh vat va cac chi tieu khac.
- Cac cdng ty Phan bdn Hda sinh Sdng Kon, Cdng ty CP lam nghiep Gia
NghTa, Xi nghiep phan bdn Polyfa Cdng ty ea phe Budn Ma Thudt
diu
cd: Nha
xudng vdi dien tich tir 700
din
1200m'; Khu san phoi 1000-1500
m^

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

vi khuan Serratia marcescens
THIO
va mdi trudng king B, MTl cho chiing P.
chlororaphis DC29. Sau 24 gid, xae djnh mat dp vi khuan tren mdi trudng MPA.
Bdng 2: Anh hu&ng
ciia
thdnh phdn moi trucmg len sinh khoi vi khudn
Thdi gian
(gid)
20
25
28
30
35
40
Mat dp vi sinh
(CFU/ml)
Serratia marcescens TH
10
MPB
1,3.10'
3,4.10'
4,3.10*
2,4.10'
3,3.10'
2,8.10'
LB
8,5.10'
4,6.10'
5,2.10*

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

Nhu vay, da lya chpn dugc mdi trudng
MTl
la mdi trudng san xuat thich
hgp cho ca 2 chiing vi khuan cd kha nang ddi khang manh
F.oxysporum
va
Ralstonia
solanacearwn
de san xuat ehe pham.
2.2.1.2. Dieu kien len men toi
iru
cho chiing vi khuan Serratia marcenscen
THIO
va P.chlororaphls DC29
a.
Xdc djnh thong so nhiet dp len men
Cac ehung vi khuan tuyen ehgn cd nhiet do nudi eay thich hgp trong dieu
kien phdng thi nghiem la 30"C. Tuy nhien trong dieu kien len men d the tich ldn
hon (ndi len men
IOO
lit), can xac dinh lai nhiet do tdi uu cho sy phat trien cua
chimg vi khuan nghien ciru.
Vdi mdi trudng va tdc do
sue
khi nhu nhau, nhiet dp len men thay ddi nhu
sau :
25"C,
30"C,
37"C,
45"C. Liy

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'

30h
2,4.10"
1,2.10*
2,1.10'
3,1.10"
2,1x10"
2,3x10*
1,2x10'
1,8x10"
35h
4,2.10"
2,2.10'
4,2.10'
3,5.10"
1,8x10"
4,5x10'
1,1x10'
2,0x10'
40h
2,5.10"
2,8.10'
8,2.10'
3,2.10'
1,2x10'
3x10'
2,4x10'
2,1x10'
48h
1,5.10"
6,7.10*

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