NGHIÊN CỨU TUYỂN CHỌN MỘT SỐ CHỦNG VI SINH VẬT PHÂN GIẢI XELLULOZA ĐỂ XỬ LÝ THÂN CÀNH CHÈ LÀM PHÂN BÓN HỮU CƠ TRONG SẢN XUẤT CHÈ AN TOÀN

NGHIÊN CỨU TUYỂN CHỌN MỘT SỐ CHỦNG VI SINH VẬT
PHÂN GIẢI XELLULOZA ĐỂ XỬ LÝ THÂN CÀNH CHÈ LÀM
PHÂN BÓN HỮU CƠ TRONG SẢN XUẤT CHÈ AN TOÀN
MỤC LỤC
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MỞ ĐẦU
1. Tính cấp thiết của đề tài
 !"#
$%&'()#!*+,'# -
'%./0(12*34556-7(
89:-#;8-2<=%>?9
2@A<7(5,@2#59211
<?)-9BC2-9;#-30
((-6DECF#18(%
82CD#!?9--?F(#-D%
" D;9;6#;F-'921
G9;(92HG(9*','%"#-892HG9
86#A''D;9;I%&,
J1?'#!K9211A#!7(
, 97J?-#(,62L9?'
A#M7(,?9(-D;9;C
ID;(-6#-(%N 2O1
-52-(2*3(-D;9;
9:D(-PCCQ;%
.R29CI7(D82Q;QK
,J2,S-)-?%.TD-?UVWXY
;Z(,S-955[8VWXY;# X(%\4
'9]#-;;2F%- 93?
8#;C?-;#^486#-
3^,+2;35?T-%&^08
:3I;92M-<?_U8DD;9;%

T9#-8;(9'MA C;CD
8M7%\4G QL,J*
#-60(@M,8-#;'%
`;UAa8S*' FbcNghiên
cứu tuyển chọn một số chủng vi sinh vật phân giải xelluloza để xử lý thân
cành chè làm phân bón hữu cơ trong sản xuất chè an toàn”.
X
2. Mục đích nghiên cứu
N M/67GC
Q=-d(MQL(2,8359?-C
I(-%
3. Ý nghĩa của đề tài
3.1. Ý nghĩa khoa học
e'PC 7(F[;B2-(/5
!,#- 7GC
G352%
3.2. Ý nghĩa thực tiễn
&(#(67GC(-
CQ;9(-D;9;9C?-D64-;%

CHƯƠNG I
TỔNG QUAN TÀI LIỆU
1. CÁC NGHIÊN CỨU VỀ ĐỐN CHÈ TRÊN THẾ GIỚI VÀ VIỆT NAM
1.1. Nghiên cứu kỹ thuật đốn chè trên thế giới
&,P(#/#-CQ;%f(-?7(0
#-CQ;8#;F9T2H
Gg%-?7'#-#^^82''5
,C99(%%%
N 2HG!"#$9C&-"U("#
hXiVXj9&TN/$HhXikYjlXm-,'7'6-

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1.2. Nghiên cứu kỹ thuật đốn chè ở Việt Nam
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2'-+?-4!+-D;(-5-,@
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Q;%
"U2'PC # 9|6*hXiqVj)(#(P#4
7(-"##^?X2+
R;XsXV92o+R;oYsoV9
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98#6590219X9;
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2'PC ),(P#4lm%"=-P#4
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&G6DYYY9CNa"y|4
hYYkjlom 2HG# ƒS\" e=-…
"1-;"#$#^!ƒS„6sƒS"/-;
"#P2HG1bX1UV
soY9, oV9uYsuV9ouVsVY-;
D;Dq9Vor-P#4
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"#2'GQ((,JD9G6,S9
D;D(-Uk9vWok9kr-
2. CÁC NGHIÊN CỨU VỀ VẬT LIỆU CHE PHỦ TRÊN THẾ GIỚI VÀ
VIỆT NAM
2.1. Nghiên cứu về vật liệu che phủ trên thế giới
#^S;]U;M#!#M%N-
,6G-?#(9#-CIz(A;
]#^;U;%w(T-?9#^?M?-;
6I35%"*P(P#4M-G
;#-;CI#!^]2M-
#^(%

"=-2'PC 7(%†%†=-hXikYm : &6I;
7(*1,y62(2=7,JV-?
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<‡(=((%"#-F22*?2ˆ-96I;4?(-
;!1=7,J<N(=#CA(=%";C*
8F5-*2*=726I
;!6iY%\-0M,@((6g(2*?2ˆ-,;
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v
D ,=7,J<-;'F8
F2CD3%
z=-2'PC 7(%†%†=-ƒ%.%…„NhXiqi)b
"#-F2=79DP(;8A2(F6
;3(*=7# #^,J,*1%
NA2,2*:3(2#M?6=7
‰uYr,FRR;%&0219D;+;2*8
5P(#Š#'6;%
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-!p `* '!\ 
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2ˆ-G';#^C,C-S#!
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,I9#9K(7(-?=,89#^Q=02O2'
'5,C9;-?8](-%‡3?
h5D;XY;Z(8M-
;Z(j9#g7%"#^=,8M,<
^#@K(D7(=,8h#/9^
((h^=j_j%"7,J'69#5#?9#^<‡ _
7Y;Z(9osVD7X%"02O#^R,L
#^Q=/Gh?9T_j982CDC?-;-;
Q(h219#/_j921#^Q=8M-oYsuY
;Z('D8%
3. NGHIÊN CỨU VỀ CÁC CHỦNG VI SINH VẬT CÓ KHẢ NĂNG
PHÂN GIẢI XELLULOZA
3.1. Nhóm vi khuẩn

N82I8G F;U
2-C'2•Xi'(%2-(/)G67
G82CDCQ=-U?<7(6G(?%
&'2•Y90(G82I'21C
Q=-%"#-*#086I(-0D2CDC
Q=-=Q=-7(82I97'8
2I'21%
.682I82CDC`=-b
s





"#-A'90(;68
'218;G(-;%N-#(:82y
XX
21G#-?<7(6G(?
%
3.2. Nhóm xạ khuẩn
`?2I682IR,9‡#(59'219',-
R#,!A9(2I5;2I21
9,-,@908RP(D#--?;%
.68Q?2ICQ=-b



 

3.3. Nhóm nấm

8F-;CQ=-?S0
7Q=-26(-!6YsoY
-
9„#-2-CU
o9Vsv9v%\4G9S07Q=-d(!(-?7(,M79
26,M7?%
.68;82CDCQ=-b



!
"*0#-8GM8(`=-p-
`=-      :( " #"      "  
%9
sN8;2((#;#6
#)#-*CQ;35/(!\N(%ƒ
35/8#6 ;2/(8
#;#-:#U,'(9:/
,#a-;#(%„(,ˆ-#z(
:/,ˆ-G-6;,#(b!9
9_%
s\*w/h\e-(/*\N(j)
 CQ;*'I/3I-;8 
p-s.% "    1   7(  p-s.   7  ;  =
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p-=ƒ"q%X82CD?-#-F2;2*?9S
C-9#46I-;#-F2y(42*8
0(98(-•7(#^9T#-
7Q(;#^#/%&Q=6CC?-;,F
3-*#0%

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5#59#?9#C35#;PC%'I/|‘.…h8(
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I\s&e7(*#g\N((*"
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*09CM*a*#0-g*7(^%
N0*?G^?T9U(
D2CD2,-#^-7(;
2"#-=#(8(#-#-%
s'I7(\w/|‘†s‹9'I(
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 %2I  %N3G# 8
•(;35#-9,:h#-=
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CHƯƠNG II
ĐỐI TƯỢNG, NỘI DUNG VÀ PHƯƠNG PHÁP NGHIÊN CỨU
2.1. Địa điểm, thời gian nghiên cứu
s&y(M1bƒ:19\"+]*8(
\e-(/2HGN*FS1(|@%
s"0(b"UXYZYXX'XZYX
2.2. Nội dung nghiên cứu
sƒG7G82CDCQ=-
s"M/7G82CDCQ=-(-

sN 6RM#!9#M7(7
GCQ=-M/
s&2CD77(7G
M/%
2.3. Vật liệu nghiên cứu
’\G
sB9;9#C7
sB;g
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2I#M%
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GGb.bXt"?bXt„

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s€yLbN(bY9kVt;bXYY%
2.4. Phương pháp nghiên cứu
2.4.1. Phương pháp phân lập các chủng vi sinh vật có khả năng phân hủy
xelluloza
7G82CD7Q=-d(G
Xu
(b
|XbXYBh--9#C9;_j--,4(
(iYL)2#g%
|bp@# @#-0(oYS9(8M@%
|ob€g=(SXy#---==-#Œ9(
-)'^6XY
sv
%
|ubp;Y9Xy!^6UXY
su
'XY
sv
<F# 
“(=#(*#0‡(d=‘„(%&M#-7;!6oq
-
#-
usuk(--8M;#Š2I?# ,%
|Vb2I?84?9@2(# 9
3#-M-1(%
2.4.2. Xác định định tính hoạt tính CMC- aza (Williams, 1983)
w27G(2*;uk@?
,<R@<Y9V-T?)I,y”# 

“(ƒ=#(*#0.Rh.bXt"?bXt„

†bX%YYYj%
p3“(?#-7;uk9(8;#(#,FR?,J
y-%
„-?1/Qy,J21:C9:
#:#-,(-P(T?hh3(021:#:#-
h€j021T?hj%
2.4.3. Xác định khả năng phân giải lignoxenlulo
|J•C#/7(B1-B%
ƒ5'b
|Xb1QuY(9)@<9;2*-
-,4(XYYY%
|b|+-T,4XYYy*;@G%
|ob&R,4#5#-7;oq
-
#-q%w(8#(?9
-?,<?;:((;2*:?(•%"•C
#/7(B1-B1=-*b
`hrj•h
-
s

jZ
-
%XYY
"#-8b
`br6C#/7(B1



b"#/2*:?7(B1

-
b"#/2*:?7(B
XV
2.4.4. Kiểm tra mật độ vi sinh vật theo phương pháp Koch
2.4.5.Xác định các điều kiện sinh trưởng, phát triển của vi sinh vật theo
phương pháp vi sinh vật thông dụng
2.4.6. Phương pháp ủ xử lý thân cành chè trong nhà lưới
$%&"'()
s"bR@XVsY9--*#-P(
6 %
s€y2G82CDCQ=-d((-bo2ZX;
%
"'bƒ#6)QLP(*#-y
*;G9C,C-G6',-\w\XYi‹>Z%
$*+',)
nN69,'6PMG#-796F7(
*1%
n.G6G+2M#(# *#0"?ys
=-%
n.G62I2M#(# *#0„(%
n.G6;2M#(# *#0d(=2%
n.G6Q?2I2M#(# *#0‡(d=%
2.5. Phương pháp phân tích và xử lý số liệu
1QL2 =-5#4•Q=%
Xv
CHƯƠNG III
KẾT QUẢ VÀ THẢO LUẬN
3.1. Phân lập các chủng vi sinh vật có khả năng phân giải xelluloza

e-C6(;(#,-#-2h,-(j# R;
Q=-d(9'oVWVYr22*2AG%";CC
I2[2-8(0=d=;,!
G%„=d=CQ=-d(0G2*-
-%" 9#-2-C0(@huk0jG#-?<
,:8MCvYWvVrQ=-d(-8M 8('iYr
Q=-d(7(T%:#-/?#a-R3
C]AG#-;9=-=8M7#-2-C0
(5%
82CD#!?9--?F(#-D%
" D;9;6#;F-'921
G9;(92HG(9*','%wCI7(
D82Q;QK,J2,S-)-?%
.TD-?UVWXY;Z(,S-955[8VWXY
;# X(%\4'9]#-;;2
F%- 93?8#;C?-
;#^486#-3^,+2;35?
T-%&^08:3I;92M-<?_U8D
D;9;%
" (07'M7
96#;1#-=7-9#C?]-
;%.6,JG8PC58
2I82CDCQ=-d(SC
35,8-;98C64 ;%
$(P#4 9UB;19#C93
5h--j9S*)GoY72IQ?2I82C
DCQ=-d(%784?921@2I
?2(%e'PC1M!,Co%Xb
Xq
Bảng 3.1: Các chủng vi sinh vật phân lập

TT Ký hiệu
chủng VSV
Nguồn phân lập
Đặc điểm khuẩn lạc phân lập trên
môi trường đặc hiệu*
X X &;#^ "#:9#@9
  &;g "#:9#@9x92*
o o ‚C7 "#:9#@9
u u &;g "#:9#@9^
V V &;g "#:9=9=
v v &;#^ "#:9^9^?98
q q &;g "#:9F-#@9=
k k &;g "#:9^?9^
i i ƒ "#:9F-#@9=
XY XY &;g "#:9#@9^
XX XX &;g "#:9=
X X ‚C7 "#:9#@98
Xo Xo &;g "#:9?98
Xu Xu ‚C7 "#:9=9=
XV XV &;#^ "#:9#@
Xv Xv &;g "#:9-9#@
Xq Xq &;g "#:9#@#-
Xk Xk ‚C7 "#:9-9=
Xi Xi &;g "#:9Q(9Q(
Y Y ƒ "#:9=9=
X X &;g "#:9-?9=
  &;g "#:9-9#@
o o ‚C7 "#:9=
u u &;g "#:9=
V V ‚C7 "#:9Q(?

v v &;g "#:9#@
q q ‚C7 "#:9?9=
k k ‚C7 "#:9#@9^
i i &; "#:9-#@9=
oY oY &;g "#:98F^Q
-$ ./0"123456
$(,Co%X-;784?921@
2I?2(%
3.2. Tuyển chọn các chủng vi sinh vật có khả năng phân giải xenlulo mạnh
|J5Qy-?1.s(d( -789:;6 
5Qy2CDC-Q=-9)/Xq7
82CDCQ=-U''?9#-88q7Q?2I
82CDCQ=-?;h21:C•oYj
Xk
b`X%v9`X%V9`u9`o9`X%q9`u%o9`X%%e'PC#4,#-
,Co%b
Bảng 3.2: Hoạt tính phân giải xelluloza của các chủng vi sinh vật phân lập
TT Ký hiệu
chủng
Nguồn phân
lập
Đường kính VPG xelluloza
(D-d,mm)
X XC1.6 Đất vùi chè 40
 XC1.5 Đất vùi chè 40
o X4 Rác thải ủ 37
u `‚X ‚5-( q
V `o ‚5-( 30
v ` ‚5-( q
q `‚o &;g q

k `u%u &;g q
i XC1.7 Đất vùi chè 33
XY `X%u &;g q
XX `V &;g k
X XC4.3 Đất vùi chè 30
Xo XC1.2 Đất vùi chè 34
Xu \X% &;g V
XV \u%o &;g Y
Xv \| &;|)|J v
Xq \| &;|)|J k
"#-Xq7G8-?1CQ=-d((-8o
7Q?2IGU;g-2'PC-021:
C(-;Uoq'uY97:?GUB;9
;#^#^9#5#?-(8021:CUVWoo
%7GA(/39,C-PC9;-
1'(%
1<8)=">"?@A*A"-/B"* 46
Xi
"'2CDC7(7
GM/,JQy6C27(B
QLGCQ=-d(-B2*
,+G(1%e(71-*P(
-?#U,y7-GC%e'PC
M!,Co%o%
Bảng 3.3: Tỷ lệ giảm khối lượng thân cành chè
trong bình ủ ở 37
0
C sau 30 ngày
TT Mẫu ủ Tỷ lệ giảm khối lượng so với mẫu đối chứng
(%)

X `X%v k9Xo–Y9X
 `X%V k9kY–Y9v
o `u q9kq–Y9X
u `o v9o–Y9V
V `X%q v9oY–Y9Y
v `u%o q9Yo–Y9X
q `X% v9vq–Y9XV
k .B s
e'PC,Co%o-;9,+7G)8PC
C2#-,4-B2*,+
G%"#-7G,+7`eo9`eV9\e9\eo
-2'PCC2(-;%&7FDMCQ;
'IGCQ=-d(%
3.3. Nghiên cứu một số đặc điểm sinh trưởng, phát triểncủa các chủng vi
sinh vật phân giải xelluloza tuyển chọn
&M?-F2G#- *;92
7G9S*)' 6RM/
bN-Q9„96%%%e'PCM!,Co%ub
Bảng 3.4: Đặc điểm sinh học của các chủng vi sinh vật tuyển chọn
Ký hiệu
chủng
Nhu cầu
oxy
Gram
Điều kiện thích hợp
pH
Nhiệt độ
(
0
C)

Môi
trường
Nồng độ NaCl
(%)
`X%V
n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
`X%v
n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
`u n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
`o n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
Y
`X%q n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
`u%o n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
`X% n n q9YWk9Y
oYsuV ‡(d=‘
Y9YVsY9v
e'PC,Co%u-;97Q?2IA(/F8
-Q9#M#-F26UoYWuV

Y
%&F2I‡#(
59y„#-2-CUq9Y'k9Y%
3.4 Đánh giá hiệu quả xử lý thân cành chè của vi sinh vật tuyển chọn
&M?-F2G-7GCQ=-d(
#!#M9#2QL7)
*#-MFK„7( 9F ?'A
#M7(;,^?#- %N-#(9M7
G81#-'IG#!#M9S*
)6#K09?# ,+ -71
U(;^],(M?--G#-'
I#!#M#-(-?9U(MFK•ZN
(--?UVZX'oYZXS-P#4C(PC
5%
7Q?2IM/`X%v9`X%V9`u9`X%V*
;@XVY:ZS# *#0‡(d=!6oq
Y
(uk0%w(8
7G#6B(=-•XbXbXbX9MI,y,+
-7%.G67G?‰XY
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