PROGRAMME DE CONTROLE DE LA QUALITE DES INCLCULIJMS ...
PROGRAMME DE CONTROLE DE LA QUALITE DES INCLCULIJMS
DE RHIZOBILJM FIU SENEGAL
z
:,:
:_
z;
.
.
8’.
.’
2 . Q U A L I T E D ’ U N I N O C U L U M
3 . C O N T R O L E D E L A f.JUALITE
DE L ’ I N O C U L U M
.
Table 1 : Nombre de Rhizobium viables et infectifs requis par graine pour une
fixation efficiente d'azote.
Taille de la graine
Nombre de Rhizobium
Légumineuses
(nombre de graines/kg)
par graine
-
petite
Trefle
200.000
103
intermédiaire
Sainfoin
30.000
200 .ooo
104
- Soja
grosses
- haricot
30.000
105
- niébé, arachide, pois
FS. 1 CONTROLE DE LA QUALITE D’UN- INOCULUti
/Tampon poptone
pH=ZO)
-.-. -_ .-..-
,/”
/
Tampon peptone
pHz7.0
.
l
t i
1’
1:625
113125
1:15625 .
Tèmoin /OI
4 UNIlES DE CROISSANCE PAR DILUTION
i:
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*
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:-
.-
. . _.
_. ._ _.
-:
w
N I
w
F..Ll I
li
n c
x
Table 2 : Composition de la solution nutritive pour la culture des plantes dans
les unités de croissance.
.
CO c12, 6H,O
0,004 mg
,5
2,86
H3B03
mg
Mn C12, 4 H20
i,al
mg
Zn S04, 7 Hz0
0,22
mg
CU S04, 5 H20
0,oa
mg
H2Mo04 > H20
0,09
mg
Mg S04, 7 Hz0
492,96
mg
K2 HP04
i74,ia
mg
KHZ P04
136,09
mg
Ca Cl2
110,99
mg
Fe Cc H5 07,
5,00
H20
mg
EDTA O,Z%
20
ml
Eau distillée
95P.
1000
ml
Ajuster le pH à 6,8 avec HC1 ou NaOh
.
5
-:.
i .- .- -7 - <:: .: -. i;: -5 _. -.
--
5: :
:
il
.-
:_
:- _’
.74
-:
--T
Fi
.--
.Y -*
.
_-
._
-.
i
.-
-- .-
.
E: x e IT$I :I F? ;.!!
___-_-
-__- - ...” -
3 . 6 . T a u x d ’ a p p l i c a t i o n de 1 ’ inoculum au c h a m p
I:; ; p [fi p 1 p
:;
.._...<._.< ..- .<...
.-
-
I
-.
2
_>
._
.- 5
..6
.i
..:i
..Y&
.*
:-
5:
!
-.
J
:;
-3
r-!
TABLE 3. Nombre le plus probable (MPN) de bactbriee calcule B partir de la distribution
des unit68 de croissance positive8 dan6 un test d’infection bat36 sur de8 s.$ries
de dilution en cinq.
1:s
1:25
1:125
1:625
1:3125
1:15625*
Estimation Limite de confiance, à ‘95%
1
0
0
0
0
0
1.1
. 0.2- 7.9
2
0
0
0
0
0
216
0.6-10.1
3
0
0
0
0
0
4.6
1.5-14.1
4
0
0
0
0
0
8.0
3.0-21-S
0
1
0
0
0
0
1.0
O.l- 7.7
1
1
0
0
0
0
2.3
0.6- 9.6
2
1
0
0
0
0
4.0
1.2-12.8
3
‘ 1
0
0
0
0
6.5
2.3-18.0
0
2
0
0
0
0
2.1
o.s- 9.2
1
2
0
0
0
0
3.5
1.1-11.9
2
2
0
0
0
0
5.5
1.9-16.0
3
2
0
0
0
0
8.7
3.3-23.0
0
3
0
0
- 0
0
3 1 0
0.9-10.6
1
3
0
0
0
0
4.9
1.6-14.6
2
3
0
0
0
0
7.2
2 ..7-19.6
3
3,
0
0
0
0
11.3
4.4729-z
.
.
:
: : .
4
1
0
0
0
0
11.4
. *. 4.4-29.5
4
2
0
0
0
0
16.2
6.2-42.4
4
.3
0
0
0
0
24.2
9 :O-64.9
4
‘4
0
0
0
0
40.4
15.3-106.6
.
4
0
1
0
0
0
10.8
4.2-28.1
4
1
1
0
0
0
15.1
5.8-39.2
4
2
1
0
0
0
21.5
8.1-57.4
-
4
3
1
0
0
0
32.8
12.2-67.9
4
0
2
0
0
0
14.1
‘5.4-36.6
4
1
2
0
0
0
19.6
7.4-51.9
4
2
2
0
0
0
28.3
10.5-76.1
4
3
2
0
0
0
43.6
16.6-114.2
4
0
3
0
0
0
18.1
.6.9-47.7
4
1
3
0
0
0
25.2
9.4-67.6
4
2
3
0
0
0
36.4
13.7-96.8
4
3
3
0
0
0
56.5
21.9-146.0
’ 3 (suite )-
-rAl;I,L
-
1:s
1:25
1:125
I:ci25
1:3125
1: 15625*
Ea$ im~tion
Limite ,de confiance à 9f
-.
4
4
1
0
0
0
5..7 x 10:
2.2-14.7 x 10:
4
4
2
0
0
0
8.1 x 10’
3.1-21.2 x 10’
4
4
3
0
0
0
12.1 x 10’
4.5-32.4 x 101
4
4
4
0
0
0
20.2 x 10’
7.6-53.3 x 10’
4
4
0
1
0
0
5.4 x 10’
2.1-14.0 x 10’
4
4
1
1
0
0
7.5 x 10’
2.9-19.6 x 10’
4
4
2
1
0
0
10.8 x 10’
4.0-28.7 x 10’
4
4
3
1
0
0
16.4 x 1ol
6.1-43.9 x 10’
4
4
0
2
0
0
7.1 x 10’
2.7-18.3 x 10’
4
4
1
2
0
0
9.8 x 10’
3.7-26.0 x 10’
4
‘ 4
2
2
0
0
14.1 x 10’
5.3-38.1 x 10’
4
4
3
2
0
0
21.8 x 10’
s-3-57.1 x 10’
4
4
0
3
0
0
9.1 x 10’
3.4-23.8 x 10’
4
4
1
3
0
0
12.6 x l(B*
4.7-33.8 x 10’
4
4
2
3
0
0
18.2 x 10’
6.9-48-4 x 10’
4
4.
3-
3
0
0
28.2 x 10’
10.9-73.0 x 10’
4
4
4
1
0
0
2.9 x 102
l.l- 7.3 x 102
4
4
4
2
0
0
4.1 x 102
1.6-10.6 x 102.
4
4
4
3
0
0
6.0 x 10’
2.3-16.2 x 102
4
4
4
4
0
0
10.1 x 102
3.8-26.6 x 102
4
4
4
0
1
0
2.7 x 102
1 .o- 7.0 x 102
4
4
4
1
1
0
3.8 x 10’
l.S- 9.8 x 102
.
4
4
4
2
x
0
5.4 x 1Q2
2.0-14.4 x 102
4
4
4
3
1
0
8.2 x 102
3.1-22.0 x 102
4
4
4
0
2
0
3’.5 x 102
1.4- 9.2 x 102
4
4
4
1
2
0
4.9 x 102
1.8-13.0 i 102
4
4
4
2
2
0
. 7.1 x 102
2.6-19.0 ,x 10~
: 4
4
4
3
2
0
10.9 x 102
+2-28.6 x 102
-
4
4
4
0
3
0
4.5 x 102
1.7-11.9 x 10*
4
4
4
1
3
0
6.3 x 102
2.3p16.9 x xo2
4
4
‘4
2
3
0
9.1 x 102
3.4-24.2 x la2
4
4
4
3
3
0
14.1 x 102
5.4-36.7 x 102
4
4
4
4
1
0
14.3 x 102
5.5-36.9 x 10~
4
4
4
4
2
0
20.3 x lO*
7.8-53.0 x 10~
4
4
4
4
3
0
30.2 x lO*
11.2-81.3 x 1cï2
4
4
4
4
4
0
SO.5 x 10*
19.0-1.33.8’ x lO*
‘4
4
4
4
0
1
13.5 x 102
5.2-35.3 x 102
4
4
4
4
1
1
18.8 x 10f
7.2-49.0 x 102
4
4
3
4
2
1
26.9 x 102
10.1-71.8 x 102
4
4
4
4
3
1
41.0 x 102
15.3-110.2 x 10’
4
4
4
4
0
2
17.7 x 102
6.8-4S.9 x 102
4
4
4
4
1
2
’ 24.5 x BO2
9.2-65.0 x 102
4
4
‘4
4
2
2
35.3 x 102
13.1-95.4 x 102
4
4
4
4
3
2
54.4 x 102
20.6-143.8 x 102
4
4
4
4
0
3
22.6 x 102
8.6-59.7 x 10’
” 4
4
4
4
1
3
31.4 x 10*
11.7-84.7 x lo2
fl
4
4
. 4
2
3
45.5 x 10*
17.0-121.4 x 102
4
4
4
4
3
3
70.6 x 102
27.1-184.2 x 102
TABLE 3 ( suite)
__
1:s
1:25
I:l25
1:625
I :3125
1:15625* Estimation
Limite de confiance ii '
-
4
4
4
4
4
1
7.1 x BO3
2 . 7 - 1 8 . 6 x 10’
4
4
4
4
4
2
10.1 x x0’
3 . 8 - 2 7 . 0 x 10’
4
4
4
4
4
3
15.1 x ILO’
5 . 4 - 4 2 . 6 x 10’
4
4
4
4
4
4
2 5 . 2 x ILO’
8 . 6 - 7 4 . 0 x 103
4
4
4
4
4
5
> 35:5 x 10’
.
TABLE 4.
Important legumes cultivatcd in Canada;
scicntific name arrnnged
alylrahetically, common name, number of sccd pcr kilogrnn and
Rhizobium specics for inoculatjon
-
Scicntific and common name
Number of seed per kg*
Rhi zobium
inoculation species
s
Arachis hypogaea, L.
peanut
Astragalus ci&, L.
286,650
citer milkvetch
Coronilla varia, L.
242,550
R-
?
crownvetch
6,667
5. japonicum
Lathyrus spp.
t--r
R. leguminosarum
flat peas
Lens
R. Leguminosarum
lentil
Lcspedeza cuneata, Don.
771.,750
R . “cowpea”
perennial sericca lespedeza
Lespedeia striata, Hook 6 Arn.
418,850
R- “cobpea”
annual common lcspedeza
Lotus corniculatus, L. (tenuis)
826; 875
R. loti
-
-
birdsfoot trefoil
a n g u s
Lupinus t i f o l i u s
----
R. lupini
lupins
441,000
.R. meliloti
- --
Helilotus albn, Desr.
573,340
5. meliloti
whitc sweetclover
Irlclilotus officinalis, Lam.
573,300
R. meliloti
yellow sweetclovcr
Onobrychis viciaefolia, Stop.
66,150
R*
?
sainfoin
PhascolGs vulgaruc.
-----;- --.. - _.. - . . .-
2,461
R. phnseoli
._---
,
r
-1
TAULE 4 (suite)
-
Scientific ancl common name
Number of sccd par kg*
Rhizobium
inoculation species
Pisum arvense, L.
4,310
R. 1 cguminosarum
field or garden pcas
Stizolobium deeringianum, Bort.
2,205
!L “cowpea”
velvetbeans, field beans
Trifolium hybridum, L.
’
1,543,500
R. trifolii
alsike clover
Trifolium incarnatum, L.
308,700
R. trifolii
c r i m s o n c l o v e r
Trifolium pratense, L.
606,375
R. trifolii
red clover.
Trifolium repens, L.
3,764,OOO
R. trifolii
white clover, ladino clover
Vicia sativa, L.
15,435
R. leguminosarum
common ve tch
Vicia villosa, Roth
44,100
R. leguminosarum
hairy vetch
Vicia faba var. equina, L. .
-
-
2,326
R. 1 cguminosanxm
horsebean
Vicia faba var. major, L.
-
-
'1,103
R. leguminosarum
b r o a d b e a n
* TO comply with Table under section 10.2 C(ii) in regulations
greater than 200,000 = small; 30,000 - 200,000 = intermediate;
lcss than 30,000 = large.
** Rhizobium species designation arc unknown for these lcgumes ‘and specific
strains are requircd.
DE RHIZOBILJM FIU SENEGAL
z
:,:
:_
z;
.
.
8’.
.’
2 . Q U A L I T E D ’ U N I N O C U L U M
3 . C O N T R O L E D E L A f.JUALITE
DE L ’ I N O C U L U M
.
Table 1 : Nombre de Rhizobium viables et infectifs requis par graine pour une
fixation efficiente d'azote.
Taille de la graine
Nombre de Rhizobium
Légumineuses
(nombre de graines/kg)
par graine
-
petite
Trefle
200.000
103
intermédiaire
Sainfoin
30.000
200 .ooo
104
- Soja
grosses
- haricot
30.000
105
- niébé, arachide, pois
FS. 1 CONTROLE DE LA QUALITE D’UN- INOCULUti
/Tampon poptone
pH=ZO)
-.-. -_ .-..-
,/”
/
Tampon peptone
pHz7.0
.
l
t i
1’
1:625
113125
1:15625 .
Tèmoin /OI
4 UNIlES DE CROISSANCE PAR DILUTION
i:
._-.
*
i-:
:
:-
.-
. . _.
_. ._ _.
-:
w
N I
w
F..Ll I
li
n c
x
Table 2 : Composition de la solution nutritive pour la culture des plantes dans
les unités de croissance.
.
CO c12, 6H,O
0,004 mg
,5
2,86
H3B03
mg
Mn C12, 4 H20
i,al
mg
Zn S04, 7 Hz0
0,22
mg
CU S04, 5 H20
0,oa
mg
H2Mo04 > H20
0,09
mg
Mg S04, 7 Hz0
492,96
mg
K2 HP04
i74,ia
mg
KHZ P04
136,09
mg
Ca Cl2
110,99
mg
Fe Cc H5 07,
5,00
H20
mg
EDTA O,Z%
20
ml
Eau distillée
95P.
1000
ml
Ajuster le pH à 6,8 avec HC1 ou NaOh
.
5
-:.
i .- .- -7 - <:: .: -. i;: -5 _. -.
--
5: :
:
il
.-
:_
:- _’
.74
-:
--T
Fi
.--
.Y -*
.
_-
._
-.
i
.-
-- .-
.
E: x e IT$I :I F? ;.!!
___-_-
-__- - ...” -
3 . 6 . T a u x d ’ a p p l i c a t i o n de 1 ’ inoculum au c h a m p
I:; ; p [fi p 1 p
:;
.._...<._.< ..- .<...
.-
-
I
-.
2
_>
._
.- 5
..6
.i
..:i
..Y&
.*
:-
5:
!
-.
J
:;
-3
r-!
TABLE 3. Nombre le plus probable (MPN) de bactbriee calcule B partir de la distribution
des unit68 de croissance positive8 dan6 un test d’infection bat36 sur de8 s.$ries
de dilution en cinq.
1:s
1:25
1:125
1:625
1:3125
1:15625*
Estimation Limite de confiance, à ‘95%
1
0
0
0
0
0
1.1
. 0.2- 7.9
2
0
0
0
0
0
216
0.6-10.1
3
0
0
0
0
0
4.6
1.5-14.1
4
0
0
0
0
0
8.0
3.0-21-S
0
1
0
0
0
0
1.0
O.l- 7.7
1
1
0
0
0
0
2.3
0.6- 9.6
2
1
0
0
0
0
4.0
1.2-12.8
3
‘ 1
0
0
0
0
6.5
2.3-18.0
0
2
0
0
0
0
2.1
o.s- 9.2
1
2
0
0
0
0
3.5
1.1-11.9
2
2
0
0
0
0
5.5
1.9-16.0
3
2
0
0
0
0
8.7
3.3-23.0
0
3
0
0
- 0
0
3 1 0
0.9-10.6
1
3
0
0
0
0
4.9
1.6-14.6
2
3
0
0
0
0
7.2
2 ..7-19.6
3
3,
0
0
0
0
11.3
4.4729-z
.
.
:
: : .
4
1
0
0
0
0
11.4
. *. 4.4-29.5
4
2
0
0
0
0
16.2
6.2-42.4
4
.3
0
0
0
0
24.2
9 :O-64.9
4
‘4
0
0
0
0
40.4
15.3-106.6
.
4
0
1
0
0
0
10.8
4.2-28.1
4
1
1
0
0
0
15.1
5.8-39.2
4
2
1
0
0
0
21.5
8.1-57.4
-
4
3
1
0
0
0
32.8
12.2-67.9
4
0
2
0
0
0
14.1
‘5.4-36.6
4
1
2
0
0
0
19.6
7.4-51.9
4
2
2
0
0
0
28.3
10.5-76.1
4
3
2
0
0
0
43.6
16.6-114.2
4
0
3
0
0
0
18.1
.6.9-47.7
4
1
3
0
0
0
25.2
9.4-67.6
4
2
3
0
0
0
36.4
13.7-96.8
4
3
3
0
0
0
56.5
21.9-146.0
’ 3 (suite )-
-rAl;I,L
-
1:s
1:25
1:125
I:ci25
1:3125
1: 15625*
Ea$ im~tion
Limite ,de confiance à 9f
-.
4
4
1
0
0
0
5..7 x 10:
2.2-14.7 x 10:
4
4
2
0
0
0
8.1 x 10’
3.1-21.2 x 10’
4
4
3
0
0
0
12.1 x 10’
4.5-32.4 x 101
4
4
4
0
0
0
20.2 x 10’
7.6-53.3 x 10’
4
4
0
1
0
0
5.4 x 10’
2.1-14.0 x 10’
4
4
1
1
0
0
7.5 x 10’
2.9-19.6 x 10’
4
4
2
1
0
0
10.8 x 10’
4.0-28.7 x 10’
4
4
3
1
0
0
16.4 x 1ol
6.1-43.9 x 10’
4
4
0
2
0
0
7.1 x 10’
2.7-18.3 x 10’
4
4
1
2
0
0
9.8 x 10’
3.7-26.0 x 10’
4
‘ 4
2
2
0
0
14.1 x 10’
5.3-38.1 x 10’
4
4
3
2
0
0
21.8 x 10’
s-3-57.1 x 10’
4
4
0
3
0
0
9.1 x 10’
3.4-23.8 x 10’
4
4
1
3
0
0
12.6 x l(B*
4.7-33.8 x 10’
4
4
2
3
0
0
18.2 x 10’
6.9-48-4 x 10’
4
4.
3-
3
0
0
28.2 x 10’
10.9-73.0 x 10’
4
4
4
1
0
0
2.9 x 102
l.l- 7.3 x 102
4
4
4
2
0
0
4.1 x 102
1.6-10.6 x 102.
4
4
4
3
0
0
6.0 x 10’
2.3-16.2 x 102
4
4
4
4
0
0
10.1 x 102
3.8-26.6 x 102
4
4
4
0
1
0
2.7 x 102
1 .o- 7.0 x 102
4
4
4
1
1
0
3.8 x 10’
l.S- 9.8 x 102
.
4
4
4
2
x
0
5.4 x 1Q2
2.0-14.4 x 102
4
4
4
3
1
0
8.2 x 102
3.1-22.0 x 102
4
4
4
0
2
0
3’.5 x 102
1.4- 9.2 x 102
4
4
4
1
2
0
4.9 x 102
1.8-13.0 i 102
4
4
4
2
2
0
. 7.1 x 102
2.6-19.0 ,x 10~
: 4
4
4
3
2
0
10.9 x 102
+2-28.6 x 102
-
4
4
4
0
3
0
4.5 x 102
1.7-11.9 x 10*
4
4
4
1
3
0
6.3 x 102
2.3p16.9 x xo2
4
4
‘4
2
3
0
9.1 x 102
3.4-24.2 x la2
4
4
4
3
3
0
14.1 x 102
5.4-36.7 x 102
4
4
4
4
1
0
14.3 x 102
5.5-36.9 x 10~
4
4
4
4
2
0
20.3 x lO*
7.8-53.0 x 10~
4
4
4
4
3
0
30.2 x lO*
11.2-81.3 x 1cï2
4
4
4
4
4
0
SO.5 x 10*
19.0-1.33.8’ x lO*
‘4
4
4
4
0
1
13.5 x 102
5.2-35.3 x 102
4
4
4
4
1
1
18.8 x 10f
7.2-49.0 x 102
4
4
3
4
2
1
26.9 x 102
10.1-71.8 x 102
4
4
4
4
3
1
41.0 x 102
15.3-110.2 x 10’
4
4
4
4
0
2
17.7 x 102
6.8-4S.9 x 102
4
4
4
4
1
2
’ 24.5 x BO2
9.2-65.0 x 102
4
4
‘4
4
2
2
35.3 x 102
13.1-95.4 x 102
4
4
4
4
3
2
54.4 x 102
20.6-143.8 x 102
4
4
4
4
0
3
22.6 x 102
8.6-59.7 x 10’
” 4
4
4
4
1
3
31.4 x 10*
11.7-84.7 x lo2
fl
4
4
. 4
2
3
45.5 x 10*
17.0-121.4 x 102
4
4
4
4
3
3
70.6 x 102
27.1-184.2 x 102
TABLE 3 ( suite)
__
1:s
1:25
I:l25
1:625
I :3125
1:15625* Estimation
Limite de confiance ii '
-
4
4
4
4
4
1
7.1 x BO3
2 . 7 - 1 8 . 6 x 10’
4
4
4
4
4
2
10.1 x x0’
3 . 8 - 2 7 . 0 x 10’
4
4
4
4
4
3
15.1 x ILO’
5 . 4 - 4 2 . 6 x 10’
4
4
4
4
4
4
2 5 . 2 x ILO’
8 . 6 - 7 4 . 0 x 103
4
4
4
4
4
5
> 35:5 x 10’
.
TABLE 4.
Important legumes cultivatcd in Canada;
scicntific name arrnnged
alylrahetically, common name, number of sccd pcr kilogrnn and
Rhizobium specics for inoculatjon
-
Scicntific and common name
Number of seed per kg*
Rhi zobium
inoculation species
s
Arachis hypogaea, L.
peanut
Astragalus ci&, L.
286,650
citer milkvetch
Coronilla varia, L.
242,550
R-
?
crownvetch
6,667
5. japonicum
Lathyrus spp.
t--r
R. leguminosarum
flat peas
Lens
R. Leguminosarum
lentil
Lcspedeza cuneata, Don.
771.,750
R . “cowpea”
perennial sericca lespedeza
Lespedeia striata, Hook 6 Arn.
418,850
R- “cobpea”
annual common lcspedeza
Lotus corniculatus, L. (tenuis)
826; 875
R. loti
-
-
birdsfoot trefoil
a n g u s
Lupinus t i f o l i u s
----
R. lupini
lupins
441,000
.R. meliloti
- --
Helilotus albn, Desr.
573,340
5. meliloti
whitc sweetclover
Irlclilotus officinalis, Lam.
573,300
R. meliloti
yellow sweetclovcr
Onobrychis viciaefolia, Stop.
66,150
R*
?
sainfoin
PhascolGs vulgaruc.
-----;- --.. - _.. - . . .-
2,461
R. phnseoli
._---
,
r
-1
TAULE 4 (suite)
-
Scientific ancl common name
Number of sccd par kg*
Rhizobium
inoculation species
Pisum arvense, L.
4,310
R. 1 cguminosarum
field or garden pcas
Stizolobium deeringianum, Bort.
2,205
!L “cowpea”
velvetbeans, field beans
Trifolium hybridum, L.
’
1,543,500
R. trifolii
alsike clover
Trifolium incarnatum, L.
308,700
R. trifolii
c r i m s o n c l o v e r
Trifolium pratense, L.
606,375
R. trifolii
red clover.
Trifolium repens, L.
3,764,OOO
R. trifolii
white clover, ladino clover
Vicia sativa, L.
15,435
R. leguminosarum
common ve tch
Vicia villosa, Roth
44,100
R. leguminosarum
hairy vetch
Vicia faba var. equina, L. .
-
-
2,326
R. 1 cguminosanxm
horsebean
Vicia faba var. major, L.
-
-
'1,103
R. leguminosarum
b r o a d b e a n
* TO comply with Table under section 10.2 C(ii) in regulations
greater than 200,000 = small; 30,000 - 200,000 = intermediate;
lcss than 30,000 = large.
** Rhizobium species designation arc unknown for these lcgumes ‘and specific
strains are requircd.