~EWBLIOLIE DU SENEGAL INSTITUT SENEGALAIS D...
~EWBLIOLIE DU SENEGAL
INSTITUT SENEGALAIS
D E
MINISTERE DE LA RECHERCHIE SCIENTIFIQUE
RECHERCHES AGRICOLES
ET TECHNIOUE.
DEPARTEMENT DE RECHERCHES FORESTIE-
JES ET HYDROBIOLOGIQUES.
DEPARTEMENT DE RECHERCHES SUR LES
PRODUCTIONS VEGETALES
PRELIMINARY STUDIES ON FOUR NITROGEN FIXING TREES
FROM SENECALESE SOILS
Ousmane DIAGNE et Mamadou GUEYE
Microbiologi&es à l'Institut Sénégalais
de Rec:herches Agricoles BP. 3120 Dakar
.&illet 1984
Office de la Recherche
Centre National de
Centre National de
Scientifique et Technique
Recherche Forostièr e
Recherche
Agronomique
!Outre M e r (ORSTOM)
@NRF)
(cNRA)
PRELIMINARY CN FOUR NITROCEN FIXING TREES
FROM SENEGALESE SOILS
0. DIAGNE' and M. GUEYE2
(1) : ORSTOM L b
a oratoire de flicrobiologie BP. 1386 Dakar (Senegal)
(2) : MIRCEN/CNRA BP. 53 Bambey (Senagal)
SUMK1RY
-n-
Because of the destruction of forests for agricultural needs, there
is a continuous degradation of soils. TO reforest these soils, itis
necessary to use trees that cari thrive in urasted soils, Fortunately
such trees exist
: they are Nitrogen Fixing Trees (NFT).
We studied four species of NFT : prosopis africana, Prosopis .juli-
flrra, Albizia lebbeck and Leucaena leucocephalq.
-
-
Prosopis africana and Albizia lebbeck uere uell nodulated in Seuth
Seneqal soils uheraas PrrJposis .iuliflora and Leucaena leucocephala
-LI-
uere not nodulated in thiç area.
Rhizcbium strains luere isolated from those NFT species and their host
spectrum uras determined. Nitrogen fixation \\uas estimated in the labo-
ratory by measuring of the shnat and nodules dry weight and shoot to-
tal N content.
The four NFT species studied fell into twn categories :
1) NFT for which nitrngen fixation could significantly be in-
creased by inoculation urith the strains UKI had isolated :
Proposif ,afri,cana and Jlbizia lebbeck;. Rhizobium strains
A1,0 and pa?, were most effective strains on Proposis
africana and Albizia lebbeck.
-
-
2) NFT for urhich nitrcgen fixation could n& significantly 5s
incrcased by inoculation uJi.th wr strains : Propoaiq juli;
flnra and Leucaena leu.cocephala. The reason of thie failure
ia proberbly that straine native to sensgalesa SOL~S u1ere not
effective on Proposi,e $~Iiflhra and $eucaena leucocephala.
- 2-
1. INTRODUCTICN
Our first objective mas tn study the nodulation of four
Nitrogen Fixing Trees (NFT), Prosnpis africanq, Pr3sopis
juliflora,
Albizia 1eSbeck and Leucaena .leucocephnla, thon
to isolate Rhizobium strains from thO3ts fd1-T t%nj firj?ls}t tu asti-
mate nitrogen fixation in the laboratory.
2. MATERIALS AND METHODS
For isolating Rhizobium strains fram the fiwe diffarent sri1s8
me studied (table l), me planted germinated seeds of the four
speoies in plastic 24 x 11 cm pouches fi.Lled UP u&th the dff-
ferent smils. The seeds had been sterilized by immersion in
cancentrated sulfuric acid (table Z), a treatment mhich u1as
also necessary Cp allow the germinatian (chemical scarifica-
tirn).
Tha oxporimeht utere cr?nducted with three seedlings per pouche
for the four NFT and fifteen replicates par treatment. Rhizo-
bium uere isalated frorn ths nodules that appearod after 55 43~s.
Iaolates ~ero plated in Petri dishes containing the routina
YEMA medium (VINCENT, 1970).
Infection plant tests were carried out in GIBSON tube (1963)
crntaining agar JENSEN medium (1942). Seedlings were inocula-
ted with a 7-day-•ld cultura ef tho isolated Rhizrbium strains
and the ORS 911 and CB 7% Rhi?nbium as reference strains.
Nitrogen fixation was cstimatcd by differencs betiuesn the total
N content alf non inocufated and incculatsd plants. In addition,
nodules and shoots IJJWZ ureightod aftar bsing oven drisd.
3. RESULTS
3.1. Modulation of the NFT
Prosopis africana and Albizia labboclc nodulatod in a11 sriils
usod. The nombor of nodules was highor in Dabo soil. Proeo-
& .iuliflrra and Leucaena laucrcophala did not nodulate in
any scil (table 3).
3.2. Isclaticn of !?hirobium strain_anQ &udy of the-ir host -ramqe
Ten strains wcr~ Asol;--ttcd, a11 of thom from Prosopis africana
-
-
- 5 -
Prosopis africana and Albizia lobback and most of them
-
-
wre effective nn both hosts. Only t.tJo of these stroins
(Pal from Prosopis africana and A16 from Albizia lebback)
tirer 0 abla ta effoctively nodulats Leucaona leucocephalz.
-
-
None of the tan strains isolated uras able to nodulato
ProsoPis juliflora.
Interestingly ORS 911 (isolated from Acacia farnasiana)
was able t3 nodulate leucaena leucocaphala (table 4).
The cffectivity of the strains tuas di2terminatod only iuibh
Prossop.is africans and Albizia lebbeck, the best strains
being Al;oand pa7 (Fig. 1, 2, 3).
CONCLUSION
Our proiiminary investigations of thc four specics of NFT shn-
wed that those species fol1 into two inoculation groups accor-
ding to ths Rhizobium that nodulated thon1 :
the first group including prosopis africana and Albizia labbeck
-
-
ürhjch noduLatr;d rosdily ujith Rhizobium strains distributed in
tho fivo stinegslesc soils studied (which ara probably ubiqui-
tcu 3
strains) . Intortistingly,
sorne of tho Rhizobium strains
uùre fast grouing,
other were SLOW growing, tho tu3 catagLries
of Rhizobia. Thore was no relation boturoon tha efficiency of
these strains and their growth rate.
Tho sticond group including Prosopis Juliffora and Loucaena
loucoc:jphnlz; tuhich did not nodulatYd iuith thc native Rhizobium
strains Froiii thcsa scils, which suggsst thet thc: Rhizobium
raquiert.w?ent;s of Prosopis juliflora end LwcaeT'a ltiucoçaphala
Lzrc m o r s sPC?cificthon th2 cnss of thc first group. it aPpe3rs
necossery to initiate invastigations in th-t field.
L
=---
-
I
D.
D
IA 0
0
0
CJ
OY
Ln 0
3
0’
r
0
u 0
0
0’
0
o-
LA 0
0
0
N
133
W
fi
v- u?
Y- 0
T-
F 0
h
V
0
T-
In
0
0
0 0
0
CG- 0
CJ 0
0
0
CY
CY 0
V
r?
W
,T
h
x-- OI
T-
Lr’i
r- 0
Y-
lr,
v-
M
0
rc
M M
V
v- Ln
T- In
<j”
\\a 0
m
h
0
W
V
.---.
v- In
o*
0 c3
0
7
L”
0”
0 0
CZY
0 0
V
F--.
a
v- Ln
0”
0 0
V
h
0
v-
In
0
0 u
Table 4 - Host spectrum of ten Rhizobium strains ielated
From prosopis aFricana and Albizia lebbeck.
-3-
Straine
prosopis
Albizia
Leucaena
?roaopio
tested
africana
lebbeck
leucocephala ,iiliflorz
pa 1
fast
E
E
c1.i.
pa 3
fast
'e
e
n.i.
Pas
Sl.OW
E
n.i.
n.i.
Pa7
sl ow
E
n..i.
nri.
Pa9
sl.ow
E
n.i.
n.i.
A12
fast
e
kl
A14
fast
E
ri.i.
A16
slow
n.i,
n.i,
A18
SlOW
n.i,
E)
FI110
fabt
n.i.
r1.i.
ORS 911
f3.St
E
E
El
CB7 56
SlOW
e
n.i.
n.i.
E = effective ; e xz ineffective ; n.i.= non in?eCtiVe
*
= Plants of this treatment were contaminated.
- -
. . .,
t
F
I.,,
..s
..I
i
.,
.
,.
_
. .
1
5
REFERENCES
CORNET, F., 1982. Rscherches preliminaires sur la symbiose de deux Acacia
0.
tropicaux (A. raddisna et A. holasericea) avec Rhizobium sp, et
un chamoignon endomycorhizien (Glomus mosseas)
-
-
D.E.A. Microbio-
loqie. Eniuers. Claude Bernard, L y o n I.
+i DREYFUS, R.L., and DOMWEHGUES, Y.R., 1981. Nodulation of s p c
Acacia c i e s
b y Fast- and Slow-growing Trripical Strains of Rhizobium. APPlied
and Environ. Microb. 4 1 (11, 97-99.
GIBSON, A .H ., 1963. Physical environment in symbiotic nitrogen fixation.
The effect of root temperature on recently nodulated Trifolium
subterranax p l a n t s . Aust, J . Siol. S c i . 1 6 , 2 8 - 4 2 .
HALLIDAY, J., and SOMASEGARAN, P., 1982. Nodulation, Nitrogen Fixation
and Rhizobium strain afinities in the Ganus Leucaena. In Leucaena
-
-
research in the Asian-pacifie Region. 2:oceedinqs of a Workshop
held in Sinqapore, 23-26 Nov. 1982. p. 27-32.
JENSEN, H.L., 1942. Nitragon fixation in loguminous plants. General character
af root nodule bacteria isolated from species of Medicaqo and
Tri folium in Australia. oroceedinqs of the linnean Societv N.S.W.
67 > 98-108.
ROSKOSKI, J.P. et ai., 1980. Nitrogen Fixation by Tropical Woody Legumes :
potential !Source of Soi1 Enrichment. Biol. Nitroq. Fixat. Toch.
for Trop. Agr., An Int. Workshop,
VINCENT, J.M., 1970. A manual for the practical study of root nodules bac-
taria. I.B.P. Handbook no 15. Blackurell Sci. publications Oxford
snd EdSnburgh.
VINCENT, J.M., 1974. Root-nodule symbiosis urith zizobium : general micro-
biology. In the biology of nitrogon fixstion Ed. by Quiepsl, A,
North-Holland - publishing CO, Amsterdam, 265-341.
INSTITUT SENEGALAIS
D E
MINISTERE DE LA RECHERCHIE SCIENTIFIQUE
RECHERCHES AGRICOLES
ET TECHNIOUE.
DEPARTEMENT DE RECHERCHES FORESTIE-
JES ET HYDROBIOLOGIQUES.
DEPARTEMENT DE RECHERCHES SUR LES
PRODUCTIONS VEGETALES
PRELIMINARY STUDIES ON FOUR NITROGEN FIXING TREES
FROM SENECALESE SOILS
Ousmane DIAGNE et Mamadou GUEYE
Microbiologi&es à l'Institut Sénégalais
de Rec:herches Agricoles BP. 3120 Dakar
.&illet 1984
Office de la Recherche
Centre National de
Centre National de
Scientifique et Technique
Recherche Forostièr e
Recherche
Agronomique
!Outre M e r (ORSTOM)
@NRF)
(cNRA)
PRELIMINARY CN FOUR NITROCEN FIXING TREES
FROM SENEGALESE SOILS
0. DIAGNE' and M. GUEYE2
(1) : ORSTOM L b
a oratoire de flicrobiologie BP. 1386 Dakar (Senegal)
(2) : MIRCEN/CNRA BP. 53 Bambey (Senagal)
SUMK1RY
-n-
Because of the destruction of forests for agricultural needs, there
is a continuous degradation of soils. TO reforest these soils, itis
necessary to use trees that cari thrive in urasted soils, Fortunately
such trees exist
: they are Nitrogen Fixing Trees (NFT).
We studied four species of NFT : prosopis africana, Prosopis .juli-
flrra, Albizia lebbeck and Leucaena leucocephalq.
-
-
Prosopis africana and Albizia lebbeck uere uell nodulated in Seuth
Seneqal soils uheraas PrrJposis .iuliflora and Leucaena leucocephala
-LI-
uere not nodulated in thiç area.
Rhizcbium strains luere isolated from those NFT species and their host
spectrum uras determined. Nitrogen fixation \\uas estimated in the labo-
ratory by measuring of the shnat and nodules dry weight and shoot to-
tal N content.
The four NFT species studied fell into twn categories :
1) NFT for which nitrngen fixation could significantly be in-
creased by inoculation urith the strains UKI had isolated :
Proposif ,afri,cana and Jlbizia lebbeck;. Rhizobium strains
A1,0 and pa?, were most effective strains on Proposis
africana and Albizia lebbeck.
-
-
2) NFT for urhich nitrcgen fixation could n& significantly 5s
incrcased by inoculation uJi.th wr strains : Propoaiq juli;
flnra and Leucaena leu.cocephala. The reason of thie failure
ia proberbly that straine native to sensgalesa SOL~S u1ere not
effective on Proposi,e $~Iiflhra and $eucaena leucocephala.
- 2-
1. INTRODUCTICN
Our first objective mas tn study the nodulation of four
Nitrogen Fixing Trees (NFT), Prosnpis africanq, Pr3sopis
juliflora,
Albizia 1eSbeck and Leucaena .leucocephnla, thon
to isolate Rhizobium strains from thO3ts fd1-T t%nj firj?ls}t tu asti-
mate nitrogen fixation in the laboratory.
2. MATERIALS AND METHODS
For isolating Rhizobium strains fram the fiwe diffarent sri1s8
me studied (table l), me planted germinated seeds of the four
speoies in plastic 24 x 11 cm pouches fi.Lled UP u&th the dff-
ferent smils. The seeds had been sterilized by immersion in
cancentrated sulfuric acid (table Z), a treatment mhich u1as
also necessary Cp allow the germinatian (chemical scarifica-
tirn).
Tha oxporimeht utere cr?nducted with three seedlings per pouche
for the four NFT and fifteen replicates par treatment. Rhizo-
bium uere isalated frorn ths nodules that appearod after 55 43~s.
Iaolates ~ero plated in Petri dishes containing the routina
YEMA medium (VINCENT, 1970).
Infection plant tests were carried out in GIBSON tube (1963)
crntaining agar JENSEN medium (1942). Seedlings were inocula-
ted with a 7-day-•ld cultura ef tho isolated Rhizrbium strains
and the ORS 911 and CB 7% Rhi?nbium as reference strains.
Nitrogen fixation was cstimatcd by differencs betiuesn the total
N content alf non inocufated and incculatsd plants. In addition,
nodules and shoots IJJWZ ureightod aftar bsing oven drisd.
3. RESULTS
3.1. Modulation of the NFT
Prosopis africana and Albizia labboclc nodulatod in a11 sriils
usod. The nombor of nodules was highor in Dabo soil. Proeo-
& .iuliflrra and Leucaena laucrcophala did not nodulate in
any scil (table 3).
3.2. Isclaticn of !?hirobium strain_anQ &udy of the-ir host -ramqe
Ten strains wcr~ Asol;--ttcd, a11 of thom from Prosopis africana
-
-
- 5 -
Prosopis africana and Albizia lobback and most of them
-
-
wre effective nn both hosts. Only t.tJo of these stroins
(Pal from Prosopis africana and A16 from Albizia lebback)
tirer 0 abla ta effoctively nodulats Leucaona leucocephalz.
-
-
None of the tan strains isolated uras able to nodulato
ProsoPis juliflora.
Interestingly ORS 911 (isolated from Acacia farnasiana)
was able t3 nodulate leucaena leucocaphala (table 4).
The cffectivity of the strains tuas di2terminatod only iuibh
Prossop.is africans and Albizia lebbeck, the best strains
being Al;oand pa7 (Fig. 1, 2, 3).
CONCLUSION
Our proiiminary investigations of thc four specics of NFT shn-
wed that those species fol1 into two inoculation groups accor-
ding to ths Rhizobium that nodulated thon1 :
the first group including prosopis africana and Albizia labbeck
-
-
ürhjch noduLatr;d rosdily ujith Rhizobium strains distributed in
tho fivo stinegslesc soils studied (which ara probably ubiqui-
tcu 3
strains) . Intortistingly,
sorne of tho Rhizobium strains
uùre fast grouing,
other were SLOW growing, tho tu3 catagLries
of Rhizobia. Thore was no relation boturoon tha efficiency of
these strains and their growth rate.
Tho sticond group including Prosopis Juliffora and Loucaena
loucoc:jphnlz; tuhich did not nodulatYd iuith thc native Rhizobium
strains Froiii thcsa scils, which suggsst thet thc: Rhizobium
raquiert.w?ent;s of Prosopis juliflora end LwcaeT'a ltiucoçaphala
Lzrc m o r s sPC?cificthon th2 cnss of thc first group. it aPpe3rs
necossery to initiate invastigations in th-t field.
L
=---
-
I
D.
D
IA 0
0
0
CJ
OY
Ln 0
3
0’
r
0
u 0
0
0’
0
o-
LA 0
0
0
N
133
W
fi
v- u?
Y- 0
T-
F 0
h
V
0
T-
In
0
0
0 0
0
CG- 0
CJ 0
0
0
CY
CY 0
V
r?
W
,T
h
x-- OI
T-
Lr’i
r- 0
Y-
lr,
v-
M
0
rc
M M
V
v- Ln
T- In
<j”
\\a 0
m
h
0
W
V
.---.
v- In
o*
0 c3
0
7
L”
0”
0 0
CZY
0 0
V
F--.
a
v- Ln
0”
0 0
V
h
0
v-
In
0
0 u
Table 4 - Host spectrum of ten Rhizobium strains ielated
From prosopis aFricana and Albizia lebbeck.
-3-
Straine
prosopis
Albizia
Leucaena
?roaopio
tested
africana
lebbeck
leucocephala ,iiliflorz
pa 1
fast
E
E
c1.i.
pa 3
fast
'e
e
n.i.
Pas
Sl.OW
E
n.i.
n.i.
Pa7
sl ow
E
n..i.
nri.
Pa9
sl.ow
E
n.i.
n.i.
A12
fast
e
kl
A14
fast
E
ri.i.
A16
slow
n.i,
n.i,
A18
SlOW
n.i,
E)
FI110
fabt
n.i.
r1.i.
ORS 911
f3.St
E
E
El
CB7 56
SlOW
e
n.i.
n.i.
E = effective ; e xz ineffective ; n.i.= non in?eCtiVe
*
= Plants of this treatment were contaminated.
- -
. . .,
t
F
I.,,
..s
..I
i
.,
.
,.
_
. .
1
5
REFERENCES
CORNET, F., 1982. Rscherches preliminaires sur la symbiose de deux Acacia
0.
tropicaux (A. raddisna et A. holasericea) avec Rhizobium sp, et
un chamoignon endomycorhizien (Glomus mosseas)
-
-
D.E.A. Microbio-
loqie. Eniuers. Claude Bernard, L y o n I.
+i DREYFUS, R.L., and DOMWEHGUES, Y.R., 1981. Nodulation of s p c
Acacia c i e s
b y Fast- and Slow-growing Trripical Strains of Rhizobium. APPlied
and Environ. Microb. 4 1 (11, 97-99.
GIBSON, A .H ., 1963. Physical environment in symbiotic nitrogen fixation.
The effect of root temperature on recently nodulated Trifolium
subterranax p l a n t s . Aust, J . Siol. S c i . 1 6 , 2 8 - 4 2 .
HALLIDAY, J., and SOMASEGARAN, P., 1982. Nodulation, Nitrogen Fixation
and Rhizobium strain afinities in the Ganus Leucaena. In Leucaena
-
-
research in the Asian-pacifie Region. 2:oceedinqs of a Workshop
held in Sinqapore, 23-26 Nov. 1982. p. 27-32.
JENSEN, H.L., 1942. Nitragon fixation in loguminous plants. General character
af root nodule bacteria isolated from species of Medicaqo and
Tri folium in Australia. oroceedinqs of the linnean Societv N.S.W.
67 > 98-108.
ROSKOSKI, J.P. et ai., 1980. Nitrogen Fixation by Tropical Woody Legumes :
potential !Source of Soi1 Enrichment. Biol. Nitroq. Fixat. Toch.
for Trop. Agr., An Int. Workshop,
VINCENT, J.M., 1970. A manual for the practical study of root nodules bac-
taria. I.B.P. Handbook no 15. Blackurell Sci. publications Oxford
snd EdSnburgh.
VINCENT, J.M., 1974. Root-nodule symbiosis urith zizobium : general micro-
biology. In the biology of nitrogon fixstion Ed. by Quiepsl, A,
North-Holland - publishing CO, Amsterdam, 265-341.