The Construction of Genetic Map of Faba Bean (Viciafaba L.) U\'ing RAPD Markers
,
I
:=~]
Bul. Agron. (29) (1) 31 -33 (2001)
c',
"'"
The Construction of Genetic Map of Faba Bean (Viciafaba L.) U\'ing RAPD Markers
ii~t'c
Memen Surahmanl)
!
~:I
ABSTRACT
.,
Fifty sevenindividuals of recombinantinbred lines (RlLs) offaba bean(Viciafaba L.) descended
from the cross
between34Morocco x Kristal125 have been analyzedfor RAPD markers. A total of 77 markers were assayed
segregatingin the 57 Rl lines and grouped into 14 linkagegroups,with total length of973.2 cM
Key words.. Viciafaba, Recombinantinbred line, RAPDs,Linkagegroups
;Ii;
iZ' ,:;;j ~ I
INTRODUCTION
were self-pollinatedand advancedto the F7 generation
Preliminary analysis of genetic linkage in faba
beanincludesthe study of morphologicaltraits (Cabrera
and Martin, 1989) and was followed by the
establishmentof linkage mapsbasedon morphological,
isozyme,RFLP and RAPD markers(Van de Yen et al.~
1991). Torres et al. (1993) reported,that when two F2
populations(20 F2 individuals derived from the cross
Vf6 x Vf173 and 44 F2 plants from the cross Vf6 x
Vf35) were analyzedusing isozyme,RFLP and RAPD,
II independently assorting linkage groups were
identified in this population. Satovic et al. (1996)
analysed13 F2 families of faba beandescendingfrom a
trisomic plant. They have been analyzed for
morphological,isozymeand RAPD markers.The study
revealeda total of 10 linkage groups,six of which have
beenprecisely assignedto specific chromosomes.The
mostrecentmap in faba beanhasbeenreportedby Vaz
of 253 lines including reciprocal lines. A random
sampleof 57 lines was characterizedbasedon RAPDs.
Segregation data were analyzed with MAPMAKER/EXP 3.0 computer program (Lander et al.,
1987; Lincoln et al., 1993),with a LaD score3.0 and
with 0.4 theta (40 Haldane cM; Haldane, 1919) as
critical distancebetweentwo linked markers.
1":" ~"" .J I
'-'c.
"'~
:
using single-seed descent. The final population consists
Patto et al. (1998). The map consist of 14 linkage
groups with total length of 984.5 cM. The main
objective of the presentstudy was to constructgenetic
map.
MATERIALS AND METHODS
;f'~
~
For the constructionof a linkage map a population
of recombinant inbred lines (RILs) derived from the
cross 34Morocco x Kristal125 was used. The
34Morocco is a bean pure line BPL228/ILBI4I from
ICARDA, Syria. Kristal125,developedat Hohenheim,
Germany, is an inbred line from the German cultivar
Kristall. These parentswere chosen becauseof their
large genetic and phenotypic difference (Schill et al.,
)997).
The F2 plants from the cross 34Morocco x
Kristal125and from the cross Kristal125x 34Morocco
?'i~
:,
I)
RESULTS
The 48 selectedprimers amplified a total of 115
polymorphic bands(datanot shown)with an averageof
2.3 polymorphic bands per primer. Fragment sizes
ranged from about 550 bp to 2400 bp. With linkage
criteria LaD >3.0 and a distanceof lessthan 40 cM, 77
RAPD marker loci were classified into 14 groups
(Figure 1). There were 38 unlinked markers. These
markerswere omitted.
The'size of the 14 linkagegroupsrangedfrom 6.3
cM to 278.7 cM. The total length of map is 973.2cM.
The largest linkage group contains 16 loci (linkage
group 3). The distancebetweenmarkersrangesfrom
1.9cM to 38.5 cM, with an averageof 14.66cM.
DISCUSSION
A total of 77 markerswere found segregatingin 57
RI F7-samplelines (Figure 1). The total map distance
defined amonglinked markerswas 973.2 centiMorgan.
A comparison of the present genetic map with the
hitherto published maps shows that its total length
resemblesthe map of Vaz Pattoet al. (1998)with a map
length of984.5 cM. Howeverthesemapsare still small
when comparedto the expectedsize of the Viciafaba
Dcpartmentof Agronomy. Facultyof Agriculture,BogorAgricultural University
KampusIPH DarmagaBogor 16610,Bogor,Email:[email protected]:
(0251)635141
31
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.
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Bul. Agron. (29) (1) 31 - 33 (2001)
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!
Figure I. Primary linkage map of faba bean (Vicia faba) with 77 RAPD-markers. Number to the left of a
linkage group representsmap distancein cM calculatedwith the Haldanemapping function. All
linkagesshownhavea LaD scoreof at least3.0. Designationsto the right are locusnames.
,
.'
;
:""'!;,
i
t
32
~.
::
MemenSurachman
,
~
-'
:':i1
8ul. Agron. (29) (1) 31 - 33 (2001)
,.
f
~
$;;
genome(1500 cM; Vaz Patio et al., 1998).Besidesthe
total maplength a completemap is obtainedonly when
thenumberof linkagegroupscorrespondsto the haploid
number of chromosomes. The haploid chromosome
numberof the faba bean is six (x = 6), therefore, six
linkage groups, instead of the 14 linkage groups
detectedhere, were expected. These results reflect
problemsassociatedwith the use of relatively small
populations(here:N = 57).
I .
fth
I t. I I
.
n vIew 0 e re a Ive y arge sIze 0fth e I.mk age
.
"
group 3, It may putatIvely be assignedto chromosome
numberoneof the fababean. In the standardkaryotype,
h. h
. h I
( T
I
t IS C romosome IS t e ongest one
orres et a.
1993). It is a metacentricchromosome.andabouttwice
as long as eachof the five subtelocentrlcchromosomes
(Cabreraet al., 1989):
.
The av~rage dIstance between markers m the
presentmap IS 14.66 cM when all 77 RAPD markers
wereused. Although the density of the map was lower
thanin the faba bean map constructedby Vaz Patio et
al. (1989) which has an averagedistanceof 12.95cM,
themappresentedherecan be consideredas sufficiently
densefor a QTL analysis. In general,a markerdistance
about of 20 cM is still adequate to detect QTL
(Tanskleyetal., 1993).
REFERENCES
Cabrera,A., and A. Martin. 1989. Analysis of genetic
linkage in faba bean (Vicia faba L.). FABIS
Newslett 24:3-5.
Cabrera, A., J.I. Cubero, and A. Martin.
1989. Genetic
mapping using trisomics in Viciafaba L. FABIS
Newslett 23:5-7.
Haldane, J.B.S.
j
,
~
1919.
The combination of linkage
value and the calculation of distancebetweenthe
loci of linked factors.J. Genet. 8:299-309.
Lander,E.S., P. Green,J: Abrahamson,A. Barlow, M.J.
Daly, S.E. Lincoln, and L. Newburg. 1987.
Mapmaker: An interactive computer packagefor
constructing primary genetic linkage maps of
experimentaland natural populations. Genomics
I: 174-181.
!
Lincoln, S.E.,. M.J. Daly: and. E.S. Lander. 19~3.
ConstructIng genetIc. lInkage maps. with
MAPMAKERlEXP verslo? 3.0: A Tu~onal and
re:erenc.e manual. WhItehead InstItute for
BIomedIcalResearch.USA.
.
Satovlc, Z.,.. A.M. Torres, and J.I. Cubero.
.
. 1996.
GenetIcmappIng of new morphologIcal, Isozyme
. f,,"
and RAPD markers m
"ICla fiaba L . usIng
.
..
t 93' 1130 - 1138
tnsomlcs. Theor. A pp.I Gene..
.
Schill, B., A.E. Melchinger, E. von Kittlitz, and W.
Link. 1997. Zilchterische Brauchbarkeit von
Intrapool-und Interpool
Kreuzungen des
mitteleuropaischenund mediterranenGenpoolsbei
der Fababohne (Vicia faba L.).
Vortr.
Pflanzenzilchtung38:127-145.
Tanksley, S.D., H. Medina-Filhoet, and C.M. Rick,
Ganal., M. W., Prince, J.P., de Vicente, M.C.,
t
Bonierbale, M. W., Broun, P., Fulton, T.M.,
~
Giovannoni, J.J., Grandillo, S., Martin, G.B.,
Messeguer,R., Miller, J.C., Miller, L., Paterson,
A.H., Pineda,0., Rlider, M.S., Wing, R.A., Wu,
W. and Young, N.D.
1993. High density
moleculat linkage maps of the tomato and potato
genomes.'Genetics 132:1141-1160.
Torres,A.M., Weeden,N.F., Martin, A. 1993. Linkage
among isozyme, RFLP and RAPD markers in
Viciafaba. Theor. Appl. Genet. 85:937-945.
Van de Yen, W.T.G., R. Waugh. N. Duncan, G.
Ramsay, N. Dow, and W. Powel.
1991.
Development of genetic linkage map in Vicia faba
using molecular and biochemical techniques.
AspectsAppl. BioI. 27:49-54.
Vaz Patio, M.C., A.M. Torres, and J.I. Cubero. 1998.
Geneticmappingin Viciafaba L. using trisomics.
3rd European Conference on Grain Legumes.
Valladolid. p.64-65.
"
;:,
;;
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~~
~;:
J:
~c
TheConstruction
of GeneticMapof...
.,.,.
I
i
"~
33
.
.
~
,
I
:=~]
Bul. Agron. (29) (1) 31 -33 (2001)
c',
"'"
The Construction of Genetic Map of Faba Bean (Viciafaba L.) U\'ing RAPD Markers
ii~t'c
Memen Surahmanl)
!
~:I
ABSTRACT
.,
Fifty sevenindividuals of recombinantinbred lines (RlLs) offaba bean(Viciafaba L.) descended
from the cross
between34Morocco x Kristal125 have been analyzedfor RAPD markers. A total of 77 markers were assayed
segregatingin the 57 Rl lines and grouped into 14 linkagegroups,with total length of973.2 cM
Key words.. Viciafaba, Recombinantinbred line, RAPDs,Linkagegroups
;Ii;
iZ' ,:;;j ~ I
INTRODUCTION
were self-pollinatedand advancedto the F7 generation
Preliminary analysis of genetic linkage in faba
beanincludesthe study of morphologicaltraits (Cabrera
and Martin, 1989) and was followed by the
establishmentof linkage mapsbasedon morphological,
isozyme,RFLP and RAPD markers(Van de Yen et al.~
1991). Torres et al. (1993) reported,that when two F2
populations(20 F2 individuals derived from the cross
Vf6 x Vf173 and 44 F2 plants from the cross Vf6 x
Vf35) were analyzedusing isozyme,RFLP and RAPD,
II independently assorting linkage groups were
identified in this population. Satovic et al. (1996)
analysed13 F2 families of faba beandescendingfrom a
trisomic plant. They have been analyzed for
morphological,isozymeand RAPD markers.The study
revealeda total of 10 linkage groups,six of which have
beenprecisely assignedto specific chromosomes.The
mostrecentmap in faba beanhasbeenreportedby Vaz
of 253 lines including reciprocal lines. A random
sampleof 57 lines was characterizedbasedon RAPDs.
Segregation data were analyzed with MAPMAKER/EXP 3.0 computer program (Lander et al.,
1987; Lincoln et al., 1993),with a LaD score3.0 and
with 0.4 theta (40 Haldane cM; Haldane, 1919) as
critical distancebetweentwo linked markers.
1":" ~"" .J I
'-'c.
"'~
:
using single-seed descent. The final population consists
Patto et al. (1998). The map consist of 14 linkage
groups with total length of 984.5 cM. The main
objective of the presentstudy was to constructgenetic
map.
MATERIALS AND METHODS
;f'~
~
For the constructionof a linkage map a population
of recombinant inbred lines (RILs) derived from the
cross 34Morocco x Kristal125 was used. The
34Morocco is a bean pure line BPL228/ILBI4I from
ICARDA, Syria. Kristal125,developedat Hohenheim,
Germany, is an inbred line from the German cultivar
Kristall. These parentswere chosen becauseof their
large genetic and phenotypic difference (Schill et al.,
)997).
The F2 plants from the cross 34Morocco x
Kristal125and from the cross Kristal125x 34Morocco
?'i~
:,
I)
RESULTS
The 48 selectedprimers amplified a total of 115
polymorphic bands(datanot shown)with an averageof
2.3 polymorphic bands per primer. Fragment sizes
ranged from about 550 bp to 2400 bp. With linkage
criteria LaD >3.0 and a distanceof lessthan 40 cM, 77
RAPD marker loci were classified into 14 groups
(Figure 1). There were 38 unlinked markers. These
markerswere omitted.
The'size of the 14 linkagegroupsrangedfrom 6.3
cM to 278.7 cM. The total length of map is 973.2cM.
The largest linkage group contains 16 loci (linkage
group 3). The distancebetweenmarkersrangesfrom
1.9cM to 38.5 cM, with an averageof 14.66cM.
DISCUSSION
A total of 77 markerswere found segregatingin 57
RI F7-samplelines (Figure 1). The total map distance
defined amonglinked markerswas 973.2 centiMorgan.
A comparison of the present genetic map with the
hitherto published maps shows that its total length
resemblesthe map of Vaz Pattoet al. (1998)with a map
length of984.5 cM. Howeverthesemapsare still small
when comparedto the expectedsize of the Viciafaba
Dcpartmentof Agronomy. Facultyof Agriculture,BogorAgricultural University
KampusIPH DarmagaBogor 16610,Bogor,Email:[email protected]:
(0251)635141
31
b
il
---::c.
.
.
I
:=~]
Bul. Agron. (29) (1) 31 -33 (2001)
c',
"'"
The Construction of Genetic Map of Faba Bean (Viciafaba L.) U\'ing RAPD Markers
ii~t'c
Memen Surahmanl)
!
~:I
ABSTRACT
.,
Fifty sevenindividuals of recombinantinbred lines (RlLs) offaba bean(Viciafaba L.) descended
from the cross
between34Morocco x Kristal125 have been analyzedfor RAPD markers. A total of 77 markers were assayed
segregatingin the 57 Rl lines and grouped into 14 linkagegroups,with total length of973.2 cM
Key words.. Viciafaba, Recombinantinbred line, RAPDs,Linkagegroups
;Ii;
iZ' ,:;;j ~ I
INTRODUCTION
were self-pollinatedand advancedto the F7 generation
Preliminary analysis of genetic linkage in faba
beanincludesthe study of morphologicaltraits (Cabrera
and Martin, 1989) and was followed by the
establishmentof linkage mapsbasedon morphological,
isozyme,RFLP and RAPD markers(Van de Yen et al.~
1991). Torres et al. (1993) reported,that when two F2
populations(20 F2 individuals derived from the cross
Vf6 x Vf173 and 44 F2 plants from the cross Vf6 x
Vf35) were analyzedusing isozyme,RFLP and RAPD,
II independently assorting linkage groups were
identified in this population. Satovic et al. (1996)
analysed13 F2 families of faba beandescendingfrom a
trisomic plant. They have been analyzed for
morphological,isozymeand RAPD markers.The study
revealeda total of 10 linkage groups,six of which have
beenprecisely assignedto specific chromosomes.The
mostrecentmap in faba beanhasbeenreportedby Vaz
of 253 lines including reciprocal lines. A random
sampleof 57 lines was characterizedbasedon RAPDs.
Segregation data were analyzed with MAPMAKER/EXP 3.0 computer program (Lander et al.,
1987; Lincoln et al., 1993),with a LaD score3.0 and
with 0.4 theta (40 Haldane cM; Haldane, 1919) as
critical distancebetweentwo linked markers.
1":" ~"" .J I
'-'c.
"'~
:
using single-seed descent. The final population consists
Patto et al. (1998). The map consist of 14 linkage
groups with total length of 984.5 cM. The main
objective of the presentstudy was to constructgenetic
map.
MATERIALS AND METHODS
;f'~
~
For the constructionof a linkage map a population
of recombinant inbred lines (RILs) derived from the
cross 34Morocco x Kristal125 was used. The
34Morocco is a bean pure line BPL228/ILBI4I from
ICARDA, Syria. Kristal125,developedat Hohenheim,
Germany, is an inbred line from the German cultivar
Kristall. These parentswere chosen becauseof their
large genetic and phenotypic difference (Schill et al.,
)997).
The F2 plants from the cross 34Morocco x
Kristal125and from the cross Kristal125x 34Morocco
?'i~
:,
I)
RESULTS
The 48 selectedprimers amplified a total of 115
polymorphic bands(datanot shown)with an averageof
2.3 polymorphic bands per primer. Fragment sizes
ranged from about 550 bp to 2400 bp. With linkage
criteria LaD >3.0 and a distanceof lessthan 40 cM, 77
RAPD marker loci were classified into 14 groups
(Figure 1). There were 38 unlinked markers. These
markerswere omitted.
The'size of the 14 linkagegroupsrangedfrom 6.3
cM to 278.7 cM. The total length of map is 973.2cM.
The largest linkage group contains 16 loci (linkage
group 3). The distancebetweenmarkersrangesfrom
1.9cM to 38.5 cM, with an averageof 14.66cM.
DISCUSSION
A total of 77 markerswere found segregatingin 57
RI F7-samplelines (Figure 1). The total map distance
defined amonglinked markerswas 973.2 centiMorgan.
A comparison of the present genetic map with the
hitherto published maps shows that its total length
resemblesthe map of Vaz Pattoet al. (1998)with a map
length of984.5 cM. Howeverthesemapsare still small
when comparedto the expectedsize of the Viciafaba
Dcpartmentof Agronomy. Facultyof Agriculture,BogorAgricultural University
KampusIPH DarmagaBogor 16610,Bogor,Email:[email protected]:
(0251)635141
31
b
il
---::c.
.
.
i
-~
Bul. Agron. (29) (1) 31 - 33 (2001)
1
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!
Figure I. Primary linkage map of faba bean (Vicia faba) with 77 RAPD-markers. Number to the left of a
linkage group representsmap distancein cM calculatedwith the Haldanemapping function. All
linkagesshownhavea LaD scoreof at least3.0. Designationsto the right are locusnames.
,
.'
;
:""'!;,
i
t
32
~.
::
MemenSurachman
,
~
-'
:':i1
8ul. Agron. (29) (1) 31 - 33 (2001)
,.
f
~
$;;
genome(1500 cM; Vaz Patio et al., 1998).Besidesthe
total maplength a completemap is obtainedonly when
thenumberof linkagegroupscorrespondsto the haploid
number of chromosomes. The haploid chromosome
numberof the faba bean is six (x = 6), therefore, six
linkage groups, instead of the 14 linkage groups
detectedhere, were expected. These results reflect
problemsassociatedwith the use of relatively small
populations(here:N = 57).
I .
fth
I t. I I
.
n vIew 0 e re a Ive y arge sIze 0fth e I.mk age
.
"
group 3, It may putatIvely be assignedto chromosome
numberoneof the fababean. In the standardkaryotype,
h. h
. h I
( T
I
t IS C romosome IS t e ongest one
orres et a.
1993). It is a metacentricchromosome.andabouttwice
as long as eachof the five subtelocentrlcchromosomes
(Cabreraet al., 1989):
.
The av~rage dIstance between markers m the
presentmap IS 14.66 cM when all 77 RAPD markers
wereused. Although the density of the map was lower
thanin the faba bean map constructedby Vaz Patio et
al. (1989) which has an averagedistanceof 12.95cM,
themappresentedherecan be consideredas sufficiently
densefor a QTL analysis. In general,a markerdistance
about of 20 cM is still adequate to detect QTL
(Tanskleyetal., 1993).
REFERENCES
Cabrera,A., and A. Martin. 1989. Analysis of genetic
linkage in faba bean (Vicia faba L.). FABIS
Newslett 24:3-5.
Cabrera, A., J.I. Cubero, and A. Martin.
1989. Genetic
mapping using trisomics in Viciafaba L. FABIS
Newslett 23:5-7.
Haldane, J.B.S.
j
,
~
1919.
The combination of linkage
value and the calculation of distancebetweenthe
loci of linked factors.J. Genet. 8:299-309.
Lander,E.S., P. Green,J: Abrahamson,A. Barlow, M.J.
Daly, S.E. Lincoln, and L. Newburg. 1987.
Mapmaker: An interactive computer packagefor
constructing primary genetic linkage maps of
experimentaland natural populations. Genomics
I: 174-181.
!
Lincoln, S.E.,. M.J. Daly: and. E.S. Lander. 19~3.
ConstructIng genetIc. lInkage maps. with
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GenetIcmappIng of new morphologIcal, Isozyme
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Kreuzungen des
mitteleuropaischenund mediterranenGenpoolsbei
der Fababohne (Vicia faba L.).
Vortr.
Pflanzenzilchtung38:127-145.
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W. and Young, N.D.
1993. High density
moleculat linkage maps of the tomato and potato
genomes.'Genetics 132:1141-1160.
Torres,A.M., Weeden,N.F., Martin, A. 1993. Linkage
among isozyme, RFLP and RAPD markers in
Viciafaba. Theor. Appl. Genet. 85:937-945.
Van de Yen, W.T.G., R. Waugh. N. Duncan, G.
Ramsay, N. Dow, and W. Powel.
1991.
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Bul. Agron. (29) (1) 31 -33 (2001)
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The Construction of Genetic Map of Faba Bean (Viciafaba L.) U\'ing RAPD Markers
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Memen Surahmanl)
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ABSTRACT
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Fifty sevenindividuals of recombinantinbred lines (RlLs) offaba bean(Viciafaba L.) descended
from the cross
between34Morocco x Kristal125 have been analyzedfor RAPD markers. A total of 77 markers were assayed
segregatingin the 57 Rl lines and grouped into 14 linkagegroups,with total length of973.2 cM
Key words.. Viciafaba, Recombinantinbred line, RAPDs,Linkagegroups
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INTRODUCTION
were self-pollinatedand advancedto the F7 generation
Preliminary analysis of genetic linkage in faba
beanincludesthe study of morphologicaltraits (Cabrera
and Martin, 1989) and was followed by the
establishmentof linkage mapsbasedon morphological,
isozyme,RFLP and RAPD markers(Van de Yen et al.~
1991). Torres et al. (1993) reported,that when two F2
populations(20 F2 individuals derived from the cross
Vf6 x Vf173 and 44 F2 plants from the cross Vf6 x
Vf35) were analyzedusing isozyme,RFLP and RAPD,
II independently assorting linkage groups were
identified in this population. Satovic et al. (1996)
analysed13 F2 families of faba beandescendingfrom a
trisomic plant. They have been analyzed for
morphological,isozymeand RAPD markers.The study
revealeda total of 10 linkage groups,six of which have
beenprecisely assignedto specific chromosomes.The
mostrecentmap in faba beanhasbeenreportedby Vaz
of 253 lines including reciprocal lines. A random
sampleof 57 lines was characterizedbasedon RAPDs.
Segregation data were analyzed with MAPMAKER/EXP 3.0 computer program (Lander et al.,
1987; Lincoln et al., 1993),with a LaD score3.0 and
with 0.4 theta (40 Haldane cM; Haldane, 1919) as
critical distancebetweentwo linked markers.
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using single-seed descent. The final population consists
Patto et al. (1998). The map consist of 14 linkage
groups with total length of 984.5 cM. The main
objective of the presentstudy was to constructgenetic
map.
MATERIALS AND METHODS
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For the constructionof a linkage map a population
of recombinant inbred lines (RILs) derived from the
cross 34Morocco x Kristal125 was used. The
34Morocco is a bean pure line BPL228/ILBI4I from
ICARDA, Syria. Kristal125,developedat Hohenheim,
Germany, is an inbred line from the German cultivar
Kristall. These parentswere chosen becauseof their
large genetic and phenotypic difference (Schill et al.,
)997).
The F2 plants from the cross 34Morocco x
Kristal125and from the cross Kristal125x 34Morocco
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RESULTS
The 48 selectedprimers amplified a total of 115
polymorphic bands(datanot shown)with an averageof
2.3 polymorphic bands per primer. Fragment sizes
ranged from about 550 bp to 2400 bp. With linkage
criteria LaD >3.0 and a distanceof lessthan 40 cM, 77
RAPD marker loci were classified into 14 groups
(Figure 1). There were 38 unlinked markers. These
markerswere omitted.
The'size of the 14 linkagegroupsrangedfrom 6.3
cM to 278.7 cM. The total length of map is 973.2cM.
The largest linkage group contains 16 loci (linkage
group 3). The distancebetweenmarkersrangesfrom
1.9cM to 38.5 cM, with an averageof 14.66cM.
DISCUSSION
A total of 77 markerswere found segregatingin 57
RI F7-samplelines (Figure 1). The total map distance
defined amonglinked markerswas 973.2 centiMorgan.
A comparison of the present genetic map with the
hitherto published maps shows that its total length
resemblesthe map of Vaz Pattoet al. (1998)with a map
length of984.5 cM. Howeverthesemapsare still small
when comparedto the expectedsize of the Viciafaba
Dcpartmentof Agronomy. Facultyof Agriculture,BogorAgricultural University
KampusIPH DarmagaBogor 16610,Bogor,Email:[email protected]:
(0251)635141
31
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