Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
PEANUT (ARACHIS HYPOGAEA L.) SEED PROTEIN CHARACTERIZATION AND GENOTYPE SAMPLE CLASSIFICATION USING POLYACRYLAMIDE GEL ELECTROPHORESIS Clifton F. Savoy School of Science and Technology Florida A & M University Tallahassee, Florida 32307 Received
December
4,1975
Summary: Peanut seed protein has been characterized using polyacrylamide gel electrophoresis. The same general electropherogram pattern as regardsnumbers and kinds of protein components resolved was observed for all forty-five samples tested. Therefore, a universal standard electropherogram protein pattern appears to exist for all peanut seed samples and is presented. However, quantitative differences of individual proteins may occur. Depending on the genoBased on type examined, four to six components contain most of the protein. amounts of these major components present, all the samples can be separatedinto one of four groups. Classification by this electropherogram grouping method offers another criterion in determining genetic relatedness. INTRODUCTION Polyacrylamide in protein protein
the
seed (l-3),
well
and in
performed
to monitor
enzymes
electrophoresis
biochemistry, have
ployed
gel
as heat
the last
various
Data presented
(18,19)
phoreticallyresolved
proteinswas
in experimentation,
andseveralreports
In these, to
develop
"standards" difficult protein
Copyright All rights
seed protein
characterize
for
various
to clearly patterns,
reveal
distinguish and defined
(15-17) peanut
0 I976 by Academic Press, Inc. of reproduction in any form reserved.
between
specifically
cultivars
886
of electro-
this
was examinedinan
Overall,
to
serve
they
report
simply were
not
as
parameter
concern
patterns
patterns
15-17)
proteins.
the totalnumber
cultivars
comparisons.
standard
(11,
animportantcontrol
electrophoretic test
and content(4-11),
and varietal
usedas
has beenem-
and localizationwithin
experiments
that
of peanut
It
on the individual
often
of different
and
effects
literature
technique.
development
and environmental
used method
many researchers
this
fractionation
(12-16),
in this
using
protein
arachin/conarachin
and roasting
few years,
analyses
and characterize
and reactions
(PAGE) has become a widely
attempt as possible that
on the basis established.
point.
it
was
of the It
was
Vol. 68, No. 3, 1976
noted
though
partially tive
BIOCHEMICAL
that
some cultivars
distinguished
from
and quantitative
indicates
instead
laboratory
peanut type
would
that
be of great
value
using
the
grown
location
in another
in these
patterns
standard
one for
be
by minorqualita-
of thedifferent
pattern
each.
An anodic
experiments.
typePAGE
systemdoes
might
information molecules
to the breeder SLS PAGE system,
exist
for
all gel
Data recently
obtained
not separate
and resolve
(SLS)
and because present
cul-
non-detergent
the sodiumlaurylsulfate
of this
could
the patterns.
a universal
of protein
geographic
protein
as wellas
In light
and kinds
accumulated
in
this
components (20).
the numbers
that
however,
reveal
protein system
the same type
of a different
system was utilized, this
in one
of electropherogram
(15-17)
cultivars
grown
differences
The similarity tivars
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
in
detergent
more knowledge
in the mature
and laboratory
investigator,
and some major
findings
of
peanut
seed
data were are
reported.
Materials and Methods: Oil was removed from all seed samples by treatment (20 mg ground seed in 100 ml solvent) for 30 min in cold (4'C) 90%acetone containing ammonium hydroxide (50 pl/lOO ml). This process was repeated, and deoiled (stripped) sample was collected by sedimentation (20,OtOg for 30 min at 4 C), quick-frozen, lyophilized to dryness and stored at -20 C until analyzed. PAGE. Detergent (SLS) gel electrophoresis was performed in 10% gels utilizing a combination of the basic systems of Ornstein and Davis (21,22) and Shapiro, Vinu'ela and Maize1 (23) as refined by Weber and Osborn (24) and Grula and Savoy (25). The capability of this procedure toseparate and resolve plant proteins has recently been reported and the methodology given in detail (20). Molecular weight (MW) determinationof resolved components was accomplished as previously described (23,24). The proteins of known MW used were: bovineserum albumin, DNase, concanavalin A, lysozyme and cytochrome c. Calibrationcurves were constructed using protein concentrations varying from 2.5 to 10 pg/gel. Results pleted
and Discussion: in this
C37, Florunner,
Jenkins PS1539
Comparison twelve
genotypes
components
analyzed
present
consisted Jumbo,
C-l-29-l
market
except
seed proteinhave
Senegal41-95 C-3,
that for
types
all
performed were
P.I.
are
quantitative
887
alike
com-
in each study,
GA 98-3-5,GA157-1, 196623,
PS 1555 C-2-33-lD, protein
been
to monitorex-
examined
of thefollowing:
of the electropherogram revealed
of peanut
SLS PAGE testingwas
One or more different
and the genotypes
229553,
studies
laboratorywherein
perimentation.
P.I.
Several
P.I.
268771B,
Bassee
NC 4144 and NC 5
patterns
obtained
as regards
numbers
differences
GA
from
(26). these
and kinds
of some individualpro-
of
Vol. 68, No. 3,1976
BIOCHEMICAL
(35.500
AND BIOPHYSICAL
MW)
12 16 CO. 500
L~~~~~~~l~~~~~l~l~i~I 00 01 0.2 0.3
RESEARCH COMMUNICATIONS
04
0.5
Retardation
0.6
MW)
07
08
09
1.0
coefflclent
Figure 1. Electropherogram component pattern of stripped peanut protein meal (Jenkins Jumbo) after PAGE in the SLS system. Detection was accomplished using CBB G250, and 60 pg meal (contains 35 to 50 pg protein) were analyzed. Malachite Green in solvent served as the marker dye, and retardation coefficients were calculated by setting the migration distance (leading edge) of the dye to one.
teins.
Fig.
scribe
1 shows
the major
tain
at least
pattern
were
protein
determined
a few on the
same gel
are
The number
type mobility
lengths molecule
This vary during
logarithm
the majority
to 74,000
considered
detergent
uniquely
with
SLS PAGE is of a globular
all
reported
approximate
converts
daltons.
samples could
MW (27). also
molecule
888
is
linear
compo-
retardation with
since
the
detection.
of the expecteddeviation
proteins
with
These
be minimal for
into
The logarithm
linear
to con-
simultaneously,
of the resolved
most
to de-
ofproteinshavl
by comparing
because
properties
can be used
Each was found
may be insufficient
of up to 10% (24) and the structural
whose
with
of 13,000
molecules
it
of the others.
analyzing
column.
of some protein
the SLS solvent.
and
to be the same in eachgenotype
andelectrophoretically
The MWvalues
Jumbo,
components,
MW distribution
coefficients
quantity
of Jenkins
characteristics
twenty-one
ing an approximate nents
the pattern
respect with
components
in
rod-like
particles
of mobility
of this
to MW; whereas the square
of
the its
ra-
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
TABLE I CLASSIFICATION OF PEANUT MARKET TYPES AND GENOTYPES BASED UPON THE PAGE ELECTROPHERCGRAM COMPONENT PATTERNS
Market Type and Genotype*
Growth
Group
Area
A
Classification B
C
D
Spanish Set
No. SPancross
1
Gi 116 Starr GK-19 comet GA 123 Spanhorns Tifspan Set No. 2 Spancross Starr
Tifton, Holland,
Set No. 1 GA 194R
Headland,
AL
Marianna, Headland, Stephenville, II
FL AL
GA VA
X
Runner
Set No. 2 Florunner II 11 ,I 1, II II 11 11 ,I II 11 II
Tifton, ,a Yoakum, Headland, Yoakum, Marianna, Headland, Marianna,
F439-16 11 11 0 ,, Early Runner II I,
X TX II
GA II TX AL TX FL AL FL
X X X X X X X
Virginia Jenkins
Jumbo
Gainesville,
FL
--Set No. 1 1C Fla 14 ?A 72R AU-2 Florigiant
Headland, 9, II 11
Set No. 2 Florigiant II 11 II 11 P.I. 31917&55 F393-6
Tifton, GA Yoakum, TX Stephenville, Marianna, FL Headland, AL
X
AL 1, 0, II
X X
TX X X
Uncertain Set
No.
1
Goldin*Set
Headland,
number
Jenkins
one and Goldin-
Judo
in
genotype
AL
X were
1973.
889
grown
in
1974,
number
two in
1971
and
Vol. 68, No. 3, 1976
dius
(24,28).
branching rod-like ably
BIOCHEMICAL
Thus,
mobility
or globular
mobility
characteristics
in the presence
However,preliminaryevidence and therefore
the similarity supports
allpeanutgenotypes.
To test
(1istedinTable
I,
and a peanut
Regardless
of these
(test
dard
pattern
previously
individual
proteins
four
for
of the
to six
protein,
acteristic
of the genotype
cially
into using
nents
group
ispoorin
proteins group bers
three
classifications 3, 5, 8-9,
and 16 are Group ponents.
C.
Genotypes
of this
that
analyzed.
season,
was also
the
obtained
for
a universal
patterns,
3, 5, 8-9,
stan-
differences
the twelve
of
group
components.
contain
5 and 12 are present
890
could
genotypes
be espe-
has
ofminor
led
compo-
showing
the resolved
2.
Descriptions
of
of this
Group
B.
5 is present,
numbers only
char-
columns
Genotypes
Number
seemingly
components,
of other
may be seen in Fig.
components.
most
reproducibility gel
noted
contain
genotypes
these
testing
Although
Group A.
was also
majorcomponentsis
Subsequent one.
it
12 and 16)
on theintensitiesof
12 and 16 as major major
or growing
Quantitative
genotype
representative
follow.
the only
Numbers
based
groups
additional
(APREA)were
genotypes
Moreover,
1).
5 and 12.
four
area
seed proteins.
of these
by subdividing
of these
for
at the 1975 meeting
Association
(numbers
(Fig.
a photograph,
geno-
exists
thirty-three
the conclusion
peanut
number
groups
numbers
to a fourth
twelve
andthe
pattern
obtained
growth
allow
twelve
though.
components
oftheseed
separated
data
of the
shown by AES or ARS coopera-
i/8160-16)
the twelve
peanut
may occur
In comparison only
for
These
does exist
area
one or more
characteristics.
standard
further,
and Education
that
patterns
possibility
genotype,
of SLS; mostprob-
globular
a universal
sample
type,
samples.
that
this
Research
described
may demonstrate
and growninthe
flour
of the market
each
idea
possess
indicates
in electropherogram the
of theAmericanPeanut
that
of molecular
whetherpeanutproteins
strongly
pattern
by extent
it isnotknown
Overall,
tors)
affected
At present
may be a glycoprotein
samples
is directly
RESEARCH COMMUNICATIONS
and size.
become rod-like.
types
AND BIOPHYSICAL
group
contain
Numbers but
the
3, 8-9,
in trace
amounts.
Group
12
amounts.
3. 8-9 and 16 as major
in trace
num-
D.
comThis
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Ah
C
D
16-
Figure 2. Photographic representation of SLS PAGE columns containingresolved CBB G250 was used for detection. The four groups peanut protein components; of electropherogram characteristics are exemplified by the patterns of thefollowing cultivars: (A) Jenkins Jumbo, (B) GA 98-3-5 (Spanish market type), (C) Florunner and (D) AU-2 (Virginia market type).
group not
is
like
present
Group
in sufficient
To determine were
C except
if
categorized
into
Separation breeder
reveals
grouping istics possess
only
similarity.
groups
might
exist,
into
B.
market
It
listed
for
type
that
all
one,
genotypes
are were
891
these though of this found
listed
all
or is
in Table
the samples
groups
previously
descriptions
as Spanish
be stated
B characteristics
type
to be missing
the samples
It was found
the market
cannot
12 appears
electropherogram
those
Except
the Group
and Virginia
detection.
one of the four
that
of Group
for
analyzed.
of the samples
number
quantity
other
electrophoretically
component
I
can be described.
provided
by the
demonstrateelectropherogram possess
that
all
market
the
patterncharacter-
the genotypes type
to be evenly
since distributed
which
the Runner among
BIOCHEMKAL
Vol. 68, No. 3,1976
the B, C and D Groups. not
at first
lowing
appear
needs
the
APREA meeting, guishable the
group it
last
the peanut
Moreover,
often
been
This
geographic tics
locations
(examine
Table
I),
nificantly Starr,
tern
are
these
grouping
offers
data
however,
until
classification of the
Runner
As a result,
be a result are
but
analyzed.
grown
in different
group
characteris-
or Florigiant
and F439-16).
that
within
and Valen-
samegenotype
and those
Classi-
market
Virginia
could
of one genotype
would
shown Because
of Group
one sample
in
Groups B are
sig-
in each of
the
C
not be so classified.
that
classification
criterion
by electropherogram
to distinguish
in classifying evaluation
Runner,
has occurred.
similar
for
apparent
and probably
is possible
should
reveal
another
of importance
it
is
designation.
quite
emphasized lines
and cultivar
Florunner
In
varieties,
the same electropherogram
(Starr,
to these,
Overall,
Spanish,
Early
this.
of the past.
confusion botanical
supposedly
possess
and F439-16
that
why samples
characteristics
Florunner
unknown,
explain
do not
different
The extent
could
of the Spancross,
and others
and D pattern
type
samples
all
those
terms
as distin-
for
have been
terms
reached.
is not
subsequenthybrid
or Virginia
in PAGE group
a necessaryeventwhen would
for
fol-
at the 1975
may be accountable
designations
the
labeling
differences
information
the
germplasm
the
is
and breeding
methods
by the fact
used as both
nomenclature
Therefore,
reasons
nomenclature
complicated
and cultivars.
peanut
does
however,
a conclusion
genetics
that
Runner
types;
such
breeding
Thus,
between
incorrect
(29) Several
literature
cia have
on peanut
to the Spanish,
further
market before
hybridization
new cultivars.
is
types
consideration
used to be.
not be eqivalent
by electropherogramgrouping
in distinguishing
discussion
few decades,
fication
classification
was mentioned
as it
developing
not
useful
information During
Thus,
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
market
types
pat-
betweenpeanutgenotypes. and genotypes
has been made on strict
botanical
remains varieties
of A. hypogaea. ACKNOWLEDGEMENTS This study was supported by Grant 516-15-61fromthe United States DepartDr. A. .I. Norden ment of Agriculture --- Cooperative State Research Service. provided the Jenkins Jumbo seed, and Dr. R. 0. Hammons the other seed in addi-
892
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
tion to a critique of the paper. Photography and technical assistance was given by Marvin extended to the Lord for overall support.
was performed by Dee Farrell, Felder. Gratitude is especially
REFERENCES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29.
Neucere, N. J. and Ory, R. L. (1970) Plant Physiol, 45, 616-619. Cheng, J. P. (1974) J. Agr. Food Chem, 22, 723-724. Young, C. T., Young, T. G. and Cherry, J. P. (1974) Proc. Am. Peanut Res. Ed. Assoc. (PAPREA), 6, 8-16. Evans, W. J., Carney, W. B., Dechary, J. M. and Altschul, A. M. (1962) Arch. Biochem. Biophys, 96, 233-239. Tombs, M. P. (1965) Biochem. J, 96, 119-133. Dawson, R. (1968) Brit. J. Nutr, 22, 601-607. Neucere, N. J. (1969) Anal. Biochem, 27, 15-24. Dawson, R. (1971) Anal. Biochem, 41, 305-313. Neucere, N. J. (1974) Agri. Food Chem, 22, 146-148. 62, 108-120. Shetty, K.J. and Narasinga Rao, M.S. (1974) Anal. Biochem, A.D. (1973) J. Sci. Fd. Agri, 24, 597-609. Dawson, R. and McIntosh, Cherry, J. P. and Ory, R. L. (1973) Phytochem, 12, 283-289. Cherry, J. P. and Ory, R. L. (1973) Phytochem, 12, 1581-1583. St. Angelo, A.J. and Ory, R.L. (1975) J. Agr. Food Chem, 23, 141-146. Ory, R. L. and Cherry, J. P. (1972) PAPREA, 4, 21-31. Cherry, J. P., Ory, R. L. and Mayne, Ruth Y. (1972) PAPREA, 4, 32-40. Cherry, J. P., Neucere, N. J. and Ory, R. L. (1971) PAPREA, 3, 63-74. Neucere, N. J., Ory, R. L. and Carney, W. B. (1969) J. Agr. Food Chem, 17, 25-28. N. J. (1972) .J. Agr. Food Chem, 20, 252-255. Neucere, Savoy, C. F.,Biochim. Biophys. Acta, submitted for publication. Ornstein, L. (1964) Ann. N. Y. Acad. Sci, 121, 321-349. Davis, B. J. (1964) Ann. N. Y. Acad. Sci, 121, 404-427. Sapiro, A. L., Vin&la, E. and Maizel, J. V. (1967) Biochem. Biophys. Res. Commun, 28, 815-820. Weber, K. and Osborn, Mary (1969) J. Biol. Chem, 244, 4406-4412. Grula, E.A. and Savoy, C.F. (1971) Biochem. Biophys. Res. Commun, 43, 325-332. Young, C.T. and Hammons, R. 0. (1973) Ol&agineux, 28, 293-297. Reynolds, J.A. and Tanford, C. (1970) J. Biol. Chem, 245, 5161-5169. Scurzi, W. and Fishbein, W.N. (1973) Trans. N.Y. Acad. Sci, 35, 396-416. Hammons, R.O., Supervisory Research Geneticist and Technical Advisor, USDA, Agriculture Research Service, Tifton, GA, 31794.
893
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
PEANUT (ARACHIS HYPOGAEA L.) SEED PROTEIN CHARACTERIZATION AND GENOTYPE SAMPLE CLASSIFICATION USING POLYACRYLAMIDE GEL ELECTROPHORESIS Clifton F. Savoy School of Science and Technology Florida A & M University Tallahassee, Florida 32307 Received
December
4,1975
Summary: Peanut seed protein has been characterized using polyacrylamide gel electrophoresis. The same general electropherogram pattern as regardsnumbers and kinds of protein components resolved was observed for all forty-five samples tested. Therefore, a universal standard electropherogram protein pattern appears to exist for all peanut seed samples and is presented. However, quantitative differences of individual proteins may occur. Depending on the genoBased on type examined, four to six components contain most of the protein. amounts of these major components present, all the samples can be separatedinto one of four groups. Classification by this electropherogram grouping method offers another criterion in determining genetic relatedness. INTRODUCTION Polyacrylamide in protein protein
the
seed (l-3),
well
and in
performed
to monitor
enzymes
electrophoresis
biochemistry, have
ployed
gel
as heat
the last
various
Data presented
(18,19)
phoreticallyresolved
proteinswas
in experimentation,
andseveralreports
In these, to
develop
"standards" difficult protein
Copyright All rights
seed protein
characterize
for
various
to clearly patterns,
reveal
distinguish and defined
(15-17) peanut
0 I976 by Academic Press, Inc. of reproduction in any form reserved.
between
specifically
cultivars
886
of electro-
this
was examinedinan
Overall,
to
serve
they
report
simply were
not
as
parameter
concern
patterns
patterns
15-17)
proteins.
the totalnumber
cultivars
comparisons.
standard
(11,
animportantcontrol
electrophoretic test
and content(4-11),
and varietal
usedas
has beenem-
and localizationwithin
experiments
that
of peanut
It
on the individual
often
of different
and
effects
literature
technique.
development
and environmental
used method
many researchers
this
fractionation
(12-16),
in this
using
protein
arachin/conarachin
and roasting
few years,
analyses
and characterize
and reactions
(PAGE) has become a widely
attempt as possible that
on the basis established.
point.
it
was
of the It
was
Vol. 68, No. 3, 1976
noted
though
partially tive
BIOCHEMICAL
that
some cultivars
distinguished
from
and quantitative
indicates
instead
laboratory
peanut type
would
that
be of great
value
using
the
grown
location
in another
in these
patterns
standard
one for
be
by minorqualita-
of thedifferent
pattern
each.
An anodic
experiments.
typePAGE
systemdoes
might
information molecules
to the breeder SLS PAGE system,
exist
for
all gel
Data recently
obtained
not separate
and resolve
(SLS)
and because present
cul-
non-detergent
the sodiumlaurylsulfate
of this
could
the patterns.
a universal
of protein
geographic
protein
as wellas
In light
and kinds
accumulated
in
this
components (20).
the numbers
that
however,
reveal
protein system
the same type
of a different
system was utilized, this
in one
of electropherogram
(15-17)
cultivars
grown
differences
The similarity tivars
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
in
detergent
more knowledge
in the mature
and laboratory
investigator,
and some major
findings
of
peanut
seed
data were are
reported.
Materials and Methods: Oil was removed from all seed samples by treatment (20 mg ground seed in 100 ml solvent) for 30 min in cold (4'C) 90%acetone containing ammonium hydroxide (50 pl/lOO ml). This process was repeated, and deoiled (stripped) sample was collected by sedimentation (20,OtOg for 30 min at 4 C), quick-frozen, lyophilized to dryness and stored at -20 C until analyzed. PAGE. Detergent (SLS) gel electrophoresis was performed in 10% gels utilizing a combination of the basic systems of Ornstein and Davis (21,22) and Shapiro, Vinu'ela and Maize1 (23) as refined by Weber and Osborn (24) and Grula and Savoy (25). The capability of this procedure toseparate and resolve plant proteins has recently been reported and the methodology given in detail (20). Molecular weight (MW) determinationof resolved components was accomplished as previously described (23,24). The proteins of known MW used were: bovineserum albumin, DNase, concanavalin A, lysozyme and cytochrome c. Calibrationcurves were constructed using protein concentrations varying from 2.5 to 10 pg/gel. Results pleted
and Discussion: in this
C37, Florunner,
Jenkins PS1539
Comparison twelve
genotypes
components
analyzed
present
consisted Jumbo,
C-l-29-l
market
except
seed proteinhave
Senegal41-95 C-3,
that for
types
all
performed were
P.I.
are
quantitative
887
alike
com-
in each study,
GA 98-3-5,GA157-1, 196623,
PS 1555 C-2-33-lD, protein
been
to monitorex-
examined
of thefollowing:
of the electropherogram revealed
of peanut
SLS PAGE testingwas
One or more different
and the genotypes
229553,
studies
laboratorywherein
perimentation.
P.I.
Several
P.I.
268771B,
Bassee
NC 4144 and NC 5
patterns
obtained
as regards
numbers
differences
GA
from
(26). these
and kinds
of some individualpro-
of
Vol. 68, No. 3,1976
BIOCHEMICAL
(35.500
AND BIOPHYSICAL
MW)
12 16 CO. 500
L~~~~~~~l~~~~~l~l~i~I 00 01 0.2 0.3
RESEARCH COMMUNICATIONS
04
0.5
Retardation
0.6
MW)
07
08
09
1.0
coefflclent
Figure 1. Electropherogram component pattern of stripped peanut protein meal (Jenkins Jumbo) after PAGE in the SLS system. Detection was accomplished using CBB G250, and 60 pg meal (contains 35 to 50 pg protein) were analyzed. Malachite Green in solvent served as the marker dye, and retardation coefficients were calculated by setting the migration distance (leading edge) of the dye to one.
teins.
Fig.
scribe
1 shows
the major
tain
at least
pattern
were
protein
determined
a few on the
same gel
are
The number
type mobility
lengths molecule
This vary during
logarithm
the majority
to 74,000
considered
detergent
uniquely
with
SLS PAGE is of a globular
all
reported
approximate
converts
daltons.
samples could
MW (27). also
molecule
888
is
linear
compo-
retardation with
since
the
detection.
of the expecteddeviation
proteins
with
These
be minimal for
into
The logarithm
linear
to con-
simultaneously,
of the resolved
most
to de-
ofproteinshavl
by comparing
because
properties
can be used
Each was found
may be insufficient
of up to 10% (24) and the structural
whose
with
of 13,000
molecules
it
of the others.
analyzing
column.
of some protein
the SLS solvent.
and
to be the same in eachgenotype
andelectrophoretically
The MWvalues
Jumbo,
components,
MW distribution
coefficients
quantity
of Jenkins
characteristics
twenty-one
ing an approximate nents
the pattern
respect with
components
in
rod-like
particles
of mobility
of this
to MW; whereas the square
of
the its
ra-
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
TABLE I CLASSIFICATION OF PEANUT MARKET TYPES AND GENOTYPES BASED UPON THE PAGE ELECTROPHERCGRAM COMPONENT PATTERNS
Market Type and Genotype*
Growth
Group
Area
A
Classification B
C
D
Spanish Set
No. SPancross
1
Gi 116 Starr GK-19 comet GA 123 Spanhorns Tifspan Set No. 2 Spancross Starr
Tifton, Holland,
Set No. 1 GA 194R
Headland,
AL
Marianna, Headland, Stephenville, II
FL AL
GA VA
X
Runner
Set No. 2 Florunner II 11 ,I 1, II II 11 11 ,I II 11 II
Tifton, ,a Yoakum, Headland, Yoakum, Marianna, Headland, Marianna,
F439-16 11 11 0 ,, Early Runner II I,
X TX II
GA II TX AL TX FL AL FL
X X X X X X X
Virginia Jenkins
Jumbo
Gainesville,
FL
--Set No. 1 1C Fla 14 ?A 72R AU-2 Florigiant
Headland, 9, II 11
Set No. 2 Florigiant II 11 II 11 P.I. 31917&55 F393-6
Tifton, GA Yoakum, TX Stephenville, Marianna, FL Headland, AL
X
AL 1, 0, II
X X
TX X X
Uncertain Set
No.
1
Goldin*Set
Headland,
number
Jenkins
one and Goldin-
Judo
in
genotype
AL
X were
1973.
889
grown
in
1974,
number
two in
1971
and
Vol. 68, No. 3, 1976
dius
(24,28).
branching rod-like ably
BIOCHEMICAL
Thus,
mobility
or globular
mobility
characteristics
in the presence
However,preliminaryevidence and therefore
the similarity supports
allpeanutgenotypes.
To test
(1istedinTable
I,
and a peanut
Regardless
of these
(test
dard
pattern
previously
individual
proteins
four
for
of the
to six
protein,
acteristic
of the genotype
cially
into using
nents
group
ispoorin
proteins group bers
three
classifications 3, 5, 8-9,
and 16 are Group ponents.
C.
Genotypes
of this
that
analyzed.
season,
was also
the
obtained
for
a universal
patterns,
3, 5, 8-9,
stan-
differences
the twelve
of
group
components.
contain
5 and 12 are present
890
could
genotypes
be espe-
has
ofminor
led
compo-
showing
the resolved
2.
Descriptions
of
of this
Group
B.
5 is present,
numbers only
char-
columns
Genotypes
Number
seemingly
components,
of other
may be seen in Fig.
components.
most
reproducibility gel
noted
contain
genotypes
these
testing
Although
Group A.
was also
majorcomponentsis
Subsequent one.
it
12 and 16)
on theintensitiesof
12 and 16 as major major
or growing
Quantitative
genotype
representative
follow.
the only
Numbers
based
groups
additional
(APREA)were
genotypes
Moreover,
1).
5 and 12.
four
area
seed proteins.
of these
by subdividing
of these
for
at the 1975 meeting
Association
(numbers
(Fig.
a photograph,
geno-
exists
thirty-three
the conclusion
peanut
number
groups
numbers
to a fourth
twelve
andthe
pattern
obtained
growth
allow
twelve
though.
components
oftheseed
separated
data
of the
shown by AES or ARS coopera-
i/8160-16)
the twelve
peanut
may occur
In comparison only
for
These
does exist
area
one or more
characteristics.
standard
further,
and Education
that
patterns
possibility
genotype,
of SLS; mostprob-
globular
a universal
sample
type,
samples.
that
this
Research
described
may demonstrate
and growninthe
flour
of the market
each
idea
possess
indicates
in electropherogram the
of theAmericanPeanut
that
of molecular
whetherpeanutproteins
strongly
pattern
by extent
it isnotknown
Overall,
tors)
affected
At present
may be a glycoprotein
samples
is directly
RESEARCH COMMUNICATIONS
and size.
become rod-like.
types
AND BIOPHYSICAL
group
contain
Numbers but
the
3, 8-9,
in trace
amounts.
Group
12
amounts.
3. 8-9 and 16 as major
in trace
num-
D.
comThis
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Ah
C
D
16-
Figure 2. Photographic representation of SLS PAGE columns containingresolved CBB G250 was used for detection. The four groups peanut protein components; of electropherogram characteristics are exemplified by the patterns of thefollowing cultivars: (A) Jenkins Jumbo, (B) GA 98-3-5 (Spanish market type), (C) Florunner and (D) AU-2 (Virginia market type).
group not
is
like
present
Group
in sufficient
To determine were
C except
if
categorized
into
Separation breeder
reveals
grouping istics possess
only
similarity.
groups
might
exist,
into
B.
market
It
listed
for
type
that
all
one,
genotypes
are were
891
these though of this found
listed
all
or is
in Table
the samples
groups
previously
descriptions
as Spanish
be stated
B characteristics
type
to be missing
the samples
It was found
the market
cannot
12 appears
electropherogram
those
Except
the Group
and Virginia
detection.
one of the four
that
of Group
for
analyzed.
of the samples
number
quantity
other
electrophoretically
component
I
can be described.
provided
by the
demonstrateelectropherogram possess
that
all
market
the
patterncharacter-
the genotypes type
to be evenly
since distributed
which
the Runner among
BIOCHEMKAL
Vol. 68, No. 3,1976
the B, C and D Groups. not
at first
lowing
appear
needs
the
APREA meeting, guishable the
group it
last
the peanut
Moreover,
often
been
This
geographic tics
locations
(examine
Table
I),
nificantly Starr,
tern
are
these
grouping
offers
data
however,
until
classification of the
Runner
As a result,
be a result are
but
analyzed.
grown
in different
group
characteris-
or Florigiant
and F439-16).
that
within
and Valen-
samegenotype
and those
Classi-
market
Virginia
could
of one genotype
would
shown Because
of Group
one sample
in
Groups B are
sig-
in each of
the
C
not be so classified.
that
classification
criterion
by electropherogram
to distinguish
in classifying evaluation
Runner,
has occurred.
similar
for
apparent
and probably
is possible
should
reveal
another
of importance
it
is
designation.
quite
emphasized lines
and cultivar
Florunner
In
varieties,
the same electropherogram
(Starr,
to these,
Overall,
Spanish,
Early
this.
of the past.
confusion botanical
supposedly
possess
and F439-16
that
why samples
characteristics
Florunner
unknown,
explain
do not
different
The extent
could
of the Spancross,
and others
and D pattern
type
samples
all
those
terms
as distin-
for
have been
terms
reached.
is not
subsequenthybrid
or Virginia
in PAGE group
a necessaryeventwhen would
for
fol-
at the 1975
may be accountable
designations
the
labeling
differences
information
the
germplasm
the
is
and breeding
methods
by the fact
used as both
nomenclature
Therefore,
reasons
nomenclature
complicated
and cultivars.
peanut
does
however,
a conclusion
genetics
that
Runner
types;
such
breeding
Thus,
between
incorrect
(29) Several
literature
cia have
on peanut
to the Spanish,
further
market before
hybridization
new cultivars.
is
types
consideration
used to be.
not be eqivalent
by electropherogramgrouping
in distinguishing
discussion
few decades,
fication
classification
was mentioned
as it
developing
not
useful
information During
Thus,
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
market
types
pat-
betweenpeanutgenotypes. and genotypes
has been made on strict
botanical
remains varieties
of A. hypogaea. ACKNOWLEDGEMENTS This study was supported by Grant 516-15-61fromthe United States DepartDr. A. .I. Norden ment of Agriculture --- Cooperative State Research Service. provided the Jenkins Jumbo seed, and Dr. R. 0. Hammons the other seed in addi-
892
Vol. 68, No. 3, 1976
BIOCHEMICAL
AND BIOPHYSICAL RESEARCH COMMUNICATIONS
tion to a critique of the paper. Photography and technical assistance was given by Marvin extended to the Lord for overall support.
was performed by Dee Farrell, Felder. Gratitude is especially
REFERENCES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29.
Neucere, N. J. and Ory, R. L. (1970) Plant Physiol, 45, 616-619. Cheng, J. P. (1974) J. Agr. Food Chem, 22, 723-724. Young, C. T., Young, T. G. and Cherry, J. P. (1974) Proc. Am. Peanut Res. Ed. Assoc. (PAPREA), 6, 8-16. Evans, W. J., Carney, W. B., Dechary, J. M. and Altschul, A. M. (1962) Arch. Biochem. Biophys, 96, 233-239. Tombs, M. P. (1965) Biochem. J, 96, 119-133. Dawson, R. (1968) Brit. J. Nutr, 22, 601-607. Neucere, N. J. (1969) Anal. Biochem, 27, 15-24. Dawson, R. (1971) Anal. Biochem, 41, 305-313. Neucere, N. J. (1974) Agri. Food Chem, 22, 146-148. 62, 108-120. Shetty, K.J. and Narasinga Rao, M.S. (1974) Anal. Biochem, A.D. (1973) J. Sci. Fd. Agri, 24, 597-609. Dawson, R. and McIntosh, Cherry, J. P. and Ory, R. L. (1973) Phytochem, 12, 283-289. Cherry, J. P. and Ory, R. L. (1973) Phytochem, 12, 1581-1583. St. Angelo, A.J. and Ory, R.L. (1975) J. Agr. Food Chem, 23, 141-146. Ory, R. L. and Cherry, J. P. (1972) PAPREA, 4, 21-31. Cherry, J. P., Ory, R. L. and Mayne, Ruth Y. (1972) PAPREA, 4, 32-40. Cherry, J. P., Neucere, N. J. and Ory, R. L. (1971) PAPREA, 3, 63-74. Neucere, N. J., Ory, R. L. and Carney, W. B. (1969) J. Agr. Food Chem, 17, 25-28. N. J. (1972) .J. Agr. Food Chem, 20, 252-255. Neucere, Savoy, C. F.,Biochim. Biophys. Acta, submitted for publication. Ornstein, L. (1964) Ann. N. Y. Acad. Sci, 121, 321-349. Davis, B. J. (1964) Ann. N. Y. Acad. Sci, 121, 404-427. Sapiro, A. L., Vin&la, E. and Maizel, J. V. (1967) Biochem. Biophys. Res. Commun, 28, 815-820. Weber, K. and Osborn, Mary (1969) J. Biol. Chem, 244, 4406-4412. Grula, E.A. and Savoy, C.F. (1971) Biochem. Biophys. Res. Commun, 43, 325-332. Young, C.T. and Hammons, R. 0. (1973) Ol&agineux, 28, 293-297. Reynolds, J.A. and Tanford, C. (1970) J. Biol. Chem, 245, 5161-5169. Scurzi, W. and Fishbein, W.N. (1973) Trans. N.Y. Acad. Sci, 35, 396-416. Hammons, R.O., Supervisory Research Geneticist and Technical Advisor, USDA, Agriculture Research Service, Tifton, GA, 31794.
893
