Plant Breeding 130, 156—164 (2011) doi:10.1111/j.1439-0523.2010.01832.

x
 2011 Blackwell Verlag GmbH

Seed protein amino acid composition of important local grain legumes Lupinus
angustifolius L., Lupinus luteus L., Pisum sativum L. and Vicia faba L.
H . S C H U M A C H E R 1 , 2 , H . M . P A U L S E N 1 , A . E . G A U 2 , W . L I N K 3 , H . U . J Ü R G E N S 4 , O . S A S S 5 and R . D I E T E R I C H 6
1
Institute of Organic Farming, Johann Heinrich von Thünen-Institute (vTI), Trenthorst 32, 23847 Westerau, http://
www.vti.bund.de, Germany, E-mail: henrik.schumacher@botanik.uni-hannover.de; 2Institute of Botany, Gottfried Wilhelm
Leibniz University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany; 3Department of Crop Sciences, Plant Breeding,
Georg-August University, Von-Siebold-Str. 8, D-37075 Göttingen, Germany; 4Institute of Resistance Research and Stress
Tolerance, Julius Kühn-Institute (JKI), Rudolf-Schick-Platz 3, 18190 Groß Lüsewitz, Germany; 5Norddeutsche Pflanzenzucht,
Hans-Georg Lembke KG, Hohenlieth, 24363 Holtsee, Germany; 6Saatzucht Steinach GmbH, Wittelsbacher Str. 15, 94377
Steinach, Germany
With 3 figures and 5 tables
Received May 14, 2010/Accepted September 27, 2010
Communicated by R. Singh

Abstract main nutritional deficiencies of legumes are the low concentra-

Grain legumes are regarded as highly valuable protein source for
human and animal nutrition. Legume protein quality is mainly limited
by the amino acids (AAs) tryptophan and sulphur AA. Organic
farming in particular requires high seed protein quality for livestock
feeding, as chemically produced AAs must not be feeded. Breeding
attempts to increase contents of limiting AA are required. In the
present survey, the AA content of 107 cultivars of important European
grain legume species (Lupinus angustifolius, Lupinus luteus, Pisum
sativum and Vicia faba) was analysed. AA contents were related to the
requirements of growing pigs and human nutrition. Feed quality could
be enhanced by choice of high quality varieties according to ideal
protein concepts. For example for sulphur, AA feed quality for pigs
could be increased by up to 22% (e.g. for L. angustifolius: ideal
protein = 100, sample mean = 59.7, sample maximum = 72.7).
Regarding livestock nutrition, ranges of limiting AA never reached
the qualities reported for soybean seeds. However, an inclusion of high
quality legume lines would reduce the need for other high quality
components in feed compositions.
Key words: legume quality — sulphur-containing amino acids
— organic farming — plant breeding — pure on-farm diets
Legumes are regarded as a high quality feedstuff for livestock
and human nutrition. To provide pure on-farm diets for
livestock in organic farming, legumes are of particular impor-
tance as a protein source. Hereby, the nutritional quality of the
cultivated feed crops is a major concern because the use of
chemically produced amino acids (AAs) is not allowed as an
additive to the livestock diets (EC 889/2008). Enriching natural
contents of AA would reduce the need for feed components
like potato- and maize-protein that are rarely available from
organic origin (Zollitsch et al. 2004).
For legume breeders, major concerns have been the increase
of yield, protein content, disease resistance, plant growth
behaviour and the reduction of anti-nutritional components.
Up to now, the enhancement of protein composition and
quality has been of secondary interest (Ranalli and Cubero
1997, Wang et al. 2003).
Nutritional value of plant seed protein is mainly determined by
three factors: the amount of anti-nutritional components, the
content of AA and the amino acid digestibility (Gatel 1994). Most
legume species possess generally high seed protein content. The
tions of sulphur AA and tryptophan (Monti and Grillo 1983,
Graham and Vance 2003). Pea seeds, for instance, contain about
0.8% methionine and about 1% cysteine in the protein, while
growing pigs require 3.5% sulphur AA in their dietary protein, of
which 1.6% has to be methionine (Tabe and Higgins 1998). This
indicates the insufficiency of legumes as sole foodstuff.
Legume seed proteins can mainly be assigned to two types of
proteins: the globulins and the albumins (e.g. for peas, the
relation is 80:20%; Gatel 1994). Globulins are distinguished
into 7S (vicilin-like proteins) and 11S (legumin-like proteins)
globulins according to their sedimentation characteristics.
Both protein families are rich in certain AAs like arginine,
aspartate and glutamate, but only the legumins contain
sulphur AA (Böttinger 2004). The ratio of vicilin to legumin
in V. faba differs from 1 : 2 to 1 : 3.7 (Müntz et al. 1999).
Furthermore, Tzitzikas et al. (2006) observed a high variability
of legumin proportion to the overall protein content between
5.3% and 24.5% in peas.
Proteins of the albumin fraction possess a far more desirable
amino acid composition with higher contents of the primarily
limiting AAs methionine and tryptophan in most legume
species (Monti and Grillo 1983).
Because the metabolism of sulphur-containing components
like cysteine and methionine can be downregulated in plants
under sulphur-restricted conditions (Hawkesford and de Kok
2006), AA composition varies according to the nutrient supply
of sites. For example, insufficient sulphur supply leads to
proteins low in sulphur-containing AA in protein of Triticum
spp. (Wrigley et al. 1980, Byers et al. 1987, Zhao et al. 1996).
Also, strong effects of the sulphur supply on the protein
composition of legume seed storage proteins and correspond-
ing effects on the sulphur AA concentrations are reported
(summarized by Scherer 2001). A recent study revealed a
strong site-specific effect on sulphur-containing AA in proteins
of Lupinus spp. (Jansen et al. 2006).
Efforts in Enhancing the Nutritive Value by Genetic
Engineering
In the past, sulphur AA-enriched legumes were predominantly
created by genetic engineering, which has mainly been focussed

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Seed amino acid composition of grain legumes
on two approaches. One is the alteration of the biosynthesis
pathways of the sulphur AA (Krishnan 2005). Hereby, genes
of key enzymes of the biosynthesis pathways are targeted for
modification. Such a key enzyme in the methionine biosyn-
thesis is the aspartate kinase, which is responsible for the
phosphorylation of aspartate, the first precursor in the
methionine biosynthesis. Both existing isoforms are feedback
inhibited by either lysine or threonine, other possible products
of this biosynthesis pathway. Introducing a feedback-insensi-
tive aspartate kinase into the plant genome leads to an
overaccumulation of several products of this pathway, includ-
ing methionine (Tabe and Higgins 1998).
The second major approach is the transfer of genes of
methionine-rich proteins into the legume genome. These can
either be already abundant endogenous proteins (e.g. albumin
fraction proteins) or proteins from other plant species like the
maize 21-kDa zein. So far, neither strategy has been successful
and has had unforeseen consequences. So Townsend and
Thomas (1994) were able to transform soybeans with the
sulphur-rich 2S albumin derived from the Brazil nut (Bert-
holletia excelsa H.B.K.). They generated transgenic soybean
lines with an elevated methionine content by 15–40%. But
2 years later, the Brazil nut albumin was identified as an major
allergen (Nordlee et al. 1996) which abruptly ended this
previously promising approach. Up to now, no breakthrough
in improving protein quality parameters by genetic modifica-
tion of legumes has been reported.
Possibilities to Breed Plants for Organic Agriculture
As the use of genetically engineered plants is strictly forbidden
in organic farming (Ifoam norms: IFOAM 2009 and EC No
834/2007), conventional plant breeding methods must be used
for breeding organic cultivars. Breeding practices based on
chemical mutation are controversial with regard to organic
guidelines [e.g. the methods of organic plant breeding given by
the European Consortium for Organic Plant Breeding, ECO-
PB, (http://www.eco-pb.com/)] and require critical discussion
(Rahmann et al. 2009).
However, there are positive examples for successful selection
of the sulphur AA traits of legumes that are based on mutation
breeding. Imsande (2001) was able to generate soybean inbred
lines with an increased amount of sulphur AA. A darker green
leaf phenotype could be associated with methionine-enriched
individual plants. Plants were screened for this trait, and
selected individual plants were interbred. In a similar breeding
approach with important European grain legume species such
as L. angustifolius, P. sativum and V. faba supplied with
methionine in the growth medium, analogue phenotypical
reactions could be observed (Schumacher et al. 2009).
As background for those breeding concepts on protein
quality, this paper gives an overview of the AA composition of
typical home-grown legumes based on an analysis of samples
selected from main commercial European plant breeders that
are active in the further development of grain legumes.
Material and Methods
Plant material: The amino acid composition of 107 legume cultivars of
four species (L. angustifolius, L. luteus, P. sativum and V. faba) was
analysed. The cultivars were selected by two German breeding
companies (Norddeutsche Pflanzenzucht, Holtsee and Saatzucht Stein-
ach, Steinach) and the Department of Crop Sciences, Plant Breeding at
157
the University Göttingen according to their available species and by
considerations to cover a wide range of AA concentrations.
The Norddeutsche Pflanzenzucht KG (NPZ) provided 50 varieties of
P. sativum. Varieties included forage peas (36), several marrowfat peas
(8), four catch crop varieties and two winter varieties. The Saatzucht
Steinach GmbH allocated 50 seed samples of L. angustifolius and
L. luteus. The 27 L. angustifolius varieties originated from Belarus (8),
Germany (4), Hungary (1), Poland (4), Australia (4), Spain (2), Morocco
(1), Russia (1), Ukraine (1) and the former Yugoslavia (1). Plants were
grown on two different locations in Germany predominantly in 2007.
Additionally, four varieties of L. luteus were provided, originating from
Germany and grown at one location at the same time.
The Department of Crop Science of the University of Göttingen
provided 50 seed samples of 27 V. faba varieties for the analysis. Of
the 27 different varieties, 25 originated from Germany. The other two
originated from Spain and Italy, respectively. Two tannin-free culti-
vars among the German varieties supplemented the sample.
Of these varieties, 23 were grown at the same location and time
under drought stress as well as normal growing conditions to analyse
stress effects on AA composition. Furthermore, four winter cultivars of
V. faba were grown during winter season at the same location.
Chemical analyses: Chemical analysis was conducted with whole seed
material. Nitrogen and sulphur analyses were performed according to
the methods of Kjeldahl and Dumas, respectively (Marco et al. 2002,
AOAC Official Method 990.03). Crude protein content was calculated
for determined nitrogen values (%N · 6.25).
Amino acids were analysed according to the Guideline 98/64/EG
(1998) of the European Union for the measurements of AA, raw fat
and olaquindox in feeding stuff. Measurements were conducted by
HPLC, and results were calculated as g/16 g N or g/kg dry matter. All
essential AAs but tryptophan were determined.
Estimation of nutritional values: Parameters for livestock feeding:
Lysine, methionine and threonine are generally regarded as the first
limiting AAs in practical nutrition of monogastric animals. Ideal protein
concepts compare AA and protein contents in feed rations to the ideal
ratio of the essential AA and sometimes also ideal N-contents required
for maintenance and production (Boisen 2000) or AA relations are used
to define protein quality (Losand et al. 2003). For the assessment and
classification of the explored variabilities in the sample, these param-
eters are used and introduced in more detail in the following. Absolute
AA concentrations as well as AA contents relative to the protein can be
taken into account for the optimization of feed rations.
Parameters for human nutrition: AA contents are compared to
appropriate concentration standards for a mature human (FAO/
WHO, 1991). Reference values are (in g/16g N) Lys = 5.5,
Met + Cys = 3.5, Thr = 4, Ile = 4, Val = 5, Leu = 7 and
Phe + Tyr = 6. Chemical score (CS) has been determined according
to Sujak et al. (2006). It is described as the quotient of the measured
amino acid divided by the ideal protein concentration of that amino
acid (CS = ap/asx · 100). The absolute CS is generally aligned to the
most limiting essential amino acid.
Protein efficiency ratio (PER) has been calculated according to
Alsmeyer et al. (1974): PER = )1.816 + 0.435 (Met) + 0.78
(Leu) + 0.211 (His))0.944 (Tyr). Originally, PER values display the
ratio of ingested protein to newly created body protein.
Statistical analysis: Statistical analysis of results was performed with
SPSS Software (Student Version 14).
Normal distribution was analysed, and means were compared with
ANOVA or non-parametric tests. Correlation of data was analysed by
Pearson rank correlation coefficient test. Confidence interval was 5%.
Results
Crude protein content of all three species in the analysed
sample varied between 18 and 45 g per 100 g dry weight of
158 H. Schumacher, H. M. Paulsen, A. E. Gau et al.

All plant species showed a high range in the crude protein

Fig. 1: Seed crude protein content of cultivars of five legume species.
Averages according to the literature (Schuster et al. 1998, Sujak et al.
2006) are indicated as black lines. (n cultivars): Pisum sativum = 50;
V. faba = 50; L. angustifolius = 46, L. luteus = 4; Glycine
max = 119 (G. max from Shi et al. 2010 and Zarkadas et al. 2007)
seed material (Fig. 1). The highest protein content was
measured in the seeds of L. luteus, with a mean value of
44%. Sujak et al. (2006) reported a similar mean protein value
of 46.5%. The lowest amount of crude protein could be found
in the seeds of P. sativum, with a mean content of 23%.
content with a mean aberration of >20%.
The results of the AA determination are displayed in
Tables 1 and 2.
In the analysed sample, maximum values of AA contents
relative to protein were between 1% and 36% higher than the
sample mean of the specific AA. The highest positive aberation
for methionine and lysine values was found in L. angustifolius
(22% and 11%, respectively; Table 1).
Referred to the absolute AA contents (Table 2), maximum
values for the AA were between 1% and 61% higher than the
sample mean. The highest positive aberration for methionine
and lysine values was found in P. sativum (24% and 23%,
respectively) and for cysteine in L. angustifolius (26%).
For V. faba, the drought stress influence on the AA
composition was investigated (Table 3). Plants grown under
drought stress conditions accumulated more crude protein and
a higher seed sulphur content. Relative AA levels were
significantly changed for several essential AA. Arginine
concentration was elevated under stress conditions. In con-
trast, relative concentrations of lysine, valine, isoleucine and
phenylalanine decreased significantly. The concentration of
S-containing AA relative to the protein content remained
unaffected, whereas the absolute contents in seeds (g/kg seeds)
increased from 11% (methionine) to 14.8% (cysteine) under
drought stress.

Table 1: Mean ± SD and range of AA concentrations (g/16 g N) in seeds of different grain legumes and in brackets positive aberration of the
maximum value from the sample mean (%). Glycine max values according to Zarkadas et al. (2007)

Pisum sativum Vicia faba Vicia faba (winter) Lupinus angustifolius Lupinus luteus Glycine max
Amino acid in
g/16 g N n = 50 n = 46 n=4 n = 46 n=4 n = 14

Ala (mean) 4.4 ± 0.2 3.9 ± 0.1 3.8 ± 0.1 3.4 ± 0.2 3.2 ± 0 3.6 ± 0.2
(range) 4.1–4.9 (11%) 3.6–4.1 (5%) 3.7–3.9 (3%) 2.9–3.8 (12%) 3.1–3.2 (1%) 3.2–3.8 (9%)
Asp (mean) 11.1 ± 0.4 10.1 ± 0.3 10 ± 0.2 9.5 ± 0.3 9.2 ± 0.1 10.3 ± 0.5
(range) 10–11.8 (6%) 9.6–10.8 (7%) 9.7–10.2 (2%) 8.8–10.1 (6%) 9.1–9.2 (1%) 9.6–11 (8%)
Cys (mean) 1.4 ± 0.1 1.1 ± 0.1 1.1 ± 0.1 1.5 ± 0.2 1.8 ± 0.1 1.8 ± 0.1
(range) 1–1.6 (19%) 0.9–1.2 (14%) 1–1.2 (12%) 1.1–1.8 (25%) 1.7–1.9 (5%) 1.5–2.1 (12%)
Glu (mean) 16.6 ± 0.5 16 ± 0.4 15.7 ± 0.4 21.1 ± 0.6 21.6 ± 0.1 16.2 ± 1.5
(range) 15.2–17.9 (8%) 15–16.9 (5%) 15.2–16 (1%) 19.6–22.4 (7%) 21.5–21.8 (1%) 15.7–16.9 (15%)
Gly (mean) 4.4 ± 0.2 4.2 ± 0.1 4.1 ± 0 4.2 ± 0.2 3.9 ± 0.1 3.4 ± 0.2
(range) 4–4.7 (8%) 3.8–4.4 (5%) 4–4.1 (1%) 3.7–4.6 (9%) 3.8–3.9 (1%) 3.1–3.5 (7%)
Pro (mean) 3.5 ± 0.1 3.5 ± 0.1 3.5 ± 0.1 3.7 ± 0.2 3.5 ± 0.1 4.6 ± 0.8
(range) 3.3–4.2 (20%) 3.2–3.7 (5%) 3.4–3.6 (2%) 3.4–4.2 (14%) 3.5–3.7 (5%) 4.1–4.6 (58%)
Ser (mean) 4.3 ± 0.2 4.3 ± 0.1 4.2 ± 0.1 4.5 ± 0.1 4.4 ± 0 5.1 ± 0.2
(range) 3.9–4.6 (7%) 4–4.4 (4%) 4.1–4.3 (3%) 4.3–4.8 (6%) 4.3–4.4 (1%) 5–5.8 (14%)
Tyr (mean) 3.6 ± 0.2 3.5 ± 0.2 3.4 ± 0 3.8 ± 0.2 3.4 ± 0.1 3.6 ± 0.1
(range) 3.2–4 (10%) 3.3–4.2 (20%) 3.4–3.4 (1%) 3.5–4.1 (9%) 3.3–3.5 (2%) 3.5–3.8 (5%)
Arg (mean) 8.5 ± 0.9 8.8 ± 0.6 8.8 ± 0.4 10.1 ± 0.5 10.4 ± 0.2 6.9 ± 0.2
(range) 7–11.7 (36%) 7.7–10.1 (15%) 8.4–9.2 (5%) 9.2–11.3 (12%) 10.2–10.7 (3%) 6.5–7.1 (3%)
His (mean) 2.4 ± 0.1 2.4 ± 0.1 2.4 ± 0 2.7 ± 0.1 2.7 ± 0 2.5 ± 0.1
(range) 2.2–2.5 (5%) 2.3–2.6 (5%) 2.4–2.5 (1%) 2.4–2.9 (7%) 2.6–2.7 (1%) 2.3–2.6 (4%)
Ile (mean) 4.4 ± 0.2 4.2 ± 0.1 4.1 ± 0.1 4.3 ± 0.1 4 ± 0.1 4.3 ± 0.2
(range) 3.9–4.7 (8%) 3.9–4.3 (4%) 4–4.2 (2%) 4–4.6 (7%) 3.9–4.1 (2%) 4–4.6 (5%)
Leu (mean) 7.2 ± 0.2 7.1 ± 0.2 7.1 ± 0.2 7 ± 0.2 7.2 ± 0.1 6.9 ± 0.2
(range) 6.6–7.7 (8%) 6.7–7.5 (5%) 6.9–7.3 (2%) 6.3–7.4 (7%) 7.2–7.3 (1%) 6.6–7.1 (3%)
Lys (mean) 7.3 ± 0.3 6.3 ± 0.3 6.1 ± 0.1 4.8 ± 0.2 4.9 ± 0.1 5.7 ± 0.2
(range) 6.3–7.9 (9%) 5.7–6.9 (9%) 6–6.3 (3%) 4.2–5.4 (11%) 4.8–4.9 (1%) 5.5–6 (4%)
Met (mean) 1 ± 0.1 0.7 ± 0 0.7 ± 0 0.7 ± 0.1 0.7 ± 0 1.8 ± 0.1
(range) 0.8–1.1 (14%) 0.6–0.8 (12%) 0.7–0.8 (8%) 0.5–0.8 (22%) 0.6–0.7 (6%) 1.5–2.1 (13%)
Phe (mean) 4.9 ± 0.2 4.2 ± 0.1 4.1 ± 0.1 4 ± 0.1 3.9 ± 0 4.8 ± 0.2
(range) 4.2–5.3 (8%) 3.9–4.4 (6%) 4.1–4.3 (4%) 3.8–4.3 (8%) 3.9–4 (1%) 4.5–5.3 (11%)
Thr (mean) 3.8 ± 0.2 3.4 ± 0.1 3.3 ± 0.1 3.5 ± 0.2 3.3 ± 0 4.7 ± 0.4
(range) 3.4–4.2 (10%) 3.1–3.6 (6%) 3.2–3.4 (4%) 3–3.9 (12%) 3.2–3.3 (1%) 4.5–5.9 (26%)
Val (mean) 4.9 ± 0.2 4.6 ± 0.1 4.6 ± 0.1 4.2 ± 0.2 3.7 ± 0.1 4.8 ± 0.2
(range) 4.5–5.3 (8%) 4.4–4.8 (4%) 4.4–4.6 (2%) 3.7–4.6 (9%) 3.7–3.8 (2%) 4.5–5.2 (8%)

AAs, amino acids.

Seed amino acid composition of grain legumes 159

Table 2: Elemental nitrogen, sulphur (%) and amino acid concentrations (g/kg dry matter) in seeds of different grain legumes. Mean ± SD,
range and in brackets positive aberration of the maximum value from the sample mean (%). Glycine max oil cake values (crude protein 44%)
according to Kirchgeßner (1997))

Pisum sativum Vicia faba Vicia faba (winter) Lupinus angustifolius Lupinus luteus

Amino acids n = 50 n = 46 n=4 n = 46 n=4 G. max (oil cake)

N% (mean) 3.7 ± 0.7 3.5 ± 0.1 4.2 ± 0.1 4.9 ± 0.5 7.1 ± 0.5 n.a.
(range) 2.9–6.2 (67%) 3.3–3.9 (12%) 4.1–4.3 (2%) 3.9–5.9 (22%) 6.8–7.8 (10%) n.a.
S% (mean) 0.2 ± 0 0.2 ± 0 0.2 ± 0 0.3 ± 0 0.4 ± 0 n.a.
(range) 0.13–0.25 (32%) 0.13–0.23 (40%) 0.1–0.2 (9%) 0.2–0.4 (33%) 0.4–0.5 (11%) n.a.
Ala (mean) 10.3 ± 1.1 10.7 ± 0.7 10.8 ± 0.1 10.1 ± 0.7 14 ± 0.3 23.8
(range) 8.4–13.3 (30%) 9–11.8 (11%) 10.7–11 (2%) 8.9–11.8 (16%) 13.6–14.3 (2%) n.a.
Asp (mean) 26 ± 2.4 27.7 ± 2.3 28.7 ± 0.8 28.3 ± 2.7 40.5 ± 1.2 59.3
(range) 21.3–31.6 (21%) 22.1–32.1 (16%) 27.6–29.4 (2%) 23.2–34.2 (21%) 38.8–41.3 (2%) n.a.
Cys (mean) 3.2 ± 0.2 3 ± 0.3 3.1 ± 0.2 4.3 ± 0.4 8 ± 0.2 7.8
(range) 2.3–3.7 (16%) 2.3–3.5 (19%) 2.9–3.4 (12%) 3.5–5.4 (26%) 7.7–8.1 (2%) n.a.
Glu (mean) 38.9 ± 3.5 43.8 ± 3.8 45 ± 1.3 62.6 ± 7.6 95.7 ± 2.3 98.8
(range) 30.8–47.6 (22%) 34.7–51.2 (17%) 43.1–46 (2%) 51.1–78.2 (25%) 92.3–97.3 (2%) n.a.
Gly (mean) 10.2 ± 0.9 11.4 ± 0.9 11.6 ± 0.2 12.4 ± 1.1 17.1 ± 0.4 22.8
(range) 8.3–12.4 (22%) 9.7–12.8 (12%) 11.4–11.8 (2%) 10.5–14.8 (19%) 16.6–17.5 (3%) n.a.
Pro (mean) 8.2 ± 0.8 9.6 ± 1 10 ± 0.3 10.9 ± 1.4 15.7 ± 0.8 26.3
(range) 6.2–11.3 (37%) 7.4–11.1 (15%) 9.7–10.2 (2%) 8.7–13.9 (27%) 14.8–16.6 (6%) n.a.
Ser (mean) 10.1 ± 0.9 11.6 ± 1 12 ± 0.4 13.4 ± 1.3 19.3 ± 0.4 28.7
(range) 8.3–12.2 (21%) 9.3–13.2 (14%) 11.5–12.3 (3%) 11.2–16 (19%) 18.7–19.7 (2%) n.a.
Tyr (mean) 8.4 ± 0.7 9.6 ± 0.9 9.7 ± 0.1 11.2 ± 1.1 15.1 ± 0.6 17.3
(range) 7.1–10.3 (22%) 7.9–11.4 (18%) 9.5–9.8 (1%) 9.4–13.9 (24%) 14.2–15.5 (3%) n.a.
Arg (mean) 20.2 ± 3.9 24.2 ± 3.4 25.1 ± 1 30.3 ± 4.6 46.1 ± 1.9 35.4
(range) 13.3–32.5 (61%) 17.4–30.9 (28%) 24.1–26.2 (4%) 22.8–40.3 (33%) 43.4–47.8 (4%) n.a.
His (mean) 5.6 ± 0.5 6.7 ± 0.5 6.9 ± 0.1 8 ± 0.7 11.7 ± 0.3 13.3
(range) 4.5–6.7 (21%) 5.5–7.5 (13%) 6.8–7 (1%) 6.7–9.6 (20%) 11.3–12 (2.5%) n.a.
Ile (mean) 10.2 ± 0.9 11.4 ± 0.9 11.8 ± 0.3 12.7 ± 1.2 17.7 ± 0.5 24.1
(range) 8.2–12.5 (22%) 9.2–13.1 (15%) 11.4–12.1 (2%) 10.7–15 (18%) 17–18.2 (3%) n.a.
Leu (mean) 16.8 ± 1.5 19.5 ± 1.7 20.3 ± 0.6 20.7 ± 1.8 32 ± 0.6 39.5
(range) 13.4–20.7 (24%) 15.5–22.5 (16%) 19.5–20.8 (3%) 17.7–24.8 (20%) 31.2–32.5 (2%) n.a.
Lys (mean) 17 ± 1.5 17.2 ± 1 17.5 ± 0.4 14.4 ± 1.1 21.5 ± 0.3 33
(range) 14.2–20.8 (23%) 14.5–19.2 (11%) 17.1–18.1 (3%) 12.4–16.8 (17%) 21–21.9 (2%) n.a.
Met (mean) 2.3 ± 0.2 1.9 ± 0.1 2 ± 0.1 2 ± 0.1 2.9 ± 0 7.5
(range) 1.9–2.8 (24%) 1.7–2.3 (17%) 1.9–2.2 (7%) 1.8–2.4 (16%) 2.9–3 (2%) n.a.
Phe (mean) 11.4 ± 0.9 11.4 ± 0.9 11.9 ± 0.4 11.9 ± 1.1 17.3 ± 0.4 24.7
(range) 9–13.8 (22%) 9.3–13.1 (14%) 11.6–12.3 (4%) 9.8–14.2 (19%) 16.7–17.7 (2%) n.a.
Thr (mean) 8.8 ± 0.8 9.3 ± 0.7 9.5 ± 0.2 10.3 ± 0.8 14.4 ± 0.3 20.8
(range) 7.5–11.1 (26%) 7.8–10.3 (11%) 9.3–9.8 (3%) 8.8–12.1 (17%) 14.1–14.8 (2%) n.a.
Val (mean) 11.5 ± 1 12.6 ± 1 13.1 ± 0.3 12.4 ± 1 16.5 ± 0.4 24.6
(range) 9.4–14.1 (22%) 10.4–14.2 (13%) 12.6–13.3 (2%) 10.6–14.5 (17%) 16.1–17 (3%) n.a.

The nutritive value of the seed protein for human consump-
tion was calculated in regard to certain protein quality
indicators (Table 4). For CS calculation, AAs were related to
ideal protein for human consumption (FAO/WHO, 1991).
In Table 5, the AA content of the seed protein of the sample
is related to ideal protein AA for growing pigs according to
Boisen (1997). Sulphur AA was most limiting for human and
pig nutritional demands. Second limiting AAs were either
threonine or lysine. Other limiting AAs were leucine and
valine. The ratio of lysine to other limiting AA in the protein
diet is displayed in Fig. 2. None of the investigated European
legume species reached an optimal ratio of sulphur AA to
lysine (0.6), which is required in pig nutrition.
The correlation of selected AA to the crude protein content is
shown in Fig. 3. Lysine was negatively correlated to the crude
protein content for L. angustifolius (r = )0.82***), V. faba
(r = )0.67***) and P. sativum (r = )0.5***). For L. angus-
tifolius and V. faba, a significant negative correlation of methi-
onine to the crude protein content could be observed (r =
)0.83*** and r = )0.83***). A weaker but still significant
correlation of the methionine content in P. sativum varieties to
the crude protein content was calculated (r = )0.38***).
Cysteine and crude protein content were negatively corre-
lated for all three species as well, but not significantly for
V. faba (r = )0.444ns) [(L. angustifolius (r = )0.584***),
P. sativum (r = )0.704***)]. Thus, total relative sulphur AA
content was negatively correlated to the crude protein content
(data not shown; V. faba: r = )0.61**. P. sativum:
r = )0.669***. L. angustifolius: r = )0.704***).
By contrast, the semi essential amino acid arginine was
positively correlated to the crude protein content (V. faba:
r = 0.79***. P. sativum: r = 0.65***. L. angustifolius:
r = 0.75***).
Discussion
The crude protein contents of the species in the analysed
legume sample (Fig. 1) match with the existing literature
values (e.g., Wang and Daun 2004). The protein content in the
pea sample ranged from 18% to 29%. A broader range from
14% to 31% in 59 pea lines was reported by Tzitzikas et al.
(2006).
The analysis confirms that P. sativum is a comparatively low
protein-containing legume while lupins are regarded as high
protein-containing legume species (Gallardo et al. 2008).
Cultivars of L. angustifolius contained a mean protein content
of 30% per dry weight, which is slightly below the literature
value of 34% (Sujak et al. 2006). The mean protein content of
160 H. Schumacher, H. M. Paulsen, A. E. Gau et al.

Table 3: Impact of different environmental conditions on the amino acid composition (g/kg dry matter) of V. faba seeds. Difference of drought
stress site to control site is displayed in per cent; n = 18 varieties per location

Control Drought stress Aberration Control Drought stress Aberration

N (%) 3.88 4.38 12.9***

S (%) 0.15 0.17 18.9***
Crude protein (g/kg) 254 ± 18 286 ± 14 12.6***
Amino acids (g/16 g N)

Cys 2.7 ± 0.2 3.1 ± 0.2 14.8*** 1.08 ± 0.09 1.1 ± 0.07 1.3 n.s.
Asp 25.8 ± 1.9 28.7 ± 1.4 11.2*** 10.16 ± 0.22 10.05 ± 0.25 )1 n.s.
Glu 40.7 ± 3.3 45.6 ± 2.1 12*** 16.05 ± 0.37 15.98 ± 0.39 )0.5 n.s.
Ser 10.8 ± 0.8 12.1 ± 0.6 12*** 4.27 ± 0.09 4.23 ± 0.12 )0.9 n.s.
His 6.2 ± 0.4 7 ± 0.3 12.9*** 2.45 ± 0.06 2.44 ± 0.06 )0.2 n.s.
Gly 10.7 ± 0.6 12.1 ± 0.5 13.1*** 4.21 ± 0.08 4.22 ± 0.09 0.3 n.s.
Arg 21.2 ± 2.5 26.1 ± 2.3 23.1*** 8.33 ± 0.45 9.13 ± 0.43 9.5***
Thr 8.7 ± 0.6 9.6 ± 0.4 10.3*** 3.45 ± 0.09 3.38 ± 0.07 )2.2*
Ala 10 ± 0.6 11.1 ± 0.4 11*** 3.96 ± 0.1 3.89 ± 0.09 )1.7 n.s.
Met 1.8 ± 0.1 2 ± 0.1 11.1*** 0.72 ± 0.04 0.7 ± 0.03 )2.8*
Pro 8.8 ± 0.8 10.1 ± 0.6 14.8*** 3.46 ± 0.1 3.55 ± 0.08 2.7*
Tyr 9 ± 0.8 10 ± 0.6 11.1*** 3.57 ± 0.16 3.5 ± 0.21 )1.9 n.s.
Val 11.8 ± 0.8 13.1 ± 0.6 11*** 4.66 ± 0.08 4.58 ± 0.11 )1.8*
Lys 16.4 ± 0.9 17.7 ± 0.5 7.9*** 6.48 ± 0.23 6.19 ± 0.28 )4.4**
Ile 10.6 ± 0.8 11.8 ± 0.6 11.3*** 4.2 ± 0.09 4.12 ± 0.13 )1.8*
Leu 18.2 ± 1.5 20.3 ± 1 11.5*** 7.16 ± 0.18 7.09 ± 0.22 )1 n.s.
Phe 10.7 ± 0.7 11.8 ± 0.5 10.3*** 4.21 ± 0.11 4.12 ± 0.14 )2.1*

n.a., not analysed; n.s., nonsignificant. *P < 0.05 **P < 0.01 ***P < 0.001 Tukey Test resp. Mann–Whitney U Test.

Table 4: Nutritional characteristics of the seed protein of the legume varieties indicated by the chemical score of essential AA. Values were
calculated in relation to human ideal protein standards

CS Pisum sativum Vicia faba Vicia faba (winter) Lupinus angustifolius Lupinus luteus Glycine max

Lys (mean) 132.0 115.2 111.4 87.9 88.3 104.5

(range) 113.7–143.6 104.4–125.8 109.3–114.8 75.7–97.8 87.4–89.6 99.3–108.7
Ile (mean) 109.1 104.2 103.1 106.5 100.2 108.2
(range) 97.9–117.9 96.6–108.4 100.4–105.2 100.2–113.8 98.7–101.8 100.3–114
Leu (mean) 102.6 102.1 101.4 99.7 103.2 98.3
(range) 93.7–110.7 95.1–106.8 98–103.9 89.6–106.4 102.2–104.4 93.7–101.1
Phe + Tyr (mean) 141.3 128.3 125.6 129.3 121.8 139.4
(range) 123.4–151.5 121.9–140.2 123.8–128.1 122.2–139.9 120.4–123.8 133.2–150.8
Cys + Met (mean) 66.7 51.3 50.9 61.4 70.7 104.7
(range) 54.4–76.3 41.1–58.2 48–56.4 46.2–74.7 67.7–74.4 93.7–109.7
Val (mean) 98.4 92.4 91.3 83.2 74.8 167.2
(range) 89.3–105.8 87.5–95.7 88.8–92.9 74–91.1 73.6–75.9 90.2–103
Thr (mean) 94.3 85.0 82.8 87.0 81.6 116.8
(range) 84.8–103.8 78–89.8 80.9–86.1 75.7–97.2 80.2–82.6 101–146.8
PER (mean) 1.3 1.3 1.3 0.9 1.5 1.5
(range) 1.1–1.6 0.7–1.6 1.1–1.5 0.5–1.4 1.4–1.6 1–1.7

CS, chemical score; PER, protein efficiency ratio; AAs, amino acids.

the V. faba cultivars was determined with 27%, which is 4%
lower than the 31% reported by Lattanzio et al. (1983).
Amino acidsÕ contents in the protein comply with published
surveys (Table 1). Sujak et al. (2006) analysed the nutri-
tional composition of several varieties of L. angustifolius and
L. luteus. The mean concentration for all essential AAs was
even or slightly lower in the eight L. angustifolius cultivars in
their work except for histidine (3.1 g/16 g N) and glutamic
acid (23.1 g/16 g N). For L. luteus, Sujak et al. (2006) reported
congruent values with most AAs. Main differences are a
considerably lower amount of valine (3.2 g/16 g N) and
isoleucine (3.5 g/16 g N).
Values in the sample of P. sativum are close to data observed
by Wang and Daun (2004). They reported a lower mean
arginine (7.9 g/16 g N) concentration and a superior amount
of proline (4.2 g/16 g N) for five Canadian pea varieties.
Measured AA contents of V. faba were comparatively low.
The Proline contents in the analysed sample are notably lower
than the values reported in the literature (4.2 g/16 g N), while
tyrosine is considerably higher (2.8 g/16 g N) (Lattanzio et al.
1983, Lisiewska et al. 2007).
Of particular interest are the limiting sulphur AAs. The
lowest mean cysteine concentration was measured in seeds of
V. faba varieties (1.1 g/16 g N). Highest cysteine concentra-
tions were measured for seeds of L. luteus (1.8 g/16 g N) and
L. angustifolius (1.5 g/16 g N). Sujak et al. (2006) reported a
similar mean cysteine content for L. angustifolius (1.4 g/16 g
N) but a higher content for L. luteus (2.2 g/16 g N). P. sativum
cysteine levels had a mean value of 1.4 g/16 g N. Several
literature sources report homogeneous mean cysteine contents
in P. sativum of about 1.5 g/16 g N (e.g. Zdunczyk et al.
1997). The maximum mean methionine content in the sample
Seed amino acid composition of grain legumes 161

Table 5: Calculated AA quality in seed protein of five legume species relative to ideal protein for growing pigs (according to Boisen 1997)

Pisum sativum Vicia faba Vicia faba (winter) Lupinus angustifolius Lupinus luteus Glycine max

Lys (mean) 103.7 90.5 87.5 69.1 69.4 82.1

(range) 89.4–112.9 82–98.8 85.9–90.2 59.5–76.8 68.6–70.4 78–85.4
Cys + Met (mean) 64.9 49.9 49.5 59.7 68.7 101.8
(range) 52.9–74.1 40–56.6 46.7–54.8 44.9–72.7 65.8–72.4 91.1–106.7
Thr (mean) 83.8 75.6 73.6 77.4 72.5 103.9
(range) 75.4–92.3 69.3–79.8 71.9–76.6 67.3–86.4 71.3–73.5 89.8–130.4
Ile (mean) 109.1 104.2 103.1 106.5 100.2 108.2
(range) 97.9–117.9 96.6–108.4 100.4–105.2 100.2–113.8 98.7–101.8 100.3–114
Leu (mean) 89.7 89.3 88.8 87.2 90.3 86.0
(range) 82–96.9 83.2–93.4 85.8–90.9 78.4–93.1 89.4–91.4 82–88.5
Val (mean) 94.6 88.9 87.8 80.0 71.9 94.6
(range) 85.9–101.7 84.1–92 85.3–89.3 71.2–87.6 70.7–73 86.7–99
Phe + Tyr (mean) 105.9 96.2 94.2 97.0 91.3 104.6
(range) 92.5–113.6 91.4–105.2 92.9–96.1 91.7–104.9 90.3–92.8 99.9–113.1

AAs, amino acids.

(Table 3). Plants accumulated higher amounts of crude pro-

Fig. 2: Mean ratio of limiting amino acids (AAs) (Sulfur AA and
threonine) to lysine in five different legume species (data for Glycine
max from Zarkadas et al. 2007). Vertical bars represent the sample
range. Dotted line indicates optimal ratio for pig nutrition (according
to Losand et al. 2003)
was measured for P. sativum (1 g/16 g N) (Table 1). Cultivars
of L. spp. and V. faba had similar mean methionine concen-
trations, whereby the range for L. spp. seeds was greater. As
expected, the methionine concentration in seeds of Glycine
max (according to Zarkadas et al. 2007) was considerable
higher (80–157%) compared to the analysed legume species.
The same applies for the mean cysteine concentration with the
exception of L. luteus.
Lysine concentration was highest in P. sativum seeds (7.3 g/
16 g N) followed by V. faba (6.3 g/16 g N) and G. max (5.7 g/
16 g N). Data are mainly in agreement with several cited
literature sources, with P. sativum and V. faba values being
slightly lower than reported elsewhere (e.g. Lattanzio et al.
1983, Wang and Daun 2004, Lisiewska et al. 2007). Contents
of threonine were very equal between the cultivars of all
investigated legume species (range from 3 to 4.2 g/16 g N), but
lower than in G. max seeds (4.1–5.9). Measured concentrations
for P. sativum and V. faba were equal to other values reported
in the literature (Wang and Daun 2004, Lisiewska et al. 2007)
but slightly higher for L. angustifolius and L. luteus (Sujak
et al. 2006).
The analysis of the drought stress effect on the AA
composition of V. faba seed protein showed that the overall
AA spectrum was less favourable regarding the nutritive value
of the proteins in plants grown under drought stress conditions
tein, but the nutritional protein value decreased slightly [e.g.
lysine (CS): 118–113].
The nutritional quality parameters are of particular interest
for the legume species (Table 4). Chemical scores of most
essential AAs were relatively high (>90) for all species. Most
limiting AAs were the sulphur AA cysteine and methionine in
all investigated seeds, except for G. max with leucine as most
limiting AA. Lisiewska et al. (2007) calculated considerably
higher CS values for the sulphur AA (CS = 86) and valine
(CS = 137) of V. faba. A similar value for lysine (CS = 111)
was found. The limiting AAs for L. angustifolius and L. luteus
are cysteine and methionine (CS 61.4 and 70.7, respectively).
Sujak et al. (2006) calculated a similar mean value of 60.2 for
eight different cultivars of L. angustifolius, but a considerably
higher mean value for nine cultivars of L. luteus (CS = 80)
can be deduced from their data. According to Sujak et al.
(2006), the most limiting AA in L. luteus was tryptophan
(CS = 59.9). The minimal CS for P. sativum of 66.7 was also
considerably lower than in the literature (e.g. CS = 74;
Zdunczyk et al. 1997). Wang and Daun (2004) reported a CS
value of 94 for sulphur AA in Canadian pea varieties, whereby
tryptophan was the most limiting AA (CS = 82). For soybean
(calculated from data of Zarkadas et al. 2007), the most
limiting AA was leucine. The highest calculated PER was
found in G. max (1.5) and L. luteus (1.5), with still compar-
atively low values compared to casein as standard protein for
an ideal PER (2.5).
The assessment of the protein quality in regard to these
nutritional properties for human nutrition is generally favour-
able. The seed protein of the varieties analysed in the present
study can be regarded as valuable, with several highly
concentrated essential AAs (e.g. lysine, leucine and isoleucine).
The lack of methionine, generally stated for legumes, was
confirmed for all tested varieties. This is additionally expressed
in the comparatively low PER value.
In regard to livestock nutrition, AA contents were further-
more related to demands of pigs (Table 5). Under consider-
ation of the range of protein qualities determined in the
sample, it is obvious that the feed quality can be increased by
the choice of high quality varieties.
As example: Compared to ideal protein values for pigs, the
choice of the variety with the best cysteine and methionine
contents of P. sativum, V. faba, V. faba (winter), L. angus-
tifolius, L. luteus and G. max would increase the quality about
162
(a)
(b)
(c)
Fig. 3: Relationship between selected amino acids lysine, methionine
and arginine and the crude protein content in the sets of cultivars
of L. angustifolius, P. sativum and V. faba. (n: L. angustifolius = 46,
P. sativum = 50, V. faba = 18)
14%, 13%, 11%, 22% and 5% compared to the sample mean,
respectively (Table 5).
Absolute methionine concentrations differed from 24%
(P. sativum), 17% (V. faba) to 16% (L. angustifolius) accord-
ing to the species (Table 2). Because of the smaller sample, the
other examined species showed a more narrow range.
Another important AA attribute for livestock nutrition is
the ratio of lysine to other limiting AA in the protein diet
(Fig. 2). None of the investigated European legume species
reached an optimal ratio of sulphur AA to lysine (0.6), which is
required in pig nutrition. This has previously been stated by
H. Schumacher, H. M. Paulsen, A. E. Gau et al.
Losand et al. (2003). Reasons are the comparatively low
content of sulphur AA as well as the comparatively high lysine
content. According to the values of Zarkadas et al. (2007),
only G. max fulfils the requirements in pig nutrition. The
optimal ratio of threonine to lysine, which should be 0.6 as
well, is reached by all plants but P. sativum and V. faba.
If the range of the sample is regarded, it is obvious that
plants with close to optimal threonine to lysine ratio are
present for V. faba. For species rich in lysine (P. sativum and
V. faba), the gap to the optimal lysine to sulphur AA ratio
remains big throughout the sample. However, the observed
variance in the analysed sample indicates the possibility to
improve nutritionally important AA ratios in all investigated
species.
The investigated correlation of selected AA to protein
contents is in agreement with published data (Fig. 3).
Wang and Daun (2004) reported similar correlations
between protein and arginine (r = 0.82**) and protein to
lysine (r = )0.52**) contents in peas. Several studies on the
correlation between AA and protein content in legume seeds
were summarized by Monti and Grillo (1983) and support
these findings. These results confirm the assumption that
plants with high protein contents accumulate proteins with
lower nutritional qualities (e.g. globulins).
Breeding attempts that are only focussed on the enhance-
ment of protein yields may therefore reduce the nutritional
quality of seed material. On the other hand, when the limited
AAs (e.g. methionine) are used as single selection criteria, the
protein content will decrease accordingly (Monti and Grillo
1983). In accordance with this postulation, a negative
correlation of total sulphur AA to the crude protein content
could be observed in the present work of all investigated
species.
Conclusions
The data of the present study can be used as background
knowledge for breeding strategies of nutritionally improved
legume cultivars. The former may help to calculate accurate
diet demands of livestock in accordance with ideal protein
concepts (e.g. Boisen 1997). Thus, nutrient losses caused by
insufficient feed utilization could be limited. Moreover, a
promising variation could be found in the concentration of
individual AA between selected legume varieties.
However, an enhancement of sulphur AA by up to 100%
(for peas) would be necessary to use them as sole feedstuff in
livestock nutrition (Tabe and Higgins 1998). But as livestock
feed rations are composed by different ingredients with various
AA and protein contents even an increase in methionine and
cysteine in legumes by 20% would reduce the expenditures to
add synthetic AA (Imsande 2001). In particular for organic
farming, increasing the protein quality of feed components
would be an important step to overcome insufficient supply of
essential AA in livestock feeding (Sundrum et al. 2000, Hovi
et al. 2003, Zollitsch et al. 2004, Bellof et al. 2005). Concerning
the AA demands of growing pigs (AA relations in Fig. 2; ideal
protein values in Table 5), it was shown that utilizing the range
of analysed concentrations can lead to quality improvements
of 10–20% in regard to single AA. The protein value (e.g. CS),
which is already quite favourable for all legume species, would
furthermore benefit.
While the improvement of protein quality by genetic
engineering (Krishnan 2005) was not successful up to now
Seed amino acid composition of grain legumes
and the technique is under critical public debate and strictly
banned in organic farming, conventional plant breeding
approaches should be regarded. In G. max Imsande (2001)
was able to increase the sulphur AA content significantly
without reducing the protein value with conventional breeding
methodology.
Acknowledgement
This research is carried out and supported within the Federal Program
for Organic Farming, BLE, Bonn, Germany.
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