CSIRO PUBLISHING

www.publish.csiro.au/journals/ajar Australian Journal of Agricultural Research, 2005, 56, 75–83

Variation for seed coat and pod wall percentage and other traits in a
germplasm collection and historical cultivars of lupins

J. C. ClementsA,C , M. DracupB , B. J. BuirchellB , and C. G. SmithB

A Centre for Legumes in Mediterranean Agriculture, University of Western Australia, 35 Stirling Hwy,
Crawley, WA 6009, Australia.
B Agriculture Western Australia, Locked Bag 4, Bentley, WA 6983, Australia.
C Corresponding author. Email: clem@cyllene.uwa.edu.au

Abstract. Accessions totalling 1425 from the Australian Lupin Collection representing 9 Old World lupin species
and Lupinus mutabilis Sweet, a New World species, were sampled for seed coat and pod wall percentage, seed
weight, and number of seeds per pod. These traits are of importance to the breeding objective of lowering seed coat
and pod wall proportions in crop lupins. Seed coat percentage mean values among the species ranged from 12.7
in L. mutabilis to 33.7 in L. pilosus L. The 4 species that have been subject to selection (L. mutabilis, L. albus L.,
L. angustifolius L., and L. luteus L.) had lower mean seed coat percentages than the other species with little
domestication. The rough-seeded lupin species had higher seed coat percentages relative to the smooth-seeded
species except for L. micranthus Guss., which had 31.5% seed coat and small seeds. Within L. angustifolius there
was no difference between the mean seed coat percentage values for wild v. domesticated or hard v. soft-seeded
entries, although wild accessions of L. angustifolius tended to have lower seed weight, higher pod wall percentage,
and more seeds per pod than domesticated accessions. There was no correlation in L. angustifolius germplasm
between seed coat percentage and pod wall percentage, indicating that selection for one will not influence the other
character. Accessions with the lowest seed coat percentage were from Turkey, Greece, and Cyprus, and those with
the lowest average pod wall percentage were from Spain and Cyprus. Mean pod wall percentages ranged from 30.9
in L. albus to 57.1 in L. micranthus; figures that are high compared with other legumes. Large ranges in seed weight
were found particularly in L. albus, L. pilosus, and L. angustifolius.
In addition to the germplasm collection, 21 Australian cultivars, released from 1967 to 1999, were evaluated at
one site over 2 years for the same traits. There was a negative correlation between seed coat percentage and seed
weight for both L. angustifolius historical cultivars and germplasm, indicating that further reductions in seed coat
percentage could be achieved by crossing large seeded types with low seed coat types. Pod wall percentage was
negatively correlated with both year of release and yield, and positively correlated with days to flowering. These data
support other findings that breeding for the reduction of pod wall can lead to yield improvements. The germplasm
collection assessed here provides lines with lower seed coat and pod wall compared with what is available in breeding
lines or cultivars.

Additional keywords: narrow-leafed lupin, white lupin, yellow lupin, pearl lupin, podwall, hull, fibre.

Introduction has the potential to increase metabolisable energy, protein,

Very high proportions of total biomass remain in the seed
coats and pod walls of most lupin species at maturity
(Reader and Dracup 1998; Clements et al. 2002). Lupinus
angustifolius seed (Australian sweet lupin) has relatively low
metabolisable energy for pig and poultry as a consequence
of its high amount of seed coat, which is largely cellulose,
hemicelluloses, and pectins (Brillouet and Riochet 1983).
De-hulling lupins improves nutritive value for monogastric
animals such as pigs, poultry, and fish species (Edwards and
van Barneveld 1998) but the cost of this process tends to make
it uneconomical. Breeding for reduced seed coat thickness
or oil (Stombaugh et al. 2000; Clements et al. 2002), and
decreasing pod wall can potentially increase harvest index
and yield. An effort to reduce seed coat in other species has
resulted in improved seed quality. For example, in Brassica
campestris, B. juncea, B. carinata, and B. napus, yellow-
seeded types have been selected that have a thinner seed coat
and higher seed oil and protein and lower crude fibre (Rashid
et al. 1994; Li et al. 2001).
Of the agriculturally important lupin species, both
L. angustifolius and L. luteus have a high percent of
seed weight in the seed coat (24% and 25%, respectively)

© CSIRO 2005 10.1071/AR03114 0004-9409/05/010075

76 Australian Journal of Agricultural Research
(Lush and Evans 1980; Reader and Dracup 1998; Clements
et al. 2002). This compares with only 7% in soybean (Lush
and Evans 1980) and 9% in field pea (Ali Khan 1993). In
current varieties of L. angustifolius, about 32–35% of the
pod dry matter is in the pod walls (Dracup et al. 1998;
Reader and Dracup 1998). The corresponding values for
other species are 29% in L. albus (Lagunes-Espinoza et al.
1999), 42% in L. luteus (Clements et al. 2002), and about
47% in L. mutabilis (Hardy et al. 1997; Reader and Dracup
1998). By comparison, this proportion is as low as 13%
in Pisum sativum (Reader and Dracup 1998) and 24% in
Phaseolus vulgaris (Wallace and Munger 1996). Natural
genetic variability for seed coat proportion and thickness
occurs in several legume species, e.g. for pea (Ali Khan
1993), common bean (P. vulgaris) (Escribano et al. 1997),
and chickpea (Waldia et al. 1996). Previous studies also
showed genetic variability for hull proportion and thickness in
lupins (Miao et al. 2001). In both L. albus (Lagunes-Espinoza
et al. 1999) and L. angustifolius (Clements et al. 2002),
negative correlations between pod wall proportion and seed
yield have been found in a range of breeding lines. Also, in
a study of bean (P. vulgaris) varieties, the highest yielding
variety had the lowest pod wall proportion (Wallace and
Munger 1996).
The Australian Lupin Collection is one of the world’s
largest and most comprehensive collections of Old World
lupins (Gladstones 1974; Clements and Cowling 1990,
1991; Cowling et al. 1998). It provides a valuable resource
to lupin breeders and has been screened for a range of traits
(Gladstones and Crosbie 1978; Cowling and Clements 1993;
Clements and Cowling 1994; Sweetingham et al. 1994; Jones
and Cowling 1995; Clements et al. 1996; Jones and Latham
1996; Harris 2000). The objective of the current work
was to screen a large range of lupin accessions in this
collection as well as the historical collection of Australian
L. angustifolius cultivars for seed coat and pod wall
proportion and to make recommendations for use in the
breeding programs.
Materials and methods
Screening accessions of a lupin germplasm collection
Fully mature pods of accessions of 10 lupin species were sampled from
plant rows (usually 2 m long and 1 m between rows) grown out as part
of germplasm maintenance at the Australian Lupin Collection, Perth,
Western Australia, from 1995 to 2002 in screenhouses (South Perth and
Shenton Park). Samples were also drawn from germplasm grown in the
open field at Wongan Hills, WA. Plants were spaced at 10 cm and rows
were 2 m long and 0.8 m apart. Fifteen to 20 pods were taken from the
main stems of each of the accessions at the end of the growing season
but prior to pod dehiscence in the case of wild types.
Preliminary studies found that a sample of 15 pods and 10 seeds was
sufficient to provide acceptably low coefficients of variation to show
a 1% and 0.5% difference with 95% accuracy in pod wall and seed
coat proportion, respectively. Accessions grown in screenhouses were
mostly unreplicated but control cvv. Gungurru, Chittick, and Merrit
were replicated (usually 2 replicates) at each site. Most accessions
J. C. Clements et al.
were classified as wild types and landraces (70%), the remainder
being semi-domesticated or fully domesticated breeding lines
and mutants.
Screening historical L. angustifolius cultivars for seed coat and
pod wall proportion
Pod samples were taken from 21 cultivars of L. angustifolius, released
in Australia from 1967 (cv. Uniwhite) to 1999 (cv. Quilinock), grown
at Wongan Hills, WA, in 2000 and 2001. Trials were part of the
Department of Agriculture lupin-breeding program. Plots were 10 by
1 m of 4 rows and target plant densities were 70 plants/m2 . Actual plant
density was found to be consistent with the target with little variation.
Trials were sown during May and harvested in November. Trials
were usually managed according to district practice. Typical practice
for trials was: Rovral fungicide at 1 mL/kg seed treatment; fertiliser
applied as double superphosphate at 80 kg/ha; pre-sowing herbicides
Sprayseed at 1.6 L/ha, Simazine at 2.5 L/ha, Talstar at 100 mL/ha, and
Brodal at 150 mL/ha post-emergent in July, and Select at 250 mL/ha
in August.
Samples consisted of 30 healthy lower main stem pods from
separate plants from central rows from 2 or 3 replicate plots that were
physically laid out in row–column neighbour designs. Seasonal rainfall
(April–October) at Wongan Hills was 221 and 233 mm for 2001 and
2002, respectively, which was below average and considered to be
drought-affected years.
Processing of samples for pod wall and seed coat proportion
Pods were counted and any pedicel material still attached was removed.
Pods were then dried at 40◦ C for 48 h, weighed, and shelled to remove
the seed. Seeds were counted and weighed. Pod wall percentage of total
pod weight was calculated by dividing the weight of pod shells by the
total weight of the 30 pods in the sample. A random subsample of
10 healthy seeds was placed in a 90-cm Petri dish and the seeds imbibed
with a minimal amount of water at 4◦ C for 24 h. Some seeds that did not
imbibe were scarified with a scalpel and left to imbibe for a further 24 h.
The seed coat was removed with the aid of a scalpel blade, ensuring
that the radicle was intact and remained with the embryo and cotyledon
tissues. Separated seed coat and embryo tissues (including cotyledons)
were dried at 105◦ C for 48 h, then placed immediately in a desiccator
and weighed to the nearest 0.001 g.
Statistical analysis
Because of the unbalanced nature of the data, seed coat percentage,
pod wall percentage of total pod weight, weight of seed, weight of
pod, and number of seeds per pod were analysed using the method of
residual maximum likelihood (REML) (Genstat 5 Committee 1993).
The random-effects model was Year * Site * Variety + Year.Site.
This model is expressed in the Genstat notation (Genstat
5 Committee 1993), in which A * B = A + B + A.B = main effect of
A + main effect of B + A × B interaction and A.B alone = effect of all
combinations of A and B, without estimation of main effects of either
factor. Correlation coefficients were derived from further analysis of
predicted means from the REML output. Comparisons were made of
wild and domesticated lines and countries of origin for all traits, soft v.
hard seeds for seed coat percentage, and dehiscing v. non-dehiscing for
pod wall percentage of total pod weight within L. angustifolius through
t-tests or analysis of variance. Lines were classified as ‘domesticated’
if they were breeders’ lines, cultivars, or landraces (i.e. at least
some human intervention in their origins). Accessions that
were classified as ‘mutants’ (miscellaneous natural or induced
mutants) were not included in the comparisons of wild v.
domesticated groups.
Variation for seed and pod traits in lupin germplasm Australian Journal of Agricultural Research 77

Table 1. Variance components, where significant, from REML analyses of seed coat %, pod wall % of total pod weight, and weight
per seed
Data from 8 years, up to 5 sites, and 1425 genotypes of lupin representing 10 species
Seed coat % Pod wall % Weight per seed
Term Component s.e. Term Component s.e. Term Component s.e.
Year 1.12 1.06 Year 1.19 0.73 Year 128 109
Access. 12.39 0.69 Site 2.15 1.68 Access. 10237 402
Year.site 1.29 0.72 Access. 30.32 1.88 Year.site 102 62
Year.access. 1.19 0.33 Site.access. 1.44 0.96 Year.site.access. 220 26
Year.site.access. 0.98 0.23 Year.site.access. 4.44 0.84

Results data were from single replicates for many accessions.

Lupin germplasm collection: wild, semi-domesticated,
and breeders’ lines
Data were obtained for 1425 accessions from the Australian
Lupin Collection. The variance output from the REML
analysis after removing terms with insignificant variance
showed that for the 4 traits analysed, the effect of
accession dominated the variance (Table 1) and the accession
means are likely to be reliable estimates even though
For seed coat percentages there were also significant
variances associated with year, year × site, year × accession,
and year × site × accession. For pod wall percentage, year,
site, site × accession, and year × site × accession were also
significant.
Table 2 summarises the 10 lupin species for seed and pod
wall characters. The 4 species that have been subject to the
greatest domestication and selection for human consumption

Table 2. Seed and pod characteristics summarising germplasm of 10 lupin species from the
Australian Lupin Collection
T-test of difference between means of wild and domesticated lines for L. angustifolius are indicated:
*P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant
Species Mean Min. Max. n Mean Min. Max. n

Seed coat % Pod wall %

L. angustifolius
All 24.6 19.4 38.8 944 34.4 25.8 54.5 666
Wild 24.6n.s. 19.4 38.8 605 34.7* 25.9 54.5 439
Domesticated 24.6n.s. 20.8 28.4 317 33.9* 25.8 52.7 208
Mutants 25.5 19.5 28.6 22 34.3 28.7 40.3 19
L. albus 18.4 12.2 27.5 187 30.9 24.6 57.7 162
L. atlanticus 28.1 23.9 33.6 11 37.7 33.3 49.6 11
L. cosentinii 29.7 15.9 37.4 63 42.7 36.4 60.5 20
L. digitatus 32.8 32.1 33.6 2 40.1 40.1 40.1 1
L. hispanicus 27.4 23.7 33.1 13 45.2 36.6 63.3 10
L. luteus 25.6 22.1 36.2 75 46.9 41.3 72.0 62
L. micranthus 31.5 24.8 44.1 20 57.1 41.0 77.7 12
L. mutabilis 12.7 10.6 16.0 98 45.1 35.8 53.1 24
L. pilosus 33.7 28.4 41.3 22 42.5 34.9 55.5 22
Seed weight (mg) Seeds per pod
L. angustifolius
All 123 31 238 940 4.8 2.4 6.6 667
Wild 107*** 35 238 601 4.8** 2.4 6.6 439
Domesticated 152*** 31 222 317 4.7** 3.3 6.4 209
Mutants 152 116 189 22 4.2 2.8 5.2 19
L. albus 376 120 869 186 4.3 2.6 5.6 162
L. atlanticus 332 202 420 11 3.7 3.1 4.5 11
L. cosentinii 189 61 272 63 4.2 3.4 4.8 20
L. digitatus 134 122 146 2 3.7 3.7 3.7 1
L. hispanicus 76 38 141 13 5.2 1.8 6.1 10
L. luteus 111 53 149 75 4.5 3.0 5.5 62
L. micranthus 89 31 149 20 2.8 0.2 3.7 12
L. mutabilis 173 79 276 98 4.6 2.7 6.8 24
L. pilosus 412 186 682 22 3.4 1.6 4.5 22
78 Australian Journal of Agricultural Research J. C. Clements et al.

Table 3. L. angustifolius seed and pod characteristic means for and wild accessions did not differ significantly for mean
country of origin seed coat percentage, although they did differ for pod wall
Significance of the main effect of origin is indicated: **P < 0.01;
***P < 0.001
percentage of total pod weight (hereafter referred to as ‘pod
wall percentage’), seed weight, and seeds per pod (P < 0.05,
Origin Seed Pod Seed weight Seeds per
Table 2). Domesticated accessions on average had a lower
coat % wall % (mg) pod
pod wall percentage and seeds per pod and higher seed
Australia 24.8 33.7 159 4.5 weight. Lines with the lowest seed coat percentage were
Bulgaria 23.9 32.2 172 5.4 wild types or mutants. There were significant differences
Byellorussia 24.5 33.5 146 5.0
Chile 23.7 33.7 172 4.9
among countries of origin for all seed and pod traits
Cyprus 23.0 30.6 76 4.8 (Table 3). Sources of low seed coat were from Turkey,
Netherlands 23.9 33.7 170 5.2 Greece, and Cyprus. Country sources with the lowest
Algeria 23.9 36.4 133 4.2 average pod wall percentage were Spain and Cyprus. Within
Spain 25.6 34.3 113 5.0 L. angustifolius, means for soft v. hard seeded lines for
France 25.4 36.7 101 5.2
Great Britain 24.0 173
seed coat percentage were 24.3 and 24.4, respectively (not
Greece 23.0 35.5 98 4.6 significant, P < 0.05), and for non-dehiscing v. dehiscing
Hungary 24.1 31.8 181 4.9 for pod wall percentage were 33.4 and 34.5, respectively
India 23.7 139 (significant, P < 0.05).
Israel 24.9 32.1 149 4.8 Correlation coefficients among traits showed that across
Italy 24.7 36.0 72 5.0
Morocco 24.0 36.5 115 4.5
all accessions, representing the 10 lupin species, seed coat
New Zealand 24.0 37.0 201 4.6 percentage was correlated with pod wall (r = 0.35, P < 0.001)
Poland 24.9 32.0 153 4.6 and seed weight (r = –0.36, P < 0.001). However, within
Portugal 24.8 33.8 118 4.8 L. angustifolius, seed coat percentage was weakly correlated
Spain 27.1 30.7 106 4.1 with seed weight (r = –0.23, P < 0.001), mature plant height,
Russia 24.7 33.5 133 4.9
Sweden 24.1 36.1 150 4.1
and collection site altitude (r = –0.35, P < 0.001) (Table 4),
Syria 25.1 31.9 91 4.9 whereas pod wall was weakly correlated with seed weight
Turkey 22.3 34.1 113 4.8 (r = –0.20, P < 0.001), seeds per pod (r = 0.18, P < 0.001),
USA 24.3 36.0 156 4.9 and plant height (r = –0.17, P < 0.001). For L. luteus
Yugoslavia 25.2 180 accessions, seed coat percentage was correlated with seed
South Africa 23.7 34.2 180 4.5
weight (r = –0.57, P < 0.001) and pod wall percentage was
Origin *** ** *** ***
correlated with seed weight (r = –0.37, P < 0.001) and seeds
l.s.d. (P = 0.05) 0.8 2.0 16 0.3
per pod (r = –0.45, P < 0.001).
Table 5 lists the accessions of L. angustifolius with the
lowest seed coat and pod wall percentage. Accessions with
(L. mutabilis, L. albus, L. angustifolius, and L. luteus) had the lowest values include wild and domesticated accessions
lower mean seed coat percentage values than the other and mutant lines specifically selected for low seed coat.
species. The rough-seeded lupin species (L. atlanticus, One accession had both low seed coat and low pod wall
L. cosentinii, L. digitatus, L. pilosus) had higher seed coat percentage (P28590) and some others had both low seed
percentage values relative to the smooth-seeded species with coat percentage and low seed weight (P28428, P28709,
the exception of L. micranthus. L. angustifolius domesticated and P28402).

Table 4. Correlation coefficients r among means for seed, pod, and plant traits for all L. angustifolius germplasm accessions
n, Number of lines; significance of r is indicated: *P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant
Pod Seed Seeds Days to Plant height Plant Collection Collection Collection
wall % weight per pod flowering at 9 weeks height at site site site
after sowing maturity altitude rainfall soil pH
Seed coat %
r 0.07 –0.23 0.07 0.03 –0.08 0.11 0.35 0.09 0.14
n 665 940 666 893 834 882 251 110 167
P n.s. *** n.s. n.s. * ** *** n.s. n.s.
Pod wall %
r –0.20 0.18 0.01 –0.05 –0.17 –0.17 –0.03 –0.02
n 662 664 616 564 607 243 110 167
P *** *** n.s. n.s. *** * n.s. n.s.
Variation for seed and pod traits in lupin germplasm Australian Journal of Agricultural Research 79

Table 5. Selected L. angustifolius accessions with the lowest seed coat and pod wall percentage for use in breeding programs
cv. Merrit for comparison in bold
Perth no. Accession Origin Breeding Seed coat Pod wall Seed weight Seeds per
name status % % (mg) pod
Selected for low seed coat %
28713 96TUR42 TUR Wild, introduced 19.4 124
M11257 AUS Mutant 19.5 33.7 129 4.2
28428 GRC116B GRC Wild, introduced 20.2 34.4 96 4.0
28709 96TUR06 TUR Wild, introduced 20.2 98
28590 CMF 99–56 GRC Wild, introduced 20.3 27.1 115 4.7
M11255a AUS Mutant 20.3 32.0 158 4.6
28402 GCN89C GRC Wild, introduced 20.6 32.9 81 4.8
28723 GM120.B.1 MAR Wild, introduced 20.6 238
25851 412E3394 ZAF Breeding line 20.8 152
28708 96TUR04A TUR Wild, introduced 20.8 119
26500 GRC5020A GRC Wild, introduced 21.1 161
27205 MAE30A GRC Wild, introduced 21.1 44.1 81 4.5
28130 MAR6825A MAR Wild, introduced 21.2 30.0 169 4.5
28426 GRC109B GRC Wild, introduced 21.2 30.2 110 4.7
26570 GRC5041A.1.2 GRC Wild, introduced 21.3 30.8 120 4.5
Selected for low pod wall %
20641 N2793 AUS Selection (ex cultivar) 25.7 25.8 84 5.8
26404 L.O.-1723 ESP Wild, introduced 24.0 25.9 86 5.9
26369 L.O.-1390 ESP Wild, introduced 26.1 26.8 60 6.1
26434 L.O.-1786 ESP Wild, introduced 24.2 26.9 72 5.2
28590 CMF 99–56 GRC Wild, introduced 20.3 27.1 115 4.7
27475 F-1 SUN Breeding line 23.7 27.1 127 4.7
26402 L.O.-1721 ESP Wild, introduced 25.0 27.1 85 5.8
26409 L.O.-1728 ESP Wild, introduced 24.7 27.3 87 5.3
27479 M-2 SUN Breeding line 26.0 27.4 117 5.4
26576 GRC5044A.2 GRC Wild, introduced 21.8 27.8 134 4.8
26372 L.O.-1403 ESP Wild, introduced 26.1 27.9 75 5.8
26290 G84–150.1 PRT Wild, introduced 23.8 27.9 131 4.5
27467 E-1 SUN Breeding line 28.2 28.0 101 4.7
27481 M-4 SUN Breeding line 23.8 28.0 109 5.6
27473 G-3 SUN Breeding line 23.9 28.2 150 4.1
28125 Merrit AUS Cultivar 24.4 33.0 141 4.4
l.s.d. (P = 0.05) 2.5 5.3 34 0.9

Australian historical L. angustifolius cultivars
Seed coat and pod wall percentage values among 21 historical
cultivars from Australia ranged from 19.7 to 22.6 and from
28.2 to 34.2, respectively (Table 6). There was a significant
genotype effect for all traits and a significant year effect
for all traits except seeds per pod and days to flowering.
Cultivars with the lowest values for seed coat and pod wall
percentage were Yorrel and Belara, respectively. Seed coat
and pod wall percentage means from 2 seasons in the present
study were lower than in previous studies for a common
subset of genotypes (Clements et al. 2002). However, the
values correlated reasonably well for seed coat percentage
and for pod wall percentage (0.83 and 0.61 Spearman rank
correlation coefficients, respectively) for the 2 studies.
Seed coat percentage was correlated with seed weight
(r = –0.64, P < 0.001) and pod wall percentage was
correlated with year of release (r = –0.43, P < 0.05),
days to flowering (r = 0.65, P < 0.001), aborted seed
per pod (r = 0.53, P < 0.01), and mean yield (r = –0.62,
P < 0.01) (Table 7). Strong relationships were indicated
from correlations between year of release and both days to
flowering (r = –0.73, P < 0.001) and mean yield (r = 0.79,
P < 0.001), and between days to flowering and mean yield
(r = –0.91, P < 0.001).
Discussion
In this study, for both germplasm and historical cultivars,
genotypic variance for the 4 traits analysed (seed coat and
pod wall percentage, seed weight, and number of seeds per
pod) was greater than the other sources of variation including
year, site, and interaction terms. Selection for these traits
in lupin is therefore possible with reasonable reliability in
different environments as has been suggested previously
(Lagunes-Espinoza et al. 1999; Clements et al. 2002). The
mean seed coat percentages for 10 lupin species ranged from
12.7 in L. mutabilis to 33.7 in L. pilosus (Table 2). The seed
80 Australian Journal of Agricultural Research J. C. Clements et al.

Table 6. Seed and pod traits for historical L. angustifolius cultivars (1967–99)
Significance of genotype and year main effects are indicated: **P < 0.01; ***P < 0.001; n.s., not significant
Genotype Days to Seed coat Pod wall Seed weight Seeds per Pod weight Aborted
flowering % % (mg) pod (mg) seeds per pod
Belara 69 21.7 28.2 166 4.8 1120 0.03
Chittick 80 21.6 32.0 169 4.5 1143 0.16
Danja 77 21.1 30.9 162 4.7 1120 0.09
Geebung 84 21.1 30.7 153 4.7 1052 0.15
Gungurru 73 22.0 30.9 155 4.3 995 0.23
Illyarrie 74 21.4 29.7 178 4.7 1205 0.09
Kalya 72 22.1 30.0 155 4.5 1032 0.27
Marri 85 21.8 32.5 153 4.9 1103 0.24
Merrit 73 22.3 31.9 153 4.2 983 0.27
Moonah 72 20.2 30.1 169 4.3 1078 0.14
Myallie 70 21.2 31.0 163 4.9 1172 0.08
Quilinock 70 21.3 31.7 175 4.6 1203 0.04
Tallerack 72 21.4 30.1 167 4.5 1102 0.32
Tanjil 74 22.6 31.8 146 4.6 1015 0.19
Unicrop 76 21.5 29.8 169 4.7 1135 0.08
Uniharvest 85 21.7 32.8 154 4.7 1092 0.25
Uniwhite 86 21.9 34.2 143 4.0 880 0.27
Warrah 77 22.1 29.4 141 4.7 957 0.10
Wonga 78 22.5 30.9 153 4.9 1098 0.16
Yandee 75 21.5 30.2 179 4.9 1263 0.06
Yorrel 70 19.7 29.9 177 4.5 1145 0.06
l.s.d. (P = 0.05) 2 0.7 1.8 10 0.3 82 0.12
Genotype *** *** *** *** *** *** ***
Year n.s. *** ** *** n.s. *** ***

Table 7. Correlation coefficients for traits of Australian historical L. angustifolius cultivars

Significance is indicated: *P < 0.05; **P < 0.01; ***P < 0.001
Year of Days to Seed coat Pod wall Seed Seeds per Pod Aborted seed
release flowering % % weight pod weight per pod
Days to flowering –0.73***
Seed coat % –0.02 0.24
Pod wall % –0.46* 0.65*** 0.27
Seed weight 0.09 –0.52** –0.64*** –0.43*
Seeds per pod 0.11 –0.05 0.02 –0.38 0.23
Pod weight 0.07 –0.34 –0.43* –0.36 0.86*** 0.66***
Aborted seed per pod –0.15 0.40* 0.44* 0.53** –0.55** –0.49* –0.60**
Mean yield (2001–02) 0.79*** –0.91*** –0.21 –0.62** 0.51** 0.17 0.41* –0.35

coat proportion in L. mutabilis is similar to other food or
feed legumes such as Cicer arietinum, Vigna unguiculata,
Vicia faba, and some species of Pisum (Lush and Evans 1980;
van der Poel et al. 1992; Ali Khan 1993; Flinn et al. 1998).
L. mutabilis is one of several food crops that has been
cultivated in the Andean region of South America for
thousands of years (Anúnez de Mayolo 1982), and probably
only a few examples of true wild types of this species
are represented in the Australian Lupin Collection. This
species has potential as a high protein crop in Australia
and the germplasm collection provides useful variation for
seed coat percentage. The lower seed coat percentage found
in L. mutabilis appears to be due to the generally thinner
tissue layers within the seed coat (palisade, hourglass, and
parenchyma; Lush and Evans 1980) compared with other
lupin species and also to the large reduction or absence of
the subpalisade layer (J. C. Clements, unpublished data).
The 4 species that have been subject to domestication
(L. mutabilis, L. albus, L. angustifolius, and L. luteus) had
lower mean seed coat percentages than the wild species
and this supports the hypothesis made by Lush and Evans
(1980) that wild species (and wild genotypes within species)
of grain legumes generally have thicker seed coats. The
2 species L. mutabilis and L. albus that have been historically
selected for human consumption (Cowling et al. 1998) had
the lowest seed coat percentages of all the species studied.
Variation for seed and pod traits in lupin germplasm
Within L. angustifolius accessions, however, there was no
difference between the mean seed coat percentage values for
wild v. domesticated lines (Table 2) and accessions with the
lowest seed coat percentage values were wild types. Values
for these accessions were as low as for induced mutants
selected for low seed coat (Table 5; Clements and Dracup
2001; Clements et al. 2002).
Mean seed coat percentage for hard-seeded (impermeable
seed coat) accessions was not significantly different from
that for soft-seeded accessions (P < 0.05), indicating that the
mollis gene for soft-seededness (Gladstones 1970) carried
by probably all of the soft-seeded types in this collection has
not generally affected seed coat percentage. These results
contrast to that of Miao et al. (2001) who reported that
selection for soft seeds in L. angustifolius has resulted in
a reduction of seed coat thickness. Our results differ possibly
because the comparison was made here between larger
sample sizes of hard- v. soft-seeded genotypes (458 hard-
seeded and 250 soft-seeded L. angustifolius accessions).
The rough-seeded lupins had higher seed coat percentage
values relative to the smooth-seeded species. L. pilosus,
which had the highest seed coat percentage, has an extremely
thick palisade layer (Lush and Evans 1980) and this is
probably similar in the other rough-seeded species. Three
wild accessions and one induced mutant of L. pilosus have
been reported that are smooth-seeded and this trait has
potential value in the domestication of this species (Buirchell
and Cowling 1992, 1998). The seed coat percentage for one
of these lines (P27445, 29.8%) was found to be lower than the
average for rough-seeded lines (33.7%, Table 2). This agrees
with the microscopy evidence of Miao et al. (2001) showing
that smooth-seeded L. pilosus has a thinner seed coat than
rough-seeded examples.
Wild accessions of L. angustifolius did tend to have lower
seed weight, higher pod wall percentage, and more seeds
per pod than domesticated accessions (Table 2). There was a
wide range of seed weights available among L. angustifolius
accessions, with large seeds (approx. 200 mg) available from
several origins. Seed coat percentage within L. angustifolius
was weakly correlated with seed weight, mature plant height,
and collection site altitude. A stronger relationship between
seed coat percentage and seed size existed for L. luteus
accessions, indicating that larger seeded genotypes can
provide reduced seed coat but there was a lack of larger
seeded L. luteus accessions among the genotypes assessed
compared with the range available for L. angustifolius. A
very large range in seed size was observed in L. pilosus and
this trait was found to be associated with other characteristics
such as leaf size, yield, and pod set that were used to define
morphological groups in the species (Clements et al. 1996).
L. albus accessions also provided an enormous range in seed
size, from the smaller seeded var. graecus types (Gladstones
1974) to the very large seeded ‘lupini’ bean types (Luckett
2000). Seed coat percentage in a few accessions of L. albus
Australian Journal of Agricultural Research 81
reached as low as 12.2%, which approaches values found in
the exceptionally low L. mutabilis.
Pod wall percentage mean values ranged from 30.9 in
L. albus to 57.1 in L. micranthus, figures that are high
compared with other legumes such as pea, faba bean, and
common bean (Wallace and Munger 1996; Reader and
Dracup 1998). Pod wall proportions tend to be higher in
some accessions, requiring very long periods of vernalisation
and hence flowering very late under increasing temperatures
(Reader et al. 1995; Buirchell and Cowling 1998). For
L. angustifolius germplasm, pod wall percentage tended to
decrease as seed weight increased (Table 4). L. angustifolius
with non-dehiscing pods had a slightly lower pod wall
percentage than those with dehiscing pods (33.4 and 34.5,
respectively, P < 0.05), indicating that lack of dehiscence is
not necessarily associated with thicker pod wall structures.
Gladstones (1967) reported that one of the 2 genes for
reduced pod shattering, le, probably results in a reduction
of the thickness of the pod endocarp and the results in the
present study appear to support this. Crosses of a low pod
wall wild type with a non-shattering current cultivar (Tanjil)
have produced low pod wall, non-shattering progeny and this
indicates that low pod wall can be achieved without shattering
(J. C. Clements and M. Dracup, unpublished data).
The range in pod wall percentage found in L. angustifolius
should provide breeders with parental material to produce
lower pod wall proportion segregants, assuming that
the trait will have reasonable heritability in this species
(Lagunes-Espinoza et al. 1999; Clements et al. 2002). The
negative correlation between pod wall proportion and yield
found for L. angustifolius was lower than that found in
L. albus by Lagunes-Espinoza et al. (1999). The results of the
latter study were not only for a different species but also from
a European, spring-sown environment, whereas our results
were for a Mediterranean environment with autumn-sown
L. angustifolius. The results here indicate that selection for a
pod wall percentage in L. angustifolius for a Mediterranean
environment should be beneficial but will not contribute
directly to rapid yield gains. Only a narrow range of pod
wall percentage was seen among L. luteus accessions and
this species may benefit more compared with L. angustifolius
from a reduction in its very high pod wall proportion,
potentially leading to higher harvest index and yield. The
screening of larger collections held in other institutions may
reveal a broader range of variation for the traits in this
species. Also, use of mutation breeding in L. luteus could
be an additional source of variation. Significant negative
correlations between pod wall percentage and both seed
weight and seeds per pod were found here and therefore
selecting for larger seeds or more seeds per pod in L. luteus
could be one other avenue for reducing pod wall in this
species. Acquisition of larger seeded L. luteus genotypes
for improving both pod wall and seed coat percentage is
recommended.
82 Australian Journal of Agricultural Research
The 1425 accessions included in this study covered
28 countries of origin. Although it is likely that some of
the recorded origins may not be the true original source
of the seed, most of the passport data will be indicative
of geographic source. When means were calculated on a
country of origin basis it was apparent that some countries
were richer sources of accessions with lower seed coat or
pod wall percentage than other countries. Accessions of
L. angustifolius with low seed coat percentage were from
Turkey, Greece, and Cyprus and of low average pod wall
percentage were from Spain and Cyprus (Table 3).
Seed coat and pod wall percentage means were relatively
low for 21 historical cultivars from the 2-year data at Wongan
Hills compared with values reported by Clements et al.
(2002) and this could be largely due to the low rainfall in
these seasons. Positive correlations were found by Clements
et al. (2002) between seed coat thickness and growing-season
rainfall and between pod wall percentage and growing-
season length. Dracup et al. (1998) found that prolonged
irrigation could increase pod wall percentage to as high
as 44 in L. angustifolius. There were significant cultivar
differences, however, for all traits measured (Table 6). Yorrel
had the lowest seed coat percentage and Belara had the
lowest pod wall percentage. The previously reported negative
correlation between seed coat percentage and seed weight
(Clements et al. 2002) was also found among historical
cultivars (r = –0.64, P < 0.001). There was no significant
correlation between seed coat percentage and year of release.
It could be suggested that selection for yield could result
in increases in seed coat percentage because seed coat
components (and cotyledon cell walls) are energetically less
expensive to produce than protein and oil. The data here
show a very low negative correlation between yield and
seed coat percentage but it was not significant (P < 0.05).
It is feasible to propose that carbon not channelled into seed
coat could be channelled into kernel or extra seed but data
to support this are yet to be reported. Pod wall percentage
was correlated with year of release (r = –0.43, P < 0.05),
days to flowering (r = 0.65, P < 0.001), aborted seed per pod
(r = 0.53, P < 0.01), and mean yield (r = –0.62, P < 0.01)
(Table 7). Along with earlier flowering and higher yield,
modern L. angustifolius cultivars tend to have a lower pod
wall percentage (e.g. Belara). A correlation between pod wall
proportion and yield was also found in a study of cultivars
and breeding lines of L. albus (Lagunes-Espinoza et al. 1999)
and this supports the validity of this character in contributing
to yield increase.
This study demonstrated the high diversity for seed and
pod traits among the Old World lupins and L. mutabilis
in the Australian Lupin Collection. In L. angustifolius, the
most widely grown crop species in Australia, the collection
contains accessions with useful sources of low seed coat,
low pod wall, and a range in seed weights that can be used
for breeding in this species. Some useful variation for traits
J. C. Clements et al.
was identified in the other crop species L. albus; however,
wider variation for traits should be sought in L. luteus.
L. mutabilis is an under-exploited species with very good
crop potential because of its high protein and oil and thin
seed coat. Further evaluation of a wider range of germplasm
for seed and pod traits should be worthwhile. The data
presented in this paper and from a previous study on breeding
lines (Clements et al. 2002) suggest that increasing seed
weight of domesticated types can lead to reduced seed coat
percentage, and decreasing pod wall can facilitate yield
improvement. Crossing genotypes that have low seed coat
percentage with genotypes with large seed weight may further
decrease seed coat percentage. The potential benefits from
reducing seed coat include reduced de-hulling cost and
higher seed quality with respect to protein content. These
characteristics can improve the value of lupins for feed and
food applications. Assessment of the effects of lower seed
coat on seed quality, including kernel cell wall thickness and
non-starch polysaccharide content, would be worthwhile for
future studies.
Acknowledgments
The Grains Research and Development Corporation funded
this research. We acknowledge the work of Mr Mark Reader,
from 1995 to 1998, on germplasm data collection and
processing. We thank the following people for their technical
assistance: Margaret Collins, Gordon Francis, Doug Francis,
John Quealy, Ali Bhatti, Krishna Mann, Rebecca Carpenter,
Liam Cranley, and Hussein Al Edany.
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Manuscript received 30 May 2003, accepted 10 December 2004