Download Improving the physiological and agronomic

DEPARTMENT for ENVIRONMENT, FOOD and RURAL AFFAIRS
Research and Development
CSG 15
Final Project Report
(Not to be used for LINK projects)
Two hard copies of this form should be returned to:
Research Policy and International Division, Final Reports Unit
DEFRA, Area 301
Cromwell House, Dean Stanley Street, London, SW1P 3JH.
An electronic version should be e-mailed to [email protected]
Project title
Improving the physiological and agronomic basis of UK lupin production.
DEFRA project code
ARO138
Contractor organisation
and location
Rothamsted Research
West Common
Harpenden
Herts.
AL5 5TS
Total DEFRA project costs
Project start date
£ £720,000
01/04/99
Project end date
31/03/03
Executive summary (maximum 2 sides A4)
Lupins are recognised as a valuable source of protein in livestock diets. Previous attempts to produce lupins in
the UK were not successful. UK production was seen as important for reasons of import substitution (of soya),
traceability and GM crop contamination. Research was conducted for more than 10 years to understand and
overcome the limitations to lupin production in the UK. Much was funded by DEFRA (formerly MAFF), BBSRC
(formerly AFRC) and the EU also contributed significant resources.
Most of the work focussed on Lupinus albus (white lupin) and specifically autumn sown cultivars as these were
seen to offer the greatest potential to UK agriculture. In 1998 it was recognised that other species, (L.
angustifolius and L. luteus) also had a valuable role to play in the UK lupin industry. Only small scale industrial
funds were available to investigate these exclusively spring sown possibilities, however.
A major breakthrough in the cultivation of all lupin species in the UK was the identification of the suitability of
genetically more determinate cultivars. The growth of the lupin plant is modular. Modules consist of individual
stems producing leaves and ending with a terminal inflorescence. Further modules, belonging to the next
(branch) order, may emanate from the leaf axils of the previous order. Cultivars in which the number of orders
of modules to grow is under genetic control are more reliable in the mild damp summer conditions of the UK.
Such cultivars matured more reliably than those relying on an environmental trigger to bring an end to such
vegetative growth.
Autumn sowing allowed early flowering after the production of numerous leaves compared to spring sowing.
This gave a longer period of good growing conditions for seed yield to be set whilst maintaining an acceptable
maturity / harvest date.
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Studies of over-winter survival of autumn sown cultivars greatly enhanced our knowledge of cold tolerance in
L. albus. Within the autumn sown cultivars and the progenitors it was not possible to determine clear genetic
variation for cold tolerance. Clear identification of the role of the environment on plant development allowed
definition of the most cold tolerant phases in plant development. The models developed from this work were
used with historical meteorological records to define management packages or agronomy that maximised the
over-winter survival of the crop.
The over-winter survival data and models contributed to the determination of the recommended seed rate.
Initial estimates of optimal spring and summer plant densities were purely empirical. Later it was realised that
the plant density was a vital factor controlling seed yield variation. More specifically the greater proportion of
the incident light reaching the lowest pods in the canopy the greater the seed yield attained (all other things
being equal). Canopy architecture and light penetration were controlled by plant density and the number of
branches or modules in the first order component.
Studies of the effects of the environment on plant architecture (number of modules in each branch order) led to
the development of models for these processes. The interaction of plant architecture with plant density
(canopy architecture) was more empirical. These models were used in combination with the over-winter
survival model and historical meteorological records to generate site specific management recommendations,
particularly sowing windows.
L. albus produces a large pod wall (thick with a large dry weight), larger than most other grain legumes. This
was seen as a waste of resources as it was dry matter that was not harvested as economic yield (a detrimental
affect on harvest index). Selecting for genotypes with smaller pod walls (lower dry weight and thickness)
appeared to have a generally positive affect on seed yield. In addition the pod wall of L.albus is known to be a
photosynthetic organ. In our experimentation much of that photosynthetic activity was internal re-fixation of
respiratory CO2. Net gas exchange with the environment was low compared to other photosynthetically active
organs. It was hypothesised that smaller pod walls would increase net gas exchange to the benefit of seed
yield. Comparison of two genotypes showing similar growth except for the size of the pod wall only
demonstrated the large variance encountered in making replicate measurements of net gas exchange by such
organs.
Wide-scale testing of the first autumn sown determinate cultivars showed that the time of maturity was not as
reliable as at first thought. Studies revealed that the availability of soil water was the dominant factor
controlling the rate of maturation in late summer and that air temperature and to a lesser extent humidity also
had an influence. The smaller pod wall character (and hence potentially greater rates of water loss) showed no
benefit in terms of earlier maturity times.
A number of fungal diseases are known to infect cultivated lupins. Most proved relatively easy to control.
Pleiochaeta setosa has not been common in UK crops despite being a serious disease of lupins elsewhere in
the world. The seedling disease caused by the pathogen has not been recorded in the UK and later in the life
of the plant infection of stem and pod is easily controlled by the same fungicides that control rust (see below).
Genetic resistance to Fusarium spp. has proved sufficient for most situations where the plant has not been
previously physically damaged. Botrytis cinerea was much less common following the recommendation that
crops be grown with more open canopies to improve light penetration to depth. Rust (Uromyces lupinocolus) is
probably the most common fungal pathogen of L. albus, autumn sown crops require 2 fungicide sprays on
most sites and in most seasons. Fortunately the rust is susceptible to low rates of low cost fungicides.
Anthracnose is a potentially devastating fungal disease of all cultivated lupins. It does not survive well in the
environment. In any one year background infection from the environment is unlikely to harm a crop seriously.
However, if seed is re-sown from such a crop, even with low levels of infection, and weather conditions are
suitable the effect on the second crop can be devastating (total crop loss is a possibility). Fungicidal control
proved costly and a little unreliable. Therefore the responsibility lies with the seed producers to ensure through
attention to crop monitoring and general hygiene that seed is free of infection. Conventional seed testing
techniques proved inadequate to detect the very low levels of infection considered capable of causing crop
loss (1 infected seed in 10,000).
Rothamsted Research made a collection of genotypes of the causal fungus (Colletotrichum acutatum or C.
gleosporoides) and tested them for pathogenicity on L. albus cv. Lucille (a commonly grown cv.). There was
CSG 15 (Rev. 6/02)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
some debate about the taxonomy of the genotypes within the two species and conventional (fungicide
sensitivity) and molecular (primer sequences) were used to distinguish the species. rDNA from the genotypes
was treated with four enzymes in the search for a banding pattern common to pathogenic genotypes that could
be used as the basis of a molecular test sensitive enough to detect infection at the 1 in 10,000 level. It was not
possible to develop this work further in the timescale of the projects.
All cultivated lupins are sensitive to alkaline soils which restricts the extent of cultivation in the UK. The soil pH
limits (4.8 to 7.2) of cultivation were defined in order to inform the industry. L. albus showed the greatest
genetic variation for tolerance to such soils. The bio-chemical and physiological basis for intolerance to
alkaline soils were defined as; an inability to maintain the concentration of iron II (Fe II) in the sap when faced
with large quantities of bicarbonate flowing into the plant from the soil (most Fe being Fe III under such
circumstances) and an inability to control the quantity of soluble Ca in the leaf.
Genotypes of L. albus more tolerant of alkaline soils and displaying indicators of possible mechanisms to
control Fe II and soluble Ca were identified. These were used to develop non-destructive repeatable screens
for tolerance to be used in the breeding of tolerant cultivars.
The effect of the environment on the tolerance of a genotype to an alkaline soil was investigated. It was
hypothesised that the partial pressure of CO2 in the soil influenced the quantity of bicarbonate entering the
plant and therefore the potential for the plant to maintain internal Fe II concentrations. The partial pressure of
CO2 in the soil is controlled by soil aeration which in turn is controlled by soil texture, structural condition and
water status. The potential soil bicarbonate concentration is primarily a function of the soil carbonate content,
modified by the partial pressure of CO2.
The project aimed to quantify these factors with respect to the intensity of stress experienced by a standard L.
albus genotype. This would allow a more sophisticated prediction of land suitability for L. albus production and
would be useful in defining the test conditions when screening breeding material.
A considerable effort was put into Knowledge Transfer alongside the commercial introduction of the first
autumn sown L. albus cultivars. Commercial introduction struggled due to the over dependence on the sowing
window (despite an in depth understanding of the processes and detailed guidance) and an industry
perception that the crop required a large number of inputs (herbicides and fungicides). Both factors are to be
considered in comparison with many spring sown cultivars of lupin which are easy to grow and produce similar
yields to the autumn sown cultivars.
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Scientific report (maximum 20 sides A4)
Project AR0138 is covered in the report as well as other projects on related work covering in total a 10 year
period, from 1993 to 2003 (see Annexe 1). The report is in the form of a review document, considering the
main agronomic aspects in sequence.
Report on DEFRA funded research on autumn sown determinate lupins.
1. Crop Establishment
1.1. Sowing Date.
From the very earliest field experiments (1992 to 1994) conducted it was obvious that the sowing date had a
very strong effect upon the probability that a crop would survive the winter and upon the structure of the
individual plants in the following spring and summer (Shield et al. 1996).
The first autumn sown determinate cultivar, Lucyanne, was extensively tested at Rothamsted and at the INRA
station at Lusignan, western France. Basic models were constructed that described the relationship between
sowing date, autumn weather and plant development. The models used accumulated thermal time (above a base
temperature of 3°C) to predict the development and the extent of lignification of the root parenchyma (crucial
to cold tolerance, Huyghe and Papineau 1990) and leaf production (Huyghe 1991 and 1993). A vernalisation
model used accumulated thermal time (between temperature limits of 1 and 14°C) to determine when the main
stem apex turned floral (Huyghe 1991 and 1993). It was demonstrated that the number of main stem leaves
initiated prior to floral initiation was closely related to the structure of the whole plant (Julier and Huyghe 1993,
Shield et al. 1996). These underlying principles were behind the modified models of the effect of the
vernalisation response on plant architecture proposed by Shield et al. (in press), and shown in Annexe 2.
The models were consequently re-worked using long term temperature records to predict optimal sowing
windows for the different regions of the UK (Milford et al. 1996). The recommended calendar dates varied with
regional climate, late August and early September in cooler northern England and Scotland and late September
and early October in warmer southern and south-western England.
Those sowing windows formed part of the management guidelines for a series of experiments testing the
geographic range of cvs Lucyanne and Ludet (a sister of Lucyanne) conducted between 1994 and 1996. Despite
the results showing that these cvs were suited to most arable sites in England and Wales they were rapidly
replaced by the dwarf determinate cvs Lucille and Lunivers. The dwarf determinates showed greater cold
tolerance and resistance to lodging.
It was demonstrated that the dwarf determinate cvs behaved similarly to Lucyanne and that the same sowing
windows could be recommended (Shield et al. in press). Latterly (from 2001 onwards) it was observed (not
measured from a designed experiment) that sowing dwarf determinate cvs on clay soils produced smaller plants
than sowing on sandy soils. These plants had similar structure in terms of leaf and branch number but with
shorter inter nodes and slightly smaller leaflets. This lead to the recommendation that crops of dwarf
determinate cvs could be safely sown earlier in the autumn than originally recommended via the sowing
windows without fear of large over winter losses or lodging the following summer.
1.2. Row Spacing and Seed Rate.
Early field experiments (1991 to 1994) were sown on wider rows than usual for combinable crops in the UK
(35 cf. 8-12 cm), as this was the practice in France. Experiments conducted by ADAS in 1995 and 1996 (at
CSG 15 (Rev. 6/02)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Rosemaund and Arthur Rickwood, unpublished data) and by Rothamsted and INRA (1996-98, Shield et al.
2002) demonstrated that the use of wide rows was unnecessary and possibly undesirable. Herbicide efficacy
was greater and light distribution potentially better (see canopy architecture below) in crops grown on 12 cm
rows.
The original seed rate recommendation for the UK (of 40 seeds m-2) was derived empirically from early
experience, but stood the test of much subsequent detailed investigation. Over winter plant losses (see below) in
crops sown within the sowing window were approximately 40%, leaving 24 plants m-2 in spring. If sown early
in the sowing window and resulting in large individual plants this could be too many plants for optimal canopy
architecture (see below). From 1999 onwards it was recommended that in good sowing conditions, such as
early sowings where over winter plant losses were predicted to be low, as few as 30 seeds m-2 may be sown.
2. Over-winter losses.
Over-winter plant losses have typically been 40% of the sown seeds in latter years, the causes have varied with
season. The three principal causes were frost, bean seed fly (Delia platura) larvae and slugs. Fungal pathogens
were only occasionally the primary cause of plant loss, but were able to infect sites of damage caused by frost
or invertebrates. Rooks and crows pulled emerging seedlings from the ground but were unable to remove
seedlings with two or more true leaves emerged. Damage to lupin crops by other birds was rare. There was
occasional grazing by deer and rabbits. Hares could be a particular problem in spring when they grazed the
extending main stem.
2.1. Cold tolerance.
Early experiments (1991 to 1993) with Lucyanne demonstrated that young seedlings were sensitive to relatively
modest frosts (Shield et al. 1996). Prior to lignification of the root parenchyma the effect of frost penetrating to
4 or 5 cm soil depth could be to kill the plant (Huyghe and Papineau, 1990). Three or more consecutive nights
with air minima of <–3°C and intervening cold days could result in severe crop damage (Shield et al. 2000).
Once the root parenchyma was fully lignified the plants could withstand air temperatures of –12°C (Shield et
al. 2000, but see below).
The autumns of 1994 and 1995 were unusually warm. All autumn sown lupin crops were at an advanced stage
of development when the first frosts were experienced. Stem extension was observed in cvs Lucyanne and
Ludet when sown at the beginning of the sowing window for a region. These extending plants suffered damage
from relatively mild frosts of –3°C.
CSG 15 (1/00)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
100
90
% of plants established lost to frost
80
70
60
Lu cya n e 1993
50
Lu cya n e 1994 & 95
Lu det 1994 & 95
40
30
No fr ost AccDDA 3°C t o 31/12
P olyn om ia l (a ll da t a except n o fr ost )
20
10
0
0
100
200
300
400
500
600
700
800
900
1000
AccDDA 3°C fr om sowin g t o fir st sever e fr ost
Figure 1. The relationship between thermal time from sowing to the first severe frost (<-3°C) and the
percentage of plants killed by that frost for two cultivars of autumn sown lupin at multiple sites in contrasting
seasons (from Shield et al. 2000).The polynomial model fitted was y = 0.0004008 x2 - 0.425 x + 137.1 which
explained 55% of the variance in y.
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
The autumn of 1993 was a complete contrast to 1994 and 1995 and autumn sown crops experienced cold
weather between sowing and the first frost. The network of experimental sites sown to define the geographic
range of the crop provided excellent data to demonstrate frost tolerance in the first autumn sown determinate
cultivars (Shield et al. 2000 and Figure 1.). The lupin plant became progressively more frost hardy with time
after sowing (as the root parenchyma became lignified) until a turning point was reached after which the plant
became progressively more sensitive to frost (as stem extension began).
The percentage of emerged plants lost to frost was variable. However, the data in Figure 1 were in agreement
with controlled environment studies reported by Leach et al. (1997). These studies showed that lignification of
the root parenchyma and adequate frost tolerance was achieved approximately 400 DDA3°C after sowing
2.1.1. Sowing late in the autumn.
In the autumn of 1996 a demonstration area was sown at Rothamsted (Figure 2). It included three cultivars
(Lumineaux, an old indeterminate, Lucyanne and Lunivers) sown at 6 dates. The objective was to demonstrate
our work on Genotype x Environment interactions and the effect canopy architecture. The ‘treatments’ were
organised for visual effect and it was in no way a designed experiment. However, it was quite clear that the cv
Lumineaux survived the winter from a later sowing date than either of the other two cvs. This suggested that
lignification of the root parenchyma occurred earlier in the life of the plant.
Figure 2. a).
Figure 2.b).
Figure 2. a) A field demonstration of the effects of sowing date and genotype. From left to right are 6 sowing
dates, 14 days apart from 12th August (left) to 22nd October (right) and from front to back are 3 genotypes
Lunivers (foreground blue flowering), Lucyanne (middle ground) and Lunineaux (background, both white
flowering). b) The same demonstration photographed from the bottom left in a) above.
Lumineaux was able to survive when sown on 7th October, whereas Lucyanne and Lunivers were not able to
survive.
CSG 15 (1/00)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Two years of field experiments (Table 1) failed to confirm this observation. The field experiments were
coupled with controlled environment experiments following the protocols described by Leach et al. (1997).
Lunivers was replaced with a new breeding line (CH 1391) thought to be more frost tolerant than many other
genotypes (Huyghe and Harzic, INRA Lusignan, personal communication). Plants were grown in soil filled
boxes through which a glycol cooling pipe passed. This allowed controlled freezing of the soil in the root and
hypocotyl zone when plants were different ages. Plants were removed from the soil tanks immediately before a
freezing treatment was applied and sections of root stained and examined under a low power light microscope
to determine the extent of lignification of the root parenchyma. It was not possible to detect any differences
between genotypes in their ability to survive soil freezing or in the timing or extent of lignification of the root
parenchyma.
Table 1. The percentage of established plants in January dying during the winter. Four sowing dates of three
genotypes in two years at Rothamsted. SED for 1998-99 = 13.02 (22 df) and for 1999-00 = 12.64 (24 df).
1998-99
Lucyanne
Lumineaux
CH 1391
11 Sept
3.1
8.2
6.0
Sowing date
25 Sept
9 Oct
15.0
21.4
14.8
23.0
14.6
24.0
1999-00
Lucyanne
Lumineaux
CH 1391
1 Sept
16.4
11.0
6.9
16 Sept
1.2
24.4
1.8
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28 Sept
12.2
40.9
24.1
8
23 Oct
52.3
50.7
53.3
19 Oct
44.5
73.8
22.6
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
2.1.2. Sowing early in the autumn.
Autumn 1995 was the first time that the dwarf determinate cultivars Lucille and Lunivers were sown on a large
scale at Rothamsted. In warm conditions they were shown to remain in the rosette stage longer than Lucyanne
or Ludet, and therefore to tolerate frost that caused severe damage to those non-dwarf cvs (Figure 3).
No. frost damaged plants m
-2
35
30
25
20
15
10
5
Dwarf
No n dwarf
0
0
2
4
6
8
10
12
14
Me an plan t h e igh t c m
Figure 3. The effect of mean plant height (from soil surface to top of plant) 655 DDA3°C (November) from
sowing on the number of frost damaged plants m-2 in spring. Cultivars Lucyanne, Ludet (non-dwarf) Lucille
and Lunivers (dwarf) sown at 40 seeds m-2 in autumn 1995.
In several cases of plants that began stem extension in autumn the frost damage occurred above the basal rosette
of leaves, killing the main stem but not the whole plant. Branches grew from the basal leaf axils and contributed
to yield. A comparison of seed yield per plant showed that such plants produced on average 70% of the yield of
undamaged plants (unpublished data).
Generally the stage of development of the lupin plants when the first frosts of <-3°C (first severe frost in Figure
1) of each winter were experienced determined the fate of the crop. In most years the plants then remained at
that stage of development until spring (Shield et al. 2000).
It was possible to detect an effect of the duration of the cold temperatures on plant survival. An extended cold
period could damage plants more than short duration extreme temperatures. In the winter 1996-97 at INRA
Lusignan in France the minimum temperature was –12.5°C, the temperature remained <0°C for 12 days and
almost all autumn sown lupins were killed, except for a small number of potentially very valuable breeding
lines. One of those lines, CH 1391, was tested alongside Lumineaux and the control cv Lucyanne in the
experiments described above, where all three failed to show any notable differences in cold tolerance.
In Herefordshire in the winter of 1999-2000 the minimum temperature was –14.5°C, but as part of a cold spell
that only lasted 3 days. Crops of Lucille in the area survived undamaged. The following year temperatures fell
to similar values but as part of a longer cold spell and crops of Lucille were damaged.
It is recognised that snow cover forms a useful insulator for autumn sown crops. At Ancenis in France, during
the same cold period described at Lusignan above, 10 cm depth of snow was sufficient to prevent the severe
damage to lupins recorded at Lusignan (Shield et al. 2002).
CSG 15 (1/00)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
2.2. Pests.
Bean seed fly larvae were the most serious insect pest of autumn sown lupins (Bateman et al. 1997). They were
present at all sites where the crop was sown, but in variable numbers from season to season. In the warm
autumns of 1994, 1995 and 1997 they were active in October, but more normally only in September around the
time of sowing and emergence. Normally the larvae tunnelled into the root / hypocotyl from the soil resulting in
plant death, more unusually surface activity resulted in damage to the plant apex. Experiments were conducted
at Rothamsted between 1997 and 1999, and at ADAS Rosemaund and Starcross (Devon) in 1998 and 1999, to
evaluate potential control methods. Unfortunately there was very little insect activity in the latter two years.
Dimethoate
@ 2-4 lvs
% plants established to be attacked by bean seed flies
30
25
No control
Chlorpyrifos
post sowing
Deltamethrin
@ 2-4 lvs
20
Imidacloprid
Seed treatment
Stale
seedbed
15
10
Bendiocarb
Seed treatment
5
Chlorpyrifos
incorporated
Sown 2nd Sept' 97
0
% root damge by BSF
% Apical damage by BSF
Figure 4. The effect of potential control methods for bean seed fly (Delia platura) larvae in autumn sown lupins,
26th September 1997.
Figure 4 shows the results from Rothamsted in autumn 1997, when bean seed fly larvae were abundant. The
data clearly shows that it is not possible to react to visual signs of damage. The treatments applied at the 2-4
true leaf stage showed no advantage over the ‘no control’ treatment. It was vital to incorporate chlorpyrifos into
the soil to prevent rapid breakdown, when incorporated it was very effective. Bendiocarb was the only
insecticide seed treatment available in the UK with a label recommendation for bean seed fly control, but not
for use in lupins. Ultimately, due to pesticide registration costs, seed was imported from France with an
alternative carbamate insecticide seed treatment applied (furathiocarb).
Several cultural control measures were tested including sowing old seed that had been stored for a number of
years. Of these the only successful method was the stale seedbed technique. If a seedbed were created 3 weeks
prior to seed sowing, and the only soil movement on the day of sowing was caused by the drill, damage could
be lessened. It is thought that this was due to the release of volatiles from soil organic matter during and
immediately following cultivations. These volatiles attract the female fly to lay her eggs. If seed sowing is then
delayed for 3 weeks that batch of eggs have completed their development and have pupated and migrated as
adults. Minimal soil disturbance by the drill prevents new egg laying from taking place.
The limitations of such a technique were demonstrated the following year on the silt soils at ADAS Rosemaund
where the soil structure was not able to sustain good seedbed conditions for three weeks after cultivations.
CSG 15 (1/00)
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
However, at Lusignan in France, in 1999 the technique was again demonstrated successfully as part of a larger
experiment on bean seed fly control. A stale seedbed was also successfully created by ADAS in Devon, but
unfortunately there was very little bean seed fly activity. The soils at Rothamsted and Lusignan are clay loams
and in Devon the soil had a relatively coarse texture. All three soils had greater natural structure and were able
to sustain seedbed conditions for three weeks.
Slugs have also been serious pests of autumn sown lupins. They often fed below the soil surface and therefore
could not be controlled with surface applied pellets. Early in the life of the plant small scale grazing of the
hypocotyl could result in death, later the plants appeared more able to withstand grazing. It is thought possible
that this could coincide with the lignification of the parenchyma, as described for the root above. Above
ground, plants damaged by bean seed fly larvae or by slugs, were observed to wilt. It was necessary to dig them
up in order to determine the cause of wilting. When bean seed fly was the cause the hypocotyl and root were
usually wet and rotten. Often it was possible to recover a larva from the hypocotyl. When slugs were the cause
the plant material remained intact except for the immediate point of damage.
Occasionally plants were damaged above ground by Thrips angusticeps. Affected plants produced
progressively smaller and thickened leaves until no further leaves emerged and the meristem appeared black
and dead. Very occasionally such plants could produce a basal branch from the axil of an early emerging leaf
and survive the damage. More typically they died. Generally thrip damage was considered insufficient to justify
a pesticide application. However, two sowings were completely destroyed by thrips, one of the first commercial
crops sown in conjunction with Dalgety in Herefordshire in 1993-94, and an experiment at Rothamsted in
1995-96. On the second occasion only the middle sowing date of three (each 14 days apart) was affected.
2.3. Diseases.
Plant losses during the winter months caused by fungal diseases were relatively uncommon. Fungi infected the
sites of damage by animals (principally slugs and bean seed flies) and by frost. Fusarium spp. were commonly,
but not exclusively, found on plant parts below the soil surface (Bateman 1997). Botrytis was more common
above the soil surface. Etheridge and Bateman (1999) working in controlled conditions demonstrated that an
artificial damage site on the hypocotyl increased the number of plants infected with these fungi and the
numbers ultimately dying.
Occasionally Botrytis was pathogenic in its own right, especially in late winter and spring, when Botrytis
cinerea infected the newly extending stem tissue. In a series of experiments studying canopy architecture and
light interception it was found that Botrytis infection was least in the more open canopies. Coincidentally these
were the canopies that produced the greatest seed yield and were therefore recommended in our agronomic
advice to farmers and growers.
In Australia intensive lupin growing has increased the incidence of Pleiocheata root rot, a seedling disease. To
date this disease has not been recorded in the UK. When causing Pleiocheata root rot the fungus infects the
hypocotyl beneath the soil surface. It can be transmitted on lupin seed or survive for a number of years (up to 5)
on crop residues in the soil. Despite not having been recorded in the UK, prevention of Pleiocheata root rot is
the principal reason for the recommendation that lupin crops are grown no more frequently than once every 5
years on a piece of land.
The causal fungus, Pleiocheata setosa, has caused ‘brown spot’ symptoms on pods and branches in late
summer in the UK. It was the subject of a number of field experiments at Rothamsted in the early 90s, but in
subsequent years was not considered sufficiently detrimental to the seed yield to require fungicide application.
In the production of seed for re-sowing control of Pleiocheata setosa infection is very much more important to
prevent seed transmission of disease. Fortunately it is very sensitive to applications of tebuconazole used
primarily to control rust (see summer diseases below).
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
3.0. Weed control.
The combination of the difficulties of registering pesticides for additional uses and the sensitivity of lupins to
herbicides generated a large amount of work to identify suitable herbicides and sequences to achieve weed
control. Fortunately products with label approval for use on combining peas and beans were given automatic
off label approval for use on lupins. However lupins are sensitive to a number of herbicides commonly used in
pea and bean crops. An added complication was that certain herbicides are no longer approved or were not
submitted for re-registration.
ADAS, PGRO and one or two agrochemical distributors assisted Rothamsted Research by also carrying out
herbicide screens (Knott 1996 and Shield et al. 2000). Most herbicides available in the mid 1990s were
screened by one or another of these partners. Many of the new introductions since that time have also been
screened.
As with many broad-leafed crops the selective control of grass weeds did not prove to be difficult. However,
from the point of view of herbicide resistance amongst the grass weeds the narrow range of chemical groups
(mainly ACC inhibitors, the Fops and Dims, and one amide) available for grass weed control in lupins is a
concern.
At the present time only the following broad leafed herbicides can be recommended to growers of winter
lupins, pre crop emergence; clomazone, simazine, simazine + trietazine, terbuthylazine + terbutryn, and
trifluarlin, post crop emergence; simazine. This list contains exclusively residual acting herbicides, which
means that they must be applied pre weed emergence and rely on good seedbed conditions and adequate soil
moisture. They are largely unsuitable for use on high organic matter soils.
It has also been clearly demonstrated that the following herbicides could be safely used on lupins were the
relative approvals to be put in place; diflufenican, isoxaben, metribuzin, quinmeric, pendimethalin pyridate and
triasulfuron. All except pyridate and triasulfuron (post-emergence only) could be applied pre and post crop
emergence.
4.0. Disease control in summer.
There are two major diseases of white lupin in summer – rust (Uromyces lupinocolous) and anthracnose
(Colletotrichum spp.).
4.1. Rust.
Rust is endemic in the UK, in most seasons most crops become infected. Left uncontrolled the effect can be
crop loss as demonstrated in 1997 at Rothamsted when omitting a sequence of two applications of tebuconazole
resulted in only 10% of the treated yield. The disease defoliates the crop during the period of rapid seed growth.
Control with fungicides is simple, but requires management effort and incurs a cost.
In 1998 an experiment was designed to study fungicide options and timing of application (Etheridge and
Bateman 1999). Early applications of all three fungicides failed to control the rust, but later application of
tebuconazole significantly increased the seed yield by controlling the foliar symptoms (Table 2). Where all
combinations of date of application were tested only those involving tebuconazole on the third date controlled
the rust.
CSG 15 (1/00)
12
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Table 2. Selected data on the effects of fungicide choice and the timing of application on seed yield (t ha-1 at
15% moisture content) in white lupin cv. Lucille at Rothamsted in 1998. The complete experiment included all
combinations of the three dates of application.
Iprodione and thiophanate-methyl
(Compass at 3 l ha-1)
Tebuconazole (Folicur at 1 l ha-1)
Prochloraz (Sportak 45 at 1 l ha-1)
No fungicide (replicated 3 times
per block)
7th April
1.45
Application date
5th May
1.36
1.51
1.38
1.39
1.30
1st June
1.60
2.97
1.66
1.46
S.E.D. for whole experiment = 0.138, 48 D.F.
Experience from commercial crops suggested that cyproconazole may be a more effective fungicide for rust
control under certain conditions, particularly in the west of England and in Ireland. In 2000-01 at Rothamsted
and in 2001-02 at Rothamsted and Taunton (managed by Arable Research Centres) experiments were sown to
investigate the theory that this may be due to the presence of different pathotypes in the geographic regions.
In 2000-01 rust did not develop to any great extent, the only statistically significant result from the experiment
was that any application of either fungicide increased yield (2.97 t ha-1) over the untreated control (2.40 t ha-1).
This included applications of tebuconazole as only 0.25 l ha-1 Folicur and cyproconazole as only 0.16 l ha-1
Alto.
In 2001-02 rust developed at Rothamsted to a greater extent than recorded previously. It was also present in the
crop on the earliest date recorded (mid April).
Despite the early presence of disease in the crop the early application of fungicide had only a small (nonsignificant) effect on seed yield. As in previous years the later application gave the greater benefit. However in
this experiment the combination of application at both dates was beneficial in the case of Alto (Table 3.).
At Taunton the result was a smaller (non significant) yield advantage to using Alto (3.46 t ha-1) compared to
Folicur (3.31 t ha-1).
Table 3. The effect of fungicide, rate of application and timing on seed yield (t ha-1 at 15% moisture content) of
cv. Lucille at Rothamsted in 2001-02.
th
16 April
Tebuconazole (Folicur at 0.25 l ha-1)
Tebuconazole (Folicur at 0.5 l ha-1)
Cyroconazole (Alto at 0.16 l ha-1)
Cyroconazole (Alto at 0.3 l ha-1)
No fungicide (replicated 3 times per
block)
1.18
1.26
1.26
1.29
Application date
19th June
16th April and
19th June
2.26
2.40
2.52
2.63
2.37
2.70
2.28
2.94
1.16
S.E.D. = 0.146, 27 D.F.
It was concluded that this small advantage to using Alto was common across the country and that no further
effort would be expended characterising pathotypes from the regions.
CSG 15 (1/00)
13
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
4.2. Anthracnose.
Although present in the UK, anthracnose was not considered endemic. It first appeared in a commercial crop of
lupins in 1999, since when it has become very much more common as more crops have been sown. The disease
does not spread easily and is not long lived in the soil. The principal transmission route is on seed. Low levels
of initial infection can rapidly increase if seed multiplication is not handled carefully. Under suitable
environmental conditions the fungus can multiply so rapidly as to destroy a crop from initial infection rates of 1
seed in 10,000 (Peter Römer, Südwestsaat GbR, Germany, personal communication).
Such low rates of infection are not detectable by standard seed testing protocols and require a PCR based test to
objectively determine presence or absence in a sample of 10,000 seeds. In order to develop such a test it was
necessary to determine which genotypes of Colletotrichum were pathogenic on lupins.
The Rothamsted Colletotrichum collection now numbers 75 genotypes, although we can not be certain that
there are no duplicates. Of these 22 genotypes were not pathogenic on Lupinus albus and 2 have not been
tested.
There is some doubt about the taxonomy of Colletotrichum. We used a benomyl sensitivity test based on
growth of isolates on media amended with 5µg/ml benomyl. Literature reports that C. acutatum is insensitive to
benomyl and that C. gleosporoides is sensitive (Freeman et al. 1998). In addition we used the primer
CaInt2/ITS4 which positively identifies C. acuatatum (Sreenivasaprasad et al. 1996).
Of the 58 isolates tested using both methods; 37 were confirmed as C. acutatum, 3 as C. gloeosporoides and 18
gave unclear results. In pathogenicity tests; 7 of the C. acutatum and all 3 C. gloeosporoides isolates were non
pathogenic on white lupin cv. Lucille. Of the 73 isolates tested for pathogenicity 22 were non-pathogenic.
Of the 5 isolates collected from strawberry plants 2 proved pathogenic on white lupin. It had previously been
believed that the isolates infecting strawberry were different to those infecting lupins (Mark Sweetingham,
Agriculture Western Australia, personal Communication).
To study differences between the Colletrotrichum isolates ribosomal DNA (rDNA) was amplified from the
fungi by PCR (primers ITS4/5) and cut by restriction digest enzymes (AluI, RsaI, HaeIII and HhaI). The
method is described in Ward and Akrofi (1994).
Of the 75 isolates 52 were treated with all 4 enzymes and 7 were either treated with some but not all or gave
indistinguishable band patterns with some enzymes. The data show that 24 isolates had the same banding
pattern when treated with these 4 enzymes. Generally these were isolates from the same or similar sources.
However this group did include one of the isolates collected from strawberry. The only isolate to show a very
different banding pattern was the only one not collected from lupins or strawberries (collected from coconut).
Unfortunately these techniques did not distinguish between pathogenic and non pathogenic isolates on lupin
and therefore could not be used as the basis for a molecular screen for seed infection. This would require further
work. A company based in Western Australia has developed a method which it offers as a service for seed
producers and this has been utilised by the UK lupin industry. The test is objective but not quantitative. Should
a quantitative test be developed it may be possible to use seed batches with very low levels of infection
providing that the seed is treated with appropriate fungicides. Römer (Südwestsaat GbR, Germany, personal
communication) confirmed that iprodione and carbendazim was valuable in preventing crop infection when < 1
infected seed in 10,000 was sown (quantitative testing by growing untreated seeds). In Australia it has been
demonstrated that thiabendazole and thiram is effective, a result confirmed by Agrichem (manufacturer) staff in
the UK. It has also been demonstrated that storage for 1 year (Greg Thomas, Agriculture Western Australia,
personal communication) and dipping seed in hot water reduce the rate of infection (Peter Römer, Südwestsaat
GbR, Germany, personal communication). However all of these techniques are only suitable for use with seed
showing nil or very low levels of infection.
CSG 15 (1/00)
14
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
5.0. Pest control.
Mammals and birds have caused damage to lupin crops from time to time. The problems are similar to those for
many other crops. Hares, rabbits and deer are the principal grazers. Rooks and crows can pull young seedlings
from the soil around the time of emergence.
Occasionally aphids colonised lupin crops. Several species were observed but the most visible and probably
most harmful was Macrosiphon albifrons. This is a large and immobile aphid hence populations on any one
plant grow very large and kill that plant. However they spread very slowly creating patches of dead plants by
late summer. The effect is much more severe within plots in experiments than on a whole field scale where
insecticides are rarely applied.
Myzus persicae is also commonly found in lupin crops. Serious feeding damage has not been recorded, but M.
persicae is a vector for bean yellow mosaic virus (BYMV), amongst other viruses. In 1997 the seed
multiplication of Lunivers became infected with BYMV. It was used to sow trait comparison experiments at
Rothamsted in 1997-98. The other varieties (Lucille, Lucyanne and Ludet) were sown from seed that was not
infected. Adult M. persicae were present in the experiments through much of the season, the other varieties did
not become infected to any great extent suggesting that they were largely resistant. If we assume that Lunivers
and Lucille have a broadly similar yield potential, an assumption that experiments to date support, we estimate
that the BYMV cut the seed yield of Lunivers by 0.66 t ha-1.
6.0. Time of flowering.
It is valuable to be able to predict the timing of the opening of the first florets (usually on the main stem
inflorescence) on the lupin plant because it signifies the beginning of reproductive (pod) growth or yield
formation. In the more restricted branching or determinate genotypes it broadly represents the end of vegetative
(branch) growth and is an early indicator of time of maturity (see below). In the less strongly restricted
branching types it represents the onset of the growth of potentially competitive sinks (branches and pods).
The time of flowering on the main stem is largely determined by the number of true leaves initiated and the
phylochron. In addition there are effects of time to emergence. Julier and Huyghe (1993) described a simple
linear model for a selection of autumn sown genotypes in which the thermal time to the opening of the first
floret on the main stem was a function of the number of true leaves on the main stem. i.e. the phylochron was
constant. Shield et al. (in press) demonstrated that the phylochron was modified by day length in the four
determinate cultivars (Lucyanne, Ludet, Lucille and Lunivers), but that this had a very small effect within the
narrow range of day-lengths experienced by an autumn sown genotype in Northern Europe. The number of true
leaves initiated on the main stem is controlled by the genotypic vernalisation response. See Annexe 2 where
empirical models for this and other processes are presented.
CSG 15 (1/00)
15
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
7.0. Time of maturity.
The biggest single problem encountered by those who introduced lupins to the UK in the past was the time of
maturity. The indeterminate or unrestricted branching varieties often grew late in the UK summer and would
not dry down in the deteriorating autumn conditions. The introduction of autumn sowing to allow early summer
flowering and the addition of genetic control of the branching (determinacy) offered great potential for late
summer maturity (Milford et al. 1993 a & b).
In the early years of testing the autumn sown determinate material the theory appeared to be correct. However
in the late 1990s reports were made of crops that we not sufficiently dry (seed moisture >25%) for harvesting in
early October (e.g. ADAS in SW England). In 1998 and 2000 this was experienced at Rothamsted.
A study was initiated to determine the cause of this unpredictable aspect of crop behaviour. The study is not
complete, but our understanding of the processes of maturation of the model white lupin genotypes Lucyanne
and Lucille has been improved.
Pod 5 on the main stem, the fifth pod from the base of the inflorescence, was chosen as the representative pod.
The time of maximum pod wall dry weight (DW) in the representative pod coincided with maximum plant or
crop DW and with observations of leaf canopy degeneration. At this time seed DW was less than 50% of the
maximum, the subsequent rapid seed growth is sustained by remobilisation of assimilates as the rate of new
carbon fixation slows with leaf degeneration. In addition there is a contribution of pod photosynthesis.
Initially it was thought that the thermal time from flowering to maximum pod wall DW was constant and that
the time from maximum pod wall DW to maturity varied. It is now thought that there may be relatively small
variation in the time of maximum pod wall DW from season to season and this is under investigation. However,
the greatest variation in time to maturity comes after the time of maximum pod wall DW. This variation has
been shown to be controlled primarily by the soil moisture deficit and secondarily by the temperature in the 14
days after the maximum pod wall DW is achieved. A soil moisture deficit indicating that the easily available
water capacity of the soil has been exhausted and high temperatures ensure early maturation. Dracup et al.
(1998) working in Western Australia found that maintaining easily available soil moisture with irrigation
interfered with the process of remobilisation of stored assimilates from the pod wall such that it was incomplete
at maturity. This may be the mechanism that delays maturation in the UK. In Wales, Ireland and parts of
northern and western England, these conditions do not prevail sufficiently frequently to ensure late summer
harvesting of dry lupin seed in most years. The data collected relates to the white lupin genotypes Lucyanne
and Lucille and appears to be relevant to other white lupin genotypes observed. There were genotypic
differences between Lucyanne and Lucille. Lucille matured on average 5 days later than Lucyanne.
Narrow leafed (L. angustifolius) and yellow (L. luteus) genotypes appear to behave differently and are more
reliable in cooler damper climates.
8.0. Yield and yield stability.
The breeding of genetically determinate or restricted branching varieties of white lupin resulted in greater yield
stability than experienced when growing indeterminate or unrestricted branching varieties (Julier et al. 1993).
This was largely achieved via a large and stable harvest index. When the dwarfing character was added to
prevent lodging the traits were in place to increase seed yield by increasing crop biomass (Cowling et al. 1998).
However, considerable unexplained variation in yield was recorded with the model determinate genotype
(Lucyanne) in a large data set derived from many environments between 1994 and 1996 (Figure 5). The data
were filtered to remove individual values with an obvious explanation for anomaly, such as poor weed or
disease control. A wide range of factors such as weather and soil variables during all or part of the growing
season were used to explain the variance in seed yield in the remaining data. Ultimately the numbers of
branches m-2 was identified as the variable that explained the greatest proportion of the variance in seed yield
(Shield et al.2001). Figure 5 shows that there were two distinct relationships between seed yield and the
number of branches (or axes, Shield and Milford 1995) carrying yield at harvest; one for high yielding crops
and one for more modestly yielding crops.
CSG 15 (1/00)
16
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
5
4.5
y=
4
4.16
------------------------------------------(1 + EXP (0.05458 * (x - 53.34)))
accounting for 92% of the variance in y
Seed yield t ha -1 (@85%DM)
3.5
y=
3
2.93
------------------------------------------(1 + EXP (0.032 * (x - 63.14)))
accounting for 83.2% of the variance in y
2.5
2
1.5
Lucyane high yield potential
1
Lucyane moderate yield potential
0.5
0
0
20
40
60
80
100
120
140
160
180
200
No. of principal yield bearing axes m-2
Figure 5. The relationship between the number of principal yield bearing axes (main stem and 1° branches) m-2
and seed yield for 50 crops of cv. Lucyanne grown at sites across the UK between 1994 and 1996.
Having identified the factor most closely linked to the variation in seed yield it was necessary to discover the
underlying cause, particularly as there were two distinct relationships. The component of yield varying with
total seed yield was the number of pods on the main stem. Regardless of the number of branches m-2 the same
seed yield m-2 was recorded from the branches. Some data available from Lucyanne suggested that there could
be competition for assimilate between pods on the main stem and the branches during the period immediately
following main stem flowering. This was thought to be the time when newly formed pods were most
susceptible to being aborted. This theory implied that a large number of branches per plant would lead to a
lower seed yield per plant due to increased competition (Shield et al. 2001). The alternative hypothesis was that
a large number of branches m-2 created shading of the main stem leaves and pods and resulted in fewer pods
being retained on the main stem.
In 1998-99 the model white lupin was changed as the dwarfing character became available. The model dwarf
determinate Lucille showed very similar developmental characters (numbers of leaves and branches) to
Lucyanne, but in a more compact plant form. A field experiment in 1999 demonstrated that yield stability in
Lucille was affected by the number of branches m-2 in a very similar way to Lucyanne.
Subsequent field experiments demonstrated that the number of branches per plant was not an important factor
controlling seed yield per plant. Lucille was sown early in autumn (thereby producing many leaves and
branches) at very low plant density. Total yield per plant and the yield on the main stem were large. This
suggested that the number of branches m-2 affected seed yield by shading of main stem leaves and pods.
This work clearly demonstrated that seed yield was not directly related to crop biomass in a dwarf determinate
genotype. Canopy architecture and light distribution were more important in determining seed yield above a
certain critical biomass.
The knowledge gained from this work in conjunction with our understanding of plant development allowed us
to refine our sowing date and seed rate recommendations for dwarf determinate genotypes like Lucille. When
sowing early (potentially earlier than the beginning of the normal sowing window for a site) the seed rate
CSG 15 (1/00)
17
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
should be 30 seeds m-2, this should increase with time until the end of the recommended sowing window, when
40 seeds m-2 should be sown. An additional factor to be considered when sowing genotypes such as Lucille is
the soil type, on clay soils the plants are more compact and a greater population density is recommended than
on sandy soils.
During the summer of 1998 MAFF (DEFRA) set a target for yield stability from the model dwarf determinate
genotype of 3 t ha-1 through a ROAME A. Table 4 shows the seed yield achieved from ‘standard agronomy’ in
the following seasons. The yields are measured from large plots to avoid edge effects etc. The very wet year of
2000 was a problem for most sectors of agriculture, the lupins were very late to mature and lodged badly, but it
was pleasing to see a good seed yield. It is disappointing that in the last two years of the sequence seed yield
was notably lower than in previous years.
These latter two years coincided with a deliberate policy to place the lupin crop on land that had grown lupins
in the recent past (1996 and 1998 for the sites used in 2002 and 2003 respectively). Previously all lupin
experimentation (except certain pathology experiments) had been placed on new sites. However, close
monitoring failed to show any problems associated with cropping history. In 2002 the pressure on the crop from
rust was the greatest that had been observed, the crop was given a ‘standard’ two fungicide spray protection.
The crop may have responded positively to a third spray. Were that spray to have preserved the 0.5 t ha-1 that
appeared to have been lost in comparison with previous years it would have been economically justified. There
was no opportunity to test that theory at the time. In autumn of 2002 on the site of the 2003 crop the soil was
not sufficiently re-consolidated following ploughing. It has been observed in the past that over-winter plant
losses have been large in poorly re-consolidated soils at Rothamsted. In this case slugs grazing underground
and out of the reach of pesticides caused most of the damage. At harvest there were only 6.4 plants m-2 of
Lucille. The severe drought of summer 2003 is not thought to have had a great influence on the seed yield.
Table 4. ‘Standard agronomy’ applied, maturity dates, lodging and seed yield from cv. Lucille during the
period 1999-2003. m.c. = moisture content.
Harvest
year
1999
2000
2001
2002
2003
Sowing
date
14/09/98
22/09/99
21/09/00
21/09/01
24/09/02
Sowing
density
Seeds m-2
Maturity
date
30
40
35
35
35
25/08/99
25/09/00
28/08/01
09/09/02
10/08/03
Harvest
date
Lodging
% of area
27/08/99
25/09/00
09/09/01
12/09/02
14/08/03
<10
25-50
0
<5
0
Seed
yield
t ha-1 @
15% m.c.
3.58
3.77
3.68
3.01
2.01
One of the great disappointments of the autumn sown lupin crop has been the failure of so many others,
whether they are farmers or researchers, to reproduce our yield results. Yield results from our trial partners
(ADAS, ARC and Reading University) are presented in Table 5 and reflect this variability. Very acceptable
yields have been achieved but must be viewed alongside complete crop failures. As in Table 4 data are selected
from the management regime most similar to our recommended agronomy.
Each crop was recorded carefully and reasons attributed to the failure or success. Often the reasons for poor
crop performance lay with operator error, such as poor site selection or drill calibration, failure to achieve the
recommended sowing date. However, the poor performance of all crops in 1998 is associated with the large
numbers of plants that survived the very warm autumn and winter, and the large vegetative structure of those
plants. The maximum yield achieved at Rothamsted in 1998 was 3.50 t ha-1 from a heavy clay soil. On a lighter
silty clay soil only 2.72 t ha-1 was recorded. This is a good example of the ability of the clay soils to produce
good seed yields from early sowings or, as in this case, in a warm autumn.
CSG 15 (1/00)
18
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Table 5. Seed yield(t ha-1 @ 15% m.c.) achieved by trial partners in the regions of the UK 1997 – 2002. The
data reflect the very variable results achieved by farmers.
– no crop sown, 0.00 crop sown but failed.
*The data from Reading University were generated by hand harvesting an area of the plot. This technique is
known to produce large values.
ADAS Devon
ADAS Rosemaund
ADAS Arthur Rickwood
ADAS High Mowthorpe
ARC Kettering
ARC Taunton
Reading University*
1997
4.23
3.34
2.31
3.22
-
1998
1.52
1.27
1.10
1.40
-
1999
2.28
2.80
2.78
-
2000
4.41
0.00
4.24
-
2001
0.00
6.61
2002
3.38
4.33
9.0. Further improvement of yield.
It was considered probable that further improvements to yield were to be gained by increasing the harvest index
in the reproductive compartment (Lagunes-Espinosa et al. 1999). Lupin species are generally known to produce
large pod walls (thick with a large dry weight) when compared to other grain legumes. In modern cultivars of
peas (Pisum sativum) the pod wall is only 13% of the pod dry weight at harvest (pod wall proportion), in white
lupin that value is often >30% (Mark Reader & Miles Dracup, Agriculture Western Australia, personal
communication). The large pod wall was considered a waste of dry matter, as it was considered to only be
necessary as packaging for the seed, and that the DM should be incorporated into the seed.
Lagunes-Espinosa et al. (1999) were able to clearly demonstrate that when a wide range of genotypes
exhibiting a range of pod wall proportions (18-32%) were grown the seed yield was negatively correlated with
the pod wall proportion. Our unpublished work (Figure 6) with one genotype (cv. Lucille) showed that the
environmental conditions that lead to the greatest seed growth also lead to the greatest pod wall DW at harvest,
and therefore very little change in pod wall proportion.
CSG 15 (1/00)
19
Improving the physiological and agronomic basis of UK lupin
production.
Project
title
DEFRA
project code
ARO138
0.8
0.7
DW Pod wall per pod (g)
0.6
0.5
0.4
y = 0.4484x + 0.0963
R2 = 0.8565
0.3
1999
2000
2001
2002
Linear (All Data)
0.2
0.1
0
0
0.2
0.4
0.6
0.8
1
1.2
1.4
DW Seed per pod (g)
Figure 6. The relationship between the dry weight of seed and pod wall per pod in cv. Lucille over 4 seasons,
demonstrating that pod wall proportion is relatively constant. Data from sowing date and seed rate treatments
setting up a range of canopy architectures and growth potentials.
0.025
Main stem pod growth rate mg °Cd
0.02
0.015
0.01
y = 0.0002x + 0.0063
R2 = 0.5454
0.005
0
0
10
20
30
40
50
60
% incident PAR reaching base of main stem infloresence at main stem flowering
Figure 7. The effect of the availability of PAR on main stem pod growth rate during the most rapid pod growth
phase (approximately 6 weeks after main stem flowering). Data from cv. Lucille in 1999 and 2000 at
Rothamsted.
CSG 15 (1/00)
20
Improving the physiological and agronomic basis of UK lupin
production.
Project
title
DEFRA
project code
ARO138
Figure 7 shows that the quantity of PAR available had a large influence over the main stem pod (seed plus wall)
growth rate in the 6 weeks after flowering. The proportion of the PAR reaching the base of the main stem
inflorescence at the beginning of main stem pod growth (flowering) was used as an indicator of the PAR
available during the most rapid phase of main stem pod growth (the 6 weeks following flowering). This and
Figure 6 indicated that it would not be possible to use agronomy to minimise the pod wall proportion and
maximise seed yield.
Assuming that the only role of the pod wall was to act as packaging for the seed was a simplification. The pod
wall acts as an intermediate storage organ for assimilates destined to drive seed growth and photosynthesises in
its own right. Lagunes-Espinosa et al. (2000) were able to demonstrate that the low pod wall proportion
genotypes were able to produce large seeds with high nitrogen (protein) concentrations.
Our work to study pod wall photosynthesis was less clear as large variance was encountered between individual
measurements. Later work was carried out in the controlled environment rooms at Rothamsted to lessen the
variation due to environmental conditions. This revealed that much variation existed between plants of the same
genotype and within pods of similar age on the same plant.
It was demonstrated that lupin pods photosynthesise but that there is only a net fixation of CO2 at high light
intensities (Figure 8). Shading the pods during the light period gave an indication of the extent of the respiration
taking place. In this example a light intensity of 1500 µmoles PAR m-2 s-1 (bright summer sunshine) was
required to maintain the pod at its CO2 compensation point. This example is typical of the large number of
measurements made of field grown plants.
The pods had relatively poor gas exchange characteristics (compared to a leaf) leading to a large build up of
CO2 in the internal gas spaces during the dark period (Figure 9). This CO2 appeared to be re-fixed during the
first few hours of the light period. It was not possible to detect with any confidence differences in pod
photosynthesis between genotypes despite differences in pod wall morphology.
2500
4
2
0
X umoles PAR m-2 s-1
1500
-2
-4
1000
-6
500
O Photosynthesis / respiration (umoles CO2 m-2 s-1)
2000
-8
12:58:00
12:57:20
12:56:40
12:56:00
12:55:00
12:54:20
12:53:40
12:53:00
12:52:30
12:52:00
12:51:30
12:51:00
12:50:30
12:50:00
12:49:36
12:49:12
12:48:48
12:48:24
12:48:00
12:47:20
12:46:30
12:45:40
12:45:00
12:44:00
12:43:20
12:42:36
12:42:12
12:41:48
12:41:24
12:41:00
12:40:36
12:40:12
12:39:45
12:39:15
12:38:00
12:36:00
12:34:00
12:32:00
12:30:00
12:28:00
12:26:00
12:24:00
12:22:00
12:20:00
12:18:00
12:16:00
-10
12:14:00
0
Time (UT)
Figure 8. The response of a main stem pod (cv. Lucille) photosynthesis to Photosynthetically Active Radiation
(PAR). Measurements made in full sunlight (1500 µmoles PAR) on July 26th 2002, shading applied twice.
CSG 15 (1/00)
21
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
2000
1800
CO2 concentration mg kg-1
1600
1400
1200
1000
800
600
400
200
0
2
7
12
17
22
Time (BST) h
Figure 9. Measurement of the CO2 concentration in the internal gas spaces of a main stem pod of cv Lucille
over one day.
CSG 15 (1/00)
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Improving the physiological and agronomic basis of UK lupin
production.
Project
title
DEFRA
project code
ARO138
10.0. Seed Quality.
With the exception of LS3606, the projects listed in Annexe 1 did not include objectives to make a detailed
investigation of seed quality in lupins. However some analyses of the data gathered gave useful indicators of
factors and processes controlling attributes of seed quality. It was noted that the number of leaves per plant was
positively related to the seed nitrogen (protein) content at harvest (Figure 10) in the model determinate
genotype Lucyanne and in the generally lower seed nitrogen genotype Ludet.
80
75
y = 0.1244x + 57.663
R2 = 0.5031
g N \ kg DM in seed
70
y = 0.0699x + 57.551
R2 = 0.5713
65
60
55
Lucyanne 94
Lucyanne 95
Ludet 95
Linear (Lucyanne 94)
Linear (Lucyanne 95)
Linear (Ludet 95)
y = 0.1249x + 49.809
R2 = 0.8128
50
45
40
0
20
40
60
80
100
120
140
Total No. leaves per plant
Figure 10. The relationship between the number of leaves per plant and the seed nitrogen content. Data from
multiple sites and sowing date experiments in 1994 and 1995 with two non dwarf determinate cvs (Lucyanne
and Ludet, 1995 only).
The number of leaves per plant is a factor determining total plant photosynthesis which in turn determines the
quantity of nitrogen fixed by the rhizobia in the root nodules. The leaves also potentially represent the organs
with the greatest nitrogen concentration during growth; therefore a large number of leaves per plant could
represent a large pool of nitrogen available for re-mobilisation to the seed during maturation.
However, a large number of leaves per plant also indicates a large number of internodes, and a large mass of
stem. Pate et al. (1998) describe stem tissue as an important site for nitrogen accumulation (especially as
arginine) and the stem as an important site for nitrogen enrichment of the xylem ensuring that large quantities
of nitrogen are supplied to the sinks in the upper parts of the plant. In 1995 in South Devon an experiment was
conducted to investigate the affect of a plant growth regulator on Lucyanne and Ludet sown on different dates
(Table 6). The growth regulator applied was paclobutrazol, a triazole with fungicidal properties (observed and
recoded in a lupin experiment at Rothamsted). Application was on April 19th, which is very early in the
growing season for the fungicidal properties to have an effect on seed quality (see the effects of timing of
tebuconazole on seed yield in Summer Diseases above). The proportion of the plant that was still green at
harvest was affected by the application of growth regulator. This was an unusual result in that prior experience
(in a range of crop types) would suggest that the growth regulator may keep the plants green for longer
(especially if there was a fungicide effect). Our results were contrary to this with more of the untreated plants
remaining green. The data are representative of one site and one season, but raise the question “is stem an
CSG 15 (1/00)
23
Improving the physiological and agronomic basis of UK lupin
production.
Project
title
DEFRA
project code
ARO138
important source of nitrogen to sustain the latter stages of seed growth ?”. Alternatively “are these results
explained by later maturity that allowed a greater accumulation of nitrogen in the seed ?”
Table 6. The affect of plant growth regulator (paclobutrazol) on cv Lucyanne and Ludet sown on two dates
(early and late September) in South Devon in 1994-95.
5 June
Main stem height (cm)
Total height (cm)
Harvest (14 August)
Proportion of plants remaining green
Seed yield (t ha-1 @ 15% m.c.)
% N in seed
+
PGR
- PGR
F pr.
SED
46.6
92.9
59.0
102.4
<0.001
0.004
2.10
3.05
0.20
2.88
6.277
0.45
0.053
2.82
NS
6.370 0.036
31 DF
0.124
0.0426
24
Testa / seed coat (% msw)
22
20
18
16
y = -0.0218x + 24.664
14
R2 = 0.7032
12
10
0
100
200
300
400
500
Mean seed weight (mg)
Figure 11. Testa as a proportion of total seed dry weight as affected by total seed dry weight. Data from a
range of white (L. albus) and yellow (L. luteus) genotypes grown in several environments.
The greatest anti-nutritional factor in sweet (low alkaloid) lupins is the quantity of non starch polysaccharides
(NSP) in the seeds. Monogastric animals can not easily break these carbohydrates down and can suffer a range
of digestive tract problems if fed modest quantities of lupin seeds. The NSP is contained in the cell walls
throughout the seed but is especially concentrated in the testa. Lupins have a relatively large testa, 17 to 22% of
the seed by weight, compared to other grain legumes (Reader and Dracup, Agriculture Western Australia,
personal communication). The proportion of the seed that is testa falls with increasing overall seed size (DW,
Figure 11). Therefore breeding and selecting for large seeded genotypes is an immediate route to lessen the
proportion of NSP in the seed.
CSG 15 (1/00)
24
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
11.0 Underpinning physiology: the response of lupins to alkaline soils, comparing
the intolerance physiology of L. albus to a more tolerant species.
It was established that the roots system, specifically the cluster roots (previously termed proteoid roots), were
highly adapted to exploit a soil environment heterogeneous for lime. This response enabled greater shoot
growth and leaf greenness (Kerley 2000a and b)
This work was progressed by comparing L. albus to L. pilosus Murr., which occurs naturally in calcareous soils
(Kerley et al. 2000). However, the reasons for the tolerance of L. pilosus to these soils had not been
investigated. By comparing L. albus with this tolerant species in both homogeneous and heterogeneous soils, it
was anticipated that specific mechanisms of intolerance in L. albus would be elucidated.
The root systems of L. albus and L. pilosus responded to a patch of acid soil within a limed-soil profile through the
specific proliferation of cluster roots in the acid soil. This proliferation resulted in a higher rhizospheric citrate
concentration on a soil dry weight basis. Although individually the cluster roots were not functionally more active
in citric acid exudation, as they were so densely packed it was probable that the zones of citric acid exudation
overlapped, ensuring extraction of P and Fe from the whole of the acid soil.
As well as cluster root proliferation in the patch of acid soil, the plants also showed increased leaf
greenness, net CO2 assimilation, shoot growth and leaf emergence, indicating that the plants had derived some
Figure 12. Radiation response curves for L. albus (solid line) and L. pilosus
(dashed line). Plants grown either in a wholly limed-soil profile (●) or in a profile
containing patches of acid soil (■).
nutritional benefit from the soil. The PAR response curve demonstrated the difference between the plants (Figure
12). There was no response by L. pilosus to the presence of patches of acid soil, in contrast L. albus showed very
little ability to respond to the increasing PAR when grown in limed soil, but the response was much greater when
acid soil was present. This difference clearly demonstrates the tolerance and intolerance of both species to the
limed soil.
Although both L. albus and L. pilosus responded strongly to a heterogeneous soil environment by the proliferation
of cluster roots, differences other than PAR response were also apparent between the species. L. pilosus produced
15% of its cluster roots in the limed soil; in contrast fewer were produced by L. albus. Being adapted to calcareous
CSG 15 (1/00)
25
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
soil, L. pilosus may grow better in the heterogeneous profile and be able to exploit the acid soil more so than L.
albus. In addition, the strong secondary and tertiary lateral roots system of L. pilosus may facilitate the exploration
of soil. It is possible that the positive response of L. pilosus to an acid soil patch and the greater growth in
homogeneous acid soil profiles indicates that although the species tolerates limed soil it might preferentially grow
in acid soil.
This work demonstrated important similarities between the two species in their response to limed and
heterogeneous soil. L. pilosus however demonstrated a better root development in the limed soil, indicating better
nutrient acquisition, which was reflected in the higher photosynthetic capacity of the shoot. The response of the
root system as a whole to the stress conditions is considered in the next section
11.1 Root system architecture response to pH, Ca and bicarbonate
The poor growth of lupins on calcareous or limed soil has been attributed principally to high pH, bicarbonate
(HCO3-) concentration, or excess calcium (Ca) (e.g. White and
Robson 1989). The plant’s responses to these stresses are mediated
through the roots and may be specific to different parts of the root
system. Differences in root architecture between stressed and nonstressed plants may be important in determining the plant’s ability to
exploit calcareous or limed soils. Additionally, this knowledge might
help evaluate tolerance.
It is difficult in soil-based experiments to determine the changes in
root architecture specific to pH, HCO3- or Ca. Liquid culture system
and two-dimensional root chambers were developed to provide an
environment in which these conditions could be evaluated more
easily (Figure 13).
The liquid culture system was used to separate pH, HCO3- and Ca,
effects on the root system of the intolerant cultivar Lucyanne (Kerley
and Huyghe 2001). These changes were specific to the type of root, and
differed depending on the stress imposed. The study also discriminated
between a genotype and cultivar of L. albus and a genotype of L.
pilosus, based on quantifiable responses to solution culture pH.
Figure 13. Image of facsimiled
chamber to demonstrate the different
root structures in the root
architecture
Generalised root measures
Increasing solution culture pH had no effect on root dry weight, weight
ratio and Specific Root Length, whereas increasing the Ca
concentration affected both the dry weight and weight ratio. In the
presence of HCO3-, root dry weights and weight ratios were lower than
in its absence. The decrease in root weight ratio indicated that the root
system was proportionately smaller than the shoot when stressed by
HCO3- at pH 6.5. Although, the pH responses with HCO3- are difficult
to interpret, the results indicate that HCO3- could be the most important
stress of the three at decreasing root exploration.
11.1.1 Tap root architecture
The tap root length was shorter in the presence of higher pH and Ca and HCO3- concentrations. The retardation of
L. angustifolius L. roots exposed to HCO3- has been attributed to decreased respiration, leading to a failure to
acidify the apoplast for cell expansion. As the roots were well aerated in this study, a lack of respiration was
unlikely. However, both HCO3- and pH did decrease root length, most likely through the process of decreased cell
elongation. The role of Ca would have been similar; the binding of Ca to cell walls would have prevented
elongation.
CSG 15 (1/00)
26
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
The tortuousness of the tap root’s pathway increased under pH and Ca stresses from an unimpeded straight line
(index 0) to maximum index of 0.36 in 2 mM Ca. Although the physiology of the tortuous response is unclear, it is
likely to have been a direct attempt to avoid the unfavourable environment. The decrease in tap root length and
increase in tortuousness however, resulted in a root system that would be less efficient for nutrient and water
acquisition. Interestingly, HCO3- stress did not affect the tortuousness as in the other treatments. Although the
reason is unclear, it does demonstrate the presence of different physiological responses to the stresses.
Tap root death was a major response to pH and HCO3- stress. In contrast, Ca had no effect on root mortality. This
further indicated that a different physiological stress occurred in response to Ca, compared to pH and HCO3-, which
was related mainly to cell elongation. Death occurred mainly at the tip of the tap root, and resulted in a loss of
apical dominance, with the subsequent proliferation of many indeterminate lateral roots.
11.1.2 Lateral root architecture
Stress responses were also apparent in the lateral roots of the cultivar Lucyanne. Fewer lateral roots were
produced under all stress conditions, which resulted in a lower density along the tap root. This indicated that the
root system under the stress conditions had a much lower capacity to exploit the media than when not stressed.
Lateral roots were defined as either determinate (<3cm) or indeterminate (>3cm); their ratio (determinate :
indeterminate) decreased due to alkaline pH, was unchanged by high Ca, and was increased due to HCO3- at pH
7.5. At the alkaline pH, the proportionately fewer indeterminate roots accounted for the decreased ratio. The
implication of this response is an impaired ability to explore the environment. In the presence of HCO3- at pH 7.5,
the fewer determinate roots accounted for the increased ratio, and could have implications for the ability of the
plant to exploit its immediate surroundings. This difference in response is evidence for these stresses being
discrete. Increasing Ca concentration did not change the ratio, however both root types were equally decreased in
numbers, indicating that both were similarly affected.
Fewer cluster roots were produced under the Ca and HCO3- stress conditions, whereas alkaline pH had no effect on
cluster root production. Care must be taken in the interpretation of the solution culture results because of the
nutrient solubility, concentration homogeneity, and the regular replacement of the solution. The process of nutrient
uptake that relies on the concentration of exudates into the rhizosphere would have been absent under liquid culture
conditions. In this respect the use of liquid culture to compare cluster root activity is unreliable. However, it is
important to determine that Ca and HCO3- do affect cluster root numbers.
By using a drip culture system in which the root system of L. albus can develop observable root architecture,
changes in architecture in response to pH, HCO3- and Ca stress were quantified. These changes were present in
the tap, lateral and cluster roots, and were seen to vary between the stresses imposed. However, as with all
liquid culture analyses, care must be taken in the relating these responses to those that may occur in soil
systems, due to the complexity involved. This technique was used as the basis of a successful system to screen
genotypes (see later section).
11.2 A nutritional basis for tolerance to calcareous or limed soil
Having established that limed or calcareous soil conditions affect root architecture and physiology (specifically
that of cluster roots), it was then considered how this would affect the shoot’s nutritional status. A change in
nutrient composition in the shoot would be expected from the reduced growth responses and photosynthetic
capacity observed earlier in the soil patch study. By examining the change in nutrient composition, it was hoped
to develop a screen to compare between cultivars.
The aim of the initial study was to compare a number of nutritional and biomass analyses to evaluate the
calcareous or limed soil tolerance of L. albus genotypes, selected from previous trials.
Evaluation of the nutritional status, dry weights, and yields of genotypes of L. albus, L. pilosus and L. angustifolius
were made in a field of mildly acidic pH 5.8 to alkaline (limed to a pH maximum of 8.4) soil at Rothamsted
(Kerley et al paper in preparation). Plants were sampled for shoot biomass and nutritional content during the season
and seed yield per plant at harvest.
CSG 15 (1/00)
27
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
L. pilosus was the species most tolerant of the calcareous soil, whereas L. angustifolius was the least tolerant.
Considerable variation in tolerance was apparent between the L. albus genotypes; the cultivar Lucyanne was
comparable with L. angustifolius, whereas the genotypes La 673, 668 and 675 were more comparable with L.
pilosus.
Table 7. Total Ca (mg / g dry weight) and Fe, (μg / g dry weight) concentrations in the stem and leaves of L.
albus cv. Lucyanne and selected genotypes, and L. pilosus and L. angustifolius sampled 56 days after sowing
(DAS) in neutral or alkaline soil.
Significance
(soil pH)
L.
angustifolius
L. pilosus
La 675
La 673
La 672
La 671
La 669
La 668
soil pH Genotype
Lucyanne
Nutrient
Ca - Stem neutral 5.1 6.9 6.2 6.4 6.5 5.6 6.5 11.2 9.2 P  0.001
alkaline 6.8 7.8 7.5 7.8 7.6 6.5 7.5 12.9 9.9
Mean 5.9 7.3 6.9 7.1 7.1 6.0 7.0 12.1 9.6
Ca - Leaf neutral 10.0 11.1 11.1 11.2 11.9 11.0 12.5 22.2 31.3 P  0.001
alkaline 12.4 11.7 12.0 12.1 13.7 10.9 13.1 22.1 27.7
Mean 11.2 11.4 11.6 11.7 12.8 11.0 12.8 22.2 29.5
Fe - Stem neutral 590 540 420 550 530 480 490 510 330 P  0.001
alkaline 610 690 580 630 520 530 600 530 430
Mean 600 610 500 580 530 500 540 520 380
Fe - Leaf
neutral 550 500 480 530 460 540 510 580 570 P  0.001
alkaline 460 410 440 480 380 480 530 520 630
Mean 500 460 460 500 420 510 520 550 600
Leaf chlorosis was shown to be an unreliable measure of calcareous soil tolerance in the field (e.g. Kerley et al
2001), whereas a genotype x soil-pH interaction in the expanded leaf number indicated that this analysis might be
of use in genotype evaluations. Nutrient concentration differences were apparent between the species and between
the L. albus genotypes. This indicated the occurrence of possible tolerance mechanisms including the control of Ca
uptake and the partitioning of Fe. Clear differences were apparent between the three species in terms of tolerance to
the calcareous soil. Within L. albus important differences were apparent when specific analyses were examined.
However, variation between different analyses and at different stages of growth resulted in these differences. Taken
as a whole between the L. albus genotypes, they were not of sufficient magnitude to discriminate potentially
tolerant from susceptible genotypes. The most interesting differences were apparent in the concentrations of Ca and
Fe in the leaves and stems (Table 7).
Although the shoot (leaf + stem) Fe contents did not differ between the soil types in this study, there was
significantly less Fe in leaf and more Fe in stem in the alkaline pH soil. This might have resulted from less
translocation of Fe from the stem due to HCO3- in the xylem.
The only nutrient that increased in concentration when grown in the calcareous soil was Ca. The L. albus
genotypes Lucyanne and La 672 showed increased Ca contents in the leaves, suggesting these plants were unable
CSG 15 (1/00)
28
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
to control Ca uptake into the leaf tissue. In contrast L. pilosus showed no change in leaf Ca concentration between
the soil types, indicating that it might be able to control the Ca concentrations in the leaf tissue. Importantly, the
response of La 673 was similar, indicating one possible mechanism of tolerance in L. albus.
When the stem tissue was analysed, all the plants increased their stem Ca concentration when grown in the alkaline
pH soil. Ca was therefore taken up in greater concentrations by all plants in the alkaline pH soil. However the
maintenance of Ca concentrations in the leaf tissue of L. pilosus and La 673, demonstrated a means of nutrient
partitioning that may provide a level of tolerance. Additionally, this partitioning may provide a means of
comparing genotypes for tolerance.
The potential of Fe and Ca to be nutritional screens led to further research on both nutrients. Fe deficiency is a
major cause of lime-induced chlorosis, however despite the appearance of chlorosis, whole-plant total Fe
concentrations were not always deficient when L. albus was grown in lime soil. From literature concerning other
species, it was possible that differences in the concentration of Fe forms (FeII and FeIII) within leaf tissue might
explain the induction of lime-induced chlorosis, even in plants sufficient in Fe. Thus the partitioning of the forms
of Fe in the shoot was examined.
Ca is one of the few elements to show an increased shoot concentration when grown in limed or calcareous soil.
Within the shoot, the control of Ca is a major tolerance mechanism of calcicole species, and its partitioning might
provide another potential screen.
Table 8. The concentration of Ca form (total, insoluble, soluble and the ratio of soluble Ca : total Ca) and Fe
form (total, FeIII, FeII and ratio of FeII : total Fe) in leaflet tissue of the L. albus cultivar Lucyanne harvested
in April (215 DAS) in neutral or alkaline-pH soil.
Nutrient
(tissue dry weight basis)
Total Ca (mg g-1)
Insoluble Ca (mg g-1)
Soluble Ca (mg g-1)
Ca ratio (soluble Ca :
total Ca)
Total Fe (μg g-1)
FeIII (μg g-1)
FeII (μg g-1)
Fe ratio (FII : total Fe)
Soil pH
Neutral
15.3
12.3
3.00
0.21
P
Alkaline
36.8
32.9
4.14
0.109
<0.001
<0.001
<0.001
<0.001
1830
1751
79.5
0.046
2251
2177
73.5
0.037
ns
ns
ns
ns
Plants were grown in neutral pH or limed soil (Kerley et al in press). In April and then in June the shoot tissue
was sampled, divided into specific tissue types and analysed for FeII and FeIII, as well as soluble and insoluble
Ca fractions. Nutritional and tissue differences were seen between the April and June sampling (Tables 8 and
9).
In April the Ca response was mainly as insoluble Ca and indicated that the excess Ca taken up from the alkaline
soil was successfully insolubly sequestered. No differences were apparent in the Fe fractions possibly because
the plants were still in an over-wintering form and were less active. The total Fe concentrations in April were
considerably higher than values reported for spring sown lupin material harvested in early summer, indicating
that continued uptake of Fe occurred over winter by the autumn sown plants. This Fe uptake may explain the
observed greenness of the leaves of these plants.
When actively growing in June, the increase in total Ca in the limed soil grown plants was as both the soluble
and insoluble forms. The percentage increase was greater in the soluble compared with insoluble form,
indicating that although the Ca was being taken up and sequestered, there remained excess Ca in the soluble
form. The presence of soluble Ca can affect plant growth e.g. by reducing net assimilation rates through the
suppression of stomatal opening by the increase in Ca concentration in guard cells. Although it is unclear
CSG 15 (1/00)
29
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
whether the concentrations of soluble Ca reported are a cause of a stress response within the plant, the change
in soluble and insoluble Ca are important analyses that responded to stress conditions.
In June, the leaflet FeII concentration declined whereas that of FeIII increased. This lower FeII has been
observed in calcifuges and is considered to be due to immobilisation of Fe into the FeIII form within tissues and
is thus an important response between the calcifuge and calcicole behaviours.
Table 9. The concentration of Ca form (total, insoluble, soluble) and Fe form (total, FeIII, FeII) in leaflet tissue
of the L. albus cultivar Lucyanne harvested in June (280 DAS) in neutral or alkaline-pH soil
Nutrient
Tissue
Soil pH
Tissue Soil pH TxS
(s.e.)
(s.e.)
(s.e.)
(tissue dry weight
basis)
Neutral
Alkaline
Total Ca (mg g-1)
2.34
12.74
2.02
11.37
0.32
1.37
93
358
68
161
25
197
3.00
14.53
2.66
12.69
0.34
1.85
125
443
102
321
23
122
Stem
Leaflet
-1
Insoluble Ca (mg g ) Stem
Leaf
-1
Soluble Ca (mg g ) Stem
Leaflet
-1
Total Fe (μg g )
Stem
Leaflet
-1
FeIII (μg g )
Stem
Leaflet
FeII (μg g-1)
Stem
Leaflet
CSG 15 (1/00)
30
<0.001
(0.280)
<0.001
(0.288)
<0.001
(0.021)
<0.001
(17.7)
<0.001
(18.6)
<0.001
(6.5)
<0.001
(0.229)
<0.001
(0.235)
<0.001
(0.017)
<0.001
(14.5)
<0.001
(15.2)
<0.001
(5.3)
<0.05
(0.396)
ns
 0.001
(0.030)
<0.05
(25.1)
 0.001
(26.4)
 0.001
(9.2)
Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
11.3 New genotype identification: defining the tolerance of ‘Giza1’ and establishing its
underpinning physiology.
L. albus has long been cultivated in soils with high Ca contents along the Nile Valley, Egypt. It is an important
secondary crop for the winter season and is grown on approximately 400 ha. There is wide variation in germplasm:
that originally collected for the Australian national collection was shown not to possess marked tolerance to
calcareous soils. This material was not collected at site-specific locations, and may have been landraces adapted to
less calcareous soil. However, a collection of a range of landraces was made in 1995-96 by Danish / Egyptian
collaboration. Importantly, this material was site specific to highly calcareous fields with in soils of pH 7.5-9.4
(Figure 14). It was suggested that this material might possess better tolerance to calcareous soils than current
European cultivars (Christiansen et al 1999).
A series of experiments compared a range of Egyptian genotypes with European cultivars and genotypes on
neutral pH and limed soils. They were compared using the physiological
parameters of photosynthesis, and Fe and Ca form and partitioning established
from previous work. The results were used to determine some physiological
causes of tolerance (Kerley et al, 2002).
13.3.1 Physiological responses of the Egyptian landraces to a limed soil
The responses of the Egyptian genotypes to the limed soil differed from either of
the intolerant European cultivars Lucyanne or Lublanc. Although leaf greenness
is not always consistent within an experiment over time, the absence of chlorosis
in any of the Egyptian material was comparable with the tolerant L. pilosus, and
indicated that the plants tolerated the stress better than the European material.
The leaf light transmission meter (SPAD) analysis discriminated between L.
pilosus and the Egyptian genotypes and indicated that the Egyptian genotypes
were subjected to a sub-chlorotic level of stress that was greater than L. pilosus.
Leaf emergence has been considered a potential means of genotype evaluation,
and did effectively discriminate between the Egyptian and intolerant plants.
However, this evaluation gives no insight into the tolerance physiology, and was
Figure 14. Giza1,
not sufficiently discriminatory in a subsequent UK field trial (see later).
harvested in Egypt on
The above screens measured the results of physiological processes; more
calcareous soil.
definitive screens were needed to measure the processes themselves, e.g. Ca and
Fe partitioning and photosynthesis. Differences in the leaf soluble Ca in the first experiment confirmed lower
concentrations in landraces Egypt 99 and 121 compared with Lucyanne and Lublanc, indicating that some
control over soluble Ca was present. Unfortunately this was not apparent in Giza1, indicating that such a
process may be minimal in this landrace. However, Giza1 did generally have lower mean values for soluble Ca
than Lublanc, with La 675 intermediary; as such it may possess some form of tolerance that requires further
study in a truly calcareous soil.
A potential tolerance mechanism was apparent in the Fe response of Giza1 compared with the intolerant La 675
(Table 10). Giza1 showed a smaller partitioning of its Fe as stem Fe3+ in limed compared with neutral-pH soil
and even increased the fraction of active Fe2+ in the leaf tissue, accounting for its lack of chlorosis. This
mechanism could facilitate calcicole growth in calcareous soil and might be one explanation why the Egyptian
material is cropped in highly calcareous soils. This response also explains the maintenance of Pmax values and
the absence of a decrease in quantum yield under limed soil conditions, which were in contrast to the nonEgyptian L. albus.
Tolerance does not, however, imply optimal growth conditions. The destructive analysis of the shoot dry weight
demonstrated that both L. pilosus and the Egyptian genotypes did experience considerable limed-induced stress
and showed that all genotypes grew optimally on the neutral-pH soil.
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Improving the physiological and agronomic basis of UK lupin
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DEFRA
project code
ARO138
Table 10. The partitioning of Fe into leaf and stem tissues and form (Fe2+/3+) of the L. albus genotypes Giza1
and La 675. S.E. presented as a measure of replicate variation when genotype, soil, or interaction effects were
significant (P≤0.05).
Analysis
Soil
Neutral
Limed
Genotype
Giza1
La 675
420
378
284
362
Genotype
Soil
GxS
Significance (S.E.)
ns
<0.05
<0.05
(21.6)
(30.5)
Whole shoot Fe
(µg/g)
Stem total Fe
(µg/g)
Neutral
Limed
68
68
67
138
<0.05
(7.8)
<0.05
(7.8)
<0.05
(10.9)
Leaf total Fe
(µg/g)
Neutral
Limed
353
216
309
224
ns
<0.001
(30.1)
ns
Stem FeIII
(µg/g)
Neutral
Limed
67
67
68
137
<0.05
(7.7)
<0.05
(7.7)
<0.05
(10.9)
Stem FeIII
(% of total Fe)
Neutral
Limed
17
24
19
39
<0.05
(1.9)
<0.001
(1.9)
<0.005
(2.7)
Leaf FeII
(µg/g)
Neutral
Limed
159
139
159
127
ns
ns
ns
Leaf FeII
(% total Fe)
Neutral
Limed
38
49
41
36
<0.05
(1.8)
ns
<0.05
(2.5)
To compare the root growth of Giza1 with a range of European plant types, they were grown in the liquid
culture system and their root architectures compared. The root systems showed genotypic differences in
response to stress. Some of these could be attributed to differences in vigour or seed size; for example shoot and
root dry weight analyses. However, some measurements did show genotype x environment interactions e.g. tap
root tortuousness and the indeterminate lateral root analyses. Compared with Lublanc, Giza1 had a lower tap
root tortuousness index at pH 7.5 and in 3 mM Ca. Crucially, it also suffered a lower proportion of tap root
death in all three stresses. This response, and its longer tap root system, possibly accounted for its higher
tortuousness index when grown in HCO3- compared with Lublanc. Giza1 also had a higher lateral root number
ratio in the HCO3- and Ca treatments which was due to a higher number and greater density of indeterminate
roots. These responses indicate that Giza1 is better adapted to root growth in these stress conditions than
Lublanc, and provides one potential explanation for the improved iron physiology identified above.
Figure 15 summarises the differences found between the current intolerant cultivars such as Lublanc, and the
more tolerant Egyptian landraces such as Giza1. However, these landraces are unsuitable for the UK (e.g. see
Figure 14) and will require a breeding strategy to incorporate their tolerance mechanisms into a suitable cultivar
form for the UK.
Because there is wide variation for tolerance, and tolerance mechanisms within the Egyptian material, there is a
need to discriminate more effectively between plant types based on more than a single screening system. Once
achieved, this discrimination will facilitate breeding programs designed to develop calcareous soil tolerant
agronomic cultivars of L. albus.
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Figure 15. Summary diagram showing possible tolerance of Giza1
compared with Lublanc
11.4 New genotype evaluation: pot and field evaluation of Egyptian material and screening
strategy
It was the aim of this work to further confirm the physiological differences between limed or calcareous soil
tolerant and intolerant plants, and to determine whether such analyses could be used to evaluate plants in field
trials and ultimately more rapidly in pot trials. Three experiments were conducted; the first using soil chambers
and the others in limed (trial 1) or naturally calcareous (trial 2) fields (Kerley et al in preparation).
11.4.1 Shoot and root dry weight comparisons
Only in the soil chamber experiment did shoot dry weight effectively discriminate the tolerance of Giza1 from
an intolerant L. albus cultivar. In the limed-soil field trial the plants may not have been under sufficient stress to
show a reduction in shoot dry weight, and in the calcareous-soil trial there was no non-stressed control to
determine the potential growth of each plant type. Shoot dry weight may not be a reliable means of evaluation,
as although tolerant, in the chamber trial the difference in dry weight of L. pilosus between neutral-pH and
limed soil was comparable with Lublanc. Additionally, shoot dry weight is also of little value, as it requires the
destruction of the plant.
Analyses of the root system could only be conducted in the soil chamber experiment. No clear response to the
limed soil was seen with Lublanc, whereas the more tolerant plants showed a proliferation of cluster roots in the
limed soil. In L. pilosus the cluster roots proliferated, possibly at the expense of the shoot growth and lateral
roots. In contrast, cluster root proliferation in Giza1 was not concomitant with a reduction in the shoot and
lateral root dry weight. These responses corroborate earlier work in which cluster root proliferation of the L.
albus cultivar Lucyanne occurred, but with a reduction in shoot and non-cluster root dry weight.
11.5 Fe forms as evaluation screens
In the soil chamber experiment, differences in the partitioning of leaf Fe were apparent between the three plant
types (Giza1, Lublanc and L. pilosus) when grown in the limed soil compared with neutral-pH soil. Lublanc
maintained its leaf total Fe concentration at levels comparable with those in non-limed soil through an increase
of FeIII, whereas there was 25% less FeII. Although in L. pilosus the concentrations of both forms of Fe in
limed soil grown plants were less compared with those in neutral-pH soil, the change in FeII was small. In
contrast, although Giza1 had a small loss of total Fe concentration, this was due to FeIII, whereas the
CSG 15 (1/00)
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
concentration of the FeII was maintained. This result agrees with recent literature which concludes that
immobilization of ‘physiologically less active’ Fe (FeIII) by calcifuges is related to chlorosis (Zohlen and
Tyler 2000).
Figure 16. Fe form (II v III) expressed plotted for each plant type. L. albus cultivars Lucyanne (◊), Lublanc (□),
Primavera 106 (o) and Bardo (∆), genotypes Giza1 (■), La668 (▲), La673 (●) and La675 (♦) and landrace
Egy121 (+) and of L. pilosus genotype LD124 (x).
The plant Fe responses in this study indicate that it is the maintenance of high concentrations of FeII in the leaf
tissue that is of major importance in the tolerance to a limed or calcareous soil. The concentration of the FeIII is
less important. However it does provide a secondary measure of the Fe physiology, allowing the whole Fe
response to be evaluated. The change in the concentration of FeIII between stressed and non-stressed plants
may be more important than absolute concentrations. Analysis of Fe form is a potentially valuable screen as it
is rapid, non-destructive (to the whole plant), requires a minimal quantity of leaf tissue and can be repeated on a
single plant basis during the growing season.
Differences in the partitioning of Fe were apparent in the field trials that support the results from the soil
chambers. The plants of Egyptian origin contained the highest concentrations of FeII, whereas the lowest
concentrations were present in Lucyanne and Lublanc. In trial 1, the FeIII of the tolerant plants was less
compared with plants in the non-limed soil, whereas concentrations were maintained in the intolerant plants.
This demonstrates the need to compare FeIII in both stressed and non-stressed plants, and indicates that
important differences were present between tolerant and intolerant plants. Such analyses could not be done in
trial 2, however most FeIII was found in the intolerant plants and least in Giza1. This indicates that in the
intolerant plants, more FeII was converted to FeIII. Fe form appears important in the response of lupins to
calcareous soil. The processes that results in their partitioning in the plant is not understood, although literature
indicated that the role of bicarbonate ion may be crucial.
Expressing the field trial Fe-form data as a ratio or graphically (Figure 16) showed that Giza1 was at one
extreme of the range and Lucyanne and Lublanc were at the other. This created ‘benchmarks of tolerance’ used
to compare the other plants. This analysis indicated that Bardo responded similarly to Lucyanne and Lublanc,
whereas Primavera 106 represented an improvement over these plants, although it was not as tolerant as Giza1.
Some caution must be taken in interpreting physiological effects of the Fe concentrations, as we do not know
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Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
their critical values or how they vary between the plants. In this absence of knowledge and the comparisons
between stressed and non-stressed plants are central in defining the processes occurring.
11.6 Ca forms as evaluation screens
The partitioning of leaf forms of Ca, as a potential screen for tolerance was not as clear as it was for Fe.
Interestingly, the highest soluble and insoluble Ca concentrations were associated with the tolerant species L.
pilosus in both the soil chamber experiment and the field trials. The tolerance mechanisms are unknown, but
may involve both Ca fractions as their concentrations were maintained irrespective of soil liming. The
maintenance of the insoluble fractions indicated that some control of Ca uptake occurred, whilst the
maintenance of the soluble fractions demonstrated a regulation of the active Ca pool.
In contrast to L. pilosus, the concentrations of both the soluble and insoluble Ca fractions increased in Lublanc,
signifying little control in the uptake of Ca, and that the plant behaved in a calcifuge manner. The Ca
concentrations in Giza1 were more comparable with Lublanc than L. pilosus. However, Giza1 differed from
Lublanc in the control of the soluble Ca; in Lublanc it increased by 35% in the limed compared with neutral-pH
soil of the soil chamber experiment, whereas that of Giza1 increased by only 6% indicating a level of control
not present in Lublanc. It is probable that whilst Lublanc showed a general increase in leaf Ca, the increase in
Figure 17. Ca form (soluble v insoluble) expressed plotted for each plant
type. L. albus cultivars. Symbols as for figure 16.
insoluble Ca of Giza1 was a means to control the concentrations of the soluble fraction. However the
physiology behind the processes require further study.
As with Fe, the Ca form analysis is a repeatable, rapid and non-destructive analysis that is conservative of the
whole plant. However, we do not know the critical values for the Ca forms, or whether high concentrations of
soluble or insoluble Ca represent stress, tolerance, or sequestration.
Comparing the Ca forms from the field trials as ratios or graphically (Figure 17) did show differences between
the plants. As with Fe, Giza1 and Lucyanne were at extremes of the plant types. Interestingly, Egy121 was
comparable with Giza1, providing evidence that the Egyptian plants are tolerant of calcareous soil. Primavera
106 had not been evaluated prior to this study, it was the cultivar that was most comparable with the Egyptian
material, providing some evidence for greater tolerance.
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
11.7 Concluding remarks on the alkaline soil problem.
This objective has demonstrated the tolerance response of Giza1 and L. pilosus and the intolerance of Lublanc
to limed and calcareous soils. It has provided a physiological basis that explains both some of the causes of
intolerance in many lupins and provides an explanation for the observed tolerance in some landraces. Analysis
of soluble and insoluble Ca and of FeII and FeIII in leaf tissue effectively discriminated between tolerance and
intolerance in both field and soil chamber based screens.
By not comparing root systems, these protocols can be used in short term (30 day) pot experiments using limed
and non-limed to provide and effective, rapid non-destructive screening systems for evaluating plant material
either in collections or during breeding projects. It is also the basis up on which a project investigating the
molecular basis for tolerance should be reliably and soundly based.
Using these plant types and analyses as evaluation parameters, Egy121 and Primavera 106 were shown to
possess levels of tolerance greater than the cultivars Bardo or Lublanc. The magnitude of the difference in Ca
concentrations between L. pilosus and L. albus must be considered when using the tolerant species as a control
for L. albus. Very different processes may be occurring in L. pilosus that are absent in tolerant L. albus plant
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
12.0. Knowledge Transfer.
Knowledge transfer requires a special skill that often has to be acquired and may be poorly recognised within
research institutes. However, the development of the lupin crop has been accompanied by a large effort to
disseminate the knowledge gained. Between 1995 and 2003 a total of 50 articles were published in the trade
and popular press including two radio interviews and two television reports.
In the autumn of 2000 an extensive web site was introduced that contained all of the agronomic information
gained from the work and a recommended management plan for autumn sown lupins. During the first year after
the launch the site received an average of 9 hits per day, approximately 67% new comers and 33% returning
viewers. Only about 15% of hits came from outside of the UK. By the end of 2003 the number had fallen to 5
hits per day, it is assumed that most people who required the information had acquired it.
From 1999 commercial development of the crop began. Two seed supply companies entered the market with
varieties of autumn sown white lupin. Rothamsted Research assisted both with expert advice. Goreham and
Bateson marketed seed nationally but were assisted in a development project based in the Welsh Marches, an
ideal target area of traditional mixed farms where beef production was struggling to maintain farmer income.
The production of all animal feed on farm was seen as directly economically beneficial and ultimately seen as a
route to add value through a premium brand. ADAS, Countrywide Farmers and Meadow Valley Livestock
formed a strong consortium to push this concept forward. The project undoubtedly had successes, but also
suffered some problems largely because the silt soils of the area are not well suited to autumn sown lupins.
Cebeco Seed Innovations also marketed seed of autumn sown varieties. Rothamsted Research assisted them in a
national programme of field days based around demonstration crops. The programme began with two large
scale days at Rothamsted during 1999. In 2000 this expanded to 5 regional events across England. All were
well attended.
All new crop types are introduced with imperfect knowledge and commercial introduction is part of the
learning process. Autumn sown white lupins were introduced with one of the best possible knowledge packages
yet struggled in commercial agriculture. The principal reason was the complete reliance on the achievement of
the correct sowing date for the site. This was compounded by the fact that unusual autumn weather could render
the recommended sowing date inappropriate, as happened in 1997-98 and 2000-01. The introduction of dwarf
determinate varieties such as Lucille was intended to lessen the reliance on the sowing date; however these
varieties were blighted by late delivery of seed from France. This is perhaps understandable as lupin is a low
volume crop demanding attention at a seed processing plant at the busiest time of year. It is clear that a number
of commercial crops failed because late delivery of seed forced late autumn sowing. The problem can be
overcome by over year storage of seed, but this is difficult with new varieties where seed is scarce and demand
is high. It also has physical and financial implications for the seed trade.
An additional problem was the lack of suitable approved post emergence broad leafed weed herbicides, and the
requirement for 2 summer fungicides. Despite the summer fungicides only being applied at low rates the crop
now has an image as requiring too many inputs.
The introduction of a number of diverse spring sown lupin varieties during the period 2000-2003 which were
cheap and easy to grow and produced seed yields similar to the winter varieties finally dissuaded farmers from
growing winter varieties.
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
14.0 References.
ATKINS, C.A. and FLINN, A.M. (1978) Carbon dioxide fixation in the carbon economy of developing seeds
of Lupinus albus (L.). Plant Physiology 62, 486-490
BATEMAN, G.L., FERGUSON, A.W. & SHIELD, I. (1997). Effects of sowing date and pesticides on survival
during the winter of the florally determinate white lupin cv. Lucyane. Annals of Applied Biology 130, 349359.
BATEMAN, G.L. (1997) Pathogenicity of fungi associated with winter loss and injury in white lupin. Plant
Pathology 46, 157-167.
CHRISTIANSEN, J.L., RAZA, S, ORTIZ, R. (1999). White lupin (Lupinus albus L.) germplasm collection and
preliminary in situ diversity assessment in Egypt. Genetic Resources and Crop Evolution 46: 169-174.
COWLING, W.A., HUYGHE, C. & SWIECICKI, W. (1998). Lupin Breeding. In Lupins as Crop Plants: Biology,
Production and Utilisation (Eds J.S. Gladsones, C.A. Atkins & J. Hamblin) pp. 93-120 UK: CAB
International
DRACUP, M., READER, M.A. and PALTA, J.A. (1998) Variation in yield of narrow-leafed lupin caused by
terminal drought. Australian Journal of Agricultural Science. 49, 799-810
ETHERIDGE, J.V. & BATEMAN, G.L. (1999) Fungicidal control of foliar diseases of white lupin (Lupinus
albus). Crop Protection 18, 349-354.
FREEMAN, S. KATAN, T & SHABI E. (1998). Characterisation of Colletotrichum species responsible for
Anthracnose diseases of various fruits. Plant Disease 82, 596-601
HUYGHE, C. (1991). Winter growth of autumn-sown white lupin (Lupinus albus L.): main apex growth model.
Annals of Botany 67, 429-434.
HUYGHE, C. (1993). Growth of white lupin seedlings during the rosette stage as affected by seed size. Agronomie
13, 145-153.
HUYGHE, C. & PAPINEAU, J. (1990). Winter development of autumn sown white lupin: Agronomic and
breeding consequences. Agronomie 10, 709-716.
JULIER, B., HUYGHE, C., PAPINEAU, J., MILFORD, G.F.J., DAY, J.M., BILLOT, C. & MANGIN, P. (1993).
Seed yield and yield stability of determinate and indeterminate autumn-sown white lupins (Lupinus albus
L.) grown at different locations in France and the UK. Journal of Agricultural Science, Cambridge. 121,
177-186.
JULIER, B. & HUYGHE, C. (1993). Description and model of the architecture of four genotypes of
determinate autumn-sown white lupin (Lupinus albus L.) as influenced by location, sowing date and
density. Annals of Botany 72, 493-501.
KERLEY S.J., (2000a). Changes in root morphology of white lupin (Lupinus albus L.) and its adaptation to soils
with heterogeneous alkaline/acid profiles. Plant & Soil. 218: 197-205.
KERLEY S.J., (2000b). The effect of soil liming on shoot growth and development and on root growth and cluster
root activity of white lupin (Lupinus albus L.) Biology & Fertility of Soils. 32: 94-101
KERLEY S.J., LEACH J.E., SWAIN J.L. & HUYGHE C. (2000). Investigations into the exploitation of
heterogeneous soils by Lupinus albus L. and L. pilosus Murr. and the effect upon plant growth. Plant &
Soil. 222: 241-253.
KERLEY S.J., SHIELD I.F. & HUYGHE C. (2001). Specific and genotypic variation in the growth and nutrient
content of white lupin (Lupinus albus L.) in soils of neutral and alkaline pH. The Australian Journal of
Agricultural Research. 52 (1): 93-102.
KERLEY S.J. & HUYGHE C. (2001). Comparison shoot and root growth of white lupin (Lupinus albus L.)
genotypes in neutral and alkaline-pH soil-based and liquid culture growth conditions. Plant & Soil 236 (2):
275-286.
KERLEY S.J., NORGAARD C., LEACH J.E., CHRISTIANSEN J.L., HUYGHE C. & RÖMER P. (2002). The
tolerance to limed soils of Egyptian genotypes of white lupin (Lupinus albus L.) and the development of
calcium and iron based tolerance screens. Annals of Botany 89: 341-349
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KERLEY S.J. & SHIELD I.F. (2001). The response to soil heterogeneity through cluster root proliferation.
Structure and Functioning of cluster roots and plant responses to phosphate deficiency. Workshop at
University of Western Australia, Perth, WA, Australia.
KNOTT, C.M. (1996) Test of Agrochemicals and Cultivars No. 17 (Annals of Applied Biology 128,
Supplement) pp. 52-53.
LAGUNES-ESPINOZA, L.C., HUYGHE, C., PAPINEAU, J. and PACAULT D. (1999) Effect of genotype
and environment on pod wall proportion in white lupin: consequences to seed yield. Australian Journal
of Agricultural Research, 50, 575-582.
LAGUNES-ESPINOZA, L.C., HUYGHE, C., PAPINEAU, J. and SHIELD, I.F. (2000) Dry matter and
nitrogen accumulation during pod wall development of white lupin genotypes differing in proportion of
pod walls. Journal of Agricultural Science 135, 389-397.
LEACH, J.E., STEVENSON, H.J., SCOTT, T. & MILFORD, G.F.J. (1997). The effect of soil freezing on the
survival of winter-sown white lupins (Lupinus albus L.). Annals of Applied Biology 130, 561-567.
LEACH, J.E. (2001) The role of pods and leaves for photosynthetic gas exchange in determinate (restricted
branching) white lupins (Lupinus albus L.). In; E. van Santen, M. Wink, S. Weissmann and P. Roemer
(eds). Lupin, and ancient crop for the New Millenium. Proceedings of the 9th International Lupin
Conference, Klink/Muritz, 20-24 June, 1999. International Lupin Association, Canterbury, New
Zealand. 346 – 349.
MILFORD, G.F.J., DAY, J.M., LEACH, J.E., STEVENSON, H.J., HUYGHE, C. & PAPINEAU, J. (1993a). The
effect of modifying plant structure on the yield and maturity of the white lupin Lupinus albus. Annals of
Applied Biology 122, 113-122.
MILFORD, G.F.J., DAY, J.M., HUYGHE, C. & JULIER, B. (1993b). Floral determinacy in autumn-sown white
lupin (Lupinus albus) : the development of varieties for cooler European climates. Aspects of Applied
Biology 34, 89-97.
MILFORD, G.F.J., SHIELD, I.F., SIDDONS, P.A., JONES, R.J.A. & HUYGHE, C. (1996). Simple physiological
models of plant development for the white lupin (Lupinus albus) and their use in agricultural practice.
Aspects of Applied Biology 46, 1996 Modelling in applied biology: Spatial aspects. 119 -123.
PATE, J.S., EMERY, R.J.N. & ATKINS, C.A. (1998). Transport Physiology and Partitioning. In Lupins as Crop
Plants: Biology, Production and Utilisation (Eds J.S. Gladsones, C.A. Atkins & J. Hamblin) pp. 93-120
UK: CAB International.
SHIELD, I.F. & MILFORD G.F.J. (1995) The performance of autumn-sown determinate genotypes of the white
lupin (Lupinus albus) in different regions of the U.K. Proceedings of the 2nd European Grain Legume
Conference, Copenhagen, 1995. p. 144.
SHIELD, I., STEVENSON, H.J., LEACH, J.E., SCOTT, T., DAY, J.M. & MILFORD, G.F.J. (1996). Effects of
sowing date and planting density on the structure and yield of autumn-sown, florally determinate white
lupins (Lupinus albus) in the United Kingdom. Journal of Agricultural Science, Cambridge 127, 183-191.
SHIELD, I.F., STEVENSON, H.J., SCOTT, T., LEACH, J.E., & CRISPIN M. (2000). Testing of herbicides for
crop safety on autumn sown white lupin. Tests of Agro-chemicals and Cultivars No. 21 pp11-12.
SHIELD. I.F., SCOTT. T., STEVENSON. H.J., LEACH J.E. & TODD. A.D. (2000). The causes of over-winter
plant losses of autumn-sown white lupins (Lupinus albus) in different regions of the UK over three
seasons. Journal of Agricultural Science, Cambridge 135, 173-183.
SHIELD, I., STEVENSON, H.J., SCOTT, T., & LEACH, J.E. (2001). The causes of, and potential solutions to,
seed yield instability in autumn-sown, determinate (restricted branching), white lupins (Lupinus albus L.)
In; E. van Santen, M. Wink, S. Weissmann and P. Römer (eds). Lupin, and ancient crop for the New
Millennium. Proceedings of the 9th International Lupin Conference, Klink/Muritz, 20-24 June, 1999.
International Lupin Association, Canterbury, New Zealand. pp 339 - 345.
SHIELD. I.F., SCOTT. T., HUYGHE, C., BRUNEAU, M., PARRISSEAU, B., PAPINEAU, J, HARZIC, N,
STEVENSON. H.J., & LEACH J.E. (2002). The effects of seed rate and row spacing on light
interception, dry matter accumulation and seed yield in non-dwarf and dwarf genotypes of autumn-sown
determinate white lupins (Lupinus albus L.) in north west Europe. Journal of Agricultural Science,
Cambridge 138, 39-55.
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Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
SHIELD. I., SCOTT. T., STEVENSON. H.J., HUYGHE, C., PARRISSEAU, B., PAPINEAU, J., & HARZIC, N.
(2003). The effect of the environment on leaf and branch number in non-dwarf and dwarf genotypes of
autumn-sown determinate white lupins (Lupinus albus L.) in north-west Europe. Journal of Agricultural
Science (in review).
SREENIVASAPRASAD, S., SHARADA, K., BROWN, E.A., and MILLS, P.R. (1996). PCR-based detection
of Colletotrichum acutatum on strawberry. Plant Pathology 45, 650-5.
WARD E. & AKROFI A.Y. (1994). Identification of fungi in the Gaeumannomyces-Phialophora complex by
RFLPs of PCR-amplified ribosomal DNAs. Mycological Research 98, 219-224.
WHITE, P.F., & ROBSON A.D. (1989). Poor soil aeration or excess soil CaCO3 induces iron deficiency in
Lupinus angustifolius L. Australian Journal of Agricultural Research, 40: 75-84.
ZOHLEN A. & TYLER G. (2000). Immobilization of tissue iron on calacareous soil: differences between
calcicole and calcifuge plants. Oikos 89: 95-106.
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title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Annexe 1
Projects at Rothamsted Research,
DEFRA AR0138 Improving the physiological and agronomic basis of UK lupin production. April 1999 to
March 2003.
DEFRA LS3606 Improve the traceability of protein sources for the livestock sector through research into the
nutritional quality of lupins. June 2000 to October 2001.
DEFRA AR0118 Improved genotypes and agronomy of lupins for UK agriculture. April 1996 to March 1999.
DEFRA AR0117 Agronomic packages for the cultivation of lupins in the UK. April 1992 to March 1996
DEFRA AR0116 Physiological constraints to the yield and early maturity of lupins in the UK. April 1992 to
March 1996
Commission of the European Communities, Agriculture and Fisheries (FAIR) specific RTD programme, CT961965, “Creation of varieties and technologies for increasing production and utilisation of high quality protein from
white lupin in Europe”. January 1997 to March 2000.
BBSRC Adapting new crops for the UK climate: genotype and environment … to February 1999.
BBSRC Internal nitrogen resources and the regulation of pod growth, crop ripening … to February 1999.
BBSRC CWS (Collaboration with Industry Scheme) and Dalgety. Defining the geographic range of autumn
sown white lupins. January 1994 to December 1996.
Affiliated projects,
DEFRA AR0132 Lupin Agronomy. September 1994 to March 1996. ADAS.
DEFRA AR0127 Land suitability for autumn-sown determinate lupins. Soil Survey and Land Resource Centre
(Cranfield University) and Rothamsted Research.
DEFRA and EU Objective 5b. MAP (Marches Alternative Protein). ADAS.
PGRO levy funded trials, intermittent from 1994 to 2003.
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Project
title
Improving the physiological and agronomic basis of UK lupin
production.
DEFRA
project code
ARO138
Annexe 2. Towards a whole crop model for autumn sown white lupin.
The following are the empirically derived equations (models) describing the effects of the environment on plant
and crop architecture and behaviour. They have been used to predict crop behaviour in the field and to
formulate crop production guidelines or agronomy.
Over winter survival (% of seeds sown)
= 0.0004008 x12 - 0.425 x1 + 137.1
Calculated at the time that the soil temperature at 10cm first falls below 2°C
No. leaves on the main stem (x3a)
= 0.03977x1 + 0.918x2 -7.23
Vernalisation requirement satisfied (AccDDA 1-14°C)
= 1339.4 * 0.9741 x3a
No. first order branches per plant
= 0.125x3b + 1.42
Time of first floret open on main stem (AccDDA 3°C)
= 20.55x3 – 46.97x2b + 1084.9
where x1 = accumulated thermal time from sowing (AccDDA 3°C), x2 = mean day length from sowing (h) and x3 =
number of leaves on the main stem (a pre-vernalisation, b post vernalisation).
No. yield bearing axes m-2 (a)
= ((seed rate (seeds m-2) * Over winter survival ) + No. first order
branches per plant) * (seed rate (seeds m-2) * Over winter survival)
Yield potential (y t ha-1 @ 15% m.c.)
If PAR approx. >20%
y=
4.16
------------------------------------------(1 + EXP (0.05458 * (a - 53.34)))
If PAR approx. <20%
y=
2.93
------------------------------------------(1 + EXP (0.032 * (a - 63.14)))
where a = No. yield bearing axes m-2, and PAR = the percentage of incident PAR penetrating the canopy to the
base of the main stem inflorescence.
Time to start of senescence (AccDDA 3°C)
= Time of first floret open on main stem + 800 (for cv.
Lucyanne)
Maturity date (days from start of senescence)
= 121.3 - 0.0635 p – 3.992 t (for cv. Lucyanne)
where p = mean Potential Soil Moisture Deficit (PSMD) and t = mean maximum air temperature in the 14 days
following the start of senescence.
Those in italics are based on an initial examination of the available data and would repay further investigation.
We do not have a robust method for modelling the proportion of incident PAR that penetrates the canopy to the
base of the main stem inflorescence.
CSG 15 (1/00)
42