Field Crops Research 270 (2021) 108211

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Field Crops Research

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Variation in traits contributing to improved use of nitrogen in wheat:

Implications for genotype by environment interaction
A.M. Manschadi a, *, A. Soltani b
a
Dep. of Crop Sciences, Institute of Agronomy, BOKU University, Vienna, Austria
b
Agronomy Group, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

A R T I C L E I N F O A B S T R A C T

Keywords: Nitrogen (N) is an essential mineral nutrient for wheat grain yield and quality. Wheat cultivars with improved N-
Wheat use efficiency (NUE, crop yield per unit availability of N from soil and fertiliser) have the potential to maximise
Nitrogen-Use efficiency farmers’ revenue and minimise environmental pollution risk. Thus, characterizing wheat cultivars for NUE traits
Trait characterisation
is important for selection of cultivars better adapted to target production environments and for season-specific N
Nitrogen harvest index
Nitrogen remobilisation efficiency
fertiliser management. The aims of this study were to examine NUE traits in a set of modern winter wheat
Environment adaptation cultivars contrasting in phenology and grain-N content, and to assess the likely consequences of genotypic
variation in N-related traits for adaptation of cultivars to specific environments. A two-year field experiment was
conducted with four widely-grown commercial winter wheat cultivars (Arnold, Aurelius, Bernstein, Emilio)
under four N application rates (0, 70, 140, 210 kg N ha− 1) at the UFT (Universitäts- und Forschungszentrum
Tulln, BOKU) in Austria. Detailed plant and soil data were collected to determine Green leaf area index (GLAI),
total shoot dry mass (TDM) and N uptake (TNU) at anthesis and harvest, grain yield (GY), N harvest index (NHI),
N remobilisation efficiency (NRE), and NUE.
The experimental factors and variation in seasonal weather conditions resulted in a wide range of GY (268
− 788 g m− 2) and TNU (6.6-22.7 g m− 2). With average values of 80 % for NHI and NRE, all cultivars were very
efficient in the use of the accumulated N for producing GY. The observed significant differences in GY were
largely due to differences in pre-anthesis N allocation to leaf and stem and post-anthesis DM production and N
uptake, as cultivars had similar amounts of TDM and TNU at anthesis. The highest-yielding cultivar Aurelius
showed consistently greater N uptake during grain filling. Cultivars also differed significantly in the partitioning
of DM and N between leaf and stem in the pre-anthesis phase, with Bernstein showing the highest GLAI and leaf-
N content in both experiments. The inter-seasonal variation in environmental conditions strongly influenced the
performance and ranking of cultivars with regard to NUE traits and GY. The implications of genotypic variation
in traits contributing to improved N use and their importance in better matching cultivar selection to target
environments and N fertiliser management are discussed.

1. Introduction
Wheat (bread wheat, Triticum aestivum and durum wheat, Triticum
turgidum) is the most widely cultivated cereal in the world with average
(i.e. for the five-year period 2014–2018) area and production of 219.0
million ha and 745.3 million tonnes, respectively (FAO, 2020). In the
same time period, the European Union produced 150.7 million tonnes of
wheat, accounting for 20.5 % of the world wheat production. With a
contribution of about 20 % to daily calories and protein, wheat is a
mainstay of today’s human diet (Shiferaw et al., 2013). Consequently,

Abbreviations: GY, grain yield; TDM, total above-ground dry mass; HI, harvest index; GNO, grain number; TNUP, total above-ground N uptake; GNU, grain N
uptake; NHI, N harvest index; %GN, grain N concentration; DM, dry mass; LFDMa, leaf DM at anthesis; ‘stem’, includes true stem leaf sheath, and spike without grain;
STDMa, ‘stem’ DM at anthesis; GLAI, green leaf area index; LFDMRE, leaf DM remobilisation efficiency; STDMRE, ‘stem’ DM remobilisation efficiency; LFNa, leaf N
content at anthesis; STNa, ‘stem’ N content at anthesis; LFNRE, leaf N remobilisation efficiency; STNRE, ‘stem’ N remobilisation efficiency; NUE, N-use efficiency;
NUpE, N-uptake efficiency; NUtE, N-utilisation efficiency.
* Corresponding author at: Dept. of Crop Sciences, Institute of Agronomy, University of Natural Resources and Life Sciences, Vienna, Konrad Lorenz Str. 24, A-
3430, Tulln, Austria.
E-mail address: manschadi@boku.ac.at (A.M. Manschadi).

https://doi.org/10.1016/j.fcr.2021.108211
Received 19 October 2020; Received in revised form 18 June 2021; Accepted 19 June 2021
Available online 25 June 2021
0378-4290/© 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
A.M. Manschadi and A. Soltani
the future productivity of wheat will have more influence on global food
security than that of any other crop species. In the course of agricultural
development, significant progress in crop yields has most often been the
result of a combination of increased availability of nitrogen and water
resources and improved genetics to take advantage of greater resources
(Sinclair and Rufty, 2012). Thus, the productivity and sustainability of
wheat production systems in the future will largely depend on the
availability and effective use of water and nitrogen.
Nitrogen (N) is an essential mineral nutrient for growth and yield
formation of crop plants that is required in greatest abundance. Conse­
quently, crop productivity and quality depend on large inputs of N into
cropping systems. Worldwide about 102 million tonnes of inorganic N
fertilisers are used in the agriculture sector. Of this, more than 50 % is
applied to three major cereals, with wheat receiving 18.2 % of global
use, followed by maize with 17.8 % and rice with 15.2 % (Heffer et al.,
2017). It is estimated that without the input of inorganic N-fertilisers
only about half of the current global population could be supplied with
sufficient food, energy and protein (Dawson and Hilton, 2011).
Increasing crop yields in many parts of the world would also require a
substantial increase in N input to cropping systems (Schils et al., 2018).
The efficiency of N use in cropping systems is, however, only about 47 %
of applied N (Lassaletta et al., 2014; Ladha et al., 2016; Swaney et al.,
2018), which means that a significant amount of N is lost from agri­
cultural fields. This unused N can cause harmful environmental impacts
through leaching of nitrate-N to groundwater and emission of nitrous
oxide to the atmosphere (Fowler et al., 2013). Therefore, it is widely
recognised that improving N-use efficiency (NUE) is crucial to reduce
the environmental impacts of crop production systems and improve
their economic and environmental performance (Raun and Johnson,
1999; Fageria and Baligar, 2005; Hirel et al., 2007).
Nutrient-use efficiency (NUE) can be defined as the crop yield per
unit availability of N from soil and fertiliser and is calculated as the
product of two subcomponents: (i) nutrient-uptake efficiency (NUpE; i.e.
total nutrient in the above-ground plant organs at maturity per unit of
nutrient supply), and (ii) nutrient-utilisation efficiency (NUtE; i.e. crop
seed yield per unit of nutrient uptake at maturity) (Moll et al., 1982;
Manschadi et al., 2014; Hawkesford, 2017). For a more quantitative
analysis of the traits affecting N use and yield of crop plants, grain yield
(GY) can be described as a function of total above-ground N uptake
(TNU), N harvest index (NHI, the proportion of TNU at harvest in the
grain), and grain N concentration (%GN) (Foulkes et al., 2009; Sinclair
and Rufty, 2012):
NHI
GY = TNU × (1)
%GN
Eq. (1) accommodates the NUtE component of NUE, because NHI can
be written as (Barraclough et al., 2010):
GNU GY × %GN
NHI = = = NUtE × %GN (2)
TNU TNU
where GNU is grain N uptake. Eqs. (1) and (2) indicate that crop yield is
directly related to total plant N uptake (TNU) and the allocation of
accumulated N to the grain (NHI). In other words, genotypic selection
for increasing GY and NUE could be focused on increasing TNU and NHI
and/or decreasing %GN. The variables determining GY as a function of
N-use in Eq. (1) – TNU, NHI, and %GN – are influenced by a number of
physiological traits governing pre-anthesis uptake and accumulation of
N in vegetative organs and N uptake and remobilisation during the
grain-filling period.
The major traits that are likely to make a significant contribution to
TNU in wheat are (i) enhancing the N storage capacity of vegetative
tissues (leaf, stem) through metabolising N into molecular forms that do
not interact with the feedback mechanisms governing the N uptake
process (e.g. ribulose bisphosphate carboxylase, Rubisco), (ii) improving
the N uptake capacity and extending the N acquisition period into the
post-anthesis phase by maintaining root growth and metabolic activity
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Field Crops Research 270 (2021) 108211
for N uptake, and (iii) delaying leaf senescence (Foulkes et al., 2009;
Barraclough et al., 2010; Gaju et al., 2011; Ciampitti and Vyn, 2012;
Reynolds et al., 2012; Sinclair and Rufty, 2012; Cormier et al., 2016;
Tian et al., 2016; Ratjen et al., 2018).
Nitrogen accumulation in vegetative tissues is limited by the vege­
tative mass of the crop available for N storage (Sinclair and Rufty, 2012).
Generally, the longer the life cycle of a crop the higher its DM produc­
tion. As wheat phenological development, and in particular the time of
flowering, is the major genetic determinant of the relative duration of
pre- and post-anthesis phases, genotypic variation in phenology can
potentially affect the amount of DM and N accumulation, with earlier
maturing cultivars tending to produce and accumulate less DM and N
(Passioura, 2006). However, DM production and N uptake are, to a large
extent, regulated by the amount of plant available water in the soil, so
that during water-deficit episodes crops can become simultaneously
water and N limited (Plett et al., 2020). Therefore, physiological pro­
cesses contributing to increased accumulation of pre-anthesis DM and N
may result in substantial risk that plants run out of water and N before
the completion of grain filling in environments with frequent
post-anthesis water deficit (Manschadi et al., 2008).
Root system characteristics are also important for increasing N up­
take. The root system of modern wheat varieties grown at high popu­
lation densities is very effective in N uptake (Aziz et al., 2017) and,
hence, does not seem to be a limiting factor for the acquisition of soil
resources from the topsoil. In fact, the critical root-length density (RLD,
root length per unit soil volume) for water and N uptake is about 1 cm
cm− 3, which is lower than actual RLDs in the topsoil of modern
commercially-grown wheat cultivars (Foulkes et al., 2009). However,
due to exponential decline of root growth with increasing soil depth,
root distribution in the subsoil (below 60 cm) are commonly lower than
the critical RLD (Hodgkinson et al., 2017). Therefore, wheat genotype
with deeper roots and greater root growth in the subsoil are capable of
exploring a larger soil volume and, hence, extracting more water and, if
available, N residuals leached during previous cropping cycles (Man­
schadi et al., 2006; Lynch and Wojciechowski, 2015; Rasmussen et al.,
2015).
Accessing more soil water post-anthesis allows retention of green leaf
area and maintenance of photosynthesis for a longer duration after
anthesis. This trait, referred to as ‘stay-green’ phenotype, has been
associated with improved grain yield of a number of cereal crop species
including wheat (Borrell et al., 2014; Christopher et al., 2016, 2018). If
N is also available in the root zone, the stay-green genotypes appear to
acquire more N during the reproductive phase. Sustained N uptake de­
lays leaf senescence and extends the period of photosynthetic activity of
leaf canopy and, thereby, providing roots with photosynthates required
as a source of energy for N uptake (Borrell et al., 2001; Sinclair and
Rufty, 2012). In addition to post-anthesis soil N uptake, remobilisation
of pre-anthesis stored N in vegetative organs represents a major source
of N supply to grains, though N remobilisation and post-anthesis N up­
take may be negatively correlated (Bogard et al., 2010). In modern
wheat varieties, 70–85 % of grain N originates from the remobilisation
of N stored in root, leaf, and stem tissues before anthesis, with little
genotypic variability in this trait (Kichey et al., 2007; Barraclough et al.,
2010; Bogard et al., 2010; Barraclough et al., 2014). Previous studies
demonstrated that NHI of modern wheat varieties is already around 80
% (Barraclough et al., 2010; Pask et al., 2012; Hitz et al., 2017) and the
scope to further improving this trait appears rather limited (Sinclair and
Rufty, 2012).
According to Eq. (1), one option to increase GY is to reduce %GN.
Breeding for reduced %GN in wheat is, however, constrained by com­
mercial demands for high grain protein content. In fact, the results of
recent retrospective analyses of the effects of breeding efforts during the
past few decades on nitrogen economy of wheat, barley (Hordeum vul­
gare L.), and maize (Zea mays L.) clearly indicate that improvements in
N-use of modern crop varieties have primarily been the result of a
reduction in %GN and to a minor degree with gains in NHI (Calderini
A.M. Manschadi and A. Soltani
et al., 1995; Barraclough et al., 2010; Gaju et al., 2011; Bingham et al.,
2012; Ciampitti and Vyn, 2012; Sadras and Lawson, 2013). Similarly, a
global analysis of published data on NUtE trends in wheat revealed that
NUtE and %GN are negatively correlated and, therefore, increasing GY
without penalizing grain protein is challenging (de Oliveira Silva et al.,
2020)
Although extensive research has been carried out on the NUE related
traits outlined above, little is known about the characteristics of
commercially-grown wheat cultivars with regard to N-related traits in
Austria. Characterisation of cultivars for attributes affecting N use and
assessing their likely interaction with environmental conditions would
enable growers to make more informed decisions about the choice of
cultivar and management of in-season N fertilisation. Therefore, the aim
of the present study was to (i) evaluate the relative contribution of NUE
traits to grain yield and quality in a selection of widely-grown bread
winter wheat cultivars contrasting in phenology and grain protein
concentration, and (ii) assess the likely consequences of genotypic
variation in N-related traits for cultivar selection and adaptation to
specific environments.
2. Materials and methods
2.1. Field experiments
Two field experiments were conducted in 2017/18 (EXP1) and
2018/19 (EXP2) at the UFT (University and Research Centre Tulln,
BOKU University) in Lower Austria (48◦ 19′ 14.68′′ N, 16◦ 04′ 07.85′′ E,
178 masl). The climate at this experimental site is characterised by the
transition of the Central European-oceanic climate and the dry, more
continental climate zone with a mean annual temperature of 8.7 ◦ C and
average annual precipitation of 673 mm (Auer et al., 2012). The soil is
classified as a chernozem with silty loam in the top soil (BFW, 2007).
In both seasons, four widely-grown Austrian winter wheat cultivars
(Arnold, Aurelius, Bernstein and Emilio) were sown on 16 October at
four levels of N fertilisation (0 (N0), 70 (N70), 140 (N140) and 210
(N210) kg N ha− 1). N levels were applied in strips and within each strip
cultivars were randomly (between and within) allocated to three blocks.
Plots were 3 m wide and 10 m long with a row spacing of 12 cm resulting
in a target population density of 300 plants m-2. The N fertiliser, calcium
ammonium nitrate (27 % N), was split in two equal doses applied as a
top-dressing on 9 April and 8 May 2018 in EXP1 and 8 March and 10
April 2019 in EXP2, corresponding to Zadoks stages Z25 and Z40 in
2018 and Z22 and Z31 in 2019. Prior to sowing, a phosphorus-potassium
fertiliser (15 % P, 40 % K) was applied at the rate of 300 kg fertiliser
ha− 1. Weeds, pests and pathogens were frequently monitored and
controlled as needed.
All four cultivars are considered as quality wheat in Austria, but they
differ in grain protein concentration (%GP, i.e. grain N concentration
multiplied by 5.7), with Arnold having on average the highest %GP
(16.1 %) followed by Aurelius and Bernstein (15.5 %) and Emilio (14.9
%) (AGES (Hrsg.), 2020). The cultivars also differ in phenological
development. In Austria, winter wheat cultivars are classified in 9
phenology groups ranging from very early (group 1) to very late (group
9) (AGES (Hrsg.), 2020). Based on the time of ear emergence (Z55) ac­
cording to the Zadoks scale (Zadoks et al., 1974), Arnold and Aurelius
belong to group 2 (very early to early), Emilio group 3 (early), and
Bernstein group 6 (medium to late). With regard to the duration of the
phase from flowering to physiological maturity (Z87), Arnold is in group
2 (very early to early), Aurelius in group 4 (early to medium), Emilio in
group 3 (early), and Bernstein in group 7 (late).
Data collection and measurement included time series of wheat
phenology, leaf canopy development, mass accumulation, and N con­
centration as well as soil mineral N (Nmin) and water content. Four
random plants were marked in each plot soon after emergence for
scoring phenology and leaf/tiller development according to the Zadoks
scale (Zadoks et al., 1974). The above-ground biomass was harvested
3
Field Crops Research 270 (2021) 108211
from a sampling area of 0.25 m2 in each plot. A final harvest was made
from 1.0 m2. The biomass samples were taken throughout the growing
season at 6 and 10 dates in EXP1 and EXP2, respectively. The biomass
sampling dates in EXP1 corresponded to wheat phenological stages of
leaf development (Zadoks 14; Z14); tillering (Z22 and Z25), stem elon­
gation (Z31− 32), anthesis (Z63− 67), and maturity (Z91). In EXP2,
biomass samples were taken at Z21, Z22, Z23, Z24, Z31, Z33, Z41− 43,
Z63− 67, Z73− 75, and 91. In this paper, however, only the data
collected at crop anthesis and physiological maturity are presented. The
pre-anthesis samples were used to parameterise a spectral index for
non-destructive estimation of crop N uptake (Palka et al., 2021). From
each biomass sample a representative subsample (ca. 30 %) was
dissected into stem (i.e. true stem, leaf sheath, and spike without grain;
thereafter referred to as ‘stem’), green leaf lamina, senesced leaf, and
grain. Green leaf area was measured using a leaf area meter (LI-3100
Area meter, LI− COR Inc., USA). Both the subsamples and the rest of the
samples were then dried in an oven (UNE 600, Memmert GmbH & Co.
KG, Germany) at 65 ◦ C for 72 h and weighed to determine the DM of
plant organs. The N concentration of plant tissues was measured using a
CN elemental analyser (Vario Macro Cube, Elementar Analysensysteme
GmbH, Germany).
Soil water content was measured gravimetrically at sowing and then
at each biomass sampling date from the plant sampling area in all plots
of two replicates. In each plot, three soil samples (one in the crop row
and two others as adjacent cores from the middle point between rows
were collected. Soil samples were taken with an auger to a depth of 120
cm divided into soil layers of 0− 10, 10− 30, 30− 60, 60− 90 and 90− 120
cm. The layer-wise samples from each plot were pooled together and
dried in an oven at 105 ◦ C for at least 3 days. Gravimetric soil water
content was converted to volumetric soil water content by multiplying it
with the dry bulk density of the respective depths. The dry bulk densities
were determined from a soil pit that was dug adjacent to the experi­
mental site. The mineral N content (Nmin) of each soil layer was
determined at crop sowing, anthesis, and physiological maturity by
analysing a fresh subsample from those collected for measuring soil
water content in the soil laboratory of Austrian Agency for Health and
Food Safety (AGES). The daily weather data (minimum and maximum
temperature, rainfall, solar radiation) were obtained from the Central
Institute of Meteorology and Geodynamics (ZAMG) weather station in
Langenlebarn located ca. 5 km from the experimental site.
2.2. Statistical analysis and calculations
Analysis of variance (ANOVA) was carried out using the GLM (gen­
eral Linear Model) procedure of SAS (Statistical Analysis System)
package (version 9.4, SAS Institute, Inc., North Carolina, United States).
As combined analysis of both experiments indicated that the interaction
of season with N supply and cultivar was significant for important crop
variables and because soil conditions (i.e. initial soil water and Nmin)
were different between seasons, statistical analysis was carried out
separately for each experiment. Since N levels were applied in strips, a
specific ANOVA model was used in which blocks were nested within N
levels. N supply, cultivar, and their interaction were considered fixed
factors. Significant differences in the mean values were determined by
LSD (least significant difference) test at a significance level of 0.05.
Graphs were produced using SigmaPlot (Version 14.0, Systat Software
Inc., San Jose, USA).
Thermal time (◦ C d, degree-day) was calculated as the summation of
daily mean temperature above a base temperature of 0 ◦ C. Plant samples
at anthesis and harvest were used to calculate post-anthesis DM and N
remobilisation from leaf and ‘stem’ to the grain, DM and N remobilisa­
tion efficiency, and post-anthesis DM production and soil N uptake.
Remobilisation of DM and N was determined using the balance approach
proposed by Ciampitti and Vyn (2013) as:
Remobilised N/DM = Vegetative N/DM – Stover N/DM,
A.M. Manschadi and A. Soltani
where vegetative N/DM and stover N/DM are the sum of N/DM in leaf
and ‘stem’ at anthesis and maturity, respectively. Remobilisation effi­
ciencies were calculated as DM/N remobilisation divided by total DM/N
at anthesis. Post-anthesis DM production was estimated as grain DM
minus DM remobilised from leaf and ‘stem’. Post-anthesis N uptake was
estimated as the difference between whole-plant N at harvest (i.e.
stover-N plus grain N) minus total N at anthesis.
For the calculation of NUE and its components, grain yield was
adjusted to 14 % water content in order to make the results of the pre­
sent study more applicable for commercially grown wheat production
systems. Nitrogen-use efficiency (NUE, kg grain yield (GY) kg− 1 N
supply) and its components N-uptake efficiency (NUpE, kg total above-
ground N uptake at harvest (TNU) kg− 1 N supply) and N-utilisation ef­
ficiency (NUtE, kg GY kg− 1 TNU) were calculated according to Foulkes
et al.(2009) as:
NUtE = GY (86 % DM) / TNU,
NUpE was calculated based on two estimates of N supply:
NUpE1 = TNU / soil N (pre-sowing Nmin plus fertiliser-N), and
NUpE2 = TNU / soil N (pre-sowing Nmin minus Nmin at harvest plus
fertiliser-N)
NUE1 = (NUpE1) * (NUtE)
NUE2 = (NUpE2) * (NUtE)
3. Results
3.1. Weather and soil conditions
Both growing seasons were slightly warmer than the long-term mean
(LTM; 1991 – 2019) from sowing to the end of December (Fig. 1a).
Temperatures in January were well above the average in 2017/18 and
very close to the LTM in 2018/19. February and March, however, were
much cooler than average in 2017/18 and warmer than the LTM in
2018/19. The critical period from stem elongation to anthesis (April –
May) was warmer in 2017/18, but temperatures during the grain-filling
phase (June – July) were similar in both seasons (Fig. 1a). The two
seasons also differed in the amount and distribution of rainfall. Although
the temporal pattern of rainfall from crop sowing to the end of March
differed between the seasons, the cumulative rainfall in this period was
9% below and 15 % above the LTM in 2017/18 and 2018/19, respec­
tively (Fig. 1b). April was extremely dry in 2017/18 and slightly wetter
than average in 2018/19. May received 122 and 170 mm of rainfall in
2017/18 and 2018/19, respectively, which were well above the LTM of
75 mm. June was much drier than average, particularly in 2018/19
(Fig. 1b). The cumulative rainfall during the growing season in 2017/18
(485 mm) was close to the LTM (500 mm), whereas it was 110 mm more
than the average in 2018/19 season. The cumulative global radiation
during the growing season in EXP1 (2644 MJ m− 2) and EXP2 (2729 MJ
m− 2) was close to the LTM (2591 MJ m− 2).
The top soil layers (0–60 cm) in EXP1 and EXP2 had similar bulk
Fig. 1. Average air temperature (a) and monthly rainfall (b) during the winter wheat growing seasons 2017/18 and 2018/19 in Tulln, Austria; dots represent long-
term mean (1991 – 2019) temperature and rainfall.
4
Field Crops Research 270 (2021) 108211
densities (BD) and organic carbon contents (OC) (Table 1). Compared to
EXP1, the subsoil (60–120 cm) in EXP2 had a higher sand content with
lower OC. In addition, the total initial soil mineral N content (Nmin) in
EXP2 (29.9 kg N ha− 1) was also about 50 % lower than that in EXP1. The
initial soil water content at sowing in EXP1 (586 mm) was higher than
that in EXP2 (364 mm) (Table 1). Measurements of volumetric soil water
content during the growing seasons from well-fertilised plots (N210) of
Bernstein were used to estimate the drained upper limit (DUL) and crop
lower limit (LL) (Table 1).
The sum of differences between DUL and LL in each soil layer was
considered as total transpirable soil water. The fraction of transpirable
soil water (FTSW) was then calculated as the ratio of actual transpirable
soil water (ATSW) to the total transpirable soil water. As depicted in
Fig. 2, the FTSW from sowing until the begin of May (1200 ◦ C d) was
higher in EXP1 but declined continuously from anthesis to physiological
maturity. The FTSW at crop maturity in EXP1 (Fig. 2) does not represent
the actual post-anthesis crop water availability because 63 % of the total
rainfall in June 2018 (53 mm) was received on 28 June (2412 ◦ C d),
which was after the plants had reached physiological maturity. The pre-
anthesis FTSW in EXP2 was lower but substantial rainfalls in May 2019
increased the FTSW at around anthesis. However, similar to EXP1, very
low rainfalls in June resulted in rapid decline in FTSW during the grain-
filling period. In both seasons, FTSW was calculated for the entire soil
profile of 120 cm without accounting for the dynamic of rooting depth.
Therefore, plants with shallower root systems had access to less soil
water and N.
3.2. Phenological development
Plants emerged 15 days after sowing (DAS) in EXP1 and 24 DAS in
EXP2. Nitrogen fertilisation did not have a significant effect on crop
phenology. The stem elongation phase (Zadoks 30; Z30) in all cultivars
started at around 184 DAS (17 April) in both seasons. The thermal time
duration of the phase from sowing to stem elongation was 961 and 1048
◦
C d in EXP1 and EXP2, respectively (Fig. 3). From the booting stage
(Z40), cultivar-specific differences in phenology started to appear, with
Arnold exhibiting the fastest and Bernstein the slowest rate of devel­
opment in both seasons. Accordingly, Arnold reached anthesis (Z65) on
18 May 2018 (1517 ◦ C d) and Bernstein 7 days later (1627 ◦ C d) in EXP1.
In the second season (EXP2), anthesis occurred on 30 May 2019 in
Arnold (1611 ◦ C d) and 6 days later in Bernstein (1756 ◦ C d). In both
growing seasons, the phenological development of Aurelius and Emilio
in the pre-anthesis phase was slightly slower than Arnold and they
reached anthesis 1–2 days later. Cultivars differed in the duration of the
reproductive phase from anthesis to physiological maturity (Z87) in
EXP1. Aurelius and Bernstein required 38 days from anthesis to Z87,
whereas the duration of post-anthesis phase in Arnold and Emilio was 4
days shorter. Expressed in thermal time since sowing, Aurelius and
Bernstein reached Z87 at 2463 ◦ C d and Arnold and Emilio 70.8 ◦ C
A.M. Manschadi and A. Soltani Field Crops Research 270 (2021) 108211

Table 1
Upper (DUL) and lower (LL) limit of plant-available soil water, bulk density (BD), water content at sowing (iniWat), organic matter (OC), and soil mineral nitrogen
content (Nmin) in the winter wheat growing seasons 2017/18 (EXP1) and 2018/19 (EXP2) at Tulln in Austria.
EXP1 EXP2

Layer Depth DUL LL BD iniWat OC Nmin DUL LL BD iniWat OC Nmin

(cm) (m3 m− 3) (m3 m− 3) (g cm− 3) (mm) (%) (kg ha− 1) (m3 m− 3) (m3 m− 3) (g cm− 3)
(mm) (%) (kg ha− 1)
1 0− 10 0.491 0.150 1.372 39.5 2.2 11.9 0.491 0.150 1.372 31.7 2.2 5.0
2 10− 30 0.531 0.270 1.497 99.9 2.2 23.8 0.531 0.270 1.497 89.3 2.2 10.3
3 30− 60 0.560 0.300 1.490 148.9 1.6 15.5 0.560 0.300 1.490 127.6 1.6 5.3
4 60− 90 0.562 0.294 1.421 151.8 1.4 7.2 0.330 0.100 1.300 84.5 0.9 3.5
5 90− 120 0.550 0.294 1.479 133.6 1.1 4.7 0.210 0.050 1.300 31.2 0.3 5.8
Sum 573.7 63.2 364.3 29.9

Fig. 2. Fraction of transpirable soil water (FTSW) vs. thermal time since sowing
(◦ C d) in the winter wheat growing seasons 2017/18 (EXP1) and 2018/19
(EXP2); S1 and S2 arrows indicate the begin of stem elongation, A1 and A2 the
time of anthesis, and M1 and M2 the physiological maturity in cultivar Arnold
in EXP1 and EXP2, respectively.

d earlier. In EXP2, plants reached physiological maturity at 2385 ◦ C

d without major differences between cultivars (Fig. 3).

3.3. Biomass production, grain yield, and nitrogen accumulation

The growing conditions and experimental treatments resulted in a

wide range of crop yield, N accumulation, and associated characteristics.
The overall mean GY (100 % DM) in EXP1 (611.7 g m− 2) was 9.2 %
higher than that in EXP2. The lowest yield (268.2 g m− 2) was obtained
from unfertilised (N0) plots of Arnold in EXP2 and the highest yield
(788.2 g m− 2) from N210 plots of Aurelius in the same experiment.
In EXP1, increasing the N rate from N70 to N210 did not affect TDM
and GY significantly. The fertilised plants produced on average 24.5 %
and 20.4 % more GY and total above-ground dry mass (TDM), respec­
tively, than unfertilised (N0) plants (Table 2). Averaged across N
treatments, Arnold was the lowest-yielding and Aurelius and Bernstein
the highest-yielding cultivars. While Arnold accumulated significantly
less TDM, the other cultivars produced similar amounts of TDM but
differed significantly in harvest index (HI), with Aurelius and Bernstein
showing higher HI and consequently greater GY than Emilio. Grain
number (GNO) was also significantly affected by N supply. In both ex­
periments, Arnold had the lowest and Emilio the highest GNO (Table 2).
In EXP2, increasing N rate resulted in significant increases in TDM,
but GY and HI were not improved from N140 to N210. Compared to
EXP1, the GY of plants at N0 and N70 was reduced by 39.4 % and 19.9
%, respectively, whereas at higher N-rates (N140 and N210) it was
increased by 6.8 %. The cultivar effect on TDM was similar to that in
EXP1 but Aurelius yielded significantly more than Bernstein and Emilio
due to higher HI (Table 2). The interaction term ‘N x C’ for GY and HI in
EXP2 was significant because, contrary to other cultivars, the GY of
Fig. 3. Phenological development (Zadoks stages) of four winter wheat culti­
vars vs. thermal time since sowing (◦ C d) grown in 2017/18 (EXP1) and 2018/
19 (EXP2) seasons; the horizontal line indicates the time of anthesis (Z65).
Bernstein did not increase by increasing the N-rate from N140 to N210
and HI showed a decreasing trend with increasing N-rate from N140 to
N210, except for Aurelius, which maintained its high HI at the highest N-
rate (data not shown).
The response of crop N accumulation to N supply was in both ex­
periments more pronounced compared to DM production and increased
significantly by every increment in N-rate, except for N140 and N210
treatments in EXP1 (Table 2). The overall mean TNU in EXP1 (18.2 g
m− 2) was higher than that in EXP2 (15.1 g m− 2). Similarly, the grain N
uptake (GNU) was significantly affected by N-rate. While Aurelius and
Bernstein accumulated significantly more N than Arnold and Emilio in
EXP1, there were no differences in TNU between cultivars in EXP2.
The mean N harvest index (NHI) across both experiments was 82.5 %
(Table 2). The significance of ‘N x C’ for NHI in EXP1 was related to NHI

5
 Table 2

A.M. Manschadi and A. Soltani

Grain yield (GY), total shoot dry mass (TDM), harvest index (HI), grain number (GNO), total N uptake (TNU), grain N uptake (GNU), nitrogen harvest index (NHI), and grain N concentration (%GN) in winter wheat grown
in 2017/18 (EXP1) and 2018/19 (EXP2) as affected by N supply level and cultivar.
GY (g m− 2) TDM (g m− 2) HI (%) GNO (m− 2) TNU (g m− 2) GNU (g m− 2) NHI (%) %GN (%)
EXP1

N supply*
N0 491.9 b 1205.9 b 40.8 b 12038.5 b 11.2 c 9.2 c 82.0 a 1.88 c
N70 639.9 a 1491.0 a 43.0 a 15416.3 a 18.4 b 15.3 b 83.4 a 2.40 b
N140 652.0 a 1535.9 a 42.4 a 16260.0 a 21.4 a 17.6 a 82.2 a 2.71 a
N210 663.1 a 1520.0 a 43.6 a 16590.3 a 21.9 a 18.2 a 83.3 a 2.76 a

Cultivar
Arnold 534.6 c 1270.6 b 42.0 b 13129.5 c 16.8 b 14.2 b 84.4 a 2.63 a
Aurelius 663.1 a 1477.3 a 44.9 a 15282.0 b 19.4 a 16.1 a 83.2 ab 2.40 b
Bernstein 657.8 a 1537.1 a 42.7 b 15428.3 b 19.2 a 15.8 a 82.3 b 2.36 b
Emilio 591.4 b 1467.7 a 40.3 c 16465.2 a 17.5 b 14.2 b 81.0 c 2.36 b
N *** *** *** *** *** *** ns ***
C *** *** *** *** *** *** *** ***
N×C ns ns ns ns ns ns * ns

EXP2

N supply*
N0 298.1 c 739.8 d 40.2 c 8052.6 d 6.6 d 5.2 d 79.1 b 1.76 d
N70 512.3 b 1146.1 c 44.6 b 13708.1 c 12.1 c 9.9 c 82.5 a 1.95 c
N140 684.1 a 1459.1 b 46.8 a 18589.4 b 18.8 b 15.7 b 83.8 a 2.31 b
N210 726.4 a 1585.2 a 45.8 ab 20178.4 a 22.7 a 19.0 a 83.6 a 2.61 a

Cultivar
Arnold 495.5 c 1141.2 b 42.6 c 13009.8 c 15.1 a 12.5 ab 82.1 a 2.42 a
6

Aurelius 599.1 a 1283.0 a 46.0 a 15260.1 b 15.6 a 13.1 a 83.2 a 2.09 b

Bernstein 564.6 b 1254.3 a 44.6 b 15860.2 a 15.0 a 12.3 b 81.5 a 2.09 b
Emilio 561.8 b 1251.7 a 44.3 b 16398.5 a 14.4 a 11.9 b 82.2 a 2.02 c
N *** *** *** *** *** *** *** ***
C *** *** *** *** ns * ns ***
N×C * ns ** ns ns ns ns ns

* N0, N70, N140 and N210 denote fertiliser rates of 0, 70, 140, and 210 kg N ha− 1.
ns: not significant; *** significant at p < 0.001; ** significant at p < 0.01; * significant at p < 0.05.
Means within each treatment with the same letter are not significantly different at p = 0.05.

Field Crops Research 270 (2021) 108211

 Table 3

A.M. Manschadi and A. Soltani

Pre- and post-anthesis total shoot dry mass accumulation (TDMpre and TDMpost), leaf and ‘stem’ dry mass at anthesis (LFDMa and STDMa), green leaf area index (GLAI), and remobilisation efficiency for leaf and ‘stem’
dry mass (LFDMRE and STDMRE) in winter wheat grown in 2017/18 (EXP1) and 2018/19 (EXP2) as affected by N supply level and cultivar.
TDMpre TDMpost LFDMa STDMa GLAI LFDMRE STDMRE
(g m− 2) (-) (%) (%)

EXP1

N supply*
N0 941.5 b 264.4 a 152.1 b 789.5 b 1.85 b 34.6 a 19.7 a
N70 1149.1 a 341.9 a 195.1 a 954.1 a 2.87 a 44.1 a 22.0 a
N140 1145.2 a 390.7 a 209.7 a 935.5 a 2.96 a 42.6 a 16.6 a
N210 1157.6 a 362.3 a 200.5 a 957.1 a 3.13 a 45.6 a 21.2 a

Cultivar
Arnold 1101.8 a 168.7 c 166.7 b 935.1 a 2.06 c 42.0 a 31.1 a
Aurelius 1034.4 a 443.0 a 183.3 b 851.1 b 2.93 b 37.6 a 15.9 b
Bernstein 1118.4 a 418.7 ab 222.3 a 896.1 ab 3.53 a 46.2 a 13.8 b
Emilio 1138.9 a 328.9 b 185.1 b 953.8 a 2.29 c 41.0 a 18.8 b
N * ns *** *** *** ns ns
C ns *** ** * *** ns **
N×C ns ns ns ns ns ns ns

EXP2

N supply*
N0 659.3 c 80.4 c 100.4 c 559.0 c 1.03 d 37.7 a 31.4 a
N70 883.7 b 262.4 b 138.5 b 745.2 b 2.05 c 39.8 a 23.8 a
N140 901.5 b 557.6 a 154.5 b 747.0 b 2.59 b 34.5 a 8.9 b
N210 1037.2 a 548.0 a 183.1 a 854.1 a 3.17 a 36.0 a 12.0 b
7

Cultivar
Arnold 856.8 a 284.4 b 137.6 a 719.1 a 1.88 b 35.1 a 21.6 a
Aurelius 872.7 a 410.3 a 143.4 a 729.3 a 2.38 a 38.3 a 19.1 a
Bernstein 848.3 a 406.1 a 159.1 a 689.2 a 2.62 a 38.2 a 13.7 a
Emilio 904.0 a 347.7 ab 136.4 a 767.7 a 1.96 b 36.3 a 21.7 a
N *** *** *** *** *** ns ***
C ns * ns ns ** ns ns
N×C ns ns ns ns ns ns ns

* for description of treatments and significance levels see Table 2.

Field Crops Research 270 (2021) 108211

A.M. Manschadi and A. Soltani
Fig. 4. Contribution of post-anthesis photosynthesis (PHOTO) and dry mass (DM) remobilisation from ‘stem’ (STDMR) and leaf (LFDMR) to grain DM as affected by
N fertiliser rate (left panel) and cultivar (right panel) in winter wheat grown in 2017/18 (a,b) and 2018/19 (c,d); N0, N70, N140, and N210 denote fertiliser rates of
0, 70, 140, and 210 kg N ha− 1; contributions with the same letter do not differ significantly (p = 0.05).
of Emilio at N140, which was 6.2 % lower than the other cultivars
(average 83.5 %). In EXP2, the effect of cultivar on NHI was not sig­
nificant and lower N availability in unfertilised plots (N0) reduced the
NHI to 79.1 % (Table 2). Grain N concentration (%GN) ranged from 1.76
% (N0 in EXP2) to 2.76 % (N210 in EXP1) and increased with N supply
following the patterns observed for TNU in both experiments (Table 2).
The lowest-yielding cultivar Arnold had the highest %GN in both
experiments.
3.4. Pre- and post-anthesis DM production and partitioning
Grain yield DM originates from post-anthesis DM production and
remobilisation of pre-anthesis DM accumulated in vegetative tissues (i.e.
leaf and ‘stem’). Table 3 summarises the effects of N and cultivar on
TDM accumulation during pre- and post-anthesis phases and partition­
ing of DM in leaf (leaf lamina) and ‘stem’ at anthesis. In both seasons, the
pre-anthesis accumulated shoot DM (TDMpre) increased with N supply.
The post-anthesis DM (TDMpost), however, was not affected by N supply
in EXP1, whereas it showed a positive response to increasing N supply
from N0 to N140 in EXP2 (Table 3). Relative to TDM, the mean fraction
of TDMpre in EXP1 (76.4 %) was higher than that in EXP2 (70.7 %).
Nitrogen supply favoured the leaf DM at anthesis (LFDMa). In EXP1, the
fraction of LFDMa to TDMpre increased from 16.2 % in unfertilised
plants to 17.5 % under N fertilisation and the effect of N supply on green
leaf area index (GLAI) followed the patterns observed for TDMpre and
LFDMa. Similar to EXP1, the unfertilised plants in EXP2 produced less
DM pre-anthesis. However, in contrast to EXP1, the fertilised plants
8
Field Crops Research 270 (2021) 108211
responded significantly to N-rate with the highest TDMpre and GLAI
observed in N210 (Table 3). The pre-anthesis fraction of leaf DM to
TDMpre increased with N rate ranging from 15.2 % in N0 to 17.7 % in
N210 treatment. Nitrogen rate affected positively the TDMpost in EXP2,
although the difference between N140 and N210 was statistically not
significant.
In both experiments, cultivar means across N levels did not differ in
TDMpre and the differences in TDMpost showed a similar pattern with
Arnold accumulating the lowest and Aurelius and Bernstein the highest
amount of DM post-anthesis (Table 3). The DM partitioning in leaf and
‘stem’ at anthesis was also significantly different among the cultivars. In
EXP1, Bernstein had the highest LFDMa and GLAI, while Arnold and
Emilio allocated more DM to ‘stem’ and, consequently, had the lowest
GLAI at anthesis. The fraction of leaf DM was 19.9 % for Bernstein and
16.4 % for other cultivars. In EXP2, cultivars did not differ significantly
in LFDMa and STDMa. However, similar to EXP1, Bernstein had the
highest GLAI and the fraction of leaf DM at anthesis was on average 17.6
% for Bernstein and Aurelius and 15.6 % for Arnold and Emilio.
Comparison of the DM data at anthesis and maturity allowed esti­
mation of the amount and efficiency of DM remobilised from leaf and
‘stem’ during the post-anthesis period. In both experiments, the effi­
ciency of DM remobilisation from leaf (LFDMRE) was not affected by N
and cultivar treatments and was on average 42.0 % in EXP1 and 37.0 %
in EXP2 (Table 3). The relative contribution of DM remobilised from
‘stem’ to grain was generally lower than that from leaf, although in
absolute terms the contribution of ‘stem’ was higher (see section 3.6,
Fig. 4). In EXP1, averaged across N levels, the remobilisation efficiency
A.M. Manschadi and A. Soltani Field Crops Research 270 (2021) 108211

Table 4
Pre- and post-anthesis total N accumulation (TNUpre and TNUpost), leaf and ‘stem’ N at anthesis (LFNa and STNa), and remobilisation efficiency of N from leaf
(LFNRE) and ‘stem’ (STNRE) in winter wheat grown in 2017/18 (EXP1) and 2018/19 (EXP2) as affected by N supply level and cultivar.
TNUpre TNUpost LFNa STNa LFNRE STNRE
(g m− 2) (%)

EXP1

N supply*
N0 10.07 c 1.17 a 3.13 b 6.93 c 80.56 a 78.17 a
N70 17.85 b 0.54 a 5.60 a 12.26 b 84.85 a 81.21 a
N140 22.11 a − 0.68 a 6.65 a 15.46 a 83.87 a 81.13 a
N210 21.58 a 0.34 a 6.51 a 15.07 a 85.09 a 81.36 a

Cultivar
Arnold 17.03 a − 0.19 a 4.34 c 12.69 a 82.13 b 84.36 a
Aurelius 17.48 a 1.91 a 5.56 b 11.92 a 83.61 ab 78.75 b
Bernstein 19.62 a − 0.41 a 7.16 a 12.47 a 86.05 a 79.68 b
Emilio 17.48 a 0.05 a 4.84 c 12.64 a 82.57 b 79.08 b
N *** ns *** *** * ns
C ns ns *** ns * *
N×C ns ns ns ns ns ns

EXP2

N supply*
N0 5.40 d 1.19 b 1.48 d 3.92 d 75.19 b 72.94 b
N70 11.13 c 0.92 b 3.25 c 7.89 c 82.64 a 79.36 a
N140 15.34 b 3.42 a 4.80 b 10.55 b 82.95 a 78.26 a
mN210 19.34 a 3.45 a 5.97 a 13.37 a 81.64 a 79.97 a

Cultivar
Arnold 12.80 a 2.27 a 3.47 bc 9.33 a 77.82 b 78.10 a
Aurelius 13.05 a 2.56 a 4.02 b 9.02 a 84.08 a 77.68 a
Bernstein 13.26 a 1.72 a 4.80 a 8.46 a 81.44 ab 75.90 a
Emilio 12.11 a 2.33 a 3.20 c 8.91 a 79.08 b 78.84 a
N *** * *** *** ** *
C ns ns *** ns ** ns
N×C ns ns ns ns ns ns

* for description of treatments and significance levels see Table 2.

of ‘stem’ DM (STDMRE) was 19.9 %, whereas in EXP2 it decreased from
27.6 % in N0 and N70 to 10.5 % in N140 and N210. Among cultivars, the
STDMRE of Arnold was significantly higher than the other cultivars in
EXP1, whereas cultivars did not differ in STDMRE in EXP2. Averaged
across all treatments, the efficiency of DM remobilisation from vegeta­
tive organs (leaf and ‘stem’) was 23.7 % and 22.1 % in EXP1 and EXP2,
respectively.
3.5. Pre- and post-anthesis N accumulation and partitioning
The wheat growing seasons differed markedly in the relative
amounts of shoot N accumulated pre-anthesis (TNUpre) and during the
post-anthesis period (TNUpost) (Table 4). Some negative mean values
for TNUpost were found in EXP1 that can be attributed to generally very
low amounts and variation in sampling area. The pre-anthesis N uptake
(TNUpre) responded positively to increasing N supply in both experi­
ments (Table 4). On average, wheat plants accumulated 97.2 % and 85.2
% of TNU pre-anthesis in EXP1 and EXP2, respectively. The very low
post-anthesis N accumulation (TNUpost) in EXP1 was not affected by N
supply (average 0.34 g N m− 2), while it increased with N supply in EXP2
reaching an average maximum of 3.43 g N m− 2 in N140 and N210
treatments. Similar to TDM, cultivars did not differ significantly in
TNUpre in both experiments. Plants on average accumulated more
TNUpre in EXP1 (17.90 g N m− 2) than in EXP2 (12.80 g N m− 2)
(Table 4). In both experiments, Aurelius had the highest TNUpost,
although the differences to other cultivars were statistically not signif­
icant (Table 4).
Leaf and ‘stem’ N contents at anthesis (LFNa, STNa) responded
positively to N supply in both seasons (Table 4). Cultivars did not differ
in STNa, but the effect of cultivar on LFNa was statistically significant,
with Bernstein showing the highest LFNa in both experiments. Overall,
about 30.7 % of total shoot N at anthesis was found in leaf. Compared to
unfertilised plants, N remobilisation from leaf and ‘stem’ were higher
under fertilisation in both growing season. In EXP1, the remobilisation
efficiency of leaf N (LFNRE) was on average 83.6 %, while in EXP2 the
fertilised plants had a similar LFNRE (82.4 %) but plants in the N0
treatment remobilised only 75.2 % of leaf N (Table 4). Similarly, the
efficiency of N remobilised from ‘stem’ (STNRE) was on average 80.5 %
in EXP1 and in EXP2 it was significantly lower in N0 (72.9 %) compared
to fertilised treatments (79.2 %) (Table 4).
In both seasons, Aurelius and Bernstein appeared to have higher N
remobilisation efficiencies from leaf (average 83.8 %) compared to
Arnold and Emilio (average 80.1 %). The cultivar effect on STNRE in
EXP2 was not significant (average 77.6 %), whereas Arnold remobilised
significantly more N from ‘stem’ (84.4 %) in EXP1 (Table 4). The overall
efficiency of N remobilisation from vegetative parts (leaf and ‘stem’) was
81.4 % and 78.5 % in EXP1 and EXP2, respectively.
3.6. Contribution of different sources to DM and N accumulation in grains
As the interaction term ‘N x C’ for contribution of different sources to
grain DM and N in both experiments was not significant, the results are
presented for the effects of N level and cultivar separately. In EXP1, the
positive response of GY to N-rate was largely due to increases in the post-
anthesis DM production (PHOTO) and remobilisation of leaf DM
(LFDMR) (Fig. 4a). Averaged across cultivars, the relative contributions
of PHOTO, STDMR and LFDMR were 53.7 %, 32.8 %, and 13.5 %,
respectively. Among cultivars, Arnold had the lowest PHOTO and the
highest STDMR (Fig. 4b). Aurelius and Bernstein had a similar contri­
bution from PHOTO (average 64.3 %) but differed in LFDMR, with
Bernstein showing a significantly higher contribution from LFDMR. In
EXP2, the mean contribution of PHOTO was higher (58.1 %), while the

9
A.M. Manschadi and A. Soltani
Fig. 5. Contribution of post-anthesis soil nitrogen uptake (SOILN) and N remobilisation from ‘stem’ (STNR) and leaf (LFNR) to grain N as affected by N fertiliser rate
(left panel) and cultivar (right panel) in winter wheat grown in 2017/18 (a,b) and 2018/19 (c,d); N0, N70, N140, and N210 denote fertiliser rates of 0, 70, 140, and
210 kg N ha− 1; contributions with the same letter do not differ significantly (p = 0.05).
relative contributions of STDMR (31.7 %) and LFDMR (10.3 %) were
lower than in EXP1.
The effect of N regime on PHOTO was strongly significant in EXP2,
with a relative contribution ranging from 25.3 % in N0 to 79.5 % in
N140 (Fig. 4c). The contribution of STDMR decreased at higher N rates,
while LFDMR showed an increasing pattern with N supply. The ranking
of cultivars with regard to PHOTO was similar to EXP1 and they did not
differ significantly in STDMR and LFDMR (Fig. 4d).
Fig. 5 depicts the contribution of post-anthesis N uptake from the soil
(SOILN) and N remobilisation from ‘stem’ (STNR) and leaf (LFNR) to
grain N. In both experiments, STNR had the largest contribution fol­
lowed by LFNR and SOILN. In EXP1, the mean relative contributions of
STNR, LFNR, and SOILN were 66.6 %, 30.4 %, and 3.1 %, respectively.
The SOILN was generally very low with no significant effects of N level
and cultivar. In EXP2, however, the relative contribution of SOILN
increased to 17.6 % largely due to increased N uptake at N140 and N210
rates, while STNR (57.4 %) and LFNR (25.0 %) were lower than in EXP1.
Nitrogen supply had a significant positive effect on STNR and LFNR in
both seasons and on SOILN in EXP2 (Fig. 5a, c). Aurelius showed the
highest SOILN in both experiments, particularly in EXP1, but the dif­
ferences to other cultivars were statistically not significant (Fig. 5b, d).
There were no significant differences in STNR between the wheat cul­
tivars but they differed in LFNR with Bernstein showing the highest
LFNR in both seasons.
10
Field Crops Research 270 (2021) 108211
3.7. Nitrogen-use efficiency
The overall efficiency of N use calculated based on N supply as the
sum of pre-sowing soil mineral N and fertiliser-N (NUE1) was in EXP1
(40.1 kg GY kg− 1 N supply) lower than that in EXP2 (45.9 kg GY kg− 1 N
supply). Nitrogen regime had a significant effect on NUE1 in both sea­
sons, with increases in N-rate resulting in decreasing NUE1 (Table 5).
The utilisation-efficiency of N (NUtE) also decreased with N supply but
the differences in EXP1 were not statistically significant. Among culti­
vars, Arnold had significantly lower NUtE than the other cultivars. In
terms of NUpE1, Aurelius showed a significantly better performance
than Arnold and Emilio in both seasons. Similar to NUtE and NUpE1, the
lowest NUE1 was observed in Arnold and the highest in Aurelius and
Bernstein in both experiments (Table 5).
When accounting for soil Nmin at harvest in the calculation of N-
uptake efficiency, there was an overall increase in NUpE2 (Table 5).
Compared to NUE1, the ranking of cultivars in EXP1 changed with
Arnold showing the highest and Bernstein the lowest NUE2. In EXP2,
however, the cultivar ranking for NUE2 was similar to NUE1 with
Aurelius showing the highest efficiency. Increases in NUpE2 resulted in
higher values of NUE2. The mean NUE2 was 62.8 kg kg− 1 and 55.2 kg
kg− 1 in EXP1 and EXP2, respectively (Table 5).
The effect of ‘N x C’ on NUE was significant except for NUE1 in EXP1.
Generally, NUE decreased with increasing N supply for all cultivars and
the interaction effect of N and cultivar was significant because the
magnitude of decrease in NUE was different among the cultivars. Only
for Aurelius, the difference in NUE2 between N70 and N140 was
A.M. Manschadi and A. Soltani Field Crops Research 270 (2021) 108211

Table 5
Nitrogen-utilisation efficiency (NUtE), N-uptake efficiency calculated based on N supply as the sum of pre-sowing soil mineral N (Nmin) and fertiliser-N (NUpE1) and
the difference between pre-sowing Nmin and Nmin at harvest plus fertiliser-N (NUpE2), and their associated N-use efficiencies (NUE1 and NUE2) of winter wheat
grown in 2017/18 (EXP1) and 2018/19 (EXP2) as affected by N supply and cultivar.
NUtE NUpE1 NUE1 NUpE2 NUE2
(Kg kg− 1)

EXP1

N supply*
N70 36.87 a 1.47 a 52.94 a 2.08 a 74.83 a
N140 32.18 a 1.19 ab 38.32 b 1.99 a 63.54 a
N210 31.00 a 0.94 b 29.09 c 1.61 a 49.90 b

Cultivar
Arnold 31.13 b 1.12 b 35.01 c 2.10 a 65.78 a
Aurelius 34.28 a 1.26 a 43.35 a 1.82 bc 62.15 b
Bernstein 33.85 a 1.27 a 42.97 a 1.74 c 58.65 c
Emilio 34.15 a 1.15 b 39.14 b 1.91 b 64.44 ab
N *** *** *** ns ***
C ** *** *** *** ***
N×C ns ns ns ns ***

EXP2

N supply*
N70 40.65 a 1.52 a 60.90 a 1.87 a 74.95 a
N140 34.70 ab 1.27 ab 43.82 b 1.50 ab 51.71 b
N210 31.31 b 1.05 b 32.90 c 1.25 b 39.97 c

Cultivar
Arnold 32.15 b 1.25 b 40.52 c 1.51 b 48.92 c
Aurelius 36.78 a 1.34 a 49.57 a 1.65 a 60.98 a
Bernstein 36.20 a 1.30 ab 47.61 ab 1.52 b 55.93 b
Emilio 37.08 a 1.23 b 45.80 b 1.47 b 55.00 b
N *** *** *** *** ***
C *** * *** * ***
N×C ns ns ** *** ***
*
For description of treatments and significance levels see Table 2.

statistically not significant (Fig. 6).
3.8. Relationships between N-related variables
Fig. 7a shows the scatter plot of GY against TNU from both experi­
ments. A non-linear log-function described this relationship well (R2 =
0.89) with the slope of the curve representing NUtE. When the model
was fitted to the data from individual cultivars, the slopes did not differ
significantly, implying that the cultivar differences in GY depended on
their TNU and not NUtE (Table 6). Similarly, grain N concentration was
very closely related to NUtE (R2 = 0.97) indicating the trade-off between
these two variables, with higher NUtE values occurring at the expense of
lower grain N concentrations (Fig. 7b). The slope of the fitted model for
Arnold was significantly lower than that for other cultivars (Table 6).
4. Discussion
The objective of the present study was to characterise a set of winter
wheat cultivars contrasting in grain protein content and phenology for
traits affecting N use and assess the implications of genotypic variation
in N-related traits for cultivar selection and adaptation to specific en­
vironments. The environmental conditions combined with N and
cultivar treatments resulted in a wide range of DM production, N
accumulation, and grain yield and quality.
4.1. Environmental conditions
Differences in soil initial water and Nmin contents together with
variation in temporal patterns of in-season rainfall and temperature in
the 2017/18 and 2018/19 growing seasons generated two contrasting
types of environment. The growing season 2017/18 can be described as
an environment type with favourable water conditions during crop
vegetative growth followed by declining water supply from flowering
through to plant maturity. This type of environment is representative for
winter wheat production systems in Pannonian north-eastern part of
Austria. The second season (2018/19), however, represented an envi­
ronment with water limitation starting during the vegetative growth but
relieved by rainfalls during the period shortly before anthesis to early
grain-filling and continuing with severe drought stress to crop physio­
logical maturity (see Fig. 2). The differences between the two environ­
ments are reflected in the observed patterns of pre- and post-anthesis
DM production. Compared to EXP2, plants in N140 and N210 treatments
of EXP1 had accumulated on average 18.8 % more DM pre-anthesis,
whereas post-anthesis DM accumulation was 46.8 % higher in EXP2
(see Table 2). The inter-seasonal variation in environmental conditions
strongly influenced the performance and ranking of cultivars with re­
gard to N use and grain yield formation (see the discussion below).
Similar strong effect of inter-annual weather variation on the response of
wheat to N fertilisation using long-term yield data (1968–2016) from
Rothamsted, UK, has been reported by Addy et al. (2020).
In addition to weather, the two seasons also differed in N supply due
to differences in soil Nmin at sowing. In both experiments, the previous
crops (winter wheat in 2016/17 and maize in 2018) were not fertilised
at all in order to reduce the soil Nmin to a similar level. However, dif­
ferences in N uptake by the previous crops and the length of the pre-
sowing fallow periods resulted in higher Nmin values in 2017/18
compared to 2018/19 (see Table 1). Thus, the overall lower mean GY in
EXP2 was related to stronger N stress in N0 and N70 treatments, as at
higher N-rates (N140 and N210) plants produced similar amounts of GY
in both experiments (Table 2). Furthermore, differences in temperature
and soil water content between the two seasons affected the amount of N
released from mineralisation of soil organic matter. The average amount
of mineralised soil N, calculated in N0 plots based on measured soil
Nmin at sowing and harvest and total crop N uptake, was 72.4 kg N ha− 1

11
A.M. Manschadi and A. Soltani
Fig. 6. Nitrogen-use efficiency calculated based on N supply as the sum of pre-sowing soil mineral N (Nmin) and fertiliser-N (NUE1) and as the difference between
pre-sowing Nmin and Nmin at harvest plus fertiliser-N (NUE2) in four winter wheat cultivars grown in 2017/18 (EXP1; a, c) and 2018/19 (EXP2; b, d); N70, N140,
and N210 denote fertiliser rates of 70, 140, and 210 kg N ha− 1.
in 2017/18 and 47.3 kg N ha− 1 in 2018/19. Consequently, the effect of
N supply on GY and TDM was less pronounced in EXP1.
4.2. Phenology, grain yield and grain protein content
The observed phenological development of the four winter wheat
cultivars in both experiments matched closely the observations from
long-term multi-environment cultivar evaluation trials reported by
AGES (Hrsg.) (2020). Arnold showed the fastest rate of development and
reached anthesis 6–7 days earlier than Bernstein. According to AGES
(Hrsg.) (2020), Aurelius and Arnold are in the same group for the timing
of anthesis but Aurelius has a longer grain-filling phase. We made
similar observations in EXP1 but in EXP2 there were no differences in
the timing of physiological maturity between the cultivars. This may be
attributed to warmer and much drier conditions in June 2019 resulting
in accelerated senescence and shortening of the grain-filling phase.
Averaged across the two seasons, the observed grain yields (86 %
DM) at the N-rate of 140 kg N ha− 1 (N140), which is comparable to the
recommended rate of 120 kg N ha− 1 in north-eastern Austria, for Arnold
(6.7 t ha− 1), Aurelius (8.2 t ha− 1), Bernstein (8.1 t ha− 1), and Emilio (7.4
t ha− 1) were on average just 3.8 % higher than the mean yields obtained
from cultivar evaluation trials conducted at 16 locations in the same two
growing season (AGES (Hrsg.), 2020). The observed %GP of the culti­
vars in EXP1 matched closely the AGES data but it was on average 15.2
% lower in EXP2, indicating that plants accumulated less N at N140 in
this season (see TNU in Table 2). This can be attributed to generally
lower N uptake in EXP2 due to unfavourable water supply. Furthermore,
12
Field Crops Research 270 (2021) 108211
the ranking of cultivars in grain yield (Arnold < Emilio, Bernstein <
Aurelius) and %GP (Arnold > Emilio, Bernstein, Aurelius) matched also
very well the AGES data. Therefore, the results of the present field study
can be considered as representative for the productivity and N use of
winter wheat in this part of Austria.
4.3. Traits contributing to improved N use
In agreement with Sinclair and Rufty (2012), the present results
clearly revealed that modern wheat cultivars are very efficient in the use
of the accumulated N for producing GY. The observed NHIs were more
than 80 %, similar to previous published data (Barraclough et al., 2010;
Gaju et al., 2011; Pask et al., 2012; Hitz et al., 2017; Liu et al., 2020).
NHI was not affected by N supply except for the unfertilised plants in
EXP2, which were exposed to severe N deficiency. The lower NHI of
N-stressed plants can be attributed to lower remobilisation efficiency of
stored N in leaf and ‘stem’ (see Table 4). The observed efficiency of N
remobilisation from vegetative parts (average 80 %) is also similar to
that reported by Barraclough et al. (2014) from a field study with twenty
elite varieties of wheat grown in the UK. While Barraclough et al. (2014)
did not observe genotypic differences in N remobilisation efficiency, our
data suggest that cultivars with greater leaf growth and N accumulation
(Bernstein and Aurelius) can also remobilise leaf N with higher effi­
ciencies (Table 4).
The observed positive relationship between GY and TNU (Fig. 7a) is
in agreement with previous work (Sinclair and Rufty, 2012). However,
similar to Savin et al. (2019), this relationship was not linear, suggesting
A.M. Manschadi and A. Soltani Field Crops Research 270 (2021) 108211

Fig. 7. Relationships between grain yield (GY) and total shoot nitrogen uptake (TNU) (a) and grain N concentration (%GN) and N-utilisation efficiency (NUtE) in
four winter wheat cultivars grown in 2017/18 and 2018/19 seasons.

Table 6
Parameters (± s.e.) of the model y = a + b ln(x) fitted to the relationships between grain yield (GY, Mg ha− 1) and total nitrogen uptake (TNU, kg ha− 1) and grain
nitrogen concentration (%GN, %) and N-utilisation efficiency (NUtE, kg kg− 1) for winter wheat cultivars grown in 2017/18 (EXP1) and 2018/19 (EXP2); all fitted
models had P < 0.0001.
Model Cultivar a b* R2

GY = a + b ln(TNU)
Arnold − 12.200 ± 0.664 3.611 ± 0.132 a 0.97
Aurelius − 12.655 ± 1.073 3.901 ± 0.021 a 0.94
Bernstein − 11.717 ± 0.771 3.690 ± 0.152 a 0.96
Emilio − 10.071 ± 1.076 3.335 ± 0.215 a 0.92
All cultivars − 11.914 ± 0.667 3.686 ± 0.132 0.89

%GN = a + b ln(NUtE)
Arnold 12.785 ± 0.446 − 2.822 ± 0.123 b 0.96
Aurelius 10.710 ± 0.286 − 2.253 ± 0.076 a 0.98
Bernstein 10.768 ± 0.233 − 2.277 ± 0.062 a 0.98
Emilio 10.043 ± 0.164 − 2.086 ± 0.043 a 0.99
All cultivars 10.876 ± 0.155 − 2.302 ± 0.042 0.97
*
Values of b for each model with the same letter are not significantly different at p = 0.05.

a diminishing return at higher N rates possibly due to limitation of water
supply. The slope of the fitted curve in Figure 7a (3.69) represents NUtE
and was lower than that calculated by Savin et al. (2019) for
non-Mediterranean environments (4.94) but similar to that for 99th yield
percentile for Mediterranean regions (3.5). This reinforces the dominant
effect of terminal drought stress on N utilisation in Pannonian
environments.
The wheat cultivars differed significantly in the TNU component of
Sinclair and Rufty (2012) equation in EXP1 but not in EXP2, indicating
the strong effect of environmental conditions on N accumulation.
Interestingly, cultivars had similar amounts of total pre-anthesis N
accumulation, so that variation in TNU was caused by differences in
post-anthesis N uptake, with the highest-yielding cultivar Aurelius
showing consistently greater N uptake during grain filling. Thus, the
ability to maintain N uptake post-anthesis appears to play a key role in
genotypic variation in N uptake and utilisation for producing GY.
Similar to our results, Bogard et al. (2010) reported that genotypic
variation in deviation from negative relationship between grain yield

13
A.M. Manschadi and A. Soltani
and grain protein concentration in 27 wheat cultivars grown in
multi-environment trials in northern France was also related to
post-anthesis N uptake independently of anthesis date and total N up­
take at anthesis. The authors hypothesized that the interacting effects of
genotypic differences in (i) accessing soil N due to differences in rooting
depth and architecture, (ii) delayed leaf senescence and prolonged de­
livery of photosynthates to roots, and (iii) N uptake regulation by plant
N status may explain the observed variability in post-anthesis N uptake
in wheat.
As N uptake is strongly dependent on soil water availability, the post-
anthesis N uptake might be reduced under conditions of water deficit,
particularly in the subsoil, during the grain-filling period. This may
explain the relatively very low post-anthesis N uptake in EXP1 compared
to EXP2. While the post-anthesis N uptake quantities in EXP2 are com­
parable with those reported by Bogard et al. (2010) from northern
France (11–35 kg N ha− 1), they are lower than the maximum values (60
kg N ha− 1) reported from higher rainfall areas in the UK (Barraclough
et al., 2014). Severini et al. (2020) also found genotypic correlation
between rooting depth and grain yield in field-grown wheat genotypes
differing in early vigour, height, tillering, and flowering time but vari­
ation in rooting depth and length caused by environmental conditions
was much larger than the genotypic effects. Under certain environ­
mental conditions, N fertilisation rate itself may also positively affect
post-anthesis N uptake (see Fig. 5c) due to stronger shoot growth leading
to enhanced root growth (Rasmussen et al., 2015).
Due to lack of post-anthesis green leaf area and rooting depth ob­
servations in the present study, it is not possible to explore the physio­
logical mechanisms underlying the superior performance of Aurelius.
However, detailed measurements of the chlorophyll content of the flag,
penultimate, and third upper leaf using a SPAD-502 chlorophyll metre
(Konica Minolta Sensing, Osaka, Japan) in a field experiment conducted
in 2019/20 with the same cultivar and N treatments clearly demon­
strated the stay-green phenotype of Aurelius resulting in significantly
higher grain yield compared to other cultivars (Manschadi et al., un­
published data).
While some studies suggest that grain yield of modern wheat culti­
vars in favourable growing conditions, such as in irrigated systems, is
determined by grain number (GNO) and, hence, is sink-limited (e.g.
Borrás et al., 2004; Fischer, 2008; Alvarez Prado et al., 2017), the GY
and GNO data from our study (see Table 2) indicate that GNO was not
the key determinant of yield because the cultivar Emilio with the highest
GNO in both seasons yielded significantly less than Aurelius. Thus,
similar to Sinclair and Jamieson (2006,2008), the present data support
the hypothesis that GNO does not determine wheat grain yield, but
rather both GNO and yield are the consequence of crop’s ability to
accumulate carbon and N, particularly during the post-anthesis phase.
Therefore, the stay-green trait seems to be a promising target for
breeding cultivars with improved N use and grain yield in rain-fed en­
vironments with frequent water and thermal stress during grain-filling,
such as the Pannonian region. Rapid screening methods such as NDVI
(Christopher et al., 2014) and spectral reflectance measurements after
anthesis may facilitate the selection of superior genotypes for the
stay-green trait.
The conventional approach for evaluating cultivar differences in N
use based on the efficiency of N uptake and utilisation reflected only to
some extent the observed genotypic variation in N-related traits.
Generally, the observed N efficiencies (NUE1) based on the common
approach of calculating N-uptake efficiency using initial soil Nmin plus
fertiliser-N were within the range reported in previous studies (30 – 60
kg kg− 1) depending on N-rate, genotype, and environment (Gaju et al.,
2011; Hitz et al., 2017). The N-uptake efficiencies in the present study
(Table 5) are substantially higher than those reported from some other
field experiments (e.g. Liu et al., 2020) due to relatively low initial soil
Nmin and high potential for N mineralisation from sol organic matter.
The NUtE and NUE1 values reflect the observed differences in physio­
logical mechanisms between the cultivars, with Arnold being least
14
Field Crops Research 270 (2021) 108211
efficient in utilising N. However, when accounting for soil Nmin at
harvest in the calculation of N-uptake (NUpE2), which provides a more
realistic estimation of actual crop N uptake and soil N mineralisation
potential, Arnold showed the highest NUE2 in EXP1, while Aurelius
became the most N-efficient cultivar in EXP2. This indicates that vari­
ation in the calculation method and prevailing environmental condi­
tions may change the ranking of genotypes with regard to N efficiency,
so that a low-yielding genotype can become the most efficient one in N
use. Therefore, assessing the efficiency of cultivars using the conven­
tional approach of NUE is less adequate compared to the framework
based on TNU, NHI, and %GN proposed by Sinclair and Rufty (2012).
4.4. Variation in genotypic strategies for N accumulation and use
The detailed quantification of patterns of DM and N accumulation
and use from the two field experiments allowed identification of con­
trasting genotypic strategies for N use among the four cultivars, which
may be beneficial in specific type of environments.
The lowest-yielding cultivar in both seasons, Arnold, is characterised
by earlier flowering, shorter grain-filling phase, and low post-anthesis
DM production (PHOTO, Fig. 4). This strategy results in high GY qual­
ity (grain protein content) and might be beneficial in environments with
severe terminal water deficit as a drought escape mechanism. Aurelius
has a similar time of flowering as Arnold but is capable of extending the
post-anthesis phase due to delayed onset of leaf senescence (stay-green
phenotype). This strategy can result in increased post-anthesis DM and N
accumulation (Fig. 4 and 5), if water is available in the subsoil late in the
season. The importance of soil moisture availability for the expression of
“stay-green” phenotype in wheat has been demonstrated in a series of
field experiments where the stay-green genotype did not exhibit the
stay-green phenotype nor a significant yield advantage over the stan­
dard variety in the absence of subsoil water (Christopher et al., 2008).
Compared to Aurelius, Bernstein has a longer pre-anthesis phase due to
delayed anthesis and allocates relatively more pre-anthesis DM to leaves
and, hence, has a higher leaf N storage capacity (Table 4). This strategy
can compensate the lower capacity for post-anthesis N uptake, achieving
a grain yield as high as Aurelius in environments with favourable water
supply conditions during the vegetative growth, such as in EXP1. But
under limited pre-anthesis water supply, such as in EXP2, Bernstein may
exhaust soil moisture and run out of water during the reproductive
phase. This explains the GY difference between Bernstein and Aurelius
in EXP2. Therefore, the vigorous phenotype of Bernstein is better
adapted to high rainfall environments with favourable temperature re­
gimes allowing a long growing season with adequate water supply.
Emilio, in contrast to Bernstein, partitions more of the pre-anthesis DM
to ‘stem’ resulting in lower stored leaf N and GLAI, which may help to
save water pre-anthesis for using it post-anthesis. This appear to have
occurred in EXP2, where Emilio yielded as high as Bernstein. But under
more favourable pre-anthesis water supply conditions, such as in EXP1,
this strategy of Emilio was not beneficial, resulting in significantly lower
GY than Bernstein.
4.5. Implications for N management
It is widely recognised that the key to optimising the trade-offs be­
tween crop yield, economic return, and environmental protection is to
achieve the best synchrony between soil N supply and crop N demand by
accounting for complex interactions between plant genetic characteris­
tics (G), management practices (M), and production environments (E).
Farmers generally tend to select an optimum combination of crop ge­
notype (G) and agronomic management (M) based on their experiences
from the past. However, the optimum G x M combination depends
strongly on seasonal weather conditions (E), which vary among sites and
growing seasons (Hammer et al., 2014).
Current N management practices are largely based on empirical
approaches representing crop demands in “average seasons”. While
A.M. Manschadi and A. Soltani
these “average” management practices have been relatively successful in
the past, increased climate variability demands a more targeted season-
specific N management in order to minimise inputs in less favourable
seasons and maximise them in seasons with high yield potential (Meza
et al., 2008; Monjardino et al., 2013). Thus, improving crop N use re­
quires a shift from empirical “average” management of N towards highly
optimised, site- and season-specific, and real-time management. Mete­
orological forecasts offer the potential to anticipate variations in crop
production early enough to adjust and optimise management decisions
(i.e. G x M) for the expected growing environment (E) in order to reduce
adverse impacts or take advantage of favourable conditions (Rodriguez
et al., 2018). Skilful forecasts of precipitation and temperature over the
next few days, weeks or months can support optimum scheduling of a
wide range of on-farm activities including N management (Asseng et al.,
2016; Klemm and McPherson, 2017).
As outlined above, winter wheat cultivars differ in N-related traits
and, therefore, matching cultivar to the seasonal environmental condi­
tions appears to be critical for improving crop N use. Linking short-range
weather forecasts with crop models and estimates of crop N status using
spectral reflection may provide an integrated tool for improving the
timing and quantity of in-season N applications. While the predictive
skill for short-range weather forecasts in Europe has been increasing
(Bauer et al., 2015), long-range forecasts are still associated with large
uncertainties. Therefore, they provide little help in the selection of the
best N-adapted winter wheat cultivar at the time of sowing in autumn to
the critical environmental conditions a few months later in the next
spring and summer months. One way to improve the decision on the
choice of winter wheat cultivar is to take into account the stored soil
water at the time of sowing because it can buffer to some extent against
low in-season rainfall (Kirkegaard and Hunt, 2010), as it was demon­
strated in the present field experiments. Therefore, modifying crop
rotation to allow for a longer fallow period and, hence, greater soil water
storage prior to winter wheat planting will allow planting of cultivars,
such as Bernstein and Aurelius, with higher GY and improved N use.
5. Conclusions
Results of the present study demonstrated significant genotypic
variation in traits contributing to improved N use even among a limited
number of elite commercial winter wheat cultivars. The four wheat
cultivars were very similar in the efficiency of using the accumulated N
for producing grain yield but differed substantially in the time of
anthesis, leaf canopy development, N storage in leaves, and post-
anthesis DM and N accumulation. The lowest-yielding cultivar
(Arnold) was characterised by a short life cycle (early flowering and
maturity), which can be beneficial in environments with severe terminal
drought stress. The highest-yielding cultivar (Aurelius) had a similar
time of anthesis as Arnold but exhibited an extended post-anthesis phase
(stay-green phenotype), resulting in increased N uptake and DM pro­
duction. This strategy can lead to higher grain yield, if water is available
in the subsoil late in the season. Bernstein was characterised by a longer
pre-anthesis phase due to delayed anthesis and allocated relatively more
pre-anthesis DM to leaves and, hence, had a higher leaf N storage ca­
pacity. This strategy can compensate the lower capacity for post-
anthesis N uptake, achieving a grain yield as high as Aurelius, in high
rainfall environments with favourable temperature regimes allowing a
long growing season with adequate water supply. Emilio, partitioned
more of the pre-anthesis DM to ‘stem’ resulting in lower stored leaf N
and leaf area, which may help to save water pre-anthesis for using it
post-anthesis, and hence is better adapted to environments with water
stress occurring during the vegetative growth.
The conventional approach for evaluating cultivar differences in N
use based on the efficiency of N uptake and utilisation reflected only to
some extent the observed genotypic variation in N-related traits and the
ranking of genotypes with regard to NUE changed depending on the
calculation method (i.e. whether or not accounting for soil Nmin at
15
Field Crops Research 270 (2021) 108211
harvest). The characterisation of winter wheat cultivars for NUE traits in
the context of environmental conditions presented here may be benefi­
cial to farmers and breeders, particularly as climate change and
increased climate variability demand a more targeted season-specific N
management in order to minimise inputs in less favourable seasons and
maximise them in seasons with high yield potential.
The details of our responses to the reviewers’ reports are presented in
the following section.
Acknowledgements
We wish to thank Prof. Hermann Bürstmayr (IFA-Tulln) for providing
the infrastructure for experiments, Mr Matthias Fidesser for his excellent
assistance in conducting the field experiments, and the graduate stu­
dents Wolfgang Fuchs, Stefan Peter Ryall, and Lukas Koppensteiner for
their contribution to data collection and analysis.
This study was conducted within the framework of the research
project FarmIT (grant number 859304). Financial support from the
Austrian Research Promotion Agency (FFG) is gratefully acknowledged.
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