全 文 ：Received 4 Jun. 2003 Accepted 19 Sept. 2003
Supported by the National Ministry of Science and Technology (G1998010208, G1998010205).
* SU Jun-Ying and YANG Guo-Hua equally contributed to this work.
** Author for correspondence. E-mail: .
http://www.chineseplantscience.com
Acta Botanica Sinica
植 物 学 报 2004, 46 (3): 302-310
Identification and Characterization of Phosphorus Use Efficiency in a
Doubled Haploid Population of Chinese Spring× Lovrin No.10
YANG Guo-Hua1, 2, SU Jun-Ying1*, LI Bin1, LIU Jian-Zhong1, LI Ming1, XIAO Yan-Mei1,
LI Ji-Yun3, TONG Yi-Ping3**, LI Zhen-Sheng1
(1. State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology,
The Chinese Academy of Sciences, Beijing 100101, China;
2. College of Chemistry and Life Sciences, Tianjin Normal University, Tianjin 300074, China;
3. Research Center for Eco-environmental Sciences, The Chinese Academy of Sciences, Beijing 100085, China)
Abstract: It is reported that chromosome 1R of rye (Secale cereale L.) convey phosphorus use efficient
gene (s), and 1RS/1BL translocation genotype Lovrin No.10 is P use efficient. So we hypothesized whether
P efficient gene(s) locate on 1RS, and the high P efficiency of Lovrin No.10 is from 1RS? To test this
hypothesis, we investigated the P use efficiency (PUE) of a doubled haploid (DH) population with 61 lines
derived from anther culture of F1 hybrid between Lovrin No.10 and phosphorus uptake inefficient genotype
Chinese Spring to see whether PUE differs between DH line with and without 1RS/1BL translocation.
Acidic polyacrylamide-gel electrophoresis (A-PAGE) of gliadin and genomic DNA in situ hybridization (GISH)
were employed to discriminate 1RS/1BL translocation DH lines from the normal 1B DH lines. Among the
61 DH lines investigated, A-PAGE analysis showed that 34 lines contained the 1RS/1BL translocation
chromosome, which was characterized by the presence of a 1RS-specific Sec-1 marker bands. Further
verification with GISH proved that 33 in the 34 lines contained a pair of homozygous 1RS/1BL transloca-
tion chromosomes, only one line was a 1RS/1BL monosomic line. A field experiment was carried out on P
deficient soil to investigate grain yield, biomass, numbers of spikes per plant (SPP), P uptake efficiency
(PUpE), and P utilization efficiency (PUtE) of the DH lines and their parents under -P (nil P applied) and +P
(60 kg P/hm2 applied) at maturity. Results showed soil P deficiency decreased the values of the first four
traits in Lovrin No.10, but were more severe for Chinese Spring. Lovrin No.10 had higher values of all the
above tested traits at both -P and +P than Chinese Spring did, but had similar PUtE with Chinese Spring.
These five traits segregated, and differed greatly among DH lines under both -P and +P conditions.
Although the variations among DH lines exceeded the difference between the two parents, the average
values of the DH lines were between the two parents. The average of the above five traits, and P deficiency
tolerance (PDT) (measured by relative grain yield of -P/+P) were not different between the DH lines with
and without 1RS/1BL translocation. This indicated that there was no association between 1RS and PUE
and PDT in Lovrin No.10, and 1RS may not have P efficient gene(s). Therefore, in the offspring of Lovrin
No.10, it is possible to combine high PUE and PDT with good quality without the negative effect of 1RS on
flour quality.
Key words: phosphorus use efficiency; wheat (Triticum aestivum); DH lines; GISH; A-PAGE; 1RS/1BL
A major constraint to crop production throughout the
world is low availability of the essential macronutrient phos-
phate in soil (Ozanne, 1980; Ae et al., 1990). Phosphorus is
one of the least available of all essential nutrients in the soil
and its concentration is generally below that of many other
macronutrients (Barber, 1963). Many soils around the world
are deficient in Pi, and even in fertile soils available Pi rarely
exceeds 10 µmol/L (Bieleski, 1973), thus P fertilizers are nec-
essary to ensure plant productivity. Because most of P
applied to the soils is immobilized and becomes
unavailable for plant uptake, and the recovery of applied P
by crop plants in a growing season is very low, large amount
of P fertilizers should be applied to overcome soil P
deficiency. During the last 40 years global P fertilization
increased 3.5-fold, even the agricultural food production
only doubled (Tilman, 1999). Now the annual input of P
fertilizers amounts to more than 30 million kg P2O5 (http://
www.fertilizer.org/ifa/). Therefore, breeding crop varieties
with greater efficiency in P acquisition from soils and effi-
cient utilization of P within plant are very desirable.
YANG Guo-Hua et al.: Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of
Chinese Spring×Lovrin No.10 303
Wheat is one of the most important food crops in the
world, while China is the world’s largest wheat producer
(115 million tons per year). It has been reported that wheat
has developed several strategies to adapt to P deficiency
stress, these include the changes of root morphologies,
and enhancing the release of phosphatases and organic
acids to rhizosphere, and up-regulated expression of phos-
phate transporters in roots (Gahoonia et al., 1999; Davies
et al., 2002; Sun and Zhang, 2002a; 2002b). Identifying
genes or QTLs responsible for these P deficiency induced
changes in wheat and its close relatives will be fundamen-
tal to P efficient wheat breeding.
Rye is one of the alien species successfully used in
wheat improvement (Baum and Apples, 1991). Particular
attention has been paid to chromosome 1R in the past de-
cades (Carrillo et al., 1992; Alconso-Blanco et al., 1993;
Orellana et al., 1993; Bush et al., 1995; McIntosh et al.,
1995), because this chromosome is the primary source for
some agronomical important traits, such as disease resis-
tance (Zeller and Fischbeck, 1971; Rajaram et al., 1983).
The short arm of rye chromosome 1R (1RS) carrying resis-
tant genes to several diseases, such as stem rust, leaf rust
and powdery mildew, was incorporated into many high-
yield wheat cultivars as 1RS/1BL translocations and widely
used in wheat production all over the world (Zeller and
Fischbeck, 1971; Rajaram et al., 1983; McIntosh et al., 1995).
Lovrin No.10, an 1RS/1BL translocation line, was intro-
duced into China in the 1970s from Europe and had played
a very important role in Chinese wheat breeding and
production. Two hundred and seventy-nine new varieties
have been selected from hybrid progeny of Lovrin No.10
crossed with other breeding lines in China (Zhang et al.,
2002). Cultivars containing the translocation chromosome
had given exceptionally high yields in fertile and high input
systems with good disease resistance characteristics for a
long time (NIAB, 1987). Our previous results using a set of
wheat-rye (imperial) addition lines demonstrated that genes
responsible for phosphate starvation tolerance and genes
encoding for phosphate starvation inducible acid
phosphatase, which catalyses the hydrolysis of organic P
were located on 1R (Liu et al., 2000). Our screening experi-
ments also proved that Lovrin No.10, an 1RS/1BL translo-
cation line, is a P efficient and P deficient tolerant variety.
In order to clarify whether the P starvation tolerance in
Lovrin No.10 is controlled by 1RS, a doubled haploid (DH)
population derived from Lovrin No.10 and P inefficient geno-
type Chinese Spring was studied in a field trial to establish
relationship between 1RS and P uptake efficiency, P-defi-
ciency tolerance in Lovrin No.10.
1 Materials and Methods
1.1 Plant materials and field trial
Sixty-one DH lines were established by anther culture
of F1 hybrid between P efficient wheat genotype, Lovrin
No.10, and P inefficient genotype, Chinese Spring. Seeds
from single plant of the DH lines were used in this study.
They were grown under field condition on a soil with pH
7.5, 2.2% CaCO3 and 1.3% organic matter. Two P fertiliza-
tion levels were employed. In the – P treatment, the soil
Olsen-P was 8 mg/kg (P), and no P was applied. In the + P
treatment, 60 kg P/hm2 was supplied as diammonium phos-
phate (DAP). Both treatments were applied 180 kg N/hm2
as urea. All fertilizers were applied before sowing. Com-
plete random block designation and four replications were
used in this field trial. In each plot, 15 seeds from each of
the DH lines and their parents were sown in a 75 cm-long
row and the row spacing was 25 cm. The wheat plants were
harvested at physiological mature stage, the biomass, grain
yield and the number of spikes per plant were investigated.
The total P contents in grains and straw were measured by
digesting plant sample with H2SO4-H2O2, then followed by
molybdate/ascorbic acid blue method for P determination
in the digestion solution.
1.2 Acid polyacrylamide-gel electrophoresis (A-PAGE)
To verify the chromosomal components of the DH lines,
each seed was cut into two parts. The part with embryo
was germinated for collecting root tips for chromosome
preparation; and the other part without embryo was used
for A-PAGE of prolamin. In each DH line, three seeds were
randomly selected and used in this study.
Prolamins were extracted as described by Metakovskey
(1991). After centrifugation at 11 000g for 10 min, the super-
natant containing gliadins or secalins was carefully trans-
ferred to a new tube for A-PAGE analysis.
The prolamin extraction solution (~100 µL) obtained by
the last step was mixed with 100 µL of a solution containing
40% (W/V) of Glycerol and 0.05% methyl green. After
centrifugation, 10 µL of the supernatant was fractionated
at pH 3.1 by A-PAGE as described by Bushuk (1978) and
Metakovskey (1991) with minor modifications. A-PAGE was
carried out in acetic acid-glycine buffer. The samples were
run with constant voltage of 500 V under 15 ℃ for 4.5 h
using Bio-Rad, Protean® Xi Cell.
1.3 Genomic in situ hybridization (GISH)
1.3.1 Chromosome preparation Another half of seed
with embryo was germinated and cultured on moist filter
paper until the root was between 1.5 to 2.0 cm long. Root
tips were pretreated in ice water for 22-24 h and then fixed
Acta Botanica Sinica 植物学报 Vol.46 No.3 2004304
in the 3:1 fixing solution (ethanol:acetic acid =3:1, V/V) for
10 h at least.
Fixed root tips were stained in 1% acetocarmine for 5-
10 min at room temperature. Meristematic regions were ex-
cised and squashed in fresh 45% acetic acid. Good slides
were immediately frozen in liquid nitrogen and the cover
slips were removed quickly. The air-dried slides were stored
at –20 ℃ for use.
1.3.2 DNA extraction and probe labeling Total genomic
DNA was extracted from rye and Chinese Spring by CTAB
method (Murray and Thompson, 1980). The genomic DNA
of rye was labeled with digoxigenin-11-dUTP (Boehringer
Mannheim) via nick translation as described in the manual
provided by manufacturer. The total DNA of Chinese Spring
was boiled for 10 min at 100 ℃ and used as blocking DNA.
1.3.3 Genomic DNA in situ hybridization Genomic DNA
in situ hybridization was performed according to the proto-
cols of Chen and Armstrong (1994) with minor modifications.
Chromosomal DNAs on the slides were treated with Pro-
teinase K (0.3 µg/mL) and RNase A (100 µg/mL, in 2×
SSC) at 37 ℃ for 10-30 min and 1 h, respectively, and then
followed by dehydration in cool ethanol (70%, 95% and
100% for 5 min each) and then dried in air. Chromosomes
on the slides were denatured at 70 ℃ for 2 min in 70%
formamide in 2×SSC followed by immediate dehydration
in cool ethanol (70%, 95% and 100% for 5 min each) then
air-dried. The hybridization mixture was prepared by add-
ing approximately 50 ng of rye probe, 10 times of wheat
blocking DNA, and 5 µg of sonicated salmon sperm DNA
in hybridization buffer containing 50% (V/V) formamide, 2
×SSC, and 10% (W/V) dextran sulphate. The probe mix-
ture was denatured at 100℃ for 10 min and quickly cooled
on ice for at least 3 min before hybridization. Ten µL of
denatured probe mixture was loaded for each slide. The
slide was covered with 22×22 mm parafilm and then placed
in a humid chamber and incubated at 60 ℃ and 37 ℃ for 10
min and 12 h respectively.
Post hybridization washing consisted of a 5-min immer-
sion in 2×SSC at 37 ℃, then by a stringent wash in 40%
(V/V) formamide in 2×SSC, 0.1×SSC, and 2×SSC at
42 ℃, respectively for 5 min followed by wash in 2×SSC
and 4×SSC (containing 0.2% Tween 20) at room tempera-
ture for 5 min each, and then drained. Detection buffer (5%
BSA in 4×SSC) was loaded onto slides and incubated at
37 ℃ for 30 min, then detection solution (containing 5%
BSA and 10 µg/mL anti-digoxigenin-fluorescein, Fab frag-
ments (Boehringer Mannheim) in 4×SSC was loaded on
the slides and kept at 37 ℃ for 1 h. After the incubation,
unbound fluorescence was washed away in 4×SSC for 5
min at RT. The slides were counterstained with 0.2 µg/mL
propidium iodide (in 1×SSC) at room temperature for 10
min, then washed in 1×SSC for 5 min at RT. Twenty µL of
fluorescence anti-fade solution (Vector laboratories) con-
taining 1 µg/mL DAPI was applied to each slide. The hy-
bridization signal was examined with Olympus BX60. Im-
ages were taken by the digital camera.
2 Results
2.1 Detection of 1RS /1BL translocation in the DH popu-
lation by A-PAGE analysis of prolamins
The short arm of 1B in Lovrin No.10 was substituted by
the 1RS. The Sec-1 proteins can be used as diagnostic
markers for the 1RS (Orellana et al., 1993; Ru et al., 2002).
The A-PAGE results showed that 34 out of the 61 lines
contained the 1RS/1BL translocation chromosome, which
was characterized by the presence of 1RS-specific Sec1
bands (Fig.1). The approximately 1:1 (34:27) ratio between
with and without 1RS/1BL translocation chromosome in-
dicated that the chromosomes in F1 hybrid (Chinese Spring
×Lovrin No.10) had been randomly segregated into male
gametes (pollen) through meiosis. Because A-PAGE can
only detect the presence or not of the 1RS, the information
about the dosages of 1RS and chromosome constitutions
of DH lines containing 1RS need to be verified by GISH.
2.2 Verification of chromosome components of the 34 DH
lines by GISH
In order to obtain the accurate information about the
dosage of 1RS/1BL translocation chromosome and chro-
mosome constitution, GISH was applied to detect the chro-
mosome components of the 1RS/1BL translocation lines
screened out by A-PAGE. Our GISH results showed that 33
out of the 34 DH lines contained a pair of homologous
1RS/1BL translocation chromosomes (Fig.2a-c). One line
was proven to be a monosomic line (2n = 41), containing
only one 1RS/1BL translocation chromosome in its nuclear
(Fig.2d, e).
2.3 P efficiencies and P deficient tolerance of the two
parents
Here P efficiencies are evaluated as PUpE (P accumula-
tion in above-ground part), PUtE (yield produced by unit P
uptake), and agronomic efficiency (yield on a given soil P
level). PDT is measured as the relative trait value of –P/+P.
As shown in Table 1, biomass and grain yield of Lovrin No.
10 are significantly higher than those of Chinese Spring at
both + P and - P treatments. For example, at – P, the biom-
ass and grain yield of Lovrin No.10 were 2.6-fold of Chi-
nese Spring. The higher yield of Lovrin No.10 was mainly
related with its higher PUpE. The PUtE of these two
YANG Guo-Hua et al.: Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of
Chinese Spring×Lovrin No.10 305
Fig.1. A-PAGE micrographs of prolamins from 61 DH lines. The arrows indicate the secaline proteins encoded by genes on 1RS(Sec-
1), which could be used as biochemical markers of 1RS. C stands for Chinese Spring, L for Lovrin No.10, 1 for DH1, 2 for DH2, etc. From
the A-PAGE micrograph, DH lines which do not contain 1RS/1BL translocation are DH: 2, 3, 4, 5, 7, 13, 17, 19, 20, 21, 23, 24, 26, 28,
29, 30, 32, 33, 38, 40, 41, 44, 54, 56, 57, 58, 66. DH lines which contain 1RS/1BL translocation are DH: 1, 6, 8, 9, 10, 11, 12, 14, 15, 16,
18, 22, 25, 27, 31, 39, 42, 43, 45, 46, 47, 48, 50, 53, 55, 59, 60, 61, 62, 63, 64, 65, 67, 68.
Fig.2. Detection of chromosomal fragments
from rye in DH population by genomic in situ
hybridization (GISH). a. Two hybridization sig-
nals detected in metaphase cell (with low concen-
tration of propidium iodide counterstaining). b.
Two hybridization signals detected in prophase
cell (without propidium iodide counterstaining).
c. Two hybridization signals detected in inter-
phase cell. d. One hybridization signal detected
in metaphase cell of DH14. e. One hybridization
signal detected in interphase cell of DH14.
Acta Botanica Sinica 植物学报 Vol.46 No.3 2004306
parents were similar. Biomass, grain yield, SPP and PUpE of
Lovrin No.10 and Chinese Spring are significantly reduced
by P deficiency, but the reductions were severer in Chinese
Spring. This indicates that Lovrin No.10 not only has high
P efficiencies, but also has strong tolerance to low P.
2.4 Characterization of DH lines on P efficiencies and P
deficient tolerance
Significant differences existed on P efficiencies in the
DH population, which were reflected by the biomass, grain
yield, PUpE and PUtE (Table 1). In the – P treatment, the
highest grain yield was 13 times of the lowest. The average
value of biomass, grain yield and P uptake of DH lines was
between those of two parents respectively. P starvation
led the biomass, grain yield and SPP to be reduced by about
20%-30%, which was revealed by analysis of the average
of –P/+P relative values. However, the average values of all
the investigated traits were not significantly different be-
tween DH lines with and without 1RS/1BL (Table 2). This
suggested that the tolerance to P starvation should not be
controlled by the 1RS in Lovrin No.10. The differences on
P uptake efficiency and P utilization efficiency in the DH
population implied that breeding for high P uptake effi-
ciency and high P utilization efficiency in wheat cultivars
was practical.
3 Discussion
Transfer of agronomical important genes from relative
species into hexaploid wheat via wild hybridization is an
important approach for wheat improvement. Rye has been
shown to be one of the most successful donors for this
purpose because of the traits it possessed (Rajaram et al.,
1983). Anther culture has been proven to play very impor-
tant roles in the transfer of genes between species and
construction of DH populations, because of its advantages
in time-saving, maintenance of maximum variations and fast
genetic stability. In this study, a DH population was con-
structed by anther culture of F1 hybrid between P efficient
genotype, Lovrin No.10 and P inefficient genotype, Chi-
nese Spring. A-PAGE results for prolamins firmly demon-
strated that the ratio between lines with and without the
1RS/1BL translocated chromosome in the DH population
was approximately 1:1 (34:27). In addition, besides the
Table 1 Comparison of P efficiency and P deficient tolerance between DH lines and their parents

Lovrin No.10 Chinese Spring
DH lines
(range value/mean（n = 61）)
+P -P -P/+P (%) +P -P -P/+P (%) +P -P -P/+P (%)
Biomass (g/plant) 21.9±0.7 18.3±1.5 83.6 9.2±0.5 7.0±0.6 76.1 3.4--22.2 2.3--18.1 48.8--103.2
13.9 10.6 75.6
Grain yield (g/plant) 8.1±0.3 7.3±0.6 90.1 3.7±0.2 2.8±0.4 75.7 1.1--8.4 0.5--7.1 48.4--111.8
5.0 4.1 80.8
Spikes/plant 5.8±0.3 4.4±0.2 75.9 3.7±0.2 2.5±0.4 67.6 1.9--7.1 1.5--5.7 48.6--89.5
4.5 3.2 71.5
P uptake 27.7±1.0 21.3±1.6 76.9 11.2±0.6 8.0±1.1 76.9 5.1--28.5 2.3--24.7 45.1--100.7
(mg p/plant) 17.6 13.6 76.3
P utilization 291.0±11.0 341.0±28.0 117.2 329.0±18.0 353.0±50.0 107.3 220.0--374.0 231.0--423.0 70.3--138.7
(g GW/g P) 287.0 304.0 106.6
Table 2 Comparison of P uptake efficiency and tolerance to P deficiency between DH lines with 1B/1R translocation (Genotype R) and
without 1B/1R translocation (Genotype C)
Genotype with Genotype without
1BL/1RS (n=34) 1BL/1RS (n=27)
+P -P -P/+P (%) +P -P -P/+P (%)
Range/mean ± SD
Biomass (g/plant) 5.5--22.2 2.7--18.1 48.8--101.8 3.4--21.5 2.3--16.1 54.9--103.2
14.0±3.7 10.7±3.5 76.0±12.3 13.9±4.2 10.8±3.4 72.4±10.5
Grain yield (g/plant) 1.9--7.8 0.9--6.8 49.7--111.8 1.1--8.4 0.5--7.1 48.4--100.1
5.0±1.4 4.2±1.4 81.3±13.8 5.2±1.6 4.2±1.4 78.5±12.2
Spikes/plant 2.1--6.1 1.5--4.9 48.6--88.2 1.9--7.1 1.6--5.7 54.6--89.5
4.4±1.0 3.2±0.9 71.9±9.4 4.6±1.3 3.3±1.0 72.5±9.3
P uptake (mg P/plant) 5.7--28.3 4.1--21.4 49.6--100.7 5.1--28.5 2.3--24.7 45.1--95.1
17.6±4.8 13.7±4.2 76.8±12.8 17.9±5.3 13.8±4.5 73.7±12.8
Ugw (g GW/g P) 232.0--374.0 231.0--423.0 70.3--138.7 221.0--352.0 237.0--369.0 83.3--127.6
289.0±38.0 305.0±45.0 106.9±11.6 288.0±35.0 304.0±34.0 106.3±10.4
YANG Guo-Hua et al.: Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of
Chinese Spring×Lovrin No.10 307
differences caused by the presence of 1RS and absence of
1BS, the band patterns for prolamins in A-PAGE diagram
differed from line to line suggested that the random recom-
binant of alleles controlling the expression of prolamins on
the other chromosomes had also occurred within the DH
population (Fig.1). Prolamins have been widely used as
reliable genetic markers for identifications of cultivar’s
identity, seed purity and hybrid descendants in cereals,
because their components are genetically stable and not
affected by environment (Kreis et al., 1985). However, the
information regarding copy number of a target gene or a
chromosome and chromosome constitutions cannot be told
using prolamins as genetic markers. In addition, it has been
reported that microspore cloning variations and changes
of both chromosome numbers and chromosome structures
can occur among anther culture-derived plants (Hu et al.,
1980; Zhang et al., 1998). Therefore, GISH is necessary in
the 34 lines to address this question. Our GISH results
showed that the majority of DH lines (33 out of 34) tested
contained a pair of homologous 1RS/1BL translocation
chromosomes with 42 chromosomes in their genomes (Fig.
2a-c), only one line was proven to be a monosomic (2n =
41) containing only one 1RS/1BL translocation chromo-
some in its genome (Fig.2d, e). These results indicated that
the majority of DH lines was quite stable in terms of chro-
mosome numbers and structures. This claim was further
supported by the microscopic examination results of chro-
mosome number in the root tip cells of the other 27 DH lines
without 1RS/1BL. Hu et al. (1980) reported that in wheat,
90% of the pollen plants derived via anther culture are hap-
loid or homozygous diploid, whereas the residual 10% are
heteroploid and aneuploid. The small size of our DH popu-
lation may probably be one of the explanations for why no
heteroploid and few aneuploids (1/61) appeared in our tested
DH lines. The fact that the signals for alien chromosome
could be detected at most cell division phases from inter-
phase to metaphase (Fig.2). This further demonstrated that
GISH is a powerful and direct technique to detect the alien
chromosome(s). However, GISH alone cannot correctly
identify the donor of alien chromatin in some cases (Ru et
al., 2002).
In a set of wheat-rye addition lines it was demonstrated
that genes responsible for PDT and genes for encoding
phosphate starvation inducible acid phosphatase, which
catalyses the hydrolysis of organic P and releases the Pi
available for plants, were located on 1R of Imperial (Liu et
al., 1997). Results in present research indicated that genes
controlling PUE and PDT are not located on 1RS in Lovrin
No. 10. They may be located on 1RL. Another explanation
is that chromosome 1R of wheat-rye (Imperial) addition lines
in the study of Liu et al. (1997) is different from the chromo-
some 1RS in Lovrin No.10 which is from Rye Petkus
(Alkhimova et al., 1999). It is well known that the 1RS brings
itself some excellent traits, such as high yield, and disease
resistance. On the other hand, it has very strong negative
effect on bread and noodle making quality. From the result
of this experiment, it is clear that PUpE and PDT are not
correlated with 1RS in Lovrin No.10 (Table 2). Therefore, it
is possible to breed varieties of high PUE and PDT without
1RS in progeny of Lovrin No. 10. In other words, it is quite
possible to combine high PUpE, PUtE, PDT and good pro-
cessing quality in the offspring of Lovrin No.10. SSR tech-
nique is being employed to map the genes controlling the
high P uptake and utilization efficiency of Lovrin No.10 in
our group.
In summary, a DH population has been constructed for
Lovrin No.10, a high P uptake efficient variety with1RS/
1BL translocation. A-PAGE and GISH analysis revealed
there were 34 lines with 1BL/1RS translocation, and 27 lines
without this translation. Performance of the two type mate-
rials under normal and starvation of phosphorus nutrient
showed that P uptake and utilization efficiency had no cor-
relation with the 1RS in Lovrin No.10.
Acknowledgements: We appreciate Prof. HU Han and
Dr. ZHANG Xue-Yong for their critical review of this
manuscript. Thanks should be given to Mrs. WEN Yu-Xiang
for her kind help in preparation of micrographs.
References:
Ae N , Arihara J, Okada K, Yoshihara T, Johansen C. 1990.
Phosphorus uptake by pigeon pea and its role in cropping
systems of the Indian subcontinent. Science, 248:477-480.
Alconso-Blanco C, Goicoechea P G, Roca A, Giraldez R. 1993. A
cytogenetic map on the entire length of rye chromosome 1R,
including one translocation breakpoint, three isozyme loci and
four C-bands. Theor Appl Genet, 85:735-744.
Alexandratos N. 1999. World food and agriculture: outlook for
the medium and longer term. Proc Natl Acad Sci USA, 96:
5908-5914.
Alkhimova A G, Heslop-Harrison J S, Shchapova A I, Vershinin A
V. 1999. Rye chromosome variability in wheat-rye addition
and substitution lines. Chromosome Res, 7:205-212.
Barber S A, Walker J M, Vasey E H. 1963. Mechanism for the
movement of plant nutrients from the soil and fertilizer to the
plant root. J Agric Food Chem, 11:204-207.
Baum M, Appels R. 1991. The cytogenetic and molecular archi-
tecture of chromosome 1R-one of the most widely used sources
Acta Botanica Sinica 植物学报 Vol.46 No.3 2004308
of alien chromatin in wheat varieties. Chromosoma, 101:1-
10.
Bieleski R L. 1973. Phosphate pools, phosphate transport, and
phosphate availability. Annu Rev Plant Physiol, 24:225-252.
Bushuk W Z. 1978. Wheat cultivar identification by gliadin
electrophoregrams Ⅰ Apparatus, method and nomenclature.
Can J Plant Sci, 58:505-515.
Cakmak I, Derici R, Torun B, Tolay I, Braun H J, Schlegel R.
1997. Role of rye chromosomes in improvement of zinc effi-
ciency in wheat and triticale. Plant Soil, 196:249-253.
Carillo J M, Vàzquez J F, Orellana J. 1992. Identification and
mapping of the Gli-R3 locus on chromosome 1R of rye (Secale
cereale L.). Theor Appl Genet, 84:237-241.
Cassman K G. 1999. Ecological intensification of cereal produc-
tion systems: yield potential, soil quality, and precision
agriculture. Proc Natl Acad Sci USA, 96:5952-5959.
Chen Q, Armstrong K. 1994. Genomic in situ hybridization in
Avena sativa. Genome, 37:607-612.
Davies T G E, Ying J, Xu Q, Li Z S, Li J, Gordon-Weeks R. 2002
Expression analysis of putative high-affinity phosphate trans-
porters in Chinese winter wheats. Plant Cell Environ, 25:
1325-1340.
Gahoonia T S, Nielsen N E, Lyshede O B. 1999. Phosphorus (P)
acquisition of cereal cultivars in the field at three levels of P
fertilization. Plant Soil, 211:269-281.
Gill B S, Friebe B, Endo T R. 1991. Standard karyotype and
nomenclature system for description of chromosome bands
and structure aberrations in wheat (Triticum aestivum).
Genome, 34:830-839.
Hu H, Xi Z Y, Ouyang J W, Hao S, He M Y, Xu Z R, Zhou M Q.
1980. Chromosome variation of pollen mother cell of pollen-
derived plants in wheat (Triticum aestivum L.). Sci Sin, 23:
905-912.
Jiang J, Friebe B, Gill B S. 1994. Recent advances in alien gene
transfer in wheat. Euphytica, 73:199-212.
Kreis M, Shewry P R, Ford B, Ford J, Miflin B J. 1985. Structure
and evolution of seed storage proteins and their genes with
particular reference to those of wheat, barley and rye. Oxford
Surv Plant Mol Cell Biol, 2:253-317.
Lawrence G J, Shepherd K W. 1981. Chromosomal location of
genes controlling seed proteins in species related to wheat.
Theor Appl Genet, 59:25-31.
Le H T, Armstrong K C, Miki B. 1989. Detection of rye DNA in
wheat-rye hybrids and wheat translocation stocks using total
genomic DNA as a probe. Plant Mol Biol Rep, 7:150-158.
Li J-Y , Liu X-D, Zhou W, Sun J-H, Tong Y-P, Liu W-J, Li Z-S ,
Wang P-T , Yao S-J . 1995. Study on the new technology of
crop breeding on available utilizing phosphate in soils. Sci
China (Ser B) , 25:41-48. (in Chinese with English abstract)
Liu J-Z , Li B, Li J-Y, Li Z-S . 1997. Genetic studies of gene
effects of rye genome on characteristics of soil phosphorus.
Act Genet Sin, 24:519-523. (in Chinese with English abstract)
Liu J Z, Li Y J, Tong Y P, Gao J W, Li B, Li J Y, Li Z S. 2001.
Chromosomal location of genes conferring the tolerance to Pi
starvation stress and acid phosphatase (APase) secretion in
the genome of rye (Secale L.). Plant Soil, 237:267-274.
Liu J-Z , Li Y-J, Li B , Yao S-J, Li J-Y, Li Z-S. 2000. Genetical
effect of different rye chromosomes on the acid phosphatase
(Acph) secretion of common wheat roots under phosphorus
starvation conditions. Act Genet Sin, 27:39-43. (in Chinese
with English abstract)
Lopez-Bucio J, de La Vega O M, Guevara-Garcia A, Herrera-
Estrella L. 2000. Enhanced phosphorus uptake in transgenic
tobacco plants that overproduce citrate. Nat Biotechnol, 18:
450-453.
Marschner H. 1995. Mineral Nutrition of Higher Plants. 2nd ed.
San Diego: Academic press.
McIntosh R A, Hart G E, Gale M D. 1995. Catalogue of gene
symbols for wheat. Li Z S, Xin Z Y . Proceedings of 8th
International Wheat Genetics Symposium. Beijing: China Ag-
ricultural Sci-tech Press. 1333-1500.
Metakovsky E V. 1991. Gliadin allele identification in common
wheat. Ⅱ. Catalogue of gliadin alleles in common wheat. J
Genet Breed, 5:325-343.
Mitsukawa N, Okumula S, Shirano Y, Sato S, Kato T, Harashima
S, Shibata D. 1997. Overexpression of an Arabidopsis thaliana
high-affinity phosphate transporter gene in tobacco cultured
cells enhances cell growth under phosphate-limited conditions.
Proc Natl Acad Sci USA, 94:7098-7102.
Ni J J, Wu P, Senadhira D, Huang N. 1998. Mapping QTLs for
phosphorus deficiency tolerance in rice (Oryza sativa L.).
Theor Appl Genet, 97:1361-1369.
Orellana J, Fernàndez-Calvin B, Vàzquez J F, Carillo J M. 1993.
Mapping of genes controlling seed storage-proteins and cyto-
logical markers on chromosome 1R of rye. Theor Appl Genet,
85:639-645.
Ozanne P G. 1980. Phosphate nutrition of plants-a general treatise.
Khasawneh E. The Role of Phosphorus in Agriculture. Madi-
son WI: American Society of Agronomy. 559-585.
Raghothama K G. 2000. Phosphate transport and signaling. Curr
YANG Guo-Hua et al.: Identification and Characterization of Phosphorus Use Efficiency in a Doubled Haploid Population of
Chinese Spring×Lovrin No.10 309
Opin Plant Biol, 3:182–187.
Rajaram S, Mann C E, Ortiz-Ferrara G, Mujeeb-Kazi A. 1983.
Adaptation, stability and high yield potential of certain 1B/
1R CIMMYT wheats. Sakamoto S. Proceedings of 6th Inter-
national Wheat Genetics Symposium. Kyoto, Japan: Plant
Germplasm Institute, Kyoto University. 613-621.
Ru Y-Y , Zhang X-Y , Li D-Y , You G-X, Yan Y-M. 2002. Risk in
explanation of GISH signals: Enlightenment from verification
of alien chromatin in a wheat translocation line (A-3) by GISH.
Acta Agric Sin , 28:6-10. (in Chinese with English abstract)
Sakano K. 1990. Proton/phosphate stoichiometry in uptake of
inorganic phosphate by cultured cells of Catharanthus roseus
(L.) G. Don. Plant Physiol, 93:479-483.
Schachtman D P, Reid R J, Ayling S M. 1998. Phosphorus uptake
by plants: from soil to cell. Plant Physiol, 116:447–453.
Sun H-G, Zhang F-S. 2002a. Effect of phosphorus deficiency on
activity of acid phosphatase exuded by wheat roots. Chin J
Appl Ecol, 13:379-381. (in Chinese with English abstract)
Sun H-G , Zhang F-S. 2002b. Morphology of wheat roots under
low-phosphorus stress. Chin J Appl Ecol , 13:295-299. (in
Chinese with English abstract)
Tilman D. 1999. Global environmental impacts of agricultural
expansion: the need for sustainable and efficient practices.
Proc Natl Acad Sci USA, 96:5995-6001.
Tilman D, Cassman K G, Matson P A A, Naylor R, Polasky S.
2002. Agricultural sustainability and intensive production
practices. Nature, 418:671-677.
Wang M L, Atkinson M D, Chinoy C N, Devos K M, Harcourt R
L, Liu C J, Rogers W J, Gale M D. 1991. RFLP-based genetic
map of rye (Secale cereale L.) chromosome 1R. Theor Appl
Genet, 82:174-178.
Wissuwa M, Wegner J, Ae N, Yano M. 2002. Substitution map-
ping of Pup1: a major QTL increasing phosphorus uptake of
rice from a phosphorus-deficient soil. Theor Appl Genet, 105:
890-897.
Wissuwa M, Yano M, Ae N. 1998. Mapping of QTLs for phos-
phorus-deficiency tolerance in rice (Oryza sativa L.). Theor
(Managing editor: HE Ping)
Appl Genet, 97:777-783.
Zeller F J, Fischbeck G. 1971. Cytologische untersuchungen zur
Identifizierung des Fremdchromosoms in der Weizensorte
Zorba (W565). Z Pfanzenzücht, 66:260-265.
Zhang X Q, Wang X P, Jing J K, Hu H, Ross K, Gustafson J P.
1998. Characterization of wheat-triticale doubled haploid lines
by cytological and biochemical markers. Plant Breeding, 117:
7-12.
Zhang X-Y, Pang B-S, You G-X, Wang L-F, Jia J-Z, Dong Y-C .
2002. Allelic variation and genetic diversity at Glu-1 loci in
Chinese wheat (Triticum aestivum L.) germplasms. Sci Agric
Sin , 35:1302-1310. (in Chinese with English abstract)
Zhu Y G, Smith S E, Smith F A. 2001. Plant growth and cation
composition of two cultivars of spring wheat (Triticum
aestivum L.) differing in P uptake efficiency. J Exp Bot, 52:
1277-1282.
Zhu Y G, Smith S E, Barritt A R, Smith F A. 2001. Phosphorus
(P) efficiencies and mycorrhizal responsiveness of old and
modern wheat cultivars. Plant Soil, 237:249-255.
Zhu Y G, Smith S E, Howes N K, Smith E A. 2001. Phosphorous
(P) uptake efficiency of doubled haploid lines of spring wheat
derived from parents with different P uptake efficiency. Horst
W J, Schenk M K, Bürkert A, Claassen N, Flessa H, Frommer
W B, Glodbach H, Olfs H W, Römheld V, Sattelmacher B,
Schmidhalter U, Schubert S, Wirén N V, Wittenmayer L.
ⅩⅣ. International Plant Nutrition Colloquium. Hannover:
Kluwer Academic Publishers. 70-71.