378 DOI: 10.1002/jpln.200800192 J. Plant Nutr. Soil Sci.

2009, 172, 378±384

Effect of phosphite±phosphate interaction on growth and quality of

hydroponic lettuce (Lactuca sativa)
Hoang Thi Bich Thao1*, Takeo Yamakawa2, and Katsuhiro Shibata1
1 Plant Nutrition Laboratory, Division of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka
812-8581, Japan
2 Plant Nutrition Laboratory, Department of Plant Resources, Faculty of Agriculture, Kyushu University, Fukuoka, 812±8581 Japan

Abstract
Although the fungicidal properties of phosphite have been recognized, its potential as a fertilizer
is still being debated. The information on how phosphite affects the growth and quality of plants
in relation to phosphate (Pi) also remains unknown. This study was conducted to investigate the
effect of phosphite in relation to Pi on growth and quality parameters of lettuce (Lactuca
sativa L.). The results showed that addition of phosphite to the nutrient solution at different rates
ranging from 0.05 to 2 mM significantly increased total P, water-extractable Pi, and phosphite in
both shoots and roots, but did not improve plant growth under various Pi supplies (0.05, 0.1,
0.15, and 0.3 mM as Pi levels for approximately 50%, 80%, 90%, and 100% of the maximum
plant growth, respectively), indicating that phosphite was well absorbed by roots and mobile
inside the plants, but did not provide any P nutrition. Also, no stimulating effect of any Pi±phos-
phite combination was observed. The effect of phosphite on plant growth was strongly depen-
dent on the level of Pi supply. In general, application of phosphite up to 2 mM did not influence
the growth of Pi-sufficient plants. However, plants fertilized with Pi for about 90% of maximum
growth were still vulnerable to phosphite at 2 mM. The negative effect of phosphite was found
even at concentrations as low as 0.2 mM, when plants were supplied with Pi adequate for about
80% of maximum growth or less. At 0.05 mM, phosphite had marginal effects on plant growth
under all the Pi levels. Although phosphite itself had little influence on the ascorbate and mineral
concentrations of lettuce, its application to Pi-deficient plants may decrease the mineral concen-
trations of plants brought about by the inhibitory effect of phosphite on root growth and hence
nutrient uptake. Since phosphite is an effective fungicide for lettuce, care should be taken on Pi
supplies prior to application of phosphite products to minimize the harmful effects.

Key words: ascorbate / phosphorus / phosphate±phosphite interaction / plant growth / quality

Accepted January 31, 2009

1 Introduction
Phosphite, a reduced form of phosphate (Pi), is widely
marketed either as fungicide or as a phosphorus (P) fertilizer
for a wide range of crops (McDonald et al., 2001; Rickard,
2000). Although it is commonly accepted that phosphite pro-
ducts are excellent fungicides (e.g., Al-fosetyl, the active
agent of Alliette), the claim regarding their potential as a fer-
tilizer is controversial. Lovatt (1990a, b) reported that phos-
phite application may replace Pi in some plants such as citrus
and avocado crops suffering from P deficiency. Rickard
(2000) summarized studies on crop responses to commercial
phosphite fertilizers and concluded that these products
caused consistent improvements in yield and quality of var-
ious crops. In contrast, there were also numerous studies
indicating that phosphite has negative effects on plant growth
and is not utilized in plants as nutrient source (Carswell et al.,
1996, 1997; Forster et al., 1998; Abel et al., 2002; Schroetter
et al., 2006). It was shown that phosphite may suppress Pi
starvation responses of plants (Ticconi et al., 2001; Varadara-
jan et al., 2002; Carswell et al., 1996, 1997). However, most
of these studies used phosphite as a sole P source or at high
rates with some of them more or less focusing on the toxicity
of phosphite to dissect P starvation responses of plants at
molecular level. The information on how phosphite affects
growth and quality of plants in relation to various Pi supplies
remained unclear.
A recent study (Thao et al., 2008a) indicated that the effect of
phosphite on growth of komatsuna was dependent on the Pi
status of the plants and suggested that those insufficiently
fertilized with Pi may become vulnerable even to a low phos-
phite concentration. The interest in using phosphite either as
a part of a total production package (Lovatt and Mikkelsen,
2006) or as a fungicide is increasing. In agricultural produc-
tion, Pi limitation is frequently encountered because P is
required in relatively high amounts but often sequestered in
chemical forms that are not readily accessible by plants.
Fertilization of P sometimes is preferred at economic opti-
mum rates, which may be lower than the rates for maximum
yield (sufficient level). The effects of phosphite on growth and
quality of plants in relation to various Pi supplies therefore

* Correspondence: Dr. H. T. B. Thao;

e-mail: tb_hoang@yahoo.com

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J. Plant Nutr. Soil Sci. 2009, 172, 378±384
merit further understanding to minimize negative effects
resulting from the use of this material, especially for crops
with higher probability of phosphite application. Recently, the
Canadian Food Inspection Agency stated that products con-
taining phosphite should not be applied when soil phosphorus
levels are suboptimal and that such products should be
applied in conjunction with a fertilizer that contains an avail-
able form of phosphate (Anonymous, 2008).
This study therefore investigated the effects of phosphite on
growth of lettuce at various Pi levels in order to examine (1)
the nutritional value of phosphite as a source of P, (2) the
effect of phosphite on plant growth in relation to Pi supplies,
(3) whether phosphite may have any stimulating effect on
plant growth, and (4) the effect of phosphite on quality para-
meters of this crop. Lettuce was selected because phosphite
is known as an effective fungicide for controlling disease of
this plant caused by Oomycetes.
2 Materials and methods
Two experiments (Pi-response experiment and phosphite
experiment) were conducted in a glasshouse at Kyushu Uni-
versity, Japan (3337 N, 13025' E, 3 m above sea level).
For both experiments, seedling preparation was conducted in
a 20C-controlled room under natural light condition. Seeds
of lettuce (Lactuca sativa L. cv. Ueaheddo) purchased from
Nichinou Co. (Saga, Japan) were germinated and grown for
2 weeks in plastic cell-trays containing peat-based compost
(Tamemaki-baido; Takii & Co. Ltd., Kyoto, Japan) with ade-
quate water supply.
2.1 Phosphate-response experiment
The experiment was carried out from October 13 to Novem-
ber 5, 2007 (day length 10.7±11.5 h, temperature range
12C±18C min., 20C±32C max.). After growing for
2 weeks, uniform seedlings were selected, carefully washed,
and transferred into 7 L hydroponic pots (six plants pot±1)
containing modified Hoagland nutrient solution (pH 6.2) with
various Pi concentrations. This basic nutrient solution con-
sisted of 0.94 mM K2SO4, 0.38 mM KCl, 6 mM NaNO3,
0.77 mM MgSO4, 1.93 mM CaCl2, 0.14 mM NaFe-EDTA,
0.29 lM ZnSO4, 3.47 lM MnSO4, 0.12 lM CuSO4, 17.62 lM
H3BO3, 0.04 lM MoO3. Phosphate was separately supplied
as H3PO4 to each pot at various concentrations of 0.025,
0.05, 0.1, 0.25, 0.5, and 1 mM. Each treatment had three
pots arranged in three randomized complete blocks. The
nutrient solutions were aerated continuously using air pumps,
and their pH was adjusted to 6.2 every 2 d by addition of 1 M
H2SO4 or 1 M NaOH as needed. Since adjustment to pH 6.2
required a very small amount of H2SO4 or NaOH, the effect
on the concentrations of Na+ and SO24ÿ is considered negligi-
ble. The nutrient solutions were renewed every 7 or 6 d.
Plants were harvested at 23 d after transplanting (DAT).
2.2 Phosphite experiment
The experiment was carried out from November 7 to 30, 2007
(day length 10.2±10.7 h, temperature range 10C±15C min.,
ã 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Phosphate±phosphite interaction effects on lettuce 379
16C±30C max.) using the same hydroponic pot system (six
plants pot±1) and the basic nutrient solution as described
above. This experiment had 16 treatments that combined
four Pi and four phosphite concentrations as a 4 ” 4 factorial
in three randomized complete blocks. The phosphite concen-
trations were 0, 0.05, 0.2, and 2 mM. The selection of Pi con-
centrations were 0.05, 0.1, 0.15, and 0.3 mM based on the
growth response of lettuce to Pi supply from the Pi-response
experiment to provide Pi for approximately 50%, 80%, 90%,
and 100% (sufficient Pi) of the maximum plant growth, re-
spectively. The Pi and phosphite used were, respectively,
phosphoric acid (H3PO4) and sodium phosphite (Na2HPO3).
In preparing stock phosphite solution, the Na was removed
by eluting the solution through a column of cation-exchange
resin (Dowex 50 W [H+]). Other experimental methods were
the same as described in the above experiment. Plants were
harvested at 23 DAT.
2.3 Plant analysis
In the Pi-response experiment, after harvesting, the fresh
weight (FW) of shoots was immediately recorded. In the
phosphite experiment, three plants per pot were harvested
for measuring FW of shoots and roots after being blotted dry
between absorbent papers. These plants then were washed
in deionized water and stored at ±80C before freeze-drying
and grinding for chemical analysis. Ascorbate and nitrate
reductase activity (NRA) assay were determined for three
levels of Pi supply (0.05, 0.15, and 0.3 mM) using the young-
est fully expanded leaves from the remaining three plants per
pot.
Total P, nitrogen (N), and potassium (K): The dried-ground
shoot samples were digested using an H2SO4-H2O2 Kjeldahl
digestion method (Ohyama et al., 1991), then analyzed for
total P (Murphy and Riley, 1962), total N (Cataldo et al.,
1994), and total K (atomic-absorption spectrophotometer).
Water-extractable Pi and phosphite: The procedure for
analysis of water-extractable Pi and phosphite in the plant
samples was described in Thao et al. (2008a). In brief, dried
plant samples (0.05 g) were extracted with 8 mL of deionized
water in a horizontal shaker (250 rpm) at room temperature
for 1 h, and then in a water bath shaker at 70C for 2 h
followed by a centrifugation at 13,000 rpm for 10 min. The
supernatants were filtered through a 0.2 lm cellulose acetate
filter and analyzed for Pi and phosphite using ion chromato-
graphy.
Calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), and
manganese (Mn): Dried-ground shoot samples were
digested in a mixture of nitric and perchloric acid (3 HNO3 : 2
HClO4), then analyzed for Ca, Mg, Fe, Zn, and Mn using the
atomic-absorption spectrophotometer.
Nitrate: Dried-ground shoot samples were extracted for 1 h
with deionized water at 70C in a water-bath shaker followed
by nitrate analysis using the salicylic acid method (Cataldo
et al., 1975).
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380 Thao, Yamakawa, Shibata
Total ascorbate assay: Fresh leaves (harvested between
9:00 and 10:00 a.m.) were washed, blotted dry, and cut into
5 mm diameter discs. The leaf discs (250 mg per sample)
were immediately stored at ±80C. Total ascorbate was
assayed in the next day using a microplate-adapted colori-
metric method (Gillespie and Ainsworth, 2007).
NRA assay: NRA was assayed in vivo by measuring NOÿ2
production in tissue that had been vacuum-infiltrated with
buffered NO3± solution. The assay is similar to the method of
Down et al. (1993) except that 0.25 M KNO3 in 0.05 M Tris-
HCl was used as the buffer solution.
2.4 Statistical analysis
The statistical analyses were performed using IRRISTAT for
Windows version 4.0 (Biometric Unit, International Rice
Research Institute). Not detectable (ND) data were considered
as zero in ANOVA analysis. Root P data were transformed tak-
ing the natural logarithm, and shoot and root phosphite were
square root±transformed (SQRT[original data + 0.5]) before
statistical analysis to reduce heteroscedasticity of variance.
Two-way ANOVA (Pi ” phosphite) showed high Pi ” phosphite
interaction for most of the measured parameters. Since unam-
biguous interpretation of the main effects on data variability was
not possible, the simple effects of phosphite were examined by
one-way ANOVA. Mean separation was performed using the
least significant difference (LSD) at p = 5%.
3 Results
3.1 Growth responses of lettuce to Pi supply
Figure 1 shows that the growth of lettuce was strongly in-
creased when the Pi supply increased from 0.025 to 0.1 mM.
100 100% (Max.) growth
Shoot fresh weight (g plant )
-1
80 90% growth
80% growth
60
50% growth
40
20
0 was no detectable phosphite in shoots or roots of plants in
0 0.2 0.4 0.6
Phosphate level (mM)
Figure 1: Growth response of lettuce to different Pi treatments in the
Pi-response experiment. Bars indicate standard errors of means (SE;
n = 3). Arrows point to the Pi levels selected for the phosphite
experiment.
ã 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
J. Plant Nutr. Soil Sci. 2009, 172, 378±384
Beyond this level, the growth response became weaker.
Lettuce reached its maximum growth at about 0.3 mM Pi, and
the growth tended to decrease when Pi supply was higher
than 0.5 mM. Based on this result, the levels of Pi supply
selected for the phosphite experiment were 0.05, 0.1, 0.15,
and 0.3 mM to reach approximately 50%, 80%, 90%, and
100% of maximum plant growth, respectively.
3.2 Effects of phosphite on shoot and root growth
of lettuce
Significant effects (p < 1%) of phosphite and Pi ” phosphite
interaction observed on both shoot and root growth (Fig. 2A
and B) indicated that the effect of phosphite on lettuce growth
was not consistent and highly dependent on the Pi status of
the plants. Under conditions of sufficient Pi supply, phosphite
up to 2 mM did not influence either root or shoot growth. How-
ever, when only 0.15 mM Pi were supplied, 2 mM phosphite
significantly decreased both shoot and root growth. Under
even more limiting Pi supply (0.05 or 0.1 mM), the negative
effect of phosphite was found even at the relatively low level
of 0.2 mM. The inhibiting effect of phosphite was more
obvious with higher phosphite level and lower Pi supply. In
comparison to the zero-phosphite treatment within the same
Pi supply, addition of 2 mM phosphite decreased the shoot
growth by 59%, 38%, and 34% (Fig. 2A) and the root growth
by 64%, 36%, and 25% (Fig. 2B) for 0.05, 0.1 and 0.15 mM
Pi, respectively. At the very low level of 0.05 mM, phosphite
had negligible effects on root or shoot growth (Fig. 2A and B)
at all Pi supplies.
3.3 Effect of phosphite on total P, Pi, and
phosphite concentrations in plant tissues
Application of phosphite significantly increased total P in both
shoots and roots (Fig. 3A and B). The increase was less
under higher Pi supply; for example, by comparison to zero-
phosphite treatment within the same Pi supply, the addition of
2 mM phosphite increased shoot total P by 220%, 122%,
83%, and 31% and root total P by 561%, 439%, 317%, and
107% for 0.05, 0.1, 0.15, and 0.3 mM Pi supplies, respective-
ly. It is interesting that application of phosphite also increased
the water-extractable-Pi concentration in both shoots and
roots (Fig. 3C and D). The increase was small and not signifi-
cant under sufficient Pi supply, but became more obvious and
statistically significant when Pi supply was reduced. Although
the total P concentrations in shoots and roots were signifi-
cantly increased by the addition of 0.05 mM phosphite, there
0.8 1 this phosphite treatment at all levels of Pi supply (Fig. 3E and
F). However, phosphite applied at 0.2 mM was detected in
both shoots and roots and phosphite concentration in shoots
or roots was greatly increased in the 2 mM±phosphite treat-
ment. There was a tendency for decreasing phosphite con-
centrations in both shoot and root with increasing level of Pi
supply.
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J. Plant Nutr. Soil Sci. 2009, 172, 378±384 Phosphate±phosphite interaction effects on lettuce 381

100 25
Two-way ANOVA A B Two-way ANOVA
90
Phosphate *** Phosphate ***
Shoot FW (g shoot )

80 20
-1

Phosphite *** Phosphite ***

Root FW (g root )
a
a aa

-1
70 Interaction *** a Interaction ***
aaa
60 aa 15 aa
50
b
b
b
c
aab
aa a Figure 2: Effects of phosphite treatments (mM) (0 [ ], &
0.05 [ ], 0.2 [ ], and 2 [j]) on shoot and root fresh weight
b a
40 aa c 10 c d c
b
30 d (FW) of lettuce grown under different Pi supplies. Pi levels of
20 c 5 0.05, 0.1, 0.15, and 0.3 mM were Pi supplies for 50%, 80%,
10 90%, and 100% of maximum plant growth, respectively (see
0 0 Fig. 1). Bars indicate SE (n = 3). Significant effects: *** p
0.05 0.10 0.15 0.30 0.05 0.10 0.15 0.30 < 0.1%. Different letters indicate significant difference
Phosphate supply (mM) Phosphate supply (mM) between means within the same Pi supply , LSD (p < 5%).

10 20
Two-way ANOVA Two-way ANOVA
9 A 18 Phosphate *** B
Phosphate ***
Root total P (mg g )

a
Shoot total P (mg g )

-1

8 16 Phosphite ***
-1

Phosphite ***
a b Interaction ***
7 Interaction *** c 14 a
a a d a a
6 12 a
b
5 c
10
b
4 b d 8 b
c c
d d
3 c 6 b
b b
2
d
4 dc
c dc
d
1 2
0 0
7 12
Shoot PO4- P (mg g )

Two-way ANOVA
-1

Two-way ANOVA
Root PO4- P (mg g )

C D
-1

6 Phosphate *** ab a 10 Phosphate ***

b ab
5 Phosphite ***
8 Phosphite ***
Interaction ns Interaction ns
4 a a
bc
ab 6 ab ab
3 a c b
ab a bab 4 a
2 c
bc b
a b
b a c bc
1 2 dc
b c c
0 0
9
Two-way ANOVA
16
Two-way ANOVA Figure 3: Effects of phosphite treatments (mM) (0 [&],
8 E 14 Phosphate ***
F
0.05 [ ], 0.2 [ ], and 2 [j]) on total P, Pi, and phosphite
Root PO3- P (mg g )

Phosphate ***
Shoot PO3- P (mg g )

-1
-1

7 Phosphite *** 12
Phosphite *** concentrations in shoots and roots of lettuce grown in
Interaction Interaction ***
6 ***
10 a different Pi supplies. Pi levels of 0.05, 0.1, 0.15, and
5 a a 0.3 mM were phosphate supplies for 50%, 80%, 90%,
8
4 a and 100% of maximum plant growth, respectively (see
6
3
a
a a Fig. 1). ND: not detectable, which were considered as
2 a
4 zero in analysis and separation of means. Bars indicate
1 b
b 2 b b SE (n = 3). Significant effects: *** p < 0.1%, ns
ND
ND

ND
ND

ND
ND

b
ND
ND

ND
ND

ND
ND

ND
ND

b
ND
ND

b b
0 0 nonsignificant. Within the same Pi supply, different letters
0.05 0.10 0.15 0.30 0.05 0.10 0.15 0.30 indicate significant difference between means by LSD (p
Phosphate supply (mM) Phosphate supply (mM) < 5%).

3.4 Concentrations of minerals (K, Ca, Mg, Fe, Mn, not influence the concentrations of the minerals in shoots
and Zn ) in shoots under all Pi supplies.

Table 1 shows that phosphite up to 2 mM did not affect the

mineral concentrations of lettuce grown in 0.3 mM Pi. How- 3.5 Nitrogen status and ascorbate concentration
ever, when only 0.15 mM Pi were supplied, 2 mM phosphite
tended to decrease the concentrations of all these nutrient As found for the other minerals, total N concentration in
elements. This effect was found even at a phosphite supply shoots of plants grown with insufficient Pi (0.05 or 0.1 mM)
as low as 0.2 mM when plants were more P-deficient (0.05 or was decreased by high-phosphite treatment (Tab. 1). The
0.1 mM Pi). By comparison to the zero-phosphite treatment maximum decrease was found at 2 mM phosphite in 0.05 mM
of the same Pi supply, maximum decreases were found at Pi (17% as compared to zero-phosphite treatment of the
2 mM phosphite in the lowest Pi supply (0.05 mM) for all the same Pi level). However, there was also a slight increase (6%
minerals (46%, 39%, 26%, 35%, 46%, and 30% for K, Ca, to 9%) in shoot N by phosphite application at very low phos-
Mg, Fe, Mn and Zn, respectively). At 0.05 mM, phosphite did phite level (0.05 mM) in low Pi supply (0.05 or 0.1 mM) or at

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382 Thao, Yamakawa, Shibata J. Plant Nutr. Soil Sci. 2009, 172, 378±384

higher phosphite level at higher Pi supply (0.2 mM phosphite 4 Discussion

in 0.15 mM Pi or 2 mM phosphite in 0.3 mM Pi supply).
The effect of phosphite on the shoot NOÿ3 concentration was
similar to that on shoot N but more pronounced (Tab. 1). For
instance, 2 mM phosphite also decreased the shoot NOÿ 3 The results in this study clearly demonstrate that the effect of
concentration in plants grown with low Pi supply; the largest
decrease was at 2 mM phosphite in 0.05 mM phosphite (56%
compared to zero-phosphite treatment of the same level).
There was also an increase in the shoot NOÿ 3 concentration
(20% to 41% compared to the zero-phosphite treatment
within the same Pi supply) by phosphite in those treatments
where the total N concentration was somehow increased.
With respect to the total ascorbate and the NRA (nitrate
reductase activity) neither the effect of phosphite alone nor its
interaction with Pi was significant at any Pi supply level (Tab.
1).
4.1 Effect of phosphite on growth and P nutrition
of lettuce
phosphite on plant growth was strongly dependent on the
level of Pi supply. In general, phosphite up to 2 mM did not
influence growth of Pi-sufficient plants, but still significantly
reduced the growth of plants supplied with Pi allowing for
90% of its maximum growth (0.15 mM Pi). Plants fertilized
with Pi sufficient for about 80% of maximum growth (0.1 mM
Pi) or lower were harmed by phosphite even at rates as low
as 0.2 mM (Fig. 2). These results support our recent study on
komatsuna (Thao et al., 2008a) suggesting that plants insuffi-
ciently fertilized with Pi may become vulnerable even to low
levels of phosphite. The great increase in total P (Fig. 3A and
B) and phosphate (Fig. 3E and F) in both shoots and roots

Table 1: Effect of phosphite treatment on quality parameters of lettuce grown under various Pi supplies.
Pi Phosphite Ascorbate NRA Shoot Shoot mineral-element concentrations
supply treatment (mg [100 g (lM NOÿ2 NO3± N K Ca Mg Fe Mn Zn
(mM) (mM) FW]±1) [g FW]±1 h±1)
(mg [g DW]±1) (lg [g DW]±1)
0.05 0.00 20.05 1.52 36.99b 43.29a 71.53a 8.95a 2.74a 72.43a 93.03a 31.51a
0.05 19.37 1.54 52.19a 46.82a 75.09a 8.40ab 2.77a 73.87a 93.19a 28.21ab
0.20 19.27 1.42 29.53c 40.48b 61.83b 7.90b 2.36b 66.22b 71.31b 26.00b
2.00 20.54 1.46 16.29d 36.06c 38.91c 5.49c 2.02c 47.32c 50.43c 22.14c
Effect ns ns *** *** *** *** ** *** *** *
0.10 0.00 38.04b 45.07a 69.62a 8.72a 2.95a 73.48a 101.64a 30.95a
0.05 51.39a 47.94a 70.40a 8.69a 2.94a 71.01a 101.80a 31.40a
0.20 33.41b 41.51ab 63.06b 8.02a 2.61b 72.37a 105.82a 28.72ab
2.00 20.91c 40.66b 53.99c 5.70b 2.16c 53.36b 85.30b 25.22b
Effect *** * *** *** *** *** * *
0.15 0.00 18.26 1.50 41.97ab 47.20b 71.56a 8.74a 3.50a 70.60 89.68a 27.11
0.05 18.33 1.49 50.24a 50.47ab 74.24a 8.88a 3.58a 72.31 98.12a 28.18
0.20 19.79 1.56 52.02a 51.23a 73.17a 7.87a 3.69a 71.23 97.51a 24.87
2.00 20.44 1.53 35.57b 47.69b 63.94b 6.60b 2.74b 69.15 71.63b 25.92
Effect ns ns *** * ** ** *** ns ** ns
0.30 0.00 18.45 1.70 40.64c 47.54b 70.77 7.78 4.14 73.38 94.37 25.66
0.05 18.11 1.68 41.49bc 48.32ab 69.89 8.64 4.29 69.75 96.20 24.34
0.20 17.65 1.59 45.71b 48.61ab 69.38 8.89 4.57 69.45 99.07 24.91
2.00 19.98 1.69 53.79a 50.82a 70.07 7.72 4.67 70.63 03.87 26.49
Effect ns ns *** * ns ns ns ns ns ns
Two-way ANOVA
Pi ns * *** *** *** *** *** *** *** ***
phosphite ns ns ** *** *** *** *** *** *** ***
Pi ” phosphite ns ns *** *** *** *** *** *** *** ***
FW, fresh weight; NRA, nitrate reductase activity.
Upper letters in italics refer to the effects of phosphite treatment as evaluated for each Pi supply separately by one-way ANOVA and LSD test
(p < 5%). Within each Pi supply, means in column followed by the same letters are not significantly different. Significant effects: ns, non-
significant, * p < 5%; ** p < 1%, *** p < 0.1%.

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J. Plant Nutr. Soil Sci. 2009, 172, 378±384
due to phosphite application under all Pi levels together with
no improvement or even significant reduction of plant growth
under low Pi supply clearly indicates that phosphite was well
absorbed by roots and mobile inside lettuce, but did not
improve the P nutrition of the plant to any extent. This is in
agreement with other studies (Forster et al., 1998; Schroetter
et al., 2006; Thao et al., 2008a). However, not all plants can
well absorb phosphite via roots as our previous study showed
that phosphite was poorly absorbed by roots of spinach
(Thao et al, 2008b).
Although it is generally known that phosphite has negative
effects on Pi-starved, but not on Pi-fertilized plants (Ticconi
et al., 2001; Varadarajan et al., 2002; Carswell et al., 1996,
1997; Schroetter et al., 2006), most of these studies used
phosphite as a sole P source or at high rates. Some
researchers (Rickard, 2000; Watanabe, 2005; Lovatt and
Mikkelsen, 2006) suggested that phosphite may be used as
P fertilizer for some plants and that phosphite, if used in
appropriate amounts, can stimulate plants. In addition, the
combination of phosphite and Pi ions is also believed to be
more effective than either ion alone in plant assimilation
(Young, 2004; Forster et al., 1998). However, our previous
studies on komatsuna (Thao et al., 2008a) and spinach
(Thao et al., 2008b) revealed that for the same total amount
of P (phosphite + Pi) application, plant growth decreased sig-
nificantly as phosphite proportion increased. The results in
the present study further indicate that for a given Pi supply,
addition of phosphite at various rates did not provide any sti-
mulation on the growth of lettuce regardless of the Pi levels,
suggesting that there was no beneficial effect of Pi±phosphite
combinations.
Studies by Carswell et al. (1996) on Brassica nigra revealed
that the addition of phosphite decreased the cellular Pi
concentration (acid-extractable Pi). The decrease in acid-
extracted Pi by phosphite treatment possibly resulted from
inhibition of Pi uptake by phosphite as indicated in Brassica
napus cell suspensions (Carswell et al., 1997). However,
Schroetter et al. (2006) indicated that application of phosphite
increased the water-extractable Pi in maize plants. Here, we
also found an increase in water-extractable Pi due to addition
of phosphite. The increase was small under high Pi supply
and became larger under lower Pi supply. This increase did
not improve plant growth at any phosphite treatment, but it
decreased the growth of Pi-deprived plants (Fig. 2A and B).
Thus, it is reasonably speculated that the increase in water-
extractable Pi by phosphite resulted from the competition of
phosphite with Pi for assimilation in the plant tissues leading
to an increase in free Pi rather than the partial oxidation of
phosphite to Pi in the plant tissues as suggested by Schroet-
ter et al. (2006). Similar results were also found for komat-
suna (Thao et al., 2008a).
4.2 Effect of phosphite on some main quality
parameters of lettuce
Although phosphite greatly reduced the growth of plants at low
Pi supply, neither a difference in ascorbate concentration nor
NRA in leaves among all treatments was observed (Tab. 1),
ã 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Phosphate±phosphite interaction effects on lettuce 383
reflecting that phosphite had no effect on ascorbate biosynth-
esis or NRA in lettuce plants grown in various Pi treatments.
Significant effects of both phosphite and phosphite ” Pi inter-
action on mineral (Ca, K, Mg, Fe, Mn, and Zn) concentrations
in shoots (Tab. 1) reflects the highly Pi-dependent effect of
phosphite. In general, the decrease in mineral concentrations
by phosphite was found only in the treatments where phos-
phite had severe inhibiting effects on both shoot and root
growth (Fig. 2, Tab. 1) with visible toxic symptoms on roots
(dark color, death of root tip, and root hair with many black
spots, data not shown). In the treatments without severe root-
growth reduction by phosphite (even at high rate such as 2
mM phosphite in 0.3 mM Pi), the effect of phosphite on shoot
minerals was negligible. The results suggest that the
decrease in mineral concentrations in shoots by phosphite
resulted from root-growth inhibition under low Pi supply,
which may impair nutrient uptake.
Although shoot NOÿ 3 was also decreased by phosphite in the
same manner as other minerals due to severe root-growth
inhibition by phosphite, there was also an increase in shoot
NOÿ 3 in the treatments where phosphite had a little or no
effect on both shoot and root growth (Tab. 1, Fig. 2). The
mechanism is unclear. No effect of phosphite on NRA was
detected in any treatment (Tab. 1). The increase in shoot
NOÿ 3 concentration was somewhat associated with the
increase in total N in shoots, suggesting that phosphite some-
how stimulated the N uptake in these cases. Further research
is needed for better understanding.
Although few of earlier works suggested that phosphite appli-
cation improved the quality of crops such as fruit size and
total soluble solids (Rickard, 2000; Lovatt, 1999; Lovatt and
Mikkelsen, 2006), it is unclear whether these effects were
due to other phosphite properties rather than its fungicidal
properties or its conversion to Pi. In this study, the experiment
was conducted under greenhouse conditions using hydropo-
nic culture to minimize the interference of pathogens as well
as the conversion of phosphite to Pi by soil microflora. The
results indicate that phosphite itself had little influence on the
ascorbate and mineral concentration of lettuce plants. How-
ever, under insufficient Pi supply, application of phosphite
may decrease the mineral concentration of plants due to an
inhibiting effect of phosphite on root growth and hence nutri-
ent uptake.
5 Conclusions
The results clearly indicate that phosphite is well absorbed by
roots of lettuce and is mobile inside the plants, while it does
not improve the P nutrition and does not have any stimulating
effect on growth of healthy plants even when used in combi-
nation with Pi. Application of phosphite to Pi-deficient plants
may decrease the mineral concentration of plants caused by
root-growth inhibition of phosphite. Phosphate fertilization
prior to phospite application can minimize the harmful effect
on plant growth.
www.plant-soil.com
384 Thao, Yamakawa, Shibata
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