Protoplasma

https://doi.org/10.1007/s00709-021-01623-3

ORIGINAL ARTICLE

Grafting improves tomato yield under low nitrogen conditions

by enhancing nitrogen metabolism in plants
Zhi Huan Zhang 1,2,3 & Ming Ming Li 4 & Bi Li Cao 1,2,3 & Zi Jing Chen 1,2,3 & Kun Xu 1,2,3

Received: 22 June 2020 / Accepted: 12 February 2021

# Springer-Verlag GmbH Austria, part of Springer Nature 2021

Abstract
To alleviate the effects of increasingly severe environmental conditions and meet the increasing demand for organic agricultural
products, this paper studied tomato grafting under low nitrogen conditions in an effort to enhance yield and improve fruit quality
by enhancing nitrogen metabolism. In this study, we screened for two tomato genotypes, a high nitrogen use efficiency genotype
(‘TMS-150’) and a low nitrogen use efficiency genotype (‘0301111’), using rootstocks from 25 tomato genotypes and studied the
effects of tomato grafting on plant yield, fruit quality, nitrogen content, activities of key nitrogen metabolism enzymes, and
nitrogen use efficiency (NUE) under different nitrogen fertilizer conditions. The results showed that the yield of the tomato
plants, the activities of key enzymes during nitrogen metabolism, the contents of different forms of nitrogen, and the efficiency of
nitrogen use were lower at low nitrogen fertilization levels and higher at higher nitrogen fertilization levels, while the measured
indicators were the highest under the N40 nitrogen fertilizer treatment. Grafting tomatoes with high-NUE tomato seedlings as the
rootstock resulted in significant increases in the nitrogen content and the activity of key enzymes, enhanced the NUE of tomato
plants, increased tomato yield, and improved fruit quality compared to those of the seedlings grafted with low-NUE rootstock.
Our results indicate that tomato plants grafted with high-NUE rootstock presented enhanced absorption and utilization of nitrogen
and increased plant yield by promoting nitrogen metabolism at different nitrogen levels.

Keywords Tomato . Grafting . Rootstock . Nitrogen fertilizer . Yield

Introduction
As an essential element of plant growth and development,
nitrogen is an important component of proteins, nucleic acids,
phospholipids, and certain growth hormones in plants and
contributes to 40–50% to the final yield of crops (Stein and
Handling Editor: Hanns H. Kassemeyer
* Kun Xu
xukun@sdau.edu.cn
1
College of Horticulture Science and Engineering, Shandong
Agricultural University, 271018 Tai’an, People’s Republic of China
2
Collaborative Innovation Center of Fruit and Vegetable Quality and
Efficient Production, Tai’an, Shandong, People’s Republic of China
3
State Key Laboratory of Crop Biology, Key Laboratory of Biology
and Genetic Improvement of Horticultural Crops in Huanghuai
Region, Ministry of Agriculture and Rural Affairs, Beijing, People’s
Republic of China
4
Taishan Property Insurance Co., Ltd., Jinan, People’s Republic of
China
Klotz 2016; Liang et al. 2018; Karki et al. 2018). Therefore,
nitrogen fertilizer plays an important role in agricultural pro-
duction. To meet the demand for crop yields, nitrogen fertil-
izer has been excessively applied in current agricultural pro-
duction, and the amount of nitrogen fertilizer applied far ex-
ceeds the needs of plants (Saleque et al. 2004; Yang et al.
2017). In fact, it has been reported that only 30 to 50% of
the nitrogen fertilizer applied to the soil can be absorbed by
plants (Hodge et al. 2000). Consequently, the excessive input
of nitrogen fertilizer does not increase the yield and quality of
crops but leads to a decrease in nitrogen use efficiency and an
increase in nitrogen losses, causing serious environmental
problems (Jensen et al. 2011; Chen et al. 2018). In particular,
excessive nitrogen fertilizer application leads to excessive ac-
cumulation of nitrogen in the soil, which increases the
leaching of nitrogen into the groundwater from soil. The high
nitrate concentration in groundwater poses a serious threat to
the safety of the groundwater environment (Lasagna et al.
2016; Martinelli et al. 2018). Therefore, given the current
increasingly serious environmental pollution conditions,
maintaining the ecological balance and simultaneously

meeting demand for organic agricultural products, achieving
the goal of reducing nitrogen fertilizer input, and improving
the nitrogen use efficiency of crops have gradually attracted
the attention of more researchers.
Nitrogen metabolism is one of the most important and ba-
sic metabolisms in plants (Wang et al. 2019). It includes the
reduction and assimilation of inorganic nitrogen and the con-
version and synthesis of organic containing nitrogen com-
pounds. There are many enzymes involved in plant nitrogen
metabolism, in which the key enzymes that play important
regulatory roles are glutamine synthetase (GS), glutamate syn-
thase (GOGAT), and glutamate dehydrogenase (GDH)
(Yamaya and Kusano 2014). GS is a multifunctional enzyme
at the center of nitrogen metabolism and it has two enzyme
activities, invertase and synthetase, and participates in the reg-
ulation of various nitrogen metabolisms (Thomsen et al.
2014). Some studies have suggested that the appropriate
amount of nitrogen application can increase the enzyme activ-
ities of GS, GOTAT, and GDH, but excessive application of
nitrogen will reduce the enzyme activity of GS and other en-
zymes (Singh et al. 2016; Wei et al. 2016). The decrease in
enzyme activity will affect the process of nitrogen assimila-
tion, which will affect the distribution of nitrogen and carbon
metabolites and carbon and nitrogen balance in plants, and
then affect the plant yield (Ge et al. 2014). Many studies have
found that there were significant differences in the activities of
related enzymes in different crop varieties and different organs
and tissues of the same crop (Qin et al. 2013; Deng et al. 2017;
Ren et al. 2017).
Tomato (Lycopersicon esculentum Mill.) is one of the most
widely cultivated vegetable crops in the world and plays an
important role in vegetable production. Studies have shown
that low-N conditions increased the concentration of phenolic
compounds and vitamin C, while the concentrations of β-
carotene and lycopene were not affected (Hernández et al.
2019). In addition, although the content of soluble sugar and
fruit dry matter in tomato fruit can be increased under moder-
ately low nitrogen conditions, the fruit commercial yield and
plant vegetative growth of tomato are reduced (Benard et al.
2009). Studies have shown that low nitrogen stress not only
seriously affects the morphological structure of crop plants but
also causes damage to their physiological metabolism, which
hinders plant growth, decreases chlorophyll content, and de-
creases photosynthesis (Wei et al. 2016; Bassi et al. 2018).
Grafting, as a cultivation technique with a long history, plays
an important role in enhancing plant resistance, increasing
yield, and improving quality (Kumar et al. 2017; Jabnoun-
Khiareddine et al. 2019). In particular, grafted plants can en-
hance the absorption of nutrients by using nitrogen-efficient
absorption rootstocks (Fernández-García et al. 2002) and im-
prove the efficiency of fertilizer and water use (Schwarz et al.
2010; Zhang et al. 2019), thereby alleviating the damage from
low nitrogen stress on plants.
Z. H. Zhang et al.
Therefore, grafting is an important way of enhancing the
adaptability of plants under low nitrogen conditions and im-
proving the efficiency of nitrogen use. Compared with other
ways of enhancing nitrogen utilization efficiency, enhance-
ment by grafting cultivation has the advantages of conve-
nience, speed, and cost savings. To this end, this paper at-
tempts to select nitrogen-efficient tomato materials by apply-
ing different nitrogen levels to different tomato genotypes and
to study the effects of different rootstocks on nitrogen absorp-
tion and utilization.
Materials and methods
Plant material and experimental design
This experiment was carried out in a solar greenhouse located
at Shandong Agricultural University in Tai’an (36° 09′ N,
117° 09′ E), eastern China, under the following environmental
conditions: natural light, photosynthetic photon flux density
(PPFD) of 1400–1800 μmol m−2 s−1 (sunny day at noon), and
temperature of 26~37/18~26 °C (day/night).
First, for the screening experiment, 25 tomato genotypes
were used as plant materials (Table 1). When the seedlings
grew to the three-leaf and one-heart stage, they were
transplanted to a water basin with a length of 37 cm, width
of 28 cm, and height of 12 cm. The liquid level was set with a
foam board with a diameter of 2 cm and used to fix tomato
seedlings, and 8 plants were planted in each pot. The plants
were first cultured in deionized water for 2 days, and then the
deionized water was replaced with nutrient solution. The nu-
trient solution was set to three nitrogen levels, namely, low
nitrogen (1.5 mmolL−1), medium nitrogen (7.5 mmolL−1), and
high nitrogen (15 mmolL−1) levels. This test used Hoagland
nutrient solution, and the nutrient solution was replaced every
6 days. CaNO3 and KNO3, which were reduced at low nitro-
gen levels, were replaced by CaCl2 and KCl, and the pH of the
nutrient solution was adjusted with HCl and NaOH. After 30
days of treatment, samples were tested for relevant indicators.
Second, for the grafting experiment, the two selected toma-
to genotypes, ‘TMS-150’ (high NUE) and ‘0301111’ (low
NUE), which were determined by first experiment, were used
as rootstocks, and the variety of ‘Jinpeng 1’ commonly used in
production as the scion. The experimental design was a split
plot: the main plot was composed of ‘Jinpeng 1’ grafted onto
‘0301111’ (T1), ‘Jinpeng1’ grafted onto ‘TMS-150’ (T2), and
self-grafted ‘Jinpeng 1’ (CK); and the subplot was different
nitrogen levels of 0, 20, 40, and 60 kg/667 m2 of urea (46% of
nitrogen). The urea was applied 4 times, of which 30% was
basal fertilizer, 20% was applied at the first fruit internode,
25% at the second fruit internode at the enlargement stage, and
25% at the fourth fruit internode. Tomatoes were planted with
a large row spacing of 90 cm, a small row spacing of 60 cm,
Grafting improves tomato yield under low nitrogen conditions by enhancing nitrogen metabolism in plants

Table 1 The material No., names and source of tomato genotypes used in the experiment

Material Material names. Material source Material Material Material source

No. No. names.

1 060114 Breeding by Shandong agricultural 14 061111-2 Sakata seed corporation of Japan

university
2 060113 Breeding by Shandong agricultural 15 0104111 Introduced from Japan
university
3 0301111 Breeding by Shandong agricultural 16 ZM606 Beijing Jingtian Seedling Company
university
4 06142 Breeding by Shandong agricultural 17 061011 Sakata seed corporation of Japan
university
5 Shadow warrior Takii seed corporation of Japan 18 0501-211 Breeding by Shandong agricultural
university
6 Block Takii seed corporation of Japan 19 060112 Takii seed corporation of Japan
7 Magnet Takii seed corporation of Japan 20 060312 Breeding by Shandong agricultural
university
8 TMS-150 Sakata seed corporation of Japan 21 0301112 Breeding by Shandong agricultural
university
9 Mikado Takii seed corporation of Japan 22 061213 Takii seed corporation of Japan
10 060911 Introduced from Japan 23 060812 Breeding by Shandong agricultural
university
11 061211 Introduced from Japan 24 08X1-11 Breeding by Shandong agricultural
university
12 060611 Sakata seed corporation of Japan 25 061212 Breeding by Shandong agricultural
university
13 061111-1 Sakata seed corporation of Japan

and plant spacing of 40 cm. Every 2 rows were treated as 1
cell, and the area of the plot was 12 m2, with 4 replicates. To
ensure the balance of nutrients, superphosphate (P2O5 16%)
150 kg/667 m2 and potassium sulfate (K2O 50%) 150 kg/667
m2 were applied to all treatments. The superphosphate was
applied once during planting, and potassium sulfate combined
with nitrogen fertilizer was applied at 20, 20, 30, and 30% at
the four fertilizer application times described above. The other
plants were managed according to the conventional grafting
cultivation techniques for tomatoes.
productivity from applied nitrogen (PFPN), and agronomic
efficiency of N applied (AEN) were as follows (Dwivedi
et al. 2016):
NUEð%Þ ¼ plant nitrogen absorption=nitrogen supply
PFPN ðgg−1Þ ¼ plant biomass=nitrogen supply
AEN ðkg kg−1Þ
¼ ðyield of nitrogen application area−yield of blank areaÞ
=nitrogen supply

Plant total nitrogen content Growth and yield analysis

Plant tissues (leaf, stem, and root) were heated at 105 °C for 20
min, dried to constant weight at 75 °C, and then ground sep-
arately in a Wiley mill to pass through a 20-mesh screen.
Dried plant tissues (0.5 g) were concentrated with H2SO4-
H2O2 and then analyzed for the total nitrogen content using
the Kjeldahl method (Matejovic 1995).
Plant nitrogen utilization
The nitrogen absorption per plant was measured as the total
nitrogen content of the whole plant. The nitrogen supply was
the supply of nitrogen in different nitrogen treatments, and the
plant biomass was the fresh weight of plants. The formulas for
calculating the nitrogen use efficiency (NUE), partial factor
Five plants were randomly selected from each treatment. The
impact on the growth of the tomato plants of the different treat-
ments was assessed by determining the leaf, stem, and root
fresh weight. When the fruit was ripe for harvesting, the weight
of a single fruit was weighed, and the yield was calculated.
Fruit quality determination
The mature fruit at the second internode was selected to de-
termine the fruit quality (Zhao et al. 2002). The content of
soluble sugar was determined by the anthrone method. The
organic acid content was determined by NaOH titration. The
vitamin C content was determined by 2,6-dichloroindophenol
titration. The soluble protein content was determined by the

Coomassie brilliant blue method. AGY-1hardness tester was
used for the fruit hardness measurement. Lycopene was mea-
sured by a petroleum ether extraction method (Choudhari and
Ananthanarayan 2007).
Determination of different forms of nitrogen
To determine the protein nitrogen content, functional leaves of
the plants were selected from each plot, and the leaves were
dried and sieved. A 0.5-g sample was weighed and extracted
with trichloroacetic acid, and then the H2SO4-H2O2 combined
digestion method was used to carry out sample digestion.
Finally, the protein nitrogen content was determined by the
Kjeldahl method.
To determine the content of nitrate nitrogen, functional
leaves of the plants were removed from each plot. Then,
0.5 g of fresh samples were placed in test tubes, 10 ml of
deionized water was added to each test tube, and the samples
were boiled in water for 30 min and cooled. The extract was
filtered into a 25-ml volumetric flask, 0.1 ml of the sample
was diluted into a test tube, 0.4 ml of 5% salicylic acid-sulfuric
acid was added, and the mixture was shaken for 20 min. Then,
9.5 ml of 8% sodium hydroxide solution was added and the
coloration was reversed at 410 nm after mixing and cooling
(Zhao et al. 2002).
To determine the ammonium nitrogen content, 0.5 g of the
fresh sample after mixing was weighed and ground with 5 ml
of water and centrifuged. An amount of 0.1 ml of the super-
natant was pipetted into a test tube, and 2.9 ml of distilled
water, 0.5 ml of phenol solution, and 0.5 ml of sodium hypo-
chlorite solution were added. The mixture was shaken for 30
min, and 0.1 ml of masking agent (400 gL-1 of sodium potas-
sium tartrate solution and 100 gL-1 EDTA disodium solution
mixed in equal volume, with 0.5 ml 10molL-1 NaOH solution
added per 100 ml of the mixture) was added. Then, 5.9 ml of
distilled water was added, and the mixture was shaken well.
Colorimetric determination was performed at 625 nm (Gips
et al. 1970).
To determine the free amino acid nitrogen content, 0.5 g of
the fresh sample was weighed after mixing, ground with 5 ml
of 10% acetic acid, diluted to 100 ml with distilled water, and
filtered into a flask. An amount of 0.1 ml of filtrate was pipet-
ted into a test tube and combined with 1 ml of distilled water,
3 ml of ninhydrin hydrate, and 0.1 ml of ascorbic acid. The
mixture was heated for 15 min in boiling water and then rap-
idly cooled. An amount of 4.9 ml of 60% ethanol was added to
the mixture, and colorimetry was performed at 570 nm (Zhao
et al. 2002).
Determination of GS, GOGAT, and GDH activity
One gram of a fresh sample was weighed and ground to obtain
a crude enzyme solution (Yin et al. 2019). Three milliliters of
Z. H. Zhang et al.
glutamine synthetase (GS) reaction solution were added to
1 ml of the crude enzyme solution and incubated at 30 °C
for 15 min. One milliliter of a mixture of trichloroacetic acid,
HCl, and ferric chloride was added and mixed for 10 min.
Colorimetry was performed at 540 nm. Both glutamate dehy-
drogenase (GDH) and glutamate synthase (GOGAT) were
measured by pipetting 0.2 ml of the crude enzyme solution
and mixing with 2.8 ml of the reaction solution, and kinetic
colorimetric analysis was performed at 340 nm, respectively.
Statistical analysis
The coefficient of variation (CV) was calculated by Excel
2007 software. Data clustering and multiple comparisons
were statistically analyzed and mapped using the DPS soft-
ware package (DPS for Windows, 2009). The differences be-
tween the means were tested by Duncan’s multiple range test
at P < 0.05.
Results
Identification and evaluation of nitrogen utilization
ability of tomato seedlings under different nitrogen
levels
After 30 days of different nitrogen treatments in the screening
experiment, 25 tomato genotypes were sampled and assayed
for the indicators of nitrogen absorption, NUE, and partial
factor productivity from applied nitrogen (PFPN) (Table 2).
The results showed that there were large differences in the
indicators of nitrogen use capacity of the different tomato
genotypes. The average nitrogen absorptions of low nitrogen,
medium nitrogen, and high nitrogen levels were 132.41,
216.80, and 281.56 mg plant-1, respectively, and the range
of variation was 78.87–194.03 mg plant -1 , 130.40–
289.52 mg plant-1, and 176.44–404.49 mg plant-1, respective-
ly. As the nitrogen level increased, the nitrogen absorption of
the tomato seedlings increased, and the range of the absorp-
tion value also increased. However, the NUE and PFPN de-
creased with increasing nitrogen levels, and the ranges of
these indicators were reduced. The CVs of the nitrogen use
capacity-related indicators were significantly different among
the different nitrogen supply levels. Both CVs were high at
low and high nitrogen levels and low at medium nitrogen
levels; all the CVs were greater than 15%. Therefore, geno-
types with different nitrogen utilization abilities could be iden-
tified from the tested tomato materials.
The clustering analysis was carried out using the NUEs of
the different tomato genotypes under three nitrogen levels as
the statistical parameters (Fig. 1a, b, c). The results showed
that the tested tomato genotypes could be divided into three
categories under the three nitrogen supply levels, which we
Grafting improves tomato yield under low nitrogen conditions by enhancing nitrogen metabolism in plants

Table 2 The correlative index of different tomato seedlings nitrogen utilization ability under different nitrogen levels

Material No. Nitrogen absorption (mg plant-1) NUE (%) PFPN (g g-1)

1.5 mmol L-1 7.5 mmol L-1 15 mmol L-1 1.5 mmol L-1 7.5 mmol L-1 15 mmol L-1 1.5 mmol L-1 7.5 mmol L-1 15 mmol L-1

1 153.23±5.62 239.93±7.76 292.54±9.46 70.05±2.57 21.94±0.70 13.36±0.43 18.09±0.66 4.67±0.15 2.60±0.08

2 149.73±5.49 234.76±7.59 297.50±9.62 68.45±2.51 21.46±0.69 13.60±0.44 18.57±0.68 4.74±0.15 2.71±0.08
3 78.87±2.89 151.84±4.91 176.44±5.70 36.06±1.32 13.88±0.44 8.06±0.26 9.52±0.34 3.07±0.09 1.62±0.05
4 108.90±3.99 180.00±5.82 243.95±7.89 49.78±1.82 16.46±0.53 11.15±0.36 12.97±0.47 3.62±0.11 2.13±0.06
5 80.10±2.94 130.40±4.21 153.35±4.96 36.62±1.34 11.92±0.38 7.01±0.22 9.64±0.35 2.81±0.09 1.50±0.04
6 157.03±5.76 246.22±7.96 335.91±10.86 71.78±2.63 22.51±0.72 15.35±0.49 18.81±0.69 4.98±0.16 2.96±0.09
7 149.59±5.49 242.82±7.85 323.27±10.46 68.38±2.51 22.19±0.71 15.18±0.49 18.92±0.69 4.81±0.15 2.98±0.09
8 194.03±7.12 289.52±9.36 404.49±13.08 88.70±3.25 26.47±0.85 18.49±0.59 22.14±0.81 5.50±0.17 3.44±0.11
9 140.63±5.16 224.28±7.25 284.03±9.19 64.29±2.36 20.50±0.66 12.97±0.41 16.79±0.61 4.45±0.14 2.50±0.08
10 108.11±3.96 196.40±6.35 246.01±7.96 49.42±1.81 17.95±0.58 11.24±0.36 13.57±0.49 3.90±0.12 2.21±0.07
11 121.14±4.44 181.13±5.86 269.10±8.70 55.38±2.03 16.56±0.53 12.30±0.39 14.28±0.52 3.60±0.11 2.33±0.07
12 142.70±5.23 237.24±7.67 281.76±9.11 65.23±2.39 21.68±0.70 12.88±0.41 17.73±0.65 4.67±0.15 2.50±0.08
13 131.79±4.83 239.23±7.74 292.91±9.47 60.24±2.21 21.87±0.70 13.39±0.43 16.31±0.59 4.69±0.15 2.63±0.08
14 122.04±4.48 192.12±6.21 244.18±7.90 55.79±2.04 17.56±0.56 11.16±0.36 14.40±0.52 3.76±0.12 2.17±0.07
15 152.24±5.58 238.26±7.70 297.22±9.61 69.59±2.55 21.77±0.70 13.58±0.43 18.21±0.66 4.74±0.15 2.69±0.08
16 142.50±5.23 209.11±6.76 273.89±8.86 65.14±2.39 19.12±0.61 12.52±0.40 17.14±0.62 4.19±0.13 2.45±0.07
17 144.30±5.29 217.63±7.04 269.51±8.72 65.97±2.42 19.89±0.64 12.32±0.39 16.78±0.61 4.21±0.13 2.35±0.07
18 150.36±5.52 233.50±7.55 320.43±10.36 68.74±2.52 21.34±0.69 14.64±0.47 18.45±0.67 4.69±0.15 1.61±0.05
19 119.20±4.37 231.66±7.49 332.14±10.74 54.49±2.00 21.17±0.68 15.18±0.49 14.52±0.53 4.62±0.14 2.89±0.09
20 122.86±4.51 216.71±7.01 263.85±8.53 56.16±2.06 19.80±0.64 12.05±0.38 15.11±0.55 4.31±0.13 2.36±0.07
21 182.80±6.71 276.12±8.93 352.19±11.39 83.57±3.06 25.24±0.81 16.09±0.52 21.30±0.78 5.48±0.17 3.01±0.09
22 84.75±3.11 158.12±5.11 227.39±7.35 38.74±1.42 14.44±0.46 10.39±0.33 9.88±0.36 3.19±0.10 1.98±0.06
23 117.86±4.32 207.60±6.71 253.75±8.21 53.87±1.97 18.97±0.61 11.60±0.37 14.04±0.51 4.05±0.13 2.23±0.07
24 120.48±4.42 223.31±7.22 317.64±10.27 55.08±2.02 20.41±0.66 14.51±0.46 14.52±0.53 4.40±0.14 2.76±0.08
25 134.98±4.95 222.19±7.18 248.78±8.04 61.70±2.26 20.31±0.65 11.37±0.36 15.71±0.57 4.29±0.13 2.25±0.07
Average 132.41 216.80 281.56 75.66 24.77 16.02 15.9 4.30 2.43
Stdev 28.12 36.60 55.33 16.19 4.17 3.16 3.28 0.68 0.47
S/A(%) 21.24 16.88 19.65 21.39 16.83 19.73 20.63 15.81 19.34

Date in the table is mean ± SD, and the mean were means of three repetitions

called high-NUE genotypes, medium NUE genotypes, and
low NUE genotypes. At low nitrogen levels, the materials
with high NUE were No. 8 and No. 21, the materials with
low nitrogen use efficiency were No. 3, No. 5, No. 22, and
the remaining 20 materials had medium NUE (Fig. 1a). The
results of the cluster analysis at the medium nitrogen level
were consistent with those for low nitrogen (Fig. 1b).
However, the results of cluster analysis at high nitrogen levels
were significantly different (Fig. 1c), with seven materials (6,
7, 8, 18, 19, 21, and 24) belonging to highly nitrogen-efficient
materials, and No. 3 and No. 5 belong to less nitrogen-
efficient materials. The higher nitrogen supply level improved
the nitrogen use efficiency of some tomato materials.
Figure 1 shows that when PFPN was used as a statistical
parameter, the clustering results at high nitrogen levels show
that only material No. 8 was a highly nitrogen-efficient geno-
type. In addition, the PFPN clustering results under low
nitrogen and medium nitrogen levels were completely consis-
tent with the results of the NUE cluster analysis. In summary,
the No. 8 material had the strongest nitrogen utilization capac-
ity, and the No. 3 and No. 5 materials had weak nitrogen
utilization capacity.
Impact of different rootstocks of grafted tomato on
plant yield and fruit quality
Subsequently, we grafted the selected tomatoes with different
NUEs and then applied different nitrogen levels for treatment.
The statistical analysis results for the fresh weight of dif-
ferent organs and the yield of grafted tomatoes are shown in
Table 3. Different rootstock grafting treatments and nitrogen
fertilizer treatments had significant effects on the growth of
the tomato plants. The fresh weight of different organs and the
yield of tomatoes in the main treatment were the lowest in CK,
 Z. H. Zhang et al.

a b c

d e f

Euclidean distance Euclidean distance Euclidean distance

Fig. 1 Clustering analysis of the NUE and PFPN of different tomato Clustering analysis tomato in the low nitrogen level; b and e clustering
genotypes under different nitrogen levels. a, b, and c Clustering analysis of tomato in the middle nitrogen level; c and f clustering analysis
analysis of the NUE; d, e, and f clustering analysis of PFPN. a and d of tomato in the high nitrogen level

those in the T2 treatment were the highest, and those in the T1
treatment were in the middle. Compared with that in the CK
treatment, the single fruit weights of T1 and T2 were increased
by 26.58 and 53.64%, respectively. The yields per plant of T1
and T2 were 24.68 and 45.46% higher than that of the CK
treatment, respectively. This indicated that the grafting of dif-
ferent rootstocks had significant effects on tomato yield in-
crease and the stronger the nitrogen utilization capacity of
the rootstock, the larger the yield increase. The relationship
among the nitrogen treatments in terms of the fresh weight and
the yield of each organ was N40 > N60 > N20 > N0, and N20,
N40, and N60 increased the yield by 47.58, 89.95, and
78.77%, respectively, compared with that in the N0 treatment.
The single fruit weight increased by 3.85, 9.50, and 9.25% in
the N20, N40, and N60 treatments, respectively, compared
with that in the N0 treatment. Table 3 also shows that the
grafting treatment and nitrogen fertilizer treatment had signif-
icant interaction effects on root fresh weight, single fruit fresh
weight, and yield.
The statistical analysis of the fruit quality of differently
treated tomatoes is shown in Table 4. The transverse diameter,
vertical diameter, firmness, and contents of soluble sugar, sol-
uble protein, vitamin C, organic acid, and lycopene in the
main treatment were the highest in the T2 treatment, followed
by those of T1 and CK. Aside from the significant differences
in soluble sugar, vitamin C, and organic acid contents among
the three grafted plants, the differences in quality index values
were not significant. In the N treatment, the contents of solu-
ble sugar, vitamin C, organic acid, and lycopene were highest
in the N0 treatment, followed by those in the N20 treatment.
Those in the N60 treatment were lower, and those in the N40
treatment were the lowest. However, the transverse diameter,
vertical diameter, firmness, and soluble protein content de-
creased in the order N40 > N60 > N20 > N0, which showed
that the larger the fruit, the lower the fruit quality content.
Furthermore, in addition to their effects on soluble protein
content, grafting and nitrogen fertilizer showed significant in-
teraction effects on fruit quality.
Grafting improves tomato yield under low nitrogen conditions by enhancing nitrogen metabolism in plants

Table 3 Statistical analysis of tomato growth and yield among different treatments at harvest time

Treatments Root FW Stem FW Leaf FW Single fruit weight Yield

(g plant-1) (g plant-1) (g plant-1) (g) (g plant-1)

Grafted treatments CK 61.28±3.32c 743.30±30.35c 767.80±23.47c 121.43±21.67b 2524.94±153.32c

T1 71.45±3.02b 928.98±10.77b 867.35±32.53b 153.71±12.84ab 3148.20±142.58b
T2 88.07±3.13a 953.63±12.68a 944.93±21.33a 168.47±16.47a 3672.84±131.64a
N treatments N0 67.17±1.44d 767.17±23.32d 825.27±4.42d 139.96±1.21c 2021.92±56.62d
N20 71.60±1.76c 863.43±14.45c 838.33±5.37c 145.35±4.47b 2984.00±112.37c
N40 79.00±1.32a 977.37±13.21a 892.53±3.25a 153.25±2.32a 3840.71±78.76a
N60 76.33±1.25b 893.23±12.63b 881.23±6.46b 152.92±3.68ab 3614.67±63.48b
P value
Grafted treatments 0.0001 0.0001 0.0001 0.0001 0.0001
N treatments 0.0015 0.0001 0.0009 0.0053 0.0064
G×N 0.0332 0.0875 0.1215 0.0291 0.0491

All data were obtained at the time of tomato harvest. The data are average of three repeated multiple comparisons ± SD. The different letters indicate a
significant difference at P < 0.05 according to Duncan’s test. The P value is obtained by multiple comparison and representing significant or nonsig-
nificant, respectively. The same as below

Changes in different forms of nitrogen content
The changes in nitrate nitrogen, ammonium nitrogen, free
amino acid nitrogen, and protein nitrogen in different grafted
tomato leaves under different nitrogen fertilizers are shown in
Figs. 2 and 3. The statistical analysis of nitrate nitrogen and
ammonium nitrogen content in grafted tomato leaves under
different N treatments is shown in Fig. 2. There were signifi-
cant differences in the three grafted tomatoes at different
growth stages. Figure 2a shows that the nitrate nitrogen con-
tent under different grafting treatments gradually increased
with the plant growth period, and the difference between dif-
ferent grafted seedlings was significant. The content of nitrate
nitrogen was the highest in the T2 grafting treatment, followed
by those in T1 and CK. Under different N treatments, except
for the N0 treatment, the nitrate nitrogen content increased
significantly with the plant growth period, while the N0 treat-
ment decreased slowly with the plant growth period (Fig. 2b).
The nitrate nitrogen contents in the N20, N40, and N60 treat-
ments were significantly higher than that of the N0 treatment,
indicating that topdressing nitrogen fertilizer contributed to
the nitrogen metabolism of the plants. In addition, the nitrate
content of tomato leaves under different nitrogen levels
showed an overall trend of N40 > N60 > N20 > N0, indicating
that the content of nitrate nitrogen increased with the increase
in the nitrogen application rate and decreased at levels less
than the optimum nitrogen supply.
The content of ammonium nitrogen, free amino acid nitro-
gen, and protein nitrogen in different grafted tomato leaves
showed a similar trend to that of nitrate nitrogen (Fig. 2b, c
and Fig. 3). The different grafting treatments showed a trend
of T2 > T1 > CK for these nitrogen contents, and under the

Table 4 Statistical analysis of tomato fruit quality among different treatments

Treatments Transverse Vertical Firmness Soluble Soluble protein Vitamin C Organic Lycopene
diameter (cm) diameter (cm) (kg cm-2) sugar (%) (mg g-1FW) (mg kg-1) acid (%) (μg g-1)

Grafted CK 5.38±0.21b 4.71±0.22b 14.43±0.52b 7.69±1.32c 0.85±0.07b 146.17±12.47c 7.26±0.84c 2.84±0.49b

treatments T1 5.78±0.15ab 5.19±0.12ab 15.20±0.30ab 9.73±0.83b 1.01±0.08ab 183.47±7.62b 9.59±0.36b 3.30±0.27ab
T2 6.73±0.86a 5.45±0.23a 15.43±0.45a 12.12±1.39a 1.19±0.12a 200.45±8.63a 11.10±0.69a 3.70±0.33a
N N0 5.62±0.15c 5.00±0.05c 14.03±0.13d 8.58±0.36c 0.96±0.03c 189.80±4.41a 9.83±0.24a 3.48±0.12a
treatments N20 5.68±0.12bc 5.00±0.09c 14.37±0.20c 9.54±0.43b 0.97±0.04c 182.38±4.85ab 9.41±0.10b 3.34±0.18ab
N40 6.47±0.21a 5.32±0.02a 15.38±0.13a 10.67±0.43a 1.10±0.03a 161.98±3.17c 8.92±0.45c 3.10±0.19b
N60 6.07±0.18b 5.15±0.08b 14.94±0.12b 10.60±0.16a 1.03±0.02b 172.61±7.04b 9.12±0.26bc 3.20±0.11b
P value
Grafted 0.0119 0.0011 0.0019 0.0005 0.0125 0.0001 0.0008 0.0011
treatments
N treatments 0.0028 0.0052 0.0001 0.0021 0.0032 0.0034 0.0092 0.0248
G×N 0.0318 0.0392 0.0483 0.0363 0.7211 0.0345 0.0387 0.0257
 Z. H. Zhang et al.

different nitrogen treatment conditions, a trend of N40 > N60 nitrogen supply, the higher the GS activity. When the nitrogen
> N20 > N0 was observed. supply exceeded the appropriate value, the GS activity
declined.
The content of GOGAT and GDH in different grafted to-
Dynamic changes in nitrogen metabolism-related mato leaves showed a similar trend to that of GS (Fig. 4). The
enzymes different grafting treatments showed a trend of T2 > T1 > CK
in GOGAT and GDH content. Under different nitrogen treat-
The changes in GS, GOGAT, and GDH in different grafted ment conditions, a trend of N40 > N60 > N20 > N0 was
tomato leaves under different nitrogen fertilizers are shown in observed.
Fig. 4. The results of statistical analysis of GS activity in
tomato leaves treated with grafting and nitrogen levels at dif-
ferent growth stages are shown in Fig. 4a and b. Over time, the Distribution of nitrogen in different organs of the
activity of GS in tomato leaves treated with different grafting grafted tomatoes
treatments showed a trend of first slowing, then increasing
significantly, and then decreasing. GS activity was low in The distribution of nitrogen in the roots, stems, leaves, and
January and peaked in March (fruit enlargement). These re- fruits of different treatments at harvest is shown in Table 5.
sults indicate that the activity of the GS enzyme is affected by The nitrogen content of tomato plants at different nitrogen
the ambient temperature and that GS exhibits higher activity supply levels showed the highest nitrogen levels in leaves
during the expansion period of the tomato fruit. In addition, and fruits. The nitrogen content was highest in the T2 grafted
there were significant differences in the activity of GS among seedlings, followed by that in T1 and CK. The nitrogen con-
grafting treatments. At different nitrogen levels, GS activity in tent in the different organs of the grafted tomato seedlings was
T2 grafted seedlings was the highest, followed by that in T1 different, and the nitrogen content in leaves and fruits was
and CK, indicating that the rootstock was the main factor significantly higher than that in roots and stems. The differ-
determining the activity of GS in the different grafted toma- ence between the different treatments was also significant.
toes. Furthermore, the GS activity of the tomato leaves under The nitrogen content of each organ in the grafting treatment
different nitrogen treatments was consistent with the grafting was the highest in T2, T1 was the second, and CK was the
treatment. There were significant differences in enzyme activ- lowest, indicating that the rootstock promoted the absorption
ity at different nitrogen levels, and enzyme activity showed a of nitrogen in the tomato plants. In the nitrogen fertilizer treat-
trend of N0 < N20 < N60 < N40, indicating that the higher the ments, the nitrogen content of the different organs increased

50 50
a CK T1 T2
b N0 N20
Nit rat e n itro gen ( m g· kg-1 )

Nitrate nitrogen (mg· kg-1 )

N40 N60
44 44

38 38

32 32

26 26

20 20

100 100
N0 N20
c CK T1 T2 d
ammonium nitrogen (mg· kg-1 )
am m o n ium n itro gen ( m g· kg-1 )

N40 N60
80 80

60 60

40 40

20 20

0 0
11-04 1-17 2-17 3-25 5-28 11-04 1-17 2-17 3-25 5-28
Date (M-d) Date (M-d)

Fig. 2 Statistical analysis of tomato leaves nitrate nitrogen and plants; d nitrate nitrogen in different nitrogen treatments. The data were
ammonium nitrogen content among the different treatments. a Nitrate analyzed by two-factor multiple comparison analysis, presented as the
nitrogen in different grated tomato plants; b nitrate nitrogen in different average of three repeated multiple comparisons ± SD
nitrogen treatments; c ammonium nitrogen in different grated tomato
Grafting improves tomato yield under low nitrogen conditions by enhancing nitrogen metabolism in plants

Fig. 3 Statistical analysis of 60 60

N0 N20
tomato leaves free amino acid a b N40 N60

Free am in o acid n it ro gen

Free amino acid nitrogen

nitrogen and protein nitrogen 48 48
content among the different
36 36

(mg· kg-1 )

(mg· kg-1 )
treatments. a Free amino acid
nitrogen in different grated 24 24
a
tomato plants; b free amino acid
nitrogen in different nitrogen 12 12
treatments; c protein nitrogen in CK T1 T2
different grated tomato plants; d 0 0
protein nitrogen in different
nitrogen treatments. The data 30 30
N0 N20
c CK T1 T2 d

Protein nitrogen (g· kg-1 )

were analyzed by two-factor P r o t ein n itr ogen ( g· k g-1 ) N40 N60
multiple comparison analysis, 25 25
presented as the average of three
20 20
repeated multiple comparisons ±
SD 15 15

10 10

5 5
11-04 1-17 2-17 3-25 5-28 11-04 1-17 2-17 3-25 5-28
Date (M-d) Date (M-d)

with the increasing nitrogen application rate. When the appli- in a certain range has a promoting effect on nitrogen absorp-
cation rate exceeded a certain level, the absorption of nitrogen tion in tomato. Moreover, grafting and nitrogen fertilizer had
decreased. The nitrogen content under the N40 treatment was significant interaction effects on nitrogen uptake by different
the highest, indicating that the application of nitrogen fertilizer organs.

Fig. 4 Statistical analysis of 15

15
tomato leaves GS, GOGAT, and a CK T1 T2
b N0 N20
GS ( µm o l· FW -1 · g-1 · m in-1 )

GS (µmol· FW -1 · g-1 · min-1 )

N40 N60
GDH activity among the different 12 12
treatments. a GS in different
grated tomato plants; b GS in
9 9
different nitrogen treatments; c
GOGAT in different grated
tomato plants; d GOGAT in 6 6
different nitrogen treatments; e
GDH in different grated tomato 3 3
plants; f GDH in different
nitrogen treatments. The data 0.6 0.6
c d
GOGAT (µm o l· FW -1 · g-1 · min -1 )

GOGAT (µmol· FW -1 · g-1 · min -1 )

CK T1 T2 N0 N20
were analyzed by two-factor 0.5 N40 N60
0.5
multiple comparison analysis,
presented as the average of three 0.4 0.4
repeated multiple comparisons ±
0.3 0.3
SD
0.2 0.2

0.1 0.1

0 0

1.2 1.2
e f
GDH (µmol· FW -1 · g-1 · min -1)
GDH ( µm o l· FW -1 · g-1 · m in-1 )

0.9 0.9

0.6 0.6

0.3 0.3
N0 N20
CK T1 T2 N40 N60
0 0
11-04 1-17 2-17 3-25 5-28 11-04 1-17 2-17 3-25 5-28
Date (M-d) Date (M-d)
 Z. H. Zhang et al.

Table 5 Statistical analysis of total nitrogen content (mg·g-1) in tomato organs the among the different treatments

Treatments Root Stem Leaf Fruit

Grafted treatments CK 18.24±1.10c 18.08±1.76c 28.40±2.56c 29.72±2.02c

T1 20.55±1.17b 21.19±1.59b 33.57±1.31b 34.27±0.97b
T2 23.61±1.15a 25.52±1.42a 37.21±1.25a 37.46±1.65a
N treatments N0 19.37±0.31d 20.14±0.47d 32.25±0.42c 32.67±0.28c
N20 20.51±0.21c 21.16±0.35c 32.67±0.43bc 33.54±0.41b
N40 22.14±0.12a 23.03±0.27a 34.07±0.44a 34.55±0.27a
N60 21.16±0.35b 22.05±0.40b 33.25±0.31b 34.52±0.32a
P value
Grafted treatments 0.0002 0.0001 0.0001 0.0001
N treatments 0.0011 0.0202 0.0191 0.0182
G×N 0.0081 0.0117 0.0424 0.0086

Effects of different rootstocks of grafted tomatoes on
nitrogen utilization efficiency
The statistical analysis of the nitrogen absorption, PFPN,
AEN, and NUE of tomato under different treatments is shown
in Table 6. At the different nitrogen supply levels, the nitrogen
absorption, PFPN, AEN, and NUE of the T1 and T2 grafting
treatments were significantly higher than those of CK and T2
> T1. The nitrogen absorption, PFPN, AEN, and NUE of the
T1 grafted seedlings were 32.97, 25.83, 44.99, and 21.04%
higher than those of the CK seedlings, respectively, and those
of the T2 grafted seedlings were 63.77, 45.47, 63.35, and
37.36% higher than those of the CK seedlings, respectively.
This indicated that the rootstock was the key factor in improv-
ing the nitrogen fertilizer utilization ability of the grafted to-
mato. The stronger the nitrogen fertilizer utilization ability of
the rootstock, the more significant the improvement in the
nitrogen fertilizer utilization ability of the grafted tomato seed-
lings was. There were also significant differences among the
different nitrogen fertilizer treatments, and the trend in
nitrogen absorption was N40 > N60 > N20 > N0. This indi-
cated that the nitrogen uptake of tomato was proportional to
the amount of nitrogen applied, up to a certain amount of
nitrogen application. When the application of nitrogen fertil-
izer exceeded a certain level, the amount of nitrogen uptake
decreased. However, the PFPN, AEN, and NUE in the differ-
ent nitrogen fertilizer treatments were highest in the N20 treat-
ment, followed by those of the N40 and N60 treatments, indi-
cating that the nitrogen use efficiency decreased with the in-
creasing nitrogen application rate. In addition, there was an
interaction effect between the grafting treatment and the nitro-
gen fertilizer treatment on nitrogen absorption and utilization.
Discussion
In the previous studies, the nitrogen efficiency evaluation
methods used for different crops or different genotypes or
growth stages of the same crop were different. It has long been
reported that plants have genotypic differences in nitrogen use

Table 6 Statistical analysis of nitrogen utilization efficiency of tomato among the different treatments

Treatments N absorption PFPN AEN NUE

(g plant-1) (kg kg-1) (kg kg-1) (%)

Grafted treatments CK 18.44±3.55c 162.59±23.02c 58.14±10.73c 38.11±4.21c

T1 24.52±2.39b 204.58±15.83b 84.29±6.52b 46.13±3.78b
T2 30.20±2.16a 236.51±12.74a 94.96±3.61a 52.35±2.14a
N treatments N0 18.40±0.97d — — —
N20 24.73±0.67c 294.86±53.18a 95.06±1.58a 62.50±8.30a
N40 28.18±0.66a 189.76±36.54b 89.86±3.41b 48.29±5.55b
N60 26.23±0.72b 119.06±33.56c 52.46±11.92c 25.80±9.12c
P value
Grafted treatments 0.0001 0.0001 0.0015 0.0051
N treatments 0.0021 0.0002 0.0001 0.0018
G×N 0.0456 0.0127 0.0332 0.0475
Grafting improves tomato yield under low nitrogen conditions by enhancing nitrogen metabolism in plants
(Chen 2006; Cui et al. 2009; Barbosa et al. 2018). The NUE of
crops is directly related to plant growth and development and
final yield (Razaq et al. 2017; Pellegrini et al. 2018; Wang
et al. 2018). Therefore, the NUE of crops is mainly determined
by their own genotypes. The growth and yield of nitrogen-
efficient genotypes are higher than those of nitrogen-
inefficient genotypes under normal or low nitrogen conditions
(Mansour et al. 2017; Zhang et al. 2018). Studies by Tirol-
Padre et al. (1996) and Fageria et al. (2010) indicated that
nitrogen uptake and utilization efficiency vary among differ-
ent genotypes of rice. Svečnjak and Rengel (2006) found dif-
ferences in nitrogen use efficiency among different varieties in
grain. These studies guided our screening of nitrogen-efficient
tomato genotypes. The results of this experiment showed that
there were large differences in genotype among the 25 tomato
materials tested under the three nitrogen supply levels. At
different nitrogen levels, the CVs of the nitrogen absorption,
NUE, and PFPN were high. These results indicate that the
three indicators are sensitive to the difference in nitrogen use
efficiency among different genotypes of tomato and can be
used as important indicators for screening for tomato geno-
types with high nitrogen utilization capacity. These results are
in line with those of Duan (2019), who compared the differ-
ences in spatial and temporal distribution properties of
different NUE maize genotypes by studying the phenotypes
of root crowns of maize at the seedling stage, bell stage, and
silking stage. The experiment also found that the higher the
nitrogen supply level, the lower the NUE and the PFPN,
which was consistent with the conclusions of Li et al. (2017).
Thus far, grafting technology has been widely used in hor-
ticultural production to improve plant resistance to various
biotic and abiotic stresses. The structure of the grafted plants
and the internal material transport system are more complicat-
ed than those of the self-rooted plants. Due to the characteris-
tics of the roots of the rootstock and the interaction between
the rootstocks, the original absorption capacity of the plants
changed. Then, the relationships among the source, the reser-
voir, and their physiological and biochemical processes were
also different from those of the self-rooted plants, resulting in
the plant growth and development, fruit yield, and quality
being affected (Rouphael et al. 2018; Ropokis et al. 2019).
This study found that compared with that of the control, the
yield of grafted seedlings with ‘TMS-150’ and ‘0301111’
rootstocks increased by 45.46 and 24.68%, respectively,
which is consistent with the conclusion that grafting cultiva-
tion can increase crop yield (Kunwar et al. 2017). The soluble
sugar, soluble protein, organic acid, vitamin C, and lycopene
contents of the different rootstock-grafted seedlings measured
in this experiment were higher than those in the control, which
is consistent with the conclusion of Kyriacou et al. (2016).
However, the differences in soluble protein and lycopene con-
tents were small, which is consistent with the findings of
Gisbert et al. (2011).
Nitrogen metabolism is one of the most basic metabolisms
in plants and plays a decisive role in crop yield and growth.
Different forms of nitrogen have different effects on plants.
The nitrogen source for plant roots is mainly inorganic nitro-
gen, such as nitrate nitrogen and ammonium nitrogen (Berge
et al. 2007). After the nitrate nitrogen is absorbed, part of it is
assimilated in the form of amino acids to be transported up-
wards, part is not assimilated but is stored in the form of nitrate
nitrogen in the root cells, and most of the ammonium nitrogen
is assimilated into amino acids and transported to the shoot
(Xu et al. 2012; Bloom et al. 2012). GS, GDH, and GOGAT
are key enzymes in nitrogen metabolism and play an impor-
tant role in ammonia assimilation in plants. Among them, GS
is a multifunctional enzyme involved in the regulation of var-
ious nitrogen metabolism processes (Robredo et al. 2011).
However, many studies have found that grafting, in addition
to increasing plant resistance to biotic and abiotic stresses, can
also increase absorptive capacity under nutrient-deficient con-
ditions (Kumar et al. 2015; Rouphael et al. 2018). In this
study, the key enzymes of nitrogen metabolism, GS,
GOGAT, and GDH; the key components nitrate nitrogen,
ammonium nitrogen, free amino acid nitrogen, protein nitro-
gen, and total nitrogen; and the plant nitrogen utilization indi-
cators PFPN, AEN, and NUE under different treatments were
measured and calculated. The results showed that the key
enzyme activities, content of key components, nitrogen con-
tent, and nitrogen utilization capacity of grafted tomato leaves
at different nitrogen fertilizer levels were higher than those of
the self-grafted control, and the nitrogen utilization rate of the
grafted plants with nitrogen-efficient rootstock was higher.
This result indicated that grafting tomatoes increased the ac-
tivity of key nitrogen metabolism enzymes, promoted the di-
gestion and absorption of nitrogen by plants, and improved
the ability of tomato to metabolize nitrogen and utilize nitro-
gen fertilizer. Moreover, the stronger the nitrogen utilization
ability of the rootstock, the more significant the improvement
in the nitrogen utilization capacity of the grafted tomato seed-
lings was.
Conclusion
Based on the nitrogen absorption, NUE, and PFPN of tomato
seedlings under three nitrogen application levels, we identi-
fied the material ‘TMS-150’ with a strong nitrogen utilization
ability and the material ‘0301111’ with a weak nitrogen utili-
zation ability from 25 tomato genotypes. Our results showed
that, in grafted tomato plants as the nitrogen fertilizer content
increased from low to high, the yield of the tomato plants, the
activity of key enzymes during nitrogen metabolism, the con-
tent of different forms of nitrogen, and the efficiency of nitro-
gen use first increased and then decreased, and these indica-
tors were highest under the N40 nitrogen fertilizer treatment.

However, under different nitrogen fertilizer conditions,
grafting tomatoes with high-NUE tomato seedlings as the
rootstock increased the nitrogen content and the activities of
key nitrogen metabolism enzymes, enhanced the nitrogen ab-
sorption, PFPN, AEN, and NUE of tomato plants, increased
the tomato yield, and improved the fruit quality. The study
also showed the complexity of nitrogen metabolism; further
investigation is still needed to understand the mechanisms by
which the interactions between the rootstock and the scion can
improve nitrogen utilization.
Authors’ contributions Kun Xu designed experiments; Zhi Huan Zhang,
Ming Ming Li, Bili Cao, and Zi Jing Chen complete the experiments; Zhi
Huan Zhang and Kun Xu wrote the manuscript.
Funding information This work was supported by the Double First-class
Discipline Construction Project of Shandong Province (No.
SYL2017YSTD06).
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