horticulturae
Article
Response of Nutritional Status and Tea Quality to the Rate and
Substitution of Chemical Fertilizers with Organic Manure
Lifeng Ma 1,2 , Yun Zhu 1,2 , Saipan Geng 1 and Jianyun Ruan 1,2, *
Citation: Ma, L.; Zhu, Y.; Geng, S.;
Ruan, J. Response of Nutritional
Status and Tea Quality to the Rate
and Substitution of Chemical
Fertilizers with Organic Manure.
Horticulturae 2022, 8, 1198. https://
doi.org/10.3390/horticulturae
8121198
Academic Editor: Miguel Guzmán
Received: 12 October 2022
Accepted: 13 December 2022
Published: 15 December 2022
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Horticulturae 2022, 8, 1198. https://doi.org/10.3390/horticulturae8121198
1 Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China
2 Key Laboratory of Tea Biology and Resource Utilization of the Ministry of Agriculture,
Hangzhou 310008, China
* Correspondence: jruan@mail.tricaas.com; Tel.: +86-571-86653938
Abstract: Proper fertilization is important to sustainable tea production. A field experiment was
conducted to investigate the response of quality components in a chlorotic tea variety (Zhonghuang-2)
to rates of fertilizers and the substitution ratio of chemical fertilizers by organic manure based on
rapeseed cake. Chlorotic tea varieties have unique metabolic characteristics and produce superior tea
containing high contents of free amino acids. Results showed that fertilization significantly increased
yield and contents of free amino acid (TFAA) but reduced contents of total polyphenol (TP) and the
ratio of TP/TFAA. Contents of TFAA and TP and the TP/TFAA ratio were closely related to nitrogen
(N) concentrations in plant tissues in response to the rate of N fertilizers. The results suggest that
the quality-related components in the chlorotic tea variety respond to fertilizers in a similar way as
normal tea varieties. The optimal rates of N, phosphorus (P), and potassium fertilizers were discussed
and recommended based on the response of quality components of tea and the contents of nutrients
in plants and soil. The full substitution of chemical fertilizers by organic manure showed no special
benefit on tea quality and had lower N and P agronomic use efficiency due to a low bioavailability of
nutrients. The partial substitution of chemical fertilizers by organic manure significantly improved
tea yield, quality, profit, and economic and environmental sustainability.
Keywords: chlorotic tea variety; organic manure; partial organic substitution; yield; free amino acids;
total polyphenol; nutrient use efficiency
1. Introduction
Tea (Camellia sinensis) is a valuable cash crop widely planted all over the word and
plays important roles in increasing farmers’ income and alleviating poverty of rural areas
in China and other tea-producing countries. The tea quality is determined by internal
chemical compositions, which is greatly affected by the supply of nutrients to tea plants [1].
Deficiency of nutrients reduces tea yield and quality [1]. Nitrogen (N), phosphorus (P),
and potassium (K) are nutrients most widely applied in tea fields. The beneficial effects
of fertilizers on yield and quality are realized only when they are properly applied [2–5].
On the other hand, the use of excessive amounts of inorganic fertilizers is well known [6,7].
The excessive application of N, P, and K fertilizers negatively affects the biosynthesis and
accumulation of free amino acids, catechins and lipid composition, and reduces tea qual-
ity [2,8–11]. In addition, the excessive application of fertilizers deteriorates soil quality and
causes serious environmental problems [12–14]. Recent studies highlighted that tea-planted
soils are global hotspots for N2 O emission as a result of the overuse of N fertilizers [15].
Organic fertilizers are important sources of nutrients. The application of organic
manure improves soil fertility and microbiological activity, mitigates soil acidification,
and decreases the runoff of nutrients [16–19]. However, compared to chemical fertilizers,
organic fertilizers have low nutrient concentrations, slow nutrient-release rates, and high
application costs. This problem can be overcome by the application of organic fertilizers
https://www.mdpi.com/journal/horticulturae
Horticulturae 2022, 8, 1198 2 of 13

in combination with chemical fertilizers. Partial substitution of chemical fertilizers with

organic manure increases nutrient use efficiency, reduces the application of chemical fertil-
izers and carbon dioxide emissions, and better sustains soil fertility without compromising
yield and quality [20,21]. However, the optimal ratios between organic and chemical fer-
tilizers are variable depending upon crop species, the application rate of fertilizers, and
the type of organic fertilizers [20]. Compared to the extensive investigations in cereal
crops [20,21], the effect of organic substitution on the yield and quality of perennial tea
plants has been tested only limitedly [17,18,22]. Synchronizing the release of nutrients from
organic fertilizers with the nutritional demand of crops is a challenge because the former is
greatly influenced by soil temperature [23]. Spring tea, especially the early spring tea, has
the best quality compared to teas of other seasons [24,25]. The growth and biosynthesis of
metabolites of spring tea might be inhibited by weak and slow release of nutrients from
organic fertilizers due to low temperatures. Previous work showed that organically grown
tea had low contents of free amino acids [26].
The flavor and quality of tea are attributed to polyphenols, alkaloids, and free amino
acids. Amino acids are the principal contributors to the mellow taste of brewed green tea.
Premium green tea has high contents of free amino acids. In recent years, several natural
mutants with chlorotic young leaves sensitive to light intensity have been cultivated
in China and Japan [27–29]. These chlorotic tea varieties have enhanced levels of free
amino acids and superior quality of made teas. They have higher economic value than
the non-chlorotic varieties. The high content of free amino acids in these tea varieties
is the result of abnormal protein degradation and weakening biosynthesis of nitrogen-
containing compounds such as chlorophyll, purines, and nucleotides rather than enhanced
biosynthesis of amino acids [27–29]. However, due to the limited photosynthetic capacity
of chlorotic leaves, these varieties generally have lower yields and contents of catechins
than normal varieties [27]. Improving the nutritional status of chlorotic tea plants through
proper fertilization is a promising approach to overcome these shortcomings. Due to unique
metabolic characteristics, these varieties might respond to fertilization differently than the
normal varieties. Compared to extensive works on the metabolism of quality components
in chlorotic mutants [27–29], the effect of fertilization has not been evaluated. In the present
work, a field experiment was conducted for three years to investigate the response of
nutrient uptake and quality parameters in a chlorotic tea variety to the application rate of
fertilizers and the substitution ratio of chemical fertilizers by organic fertilizers. The aims
are to develop proper nutrient management schemes for these cherished tea varieties to
achieve sustainable yields, good quality, and high profits with reduced environmental risks.

2. Materials and Methods

2.1. Field Experiment
A field experiment was set up in 2017 in Tiantai County, Zhejiang Province, China.
The tea cultivar was Zhonghuang-2, a native mutant with chlorotic young leaves under
natural light intensity [27]. The soil pH was 5.70. The contents of total organic carbon,
total N, available P, and available K were 9.63 g kg−1 , 1.1 g kg−1 , 10.1 mg kg−1 , and
134 mg kg−1 , respectively. There were five treatments: A control without any fertilizers
(CK), chemical fertilizers at optimized application rates (referred to as NPK), substitution
of the chemical fertilizers fully or partially by organic manure (referred to as OM and
NPKOM, respectively), and high application rates of chemical fertilizers (referred to as
NPK-H). The amounts and compositions of nutrients of treatments are presented in Table 1.
The NPK, OM, and NPKOM received the same total amount (527 kg ha−1 ) of N, P2 O5 ,
and K2 O but different rates of each nutrient. In the NPK treatment, a compound fertilizer
specially formulated for tea (N-P2 O5 -K2 O -MgO, 18-8-12-2) [30] was applied in the middle
of October as a basal fertilizer and two topdressings in early February and at the end of
April after the harvest of spring tea. The amounts of fertilizers of the three applications
were 40%, 30%, and 30% of the total, respectively. In the OM treatment, rapeseed cake
manure (N-P2 O5 -K2 O, 5.8-2.7-1.5) was applied in early October as the basal fertilizer. In
Horticulturae 2022, 8, 1198 3 of 13

NPKOM treatment, half of the total amount of nutrients was supplied from rapeseed cake
manure and another half from compound fertilizer and urea. In this treatment, all organic
manure and compound fertilizer were applied as the basal fertilizer. A certain amount
of urea was applied to supplement N to the defined rate (Table 1). Urea was applied as
two topdressings equally in early February and at the end of April. The NPK-H treatment
received a doubled amount of compound fertilizer of the NPK treatment. The application
timing of NPK-H was the same as the NPK treatment. In all treatments, the basal fertilizers
were applied to furrows with depths of 15–20 cm and the topdressing fertilizers to furrows
with depths of 5–10 cm located between rows. The furrows were covered with soil after
the application of fertilizers. The cost of fertilization including fertilizers and labor for
application ranged from 7200 to 20,250 CNY ha−1 (Table 1). The area for a plot was
20 m2 and plots were randomly arranged in the field. There were four replications for each
treatment. All other field management was the same as in the local tea plantations.

Table 1. The amount and composition of fertilizers, and the cost of fertilization including fertilizers
and application of treatments.

Nutrient Treatment
CK NPK OM NPKOM NPK-H
Total (kg ha−1 ) N 0 250 312 282 500
P2 O5 0 110 140 125 220
K2 O 0 167 75 120 334
Sum 0 527 527 527 1054
Organic (kg ha−1 ) N 0 0 312 158 0
P2 O5 0 0 140 70 0
K2 O 0 0 75 37 0
Sum 0 0 527 264 0
Cost (CNY ha−1 ) 0 7200 20,250 12,600 11,370

2.2. Samples and Measurements

Only spring tea was harvested for chlorotic mutant tea varieties for the highest price
and profit in spring compared to other seasons. Young shoots consisting of one bud with
one or two expanding leaves were harvested by hand and weighed as the fresh yield. Yield
data of 2019–2020 were presented in the present work. Samples of harvested young shoots
were dried in an electric oven at 60 ◦ C following inactivation of the oxidase enzyme in a
microwave oven (800 W) for 2 min, and finally ground into fine powders (using a ball mill
(Mixer Mill MM300, Retsch GmbH, Haan, Germany)) for the measurement of nutrients.
In 2020, young shoots of buds with one expanding leaf were sampled on 4 April
and 21 April (hereafter referred to as early and late spring, respectively). Samples were
quickly frozen in liquid nitrogen and then transported to a laboratory to measure contents
of chlorophyll. Chlorophyll content of young shoots was determined by spectrophotometer
following extraction by acetone [1]. A portion of young shoots were freeze-dried and
then ground into fine powders for the measurement of total free amino acid (TFAA) and
total polyphenol (TP). Samples of young shoots (100 mg) were extracted with 5 mL of
H2 O in a boiling water bath for 5 min. The extract was used for determination of TFAA
by spectrophotometry following reacting with ninhydrin reagent and for TP following
reacting with Fe-tartrate reagent [4]. Mature leaves on the surface canopy were taken on
21 April 2020, dried at 60 ◦ C in an electric oven, and then ground into fine powders for
the measurement of nutrients. N concentration in young shoots and mature leaves was
determined by an elemental analyzer (Vario Macro Cube, Elementar Analysensysteme
GmbH, Langenselbold, Germany). Concentrations of P and K in plant samples were
determined by Inductive Coupled Plasma-Atomic Emission Spectrometer (iCAP™ 7400
ICP-OES, Thermo Fisher Scientific, Waltham, MA, USA) after digestion at 550 ◦ C and
re-dissolved in dilute nitric acid.
Horticulturae 2022, 8, 1198 4 of 13

Soil samples were taken using an auger in October 2020 at 5 randomly selected sites of
each plot from depths of 0–20 cm and 80–100 cm. Soil from the same plot and layer was
combined to obtain a composite sample. Stones and debris of roots were removed. Soil
samples were separated into two portions after thorough mixing. One portion was tem-
porarily stored in a refrigerator at 4 ◦ C for the determination of water content and inorganic
nitrogen. Ammonium (NH4 + -N) and nitrate (NO3 − -N) in fresh soil were extracted with
2 mol L−1 potassium chloride (1/10 w/v) and determined by Discrete Chemistry Ana-
lyzer (Smartchem 140, AMS Alliance, Frepillon, France). Another portion was air-dried
and passed thorough 20 mesh sieves for determining soil pH and available P and K. Soil
pH was measured in 1:2.5 water paste by a glass electrode (Orion 3 Star, Thermo Ltd.,
Waltham, MA, USA). Soil available P and K were extracted by Mehlich-3 reagent [31] and
determined by Inductive Coupled Plasma-Atomic Emission Spectrometer (iCAP™ 7400
ICP-OES, Thermo Fisher Scientific, Waltham, MA, USA). All chemicals were purchased
from Merck & Co., Inc. (Rahway, NJ, USA) or China National Pharmaceutical Group Co.,
Ltd. (Beijing, China) with analytical or higher purity.

2.3. Calculation of Agronomic Use Efficiency and Profit

Agronomic use efficiency (AE, kg kg−1) of nutrients was calculated from the yield
(YF, kg ha−1) and the total amount (F, kg ha−1) of N, P2O5, and K2O and their sum (Table 1) in
the treatments relative to the yield of CK (YCK, kg ha−1) according to the following Equation (1):

AE = (YF − YCK )/F (1)

To compare the interaction effect on nutrient agronomic use efficiency between organic
and inorganic resources in NPKOM, the theoretical nutrient use efficiency was calculated
from the following Equation (2) according to [32]:

AEmix = (Forg × AEOM + Finorg × AENPK )/F × 100 (2)

where AEmix is the theoretical nutrient use efficiency (in kg kg−1 ) of the NPKOM treatment
and Forg , Finorg , and F are the nutrient amounts from organic and inorganic fertilizers and
total amount (kg ha−1 ), respectively; AEOM is the nutrient use efficiency (in kg kg−1 ) of the
100% organic treatment (OM), and AENPK is the nutrient use efficiency (in kg kg−1 ) of the
100% inorganic treatment (NPK).
The average price of fresh spring tea was approximately 60 CNY kg−1 according to
the local market. The value (V) of fresh shoots was calculated as the product of the average
price and the yield. The net profit of fertilization was calculated as the difference between
the values of treatments (VF ) and CK (VCK ) and the fertilization cost (CF ), according to
the following Equation (3). The cost of harvesting young shoots was not included in the
calculation of the net profit, as this provided important employment for local farmers.

Profit = (VF − VCK ) − CF (3)

2.4. Estimation of Greenhouse Gas Emissions Derived from Fertilization

The greenhouse gas emissions (GHGs) derived from fertilization were estimated ac-
cording to the default method provided by the Intergovernmental Panel on Climate Change
(IPCC) of the United Nations [33]. GHGs were divided into four parts generated from
the production and transportation of fertilizers and the direct and indirect N2 O emissions
caused by the application of N fertilizers. Details and equations for the estimation can be
found in Supplemental Table S1. The GHGs (CO2 equivalent) generated from the production
and transportation of chemical fertilizers were estimated from their application rates per
hectare and respective emission factors (EFs). The EFs of manufacture and transportation
were 8.21 and 0.09 for N fertilizers, 0.73 and 0.06 for P fertilizers, and 0.5 and 0.05 for K
fertilizers [34,35], respectively. We assumed that rapeseed cake manure was locally recycled
without long-distance transportation. The direct emission of N2 O from the application of
Horticulturae 2022, 8, 1198 5 of 13

chemical and organic N fertilizers was estimated from their application rates per hectare
and their respective emission factors specified for tea plantations (0.0175 and 0.0261) [15].
The indirect N2 O emissions caused by the application of N fertilizers were calculated from
N application rates per hectare, fractions of N loss through leaching (14.53%) and runoff
(8.20%), which were taken from recent respective field experiments under similar conditions,
and an emission factor of 0.011 [16,35,36]. Indirect N2 O emissions caused by the loss of
N through NH3 volatilization was not considered for strongly acid tea soil and covering
with soil after the application of fertilizers. The direct and indirect emissions of N2 O were
converted to CO2 equivalent greenhouse gas emissions. Total GHGs were further converted
to emissions scaled per yield (GHGA ) and per profit (GHGP ), respectively.

2.5. Statistical Analysis

One-way analysis of variance (ANOVA), combined with the least significant difference
(LSD) test, was used to compare the effect of treatments. Regression analysis was performed
to describe the relations between quality parameters with the rate of fertilizers and nutrient
concentrations in plant tissues. The best-fitting curves were selected according to their
corrected R2 -values together with visual inspection of each curve type. All analyses were
performed using SigmaStat embedded in SigmaPlot (Version 12.0, Systat Software Inc.,
Palo Alto, CA, USA).

3. Results
3.1. Yield, Nutrient Absorption, and Nutrient Use Efficiency
Compared to CK, the average yield of two years was increased by 75–111% in the
fertilization treatments (Table 2). The high rate of nutrients (NPK-H) increased yield more
significantly than full organic manure (OM) in 2020. However, there was no significant
difference within the means of two years among the four fertilization treatments, although
NPK-H and NPKOM had slightly (p > 0.05) higher yields. The profit of fertilization varied
from 43,235 to 70,261 CNY ha−1 (Table 2). Without considering the impact of tea quality
on the price of fresh shoots, OM had the lowest profit and profit/cost ratio. Profit was the
highest in NPK-H, followed by NPKOM and NPK. However, the profit/cost ratio was the
highest in NPK.

Table 2. Yield, profit, and profit/cost ratio of fertilization treatments.

Treatment Yield (kg ha−1 ) Profit Profit/Cost

2019 2020 Mean (CNY ha−1 ) Ratio
CK 569a 312a 440a 0 /
NPK 812b 898bc 855b 64,957 9.0
OM 824b 720b 772b 43,235 2.1
NPKOM 836b 981bc 908b 67,603 5.4
NPK-H 791b 1068c 929b 70,261 6.2
Different letters following data of the same line indicate significant difference among treatments.

N concentrations of young shoots were considerably higher in the early (4 April) than
in the late spring (21 April) (Figure 1a). By contrast, K and P concentrations were higher in
the late than in the early spring (Figure 1b,c). Their concentrations were also significantly
affected by the fertilization. For both the early and late spring, the trends of N, P, and
K concentrations in the different treatments are well described by quadratic regressions.
Young shoots of NPK-H had the highest N, P, and K concentrations. The concentrations of
N, P, and K were not significantly different among OM, NPK, and NPKOM.
 in the late than in the early spring (Figure 1b,c). Their concentrations were also signifi- 223
cantly affected
in the late bythe
than in theearly
fertilization. For both
spring (Figure the Their
1b,c). early and late spring,were
concentrations the trends of N, P, 223224
also signifi-
and Kaffected
cantly concentrations in the different
by the fertilization. treatments
For both areand
the early welllate
described by quadratic
spring, the regres-
trends of N, P, 224225
sions.
and Young shoots in
K concentrations of the
NPK-H had the
different highestare
treatments N, well
P, and K concentrations.
described Theregres-
by quadratic concen- 225226
Horticulturae 2022, 8, 1198 trations
sions. of N,shoots
Young P, andofKNPK-H
were nothadsignificantly
the highestdifferent among
N, P, and OM, NPK, and
K concentrations. TheNPKOM.
concen-
6 of 13 226227
trations of N, P, and K were not significantly different among OM, NPK, and NPKOM. 227
80
(a) Early spring Late spring

Young shoot N (mg g )

Young shoot N (mg g-1) -1
80 CK CK
(a) Early spring Late spring
70 2
R =0.652**** NPK NPK
CKOM CK OM
70 2
R =0.652**** NPKNPKOM NPKNPKOM
60 OMNPK-H OMNPK-H
NPKOM
Quadratic NPKOM
Quadratic
60 NPK-H
regression NPK-H
regression
50 2
R =0.732**** Quadratic Quadratic
regression regression
2
50 R =0.732****
40
0 100 200 300 400 500 600
40
0 100 200 -1
N rate300 400
(kg ha ) 500 600
7 -1 28
N rate (kg ha )
(b) (c)
Young shoot P (mg g )

Young shoot K (mg g-1)

7 28
Young shoot P (mg g-1) -1

2
(b) R =0.553*** (c)

Young shoot K (mg g-1)

2
6 R =0.553*** 24 R2=0.202
6 24 R2=0.202

5 2 20
R =0.468**
5 2 20
R =0.468** 2
R =0.438***
2
4 16 R =0.438***
4 0 50 100 150 200 250 16 0 100 200 300 400
0 50 100 150 200 250 0 100 200 300 400
P2O5 rate (kg ha-1) K2O rate (kg ha )-1

P2O5 rate (kg ha-1) K2O rate (kg ha-1)

228
228
Figure 1. Response of concentrations of N (a), P (b), and K (c) in young shoots to the rate of fertilizers 229
Figure
Figure
in the 1. Response
early (4 April) concentrations
of concentrations of N
of
and late (21 April N2020)
(a), Pspring.
(a), K (c) in
(b), andSingle young
bars shoots
in color to the rate
without fertilizers
dataofpoint
fertilizers
are LSD 229230
the early
invalues
the early (4 April) and late (21 April 2020)
2020) spring.
spring. Single bars in color without
indicative of significant difference among treatments. Quadratic regression lines are data point are
are LSD
indi- 230231
LSD
values
values indicative
indicative of
of significant
significant difference
difference among
among treatments.
treatments. Quadratic
Quadratic regression
regression
cated with R values and significance levels (*, p < 0.05; **, p < 0.01; ****, p < 0.0001; n = 20).
2 lineslines
are are indi-
indicated 231
232
catedRwith
with R2 values
2 values and significance
and significance levels levels
(**, p <(*,0.01;***,
p < 0.05;
p<**,0.001;
p < 0.01;
****,****,
p < p0.0001;
< 0.0001;
n =n20).
= 20). 232

Mature leaves of NPK-H had the highest N and P concentrations, followed by NPK 233
Matureleaves
leavesofofNPK-H
NPK-Hhad had the highest N and P concentrations, followedNPK by NPK
and 233234
andMature
NPKOM (Figure the
2a,b). Mature highest
leaves ofN OM
and Phad
concentrations,
significantlyfollowed
lower Nby and P concen-
and NPKOM
NPKOM (Figure
(Figure 2a,b).2a,b).
MatureMature
leaves leaves
of OM ofhad
OMsignificantly
had significantly
lower lower
N and N
P and P concen- 234
concentrations
trations than NPK and NPKOM. K concentrations of mature leaves in NPK-H, NPK, and 235
trations
than NPK than
andNPK and NPKOM.
NPKOM. K concentrations
K concentrations of matureofleaves
maturein leaves
NPK-H, inNPK,
NPK-H, andNPK,
NPKOMand 235
NPKOM were not significantly different but were higher than those in OM (Figure 2c). 236
NPKOM
were were not significantly
not significantly different
different but but were
were higher higher
than thosethan
in OM those in OM
(Figure 2c).(Figure 2c). 236
Similarly,
Similarly, the relation between the concentrations of N, P, and K in mature leaves with the 237
the relation between the concentrations of N, P, and K in mature leaves with the ratesthe
Similarly, the relation between the concentrations of N, P, and K in mature leaves with of 237
rates of nutrients were well described by quadratic regressions (Figure 2). The yield was 238
rates of nutrients
nutrients were well were well described
described by quadraticby quadratic regressions
regressions (Figure (Figure 2). The
2). The yield was yield was 238
closely
closely related to N concentration in mature leaves and their relation was well described 239
closely related
related to N concentration
to N concentration in mature
in mature leavesleaves and their
and their relation
relation was was
wellwell described
described by 239
byquadratic
by quadratic
quadratic
regression
regression
regression
(Figure
(Figure
(Figure
3a).
3a).3a). 240
240

45
45 (a) y=20.215+0.054x-3.92*10-5x2 CK
40 (a) y=20.215+0.054x-3.92*10
-5 2 CKNPK
Mature leaf N (mg g )

R2=0.844**** x
Mature leaf N (mg g )-1

40 R2=0.844**** NPK
-1

OM
35 OMNPKOM
35 NPKOM
NPK-H
30 NPK-H
30 Quadratic regression
Quadratic regression
25
25
20
20
15
15
10
10
0 0 100 100 200200 300
300 400400 500
500
-1
NN rate
rate (kg
(kg haha
-1 )
)
2.8
2.8 20 20 (c)
(b)(b) y=2.036+0.02x-3.27*10
-6 2
y=2.036+0.02x-3.27*10
-6 2 x
x (c)
Mature leaf P (mg g )

Mature leaf K (mg g )

2
Mature leaf P (mg g-1-1)

Mature leaf K (mg g )

-1

R2R=0.296*
=0.296*
-1

2.4 18 18
2.4

16 16
2.0
2.0
14 14
1.6
1.6 y=12.647+0.044x-9.43*10
y=12.647+0.044x-9.43*10-5 2 x
x
-5 2
12 12
R2=0.547****
R2=0.547****
1.2
1.2 10 10
00 5050 100 150
100 150 200
200 250250 0 0 100100 200200 300300 400400
-1 -1 -1
P2PO25Orate
5 rate (kg
(kg haha) ) K2ratel
K2O O ratel ha-1ha
(kg(kg ) )
241241

Figure 2. Response of concentrations of N (a), P (b), and K (c) in mature leaves to the rate of fertilizers.
Single bars in red without data point are LSD values indicative of significant difference among
treatments. Quadratic regression lines are indicated with equations, R2 values, and significance levels
(*, p < 0.05; ****, p < 0.0001). The circled data (OM treatment) were not included in the regression
analysis for the significantly low bioavailability of N and P.
 Figure 2. Response of concentrations of N (a), P (b), and K (c) in mature leaves to the rate of fertiliz- 242
ers. Single bars in red without data point are LSD values indicative of significant difference among 243
treatments. Quadratic regression lines are indicated with equations, R2 values, and significance lev- 244
Horticulturae 2022, 8, 1198 7 of 13
els (*, p < 0.05; ****, p < 0.0001). The circled data (OM treatment) were not included in the regression 245
analysis for the significantly low bioavailability of N and P. 246

1400 80
(a) (b)
CK
1200 NPK
60 OM

TFAA (mg g-1)

Yield (kg ha-1)
1000
NPKOM
800 NPK-H
40 Quadratic
600 regression
400 y=-1664.5+142.4x-1.88x2 20 y=81.02-2.45x+0.030x2
200
2
R =0.652*** R2=0.819****

0 0
15 20 25 30 35 40 45 15 20 25 30 35 40 45
400 14
(c) (d)
12 y=22.17-0.90x+0.011x2
300 R2=0.778****
10
TP (mg g-1)

TP/TFAA
8
200
6

100 2
4
y=370.74-4.10x-0.0024x
R2=0.584*** 2
0 0
15 20 25 30 35 40 45 15 20 25 30 35 40 45
-1
Mature leaf N concentration (mg g ) 247
Figure3.3.Relation
Figure Relationofoftea
teayield
yield(a),
(a),total
totalfree
freeamino
aminoacid
acid(TFAA,
(TFAA,b),
b),total
totalpolyphenol
polyphenol(TP,
(TP,c),
c),and
andthe
the 248
ratio of TP/TFAA (d) in young shoots with the N concentration in mature leaves on April
ratio of TP/TFAA (d) in young shoots with the N concentration in mature leaves on 21 April 2020. 21, 2020. 249
Quadratic regression lines are indicated with equations, R2 values, and significance levels (***, p < 250
Quadratic regression lines are indicated with equations, R2 values, and significance levels
0.001; ****, p < 0.0001). 251
(***, p < 0.001; ****, p < 0.0001).

The amounts
The amounts of of N,
N,P,P,and
andKKininyoung
young shoots were
shoots all significantly
were increased
all significantly by ferti-
increased by 252
lization and mostly by NPK-H (Table 3). The agronomic use efficiencies
fertilization and mostly by NPK-H (Table 3). The agronomic use efficiencies of N (AE of N (AE N), ),
N
P 253
(AE ), and the sum (AE ) were significantly higher in NPKOM
P (AEP ), and the sum (AEsum ) were significantly higher in NPKOM and NPK than in
P sum and NPK than in NPK- 254
H and OM.
NPK-H and OM.The agronomic
The agronomicuse efficiency of Koffertilizers
use efficiency (AEK(AE
K fertilizers ) was higher in OM and
K ) was higher in OM
255
NPKOM
and NPKOM than in NPK
than in NPKandandNPK-H
NPK-H (Table
(Table3).
3).The
Thetheoretical
theoretical agronomic
agronomic useuse efficiencies
efficiencies 256
(AEmix
(AE mix)) of
of N,
N,P,P,and
andKKand
andtheir sum
their sumforfor
NPKOM
NPKOM according
according to Equation
to Equation(2) were 1.32,1.32,
(2) were 2.97, 257
3.08, and 0.71, respectively. All AE values were
2.97, 3.08, and 0.71, respectively. All AEmix values were lower than
mix lower than their counterparts
counterparts of of 258
NPKOM presented in
NPKOM presented in Table 3. Table 3. 259

Table3.3.Amount
Table Amountofof nutrients
nutrients in in harvested
harvested young
young shoots,
shoots, agronomic
agronomic nutrient
nutrient use efficiency
use efficiency (AE),(AE),
and 260
and greenhouse gas emissions scaled per area (GHG A), yield (GHGY), and profit (GHGP).
greenhouse gas emissions scaled per area (GHGA ), yield (GHGY ), and profit (GHGP ). 261

Treatment Amount (kg ha−1) AE (kg kg−1) GHGA GHGY GHGP

Treatment Amount (kg ha−1 ) AE (kg kg−1 ) GHGA GHGY GHGP
N P K N P2O5 K2O Sum (kg ha−1)
# (kg kg−1) (kg Yuan−1)
N P K N P2 O5 K2 O Sum # (kg ha−1 ) (kg kg−1 ) (kg Yuan−1 )
CK 6.0a 0.58a 2.34a / / / / 0 0 0
CK
NPK 6.0a 0.58a 2.34a
12.8bcd 1.18bc 4.33b / 1.66b / 3.74b / 2.48a / 0.79b 0 4596 05.38 0
0.071
NPK 12.8bcd 1.18bc 4.33b 1.66b 3.74b 2.48a 0.79b 4596 5.38 0.071
OM
OM
11.9bc 1.07b
11.9bc
1.07b 4.04b
4.04b 1.06a 2.37a
1.06a
2.37a 4.42b
4.42b 0.63ab4179
0.63ab
4179 5.41
5.41
0.097
0.097
NPKOM
NPKOM 13.4cd
13.4cd 1.22bc
1.22bc 4.54b
4.54b 1.66b
1.66b 3.74b
3.74b 3.87b
3.87b 0.89b
0.89b 4401
4401 4.85
4.85 0.065
0.065
NPK-H
NPK-H 14.0d
14.0d 1.27c 4.62b
1.27c 4.62b 0.98a 2.20a
0.98a 2.20a 1.46a
1.46a 0.46a 9192
0.46a 9192 9.89
9.89 0.131
0.131
* Different
Different letters
letters following
following data of data of the
the same linesame line
indicate indicatedifference
significant significant difference
among among
treatments. treatments.
# Sum represents 262
# Sum represents N+P2O5+K2O. 263
N+P 2 O5 +K2 O.

3.2.
3.2.Tea
TeaQuality
Quality 264
Fertilization
Fertilizationhad
hadlittle
littleeffect onon
effect thethe
content of chlorophyll
content of chlorophyllin young shoots
in young (Figure
shoots 4a).
(Figure 265
Compared to CK,tothe
4a). Compared content
CK, of total of
the content free amino
total freeacid (TFAA)
amino acidin(TFAA)
young shoots
in youngwasshoots
increased
was 266
mostly by NPK-H
increased mostly byfollowed
NPK-Hbyfollowed
NPK and byNPKOM
NPK and (Figure
NPKOM 4b).(Figure
The increase in TFAA
4b). The was
increase in 267
smaller in OM than in other fertilization treatments. TFAA content was higher in the
early (mean 56.6 mg g−1 ) than in the late spring (mean 48.4 mg g−1 ). The differences
between the early and late spring were greater in CK and OM than in other treatments. For
instance, TFAA contents of the late stage were 24–30% lower than the early stage in CK and
OM, whereas such differences were only 10–12% in NPK, NPK-H, and NPKOM. TFAA
contents in the early and late spring were significantly and positively related to their N
 TFAA was smaller in OM than in other fertilization treatments. TFAA content was higher 268
268
in the early (mean 56.6 mg g−1−1) than in the late spring (mean 48.4 mg g−1 −1). The differences 269
269
between the early and late spring were greater in CK and OM than in other treatments. 270
270
Horticulturae 2022, 8, 1198
For instance, TFAA contents of the late stage were 24–30% lower than the early stage in
8 of 13
271
271
CK and OM, whereas such differences were only 10–12% in NPK, NPK-H, and NPKOM. 272
272
TFAA contents in the early and late spring were significantly and positively related to 273
273
their N concentrations of young2 shoots (R22 = 0.253, p <2 0.05 and R22 = 0.673, p < 0.0001, 274
274
concentrations of young
respectively; Figure 5). shoots
TFAA (Rcontent waspalso
= 0.253, < 0.05 and R = 0.673,
significantly p <in0.0001,
related respectively;
the quadric pattern 275
275
Figure 5). TFAA content was also significantly related
with the N concentration of mature leaves (Figure 3b). in the quadric pattern with the N 276
276
concentration of mature leaves (Figure 3b).
0.25
0.25 (a) 100
100 (b)
(a) CK
CK (b)
NPK
NPK

(mggg-1-1))
0.20
0.20 OM
OM 80
80
NPKOM

(mggg-1-1))
NPKOM
NPK-H
Chlorophyll(mg
NPK-H
0.15
0.15 60
60

TFAA(mg
Chlorophyll

0.10 40

TFAA
0.10 40

0.05
0.05 20
20

0.00
0.00 0
0
Chl-a
Chl-a Chl-b
Chl-b Chl
Chl Early
Early spring
spring Late
Late spring
spring
350
350 (c) 12
12 (d)
(c) (d)
300
300 10
10
250
250
(mggg-1-1))

8
8

TP/TFAA
TP/TFAA
200
200
6
TP(mg

6
150
150
4
TP

100 4
100
50 2
2
50
0
0 0
0
Early
Early spring
spring Late
Late spring
spring Early
Early spring
spring Late
Late spring
spring
277
277
Figure
Figure4.4.
Figure Concentrations
Concentrationsofof
4.Concentrations chlorophyll
ofchlorophyll (chl-a,
chlorophyll(chl-a, chl-b,
(chl-a,chl-b, and
chl-b,and total)
andtotal) (a),
total)(a), total
(a),total free
totalfree amino
freeamino acid
aminoacid (TFAA,
acid(TFAA,
(TFAA,b),b),
b), 278
278
total polyphenol
totalpolyphenol
total polyphenol(TP, (TP, c) and
(TP,c)c)and their
andtheir ratio
theirratio (PP/TFAA)
ratio(PP/TFAA)
(PP/TFAA)(d) (d) in young
(d)ininyoung shoots
youngshoots in the
shootsininthe early
theearly and
earlyand late
andlate spring.
latespring.
spring. 279
279
Single bars
barsinin
Singlebars
Single red
inred without
redwithout
withoutdata data above
dataabove columns
abovecolumns
columnsare are LSD
areLSD values
LSDvalues indicative
indicativeofof
valuesindicative significant
ofsignificant difference
significantdifference
difference 280
280
among fertilization
amongfertilization treatments.
fertilizationtreatments.
treatments. 281
281
among

100
100 (a)
(a) Early
Early spring
spring Late
Late spring
spring
80
80 CK
CK CK
CK
R22=0.673**** NPK OM
(mggg-1-1))

R =0.673**** NPK OM
OM
OM NPK
NPK
60
60 NPKOM NPK-H
TFAA(mg

NPKOM NPK-H
NPK-H
NPK-H NPKOM
NPKOM
40 Linear
Linear regression Linear
Linear regression
TFAA

40 2 regression regression
R
R2=0.253*
=0.253*
20
20

0
0
45
45 50
50 55
55 60
60 65
65 70
70 75
75 80
80
400
400 (b) 14
14 (c)
(b) (c)
12
12
300
300 10
10
(mggg-1-1))

TP/TFAA
TP/TFAA

2
8
8 R
R2=0.420**
=0.420**
200
TP(mg

200 R 2
R2=0.657****
=0.657**** 2 6
6
R =0.206*
R 2
=0.206*
TP

100 4
4
100 2
2 R =0.781****
R 2
=0.781****
2
0
0 0
0
45
45 50
50 55
55 60
60 65
65 70
70 75
75 80
80 45
45 50
50 55
55 60
60 65
65 70
70 75
75 80
80
-1
Young
Young shoot
shoot N
N concentration
concentration (mg
(mg g
g ))
-1
282
282
Figure
Figure5.5.
Figure Response
5. Response
Response of of total free
of total free amino
amino acid
acid (TFAA,
acid (TFAA, a),
(TFAA, a), total
a), total polyphenol
total polyphenol(TP,
polyphenol (TP,b),
(TP, b),and
b), andthe
and theratio
the ratioof
ratio of 283
283
TP/TFAA
TP/TFAA (c)
(c) to
to N
N concentration
concentration in
in young
young shoots
shoots in
in the
the early
early (open
(open
of TP/TFAA (c) to N concentration in young shoots in the early (open symbols) and the latesymbols)
symbols) and
and the
the late
late spring
spring 284
284
(closed
(closed symbols).
symbols). Linear regression
LinearLinear
regression lines are
are indicated
lines lines
indicated with R
R22 values
with with values and
and significance
significance levels (*,
(*, pp <<
levelslevels 285
285
spring (closed symbols). regression are indicated R2 values and significance
0.05;
0.05; **,
**, pp << 0.01;
0.01; ****,
****, pp << 0.0001).
0.0001). 286
286
(*, p < 0.05; **, p < 0.01; ****, p < 0.0001).

ComparedtotoCK,
Compared CK,fertilization
fertilizationsignificantly
significantlyreduced
reducedthe
thecontent
contentofoftotal
totalpolyphenol
polyphenol 287
287
(TP) in young shoots (Figure 4c). The greatest decrease was found in
(TP) in young shoots (Figure 4c). The greatest decrease was found in the NPK-H andthe NPK-H and 288
288
followed by, in decreasing order, NPK, NPKOM, and OM. TP contents in the early and late
spring were significantly and negatively correlated with their N concentrations of young
shoots (R2 = 0.206, p < 0.05 and R2 = 0.657, p < 0.0001, respectively) (Figure 5b). TP content
of the late stage was also negatively (R2 = 0.584, p < 0.001) related to N concentration in the
mature leaves (Figure 3c). The ratio of TP/TFAA in young shoots responded to fertilization
in a similar way as TP (Figure 5c). TP/TFAA was negatively related with N concentration
in mature leaves and their relation was best described by quadratic regressions (Figure 3d).
 young shoots (R2 = 0.206, p < 0.05 and R2 = 0.657, p < 0.0001, respectively) (Figure 5b). TP 291
content of the late stage was also negatively (R2 = 0.584, p < 0.001) related to N concentra- 292
tion in the mature leaves (Figure 3c). The ratio of TP/TFAA in young shoots responded to 293
fertilization in a similar way as TP (Figure 5c). TP/TFAA was negatively related with N 294
Horticulturae 2022, 8, 1198 concentration in mature leaves and their relation was best described by quadratic regres-
9 of 13 295
sions (Figure 3d). 296

3.3. Soil Properties 297

3.3. Soil Properties
The soil pH varied from 4.81 to 5.95 and was the lowest in the surface soil and in- 298
The soil pH varied from 4.81 to 5.95 and was the lowest in the surface soil and increased
creased
in in the
the deep soildeep soil (Figure
(Figure 6a). Surface
6a). Surface soil pHsoil
waspH was significantly
significantly decreaseddecreased by fertili-
by fertilization, 299

with the greatest decrease in NPK-H. OM also decreased soil pH to a lesser extentextent
zation, with the greatest decrease in NPK-H. OM also decreased soil pH to a lesser than 300
than other
other fertilization
fertilization treatments.
treatments. NPKOMNPKOM decreased
decreased soilsoil
pHpH but
but totoa asmaller
smallerextent
extent than
than 301
NPK. The content of NH +-N increased with the rate of N fertilizers (Figure 6b,c). Their
4+
NPK. The content of NH4 -N increased with the rate of N fertilizers (Figure 6b,c). Their 302
relationship could
relationship couldbe bedescribed
describedby bylinear
linearregression.
regression.NH NH+4+− −N
N was
was the highest in
the highest in NPK-H
NPK-H 303
4
and was
and wasslightly
slightlylower
lowerininOM OMthan
thaninin NPK.
NPK. NO NO −3 −N contents increased sharply in NPK-
- 304
3 –N contents increased sharply in NPK-H,
H, which were 3.5–12.5 times those in other treatments
which were 3.5–12.5 times those in other treatments (Figure (Figure
6b,c).6b,c). The response
The response of NOof3 NO
− –N3- 305
−N content
content to rate
to the the rate
of Noffertilizers
N fertilizers
could could be best
be best described
described by the
by the equation
equation of exponen-
of exponential 306
tial growth (y=a × e
x x). The ratio of NO3-− −N/NH +−N+ was significantly
growth (y = a × e ). The ratio of NO3 –N/NH4 –N was significantly different
4 different among
amongfer- 307
tilization treatments and was the highest
fertilization treatments and was the highest in NPK-H.in NPK-H. 308

Soil pH
3 4 5 6 7
CK
(a)
NPK
Depth (cm)

0-20 OM
NPKOM CK
NPK-H NPK
80-100 OM
NPKOM
NPK-H

40 200 300
(b) NH4+, R2=0.479**** (d) (f) R2=0.493***
30 NO3-, R2=0.893**** R2=0.724****
150
200
20 100

Available K (mg kg-1)

Inorganic N (mg kg )

Available P (mg kg )
-1

-1

10 50 100

0 0
0
40 20 300
+ 2
(c) NH4 , R =0.277** (e) R2=0.494**** (g)
30 NO3-, R2=0.565**** 15 R2=0.338**
200
20 10

10 5 100

0 0
0
0 100 200 300 400 500 0 50 100 150 200 250 0 100 200 300 400
-1
N rate (kg ha ) P2O5 rate (kg ha-1) K2O rate (kg ha-1)
309
Figure 6.
Figure 6. Soil
SoilpH
pH(a),
(a),contents
contentsofofNONO−N − (b,c, open symbols), NH −N (b,c, + closed symbols), avail-
3 −N (b,c, open symbols), NH4 −N (b,c, closed symbols),
3- 4+ 310
available P (d,e), and K (f,g) in the soil of 0–20 cm (b,d,f; in blue) and 80–100 cm (c,e,g;in
able P (d,e), and K (f,g) in the soil of 0–20 cm (b,d,f; in blue) and 80–100 cm (c,e,g; ingreen)
green) depths.
depths. 311
Single bars without data are LSD values indicative of significant difference among treatments. Re- 312
Single bars without data are LSD values indicative of significant difference among treatments. Regression
gression lines are indicated with R2 values and significance levels (*, p < 0.05; **, p < 0.01; ***, p < 313
lines are indicated with R2 values and significance levels (**, p < 0.01; ***, p < 0.001; ****, p < 0.0001).
0.001; ****, p < 0.0001). 314

Available P accumulated mostly in the surface soil and increased significantly with
Available
the rate P accumulated
of P fertilizers mostlyThe
(Figure 6d,e). in the surface
relations soil and
between increased
available P insignificantly
the soil andwith
the 315
the rate
rate of Pof P fertilizers
fertilizers could(Figure 6d,e).
be well The relations
described betweenequations.
by quadratic available PThein the
soilsoil andOM
of the the 316
rate of P fertilizers could be well described by quadratic equations. The
treatment had slightly lower available P contents than NPK and NPKOM regardless of its soil of the OM 317
treatment
higher ratehad
of Pslightly lower
fertilizers. availableKPcontent
Available contents than
was NPK and NPKOM
significantly affected byregardless
the rate ofof its
K 318

fertilizers (Figure 6f,g). Their relations in different soil depths could be well describedofby
higher rate of P fertilizers. Available K content was significantly affected by the rate K 319
fertilizers (Figure 6f,g). Their relations in different soil depths could be well
quadratic equations. Available K was higher in the surface soil than in the deep soil. The described by 320
quadratic in
difference equations.
availableAvailable
P among K was
soil higher
depths wasin much
the surface
largersoil
thanthan
thatinofthe deep soil.
inorganic The
N and 321
available K.

3.4. Estimation of Greenhouse Gas Emissions

Greenhouse gas emissions (GHGs) from the production and transport of chemical
fertilizers accounted for half of the total emissions in NPK and NPK-H and accounted for
about 25% of the total in NPKOM (Table S1). The GHGs ranged from 4179 to 9192 kg ha−1 ,
from 4.85 to 9.89 kg kg−1 , and from 0.065 to 0.13 kg CNY−1 for area, yield, and profit scaled
emissions, respectively (Table 3). GHGs were the highest in NPK-H compared to other
treatments. The OM treatment had lower area scaled emissions, but higher yield and profit
Horticulturae 2022, 8, 1198 10 of 13

scaled emissions than NPK and NPKOM. NPKOM had lower area, yield, and profit scaled
total emissions than NPK.

4. Discussion
4.1. Response of Tea Yield, Quality Parameters, and Soil Property to Nutrient Application Rates
The present work showed that increasing the rate of N fertilizers enhanced the N status
of tea plants and the content of TFAA in young shoots. Furthermore, the contents of TFAA,
TP, and the ratio of TP/TFAA were closely associated with the N concentrations of mature
leaves. Generally, these relations were well described by quadratic regressions. According
to these regressions, the optimal rate of N fertilizers was estimated 250–280 kg ha−1 (in
NPK and NPKOM) (Figures 2a and 3). The estimated optimal N rates were lower than
the values of 296–422 kg ha−1 recommended by a Nutrient Expert for normal tea varieties
based on yield performance [5]. The high N rate of NPK-H resulted in the highest TFAA
content but the lowest TP content and TP/TFAA ratio (Figure 4). The overall tea quality of
NPK-H was considered deteriorated because of a low TP and unbalance between TP and
TFAA, weakening the body of the infusion. These results suggest that the quality-related
components respond to N in the chlorotic tea variety in a similar way as normal tea varieties
and thus do not support the hypothesis that they might be different [8,37]. The contents of
residual NO3 − −N in soil responded to the application rate of N fertilizers in exponential
ways (Figure 6b,c), in line with previous findings with field crops [38,39]. By contrast, the
source of fertilizer had little effect on the content of residual inorganic N in the soil. The
high N application rate resulted in sharply increased residual NO3 − −N in the soil and the
deep soil (80–100 cm) below the root zone. Nitrate is prone to leaching because of abundant
rainfall in tea planting areas. According to the exponential regressions, the transition rate
of N fertilizers beyond which the residual NO3 − –N content in soil tremendously increased
was estimated around 350–400 kg ha−1 . Hence, the optimal N rates appeared safe from
risks of strong NO3 − leaching.
A soil analysis indicated that the experimental site had a low status of available
P and medium-level K in the soil at the time the present work was started. P and K
fertilization greatly improved their concentrations in plants (Figures 1 and 2). According
to the quadratic regressions, P and K concentrations in mature leaves and young shoots
no longer significantly increased when the application rates of P and K fertilizers reached
110–126 kg ha−1 and 120 kg ha−1 in NPK and NPKOM, respectively. Higher rates of P and
K fertilizers beyond these values (in NPK-H) no longer increased P and K concentrations
in tea plants and decreased remarkably the agronomic P and K use efficiency (Table 3).
Available P in the soil increased with the rate of P fertilizers in the quadratic tendency.
Available P accumulated and was affected by P fertilizers mostly in the surface soil (0–20 cm)
(Figure 6d). The impact of P fertilizers on available P of the soil was observed down to
the depth of 80–100 cm (Figure 6e), indicating somewhat downward P leaching in acidic
tea soils. Previous work proposed a change-point index of soil extractable P to predict
the leaching potential [40,41]. The critical index of Bray-1 extractable soil P (referred to as
PBray-1 ) was found to be 75.1 mg kg−1 for tea plantations [14]. The Bray-1 extractable soil P
can be converted to Mehlich-3 extractable P (referred to as PM3 ) according to a previously
established regression equation (PM3 = 1.4974 × PBray-1 − 4.4468, R2 = 0.98, p < 0.0001) [42]
for tea soils. The critical value of PM3 was estimated to be about 108 mg kg−1 , which was
surpassed in the soil receiving the high rate of P fertilizers in NPK-H and suggests a high
risk of potential P leaching. Available K in the soil responded to the rate of K fertilizers
in the quadratic mode (Figure 6f,g). Such a response was observed down to the depth of
80–100 cm, indicating significant downward K leaching in the soil profile. These results
suggest that under the condition of continuous applications, the rates of P and K fertilizers
need further optimization by soil analysis to mitigate a high accumulation in the soil and
to reduce the risk of potential leaching [41].
Horticulturae 2022, 8, 1198 11 of 13

4.2. The Efficacy of Full and Partial Organic Substitution

Substitution of chemical fertilizers with organic manure is a promising approach to
reduce the use of chemical fertilizers and promote the recycling of nutrients in agricultural
systems [20]. The present experiment demonstrated that full organic substitution did not
significantly affect tea yield of chlorotic variety Zhonghuang-2 relative to the application
of chemical fertilizers alone. However, compared to NPK and NPKOM, full organic
substitution decreased the content of TFAA but increased that of total polyphenol and the
TP/TFAA ratio in young shoots (Figure 4), suggesting a decrease in tea quality. This finding
is in line with previous results showing higher phenolic contents in organically than in
conventionally grown tea and other crops [23,26,43]. Our previous works showed that the
biosynthesis of polyphenols is upregulated while that of amino acids is downregulated in
the case of low N supply [8,37]. These findings together with the lower N concentrations in
mature leaves in OM suggest that these tea plants had insufficient N supply, even though
they received a slightly higher N rate compared to NPK and NPKOM (Figure 2a and
Table 1). Furthermore, OM had a significantly lower agronomic N use efficiency (Table 3).
Therefore, the low N status of tea plants in OM was most likely caused by a slow release
and then weak intensity of N from the organic manure. In addition, the absorption of
N by tea plants might be also competed by the microbial immobilization of mineral N
in the soil stimulated by the additional carbon from organic fertilizers. Meanwhile, the
low P agronomical use efficiency and low P concentration in mature leaves were also
observed in the full substitution treatment (Figure 2b and Table 3), likely suggesting a
low bioavailability of P from rapeseed cake manure. The present work showed that the
bioavailability of nutrients (especially N) in soils fertilized only with rapeseed cake manure
is inadequate to cover the demand necessary to obtain a high-quality spring tea [44].
Partial substitution of chemical fertilizers with organic manure had similar contents of
TFAA and TP and a similar ratio of TP/TFAA with slightly increased yield (by 6%, p > 0.05)
compared to its chemical counterpart (NPK). The greenhouse gas emissions scaled per area,
yield, and profit were all decreased by the partial substitution. Agronomic use efficiency of
N and P was improved by the positive interaction of chemical fertilizers in combination with
organic manure, as indicated by AEmix < AE(NPKOM) . The advantage of partial substitution
might be the better balance between the readily and mineralizable N and P supply to
match the nutrient demand of perennial tea plants and additionally the improvement of
soil quality (e.g., soil microbial biomass, abundance, and activity) [17,18]. The application
of specially formulated compound fertilizer alone also significantly improved tea quality
and achieved a high profit with low greenhouse gas emissions (GHGY , GHGP ). In practical
use, the specially formulated compound fertilizer might be selected for mountainous tea
plantations without convenient transport systems.

5. Conclusions
The present work demonstrated that the productivity and quality of chlorotic tea variety
Zhonghuang-2 were closely related with the nutritional status of tea plants and the rate of
fertilizers and sources. The results suggest that the quality-related components in chlorotic tea
variety Zhonghuang-2 respond to fertilizer in a similar way as normal tea varieties, despite
its unique metabolic characteristics. The full substitution of chemical fertilizers with organic
fertilizers had no advantage on tea productivity and quality due to the low bioavailability
of nutrients, especially N in organic fertilizers based on rapeseed cake manure. The partial
organic substitution better matched the nutrient demand by perennial tea plants to ensure
good tea quality, high economic profits, and reduced environmental risks.

Supplementary Materials: The following are available online at https://www.mdpi.com/article/10

.3390/horticulturae8121198/s1, Table S1: Estimation of greenhouse gas emissions.
Author Contributions: Conceptualization and experimental design, J.R. and L.M.; field investigation,
sample collection, and analysis, L.M., Y.Z. and S.G.; data curation, J.R., L.M. and Y.Z.; writing of
manuscript, J.R. and Y.Z. All authors have read and agreed to the published version of the manuscript.
Horticulturae 2022, 8, 1198 12 of 13

Funding: This work was financially supported by the National Key Research and Development
Project (2021YFD1601100), the Earmarked Fund for China Agriculture Research System (CARS 19),
the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural
Sciences (CAAS-ASTIP-TRICAAS), and the Agricultural Department of Zhejiang Province through
contract no. 2021SNLF032.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Conflicts of Interest: The authors declare no conflict interest.

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