Plant Soil (2019) 445:183–198

https://doi.org/10.1007/s11104-019-04293-6

REVIEW ARTICLE

Salicylic acid alleviates aluminum-induced inhibition

of biomass by enhancing photosynthesis and carbohydrate
metabolism in Panax notoginseng
Chunyan Dai & Lisha Qiu & Lanping Guo & Sisi Jing &
Xiaoya Chen & Xiuming Cui & Ye Yang

Received: 14 May 2019 / Accepted: 29 August 2019 / Published online: 17 October 2019
# Springer Nature Switzerland AG 2019

Abstract Methods Effects of SA on the biomass of P. notoginseng

Aims Determine whether salicylic acid (SA) can allevi-
ate the inhibitory effects of Al stress on dry matter
accumulation in Panax notoginseng, and to investigate
the mechanism by which the SA regulates photosynthe-
sis and carbohydrate metabolism.
and the concentration of Al in leaves under Al stress were
evaluated. SA and the scavenger paclobutrazol (PAC, SA
synthetic inhibitor) were used to assess the effects of SA
on photosynthesis and the metabolism of total nonstruc-
tural carbohydrates (TNC) in P. notoginseng.

Chunyan Dai and Lisha Qiu contributed equally to this work.

Responsible Editor: Miroslav Nikolic.

Electronic supplementary material The online version of this

article (https://doi.org/10.1007/s11104-019-04293-6) contains
supplementary material, which is available to authorized users.

C. Dai C. Dai : L. Qiu : S. Jing : X. Chen : X. Cui : Y. Yang

Faculty of Environmental Science and Engineering, Kunming
University of Science and Technology, Kunming 650500, China
C. Dai : L. Qiu : S. Jing : X. Chen : X. Cui (*) :
Y. Yang (*)
Yunnan Provincial Key Laboratory of Panax notoginseng|,
Kunming University of Science and Technology, No.727 South
Jingming Rd., Chenggong District, Kunming 650500, China
e-mail: sanqi37@vip.sina.com
e-mail: yangyekm@163.com
C. Dai : L. Qiu : S. Jing : X. Chen : X. Cui : Y. Yang
Key Laboratory of Panax notoginseng Resources Sustainable
Development and Utilization of State Administration of
Traditional Chinese Medicine, Kunming University of Science
and Technology, No.727 South Jingming Rd., Chenggong
District, Kunming 650500, China
Kunming Key Laboratory of Sustainable Development and
Utilization of Famous-Region Drug, Kunming University of
Science and Technology, No.727 South Jingming Rd., Chenggong
District, Kunming 650500, China
C. Dai : L. Qiu : S. Jing : X. Chen : X. Cui : Y. Yang
University Based Provincial Key Laboratory of Screening and
Utilization of Targeted Drugs, Kunming University of Science and
Technology, No.727 South Jingming Rd., Chenggong District,
Kunming 650500, China
C. Dai : L. Qiu : S. Jing : X. Chen : X. Cui : Y. Yang
Faculty of Life Science and Technology, Kunming University of
Science and Technology, No.727 South Jingming Rd., Chenggong
District, Kunming 650500, China
L. Guo
Chinese Medica Resources Center, China Academy of Chinese
Medicinal Sciences, Beijing 100700, China
184 Plant Soil (2019) 445:183–198

Results SA reduced Al by 13.23% in the leaves of Introduction

P. notoginseng under Al stress. It also promoted the
synthesis of carbohydrates in the leaves and their trans- Panax notoginseng (Burk.) F. H. Chen (P. notoginseng)
port to the roots. Additionally, SA alleviated the Al- is a perennial herb that belongs to the Araliaceae family
induced reduction of chlorophyll (Chl) by upregulating of the genus Panax. The roots and rhizomes of
the expression of Chl synthesis genes (CHLM, CHLG, P. notoginseng are famous Chinese materia medica that
HEMB1, HEMC, and HEME) and downregulating the are commonly used in the treatment of trauma and
degradation genes (CLH2 and PPH1). Concomitantly, ischemic cardiovascular diseases (Hua et al. 2012;
the rubisco activase (RCA) and rubisco enzyme activi- Zhang et al. 2003). The stem and leaf of
ties were promoted by upregulating the expressions of P. notoginseng have similar medicinal value as the root.
the RCA, rbcL, and rbcS genes via treatment with SA, It is reported in the Pents’ao Kang Mu that the stem and
thereby restoring the synthesis and metabolism of TNC. leaf are suitable for the treatment of fractures, bruises,
Furthermore, SA attenuated the decrease in photosyn- and bleeding (Wang et al. 2016). P. notoginseng typi-
thetic capacity and photosynthesis system II photo- cally grows in the acidic red soils of the Wenshan area in
chemistry efficiency in Al-stressed plants. Furthermore, Yunnan Province (Gao et al. 2001), and the plantation
SA significantly reduced the H2O2 concentration by distribution is highly coincident with that of bauxite
12.22% under Al stress. mineral resources. It is widely recognized by the scien-
Conclusions By regulating photosynthesis-related tific community that Al toxicity is a major limiting
genes and photochemical processes, SA promoted pho- factor for crop growth and yield in acidic soils. Our
tosynthesis and enhanced leaf carbohydrate synthesis previous studies have shown that the Al concentration
under Al stress. Thus, SA enhanced the production of in P. notoginseng has a significant correlation with Al in
P. notoginseng biomass, ultimately reducing the inhibi- soil, and inhibits the growth and development of
tory effects of Al. P. notoginseng (Yang et al. 2018a). With the increase
of soil acidification, the cultivation of P. notoginseng is
facing a widespread and increasingly serious problem of
Keywords Aluminum . P. notoginseng . Salicylic acid . Al toxicity. However, research in this field is scarce.
Carbohydrate . Photosynthesis Therefore, determining how to alleviate the reduction of
the yield and quality of P. notoginseng under Al stress is
Abbreviations an urgent problem that must be solved.
Chl Chlorophyll Salicylic acid (SA) serves as an important signaling
Ci Intercellular CO2 concentration molecule for stress resistance in plants, including the
Fv/Fm Maximal quantum yield of photosystem II alleviation of the decrease of biomass accumulation due
Gs Stomatal conductance to heavy metal stress. For example, exogenous SA has
NPQ Non-photochemical quenching coefficient been found to significantly alleviate the growth and
PAC Paclobutrazol yield inhibition of wheat/rapeseed under boron stress
Pn Net photosynthetic rate by regulating carbohydrate metabolism (El-Shazoly
PS II Photosystem II et al. 2019; Metwally et al. 2018). In addition, exoge-
qP Photochemical quenching coefficient nous SA has also been found to reduce the Al concen-
RCA Rubisco activase tration in rice and alleviate the inhibitory effect of
Rubisco Ribulose-1,5-bisphosphate carboxylase/ growth, thus promoting dry matter accumulation
oxygenase (Pandey et al. 2013). We previously demonstrated that
SA Salicylic acid exogenous SA reduced the Al concentration in
SC Structural carbohydrates P. notoginseng roots (Yang et al. 2018b). However,
TCA Trichloroacetic acid whether SA can alleviate the inhibitory effects of Al
TNC Total nonstructural carbohydrates stress on dry matter accumulation in P. notoginseng and
Tr Transpiration rate its corresponding mechanism remains to be determined,
TSC Total soluble carbohydrates and thus is the focus of the present study.
ΦPSII Quantum efficiency of photosystem II The accumulated biomass of plants is mainly derived
from carbohydrates produced by photosynthesis.
Plant Soil (2019) 445:183–198
Carbohydrates include structural carbohydrates (SC)
and total non-structural carbohydrates (TNC). SC in-
clude cellulose and lignin, are a constituent of plant cell
walls and pillar tissues, and are rarely involved in plant
metabolic activities. TNC includes starch and TSC (su-
crose + glucose + fructose). Through the sugar metabo-
lism pathway, TNC can directly or indirectly affect the
synthesis, transport, and accumulation of organic matter,
thereby affecting the yield and quality of crops
(Gururani et al. 2013; Nowicka et al. 2018). Drought
stress has been found to increase the activity of sucrose
synthase and decrease photosynthesis in cotton, leading
to the accumulation of TNC which exceeded its utiliza-
tion by photorespiration, and ultimately caused the in-
crease of TNC content and the decrease of plant biomass
(Zahoor et al. 2017). Rubisco (Ribulose-1,5-
bisphosphate carboxylase/oxygenase E.C. 4.1.1.39) is
the key enzyme involved in CO2 fixation in photosyn-
thesis, and is composed of large subunits (RBCL) and
small subunits (RBCS). The large subunit of rubisco in
higher plants is encoded in the chloroplast by rbcL,
while the small subunit is encoded by the nuclear rbcS
gene family. The activity of rubisco depends on the
activation degree by rubisco activase (RCA)
(Fukayama et al. 2018). The decrease of RCA activity
in tobacco under salt stress was found to result in the
decrease of rubisco activity, which impaired the photo-
synthesis of tobacco and inhibited its growth (Lee et al.
2014). Therefore, promoting CO2 fixation in plants can
promote their photosynthesis, thereby creating a mate-
rial foundation for improving plant biomass.
Chloroplast, the main plant cell organelle for photo-
synthesis, contains chlorophyll (Chl) and light systems.
The synthesis and degradation under stress of the total
Chl concentration is affected by various key genes,
including HEMA, HEMB, HEMC, HEME, CHLM,
CHLI, and CHLG in the Chl synthesis pathway. The
genes that regulate Chl degradation include PPH, PAO,
and CLH (Meng 2017). For example, Chl biosynthesis
at the Mg branch was found to be inhibited by silencing
CHLI in Pisum sativum (Luo et al. 2016). The photo-
synthetic rate can be reduced by the loss of plasma
membrane integrity and total Chl in leaves under stress
(Surapu et al. 2014; Guo et al. 2013). Additionally, the
photosynthetic system can be impaired by the
photoinhibition of the photosystem II (PSII) complex,
and the impairment level can be estimated by the reduc-
tion degree of the maximal quantum yield (Fv/Fm) or
quantum efficiency (ΦPSII) of photosystem II (Vass
185
2012). For instance, the decrease of Fv/Fm or ΦPSII in
Brassica napus L. leaves under Cu stress demonstrated
that Cu stress damaged the electron transport in photo-
synthetic systems (Zhang et al. 2019). Under the condi-
tion of sub-high temperature and high light intensity, the
photosynthesis of tomato leaves was found to be
inhibited, and the light absorbed by the tomato leaves
exceeded the utilization range of the photosynthetic
system, thus inducing the increase of the H2O2 concen-
tration, which was highly destructive to the photosyn-
thetic system and TNC metabolism (Lu et al. 2017). To
alleviate the peroxidation damage in the photosystem,
plant leaves will spontaneously consume excess light
energy through non-photochemistry dissipation or heat
dissipation, which can lead to the decrease of photo-
chemical quenching (qP) and the increase of non-
photochemical quenching (NPQ) (Zhang et al. 2015a).
Thus, alleviating the inhibition effect of stress on pho-
tosynthetic function can restore TNC metabolism,
which can be used as an effective method to maintain
or increase crop yield.
Exogenous SA can decrease the inhibitory effects of
heavy metal stress on photosynthesis by regulating PSII,
photosynthetic pigments, and rubisco enzyme activities
(Al-Whaibi et al. 2012; Noriega et al. 2012; Zhang et al.
2015b). In addition, exogenous SA has been found to
improve the photosynthetic characteristics of Oryza
sativa, Zea mays, and Phaseolus vulgaris under the
stress of Pb, Cd, and Cu, thereby alleviating the inhib-
itory effect of heavy metals on plant growth (Khan et al.
2015). Here, we hypothesized that SA could alleviate
the inhibitory effect of Al stress on photosynthesis by
reducing the Al concentration in leaves of
P. notoginseng, and as a result could alleviate the reduc-
tion of dry matter accumulation. Accordingly, the plant
growth, TNC, photosynthetic pigments, photosynthetic
parameters, Chl fluorescence, oxidative damage, and
expression of key genes of the Calvin cycle in
P. notoginseng under Al stress with or without SA
treatment were evaluated.
Materials and methods
Experimental materials
Two-year old seedlings of P. notoginseng were planted
in the greenhouse of faculty of life science and
186
technology (E 102.87, N 24.85, altitude 1895 m), Kun-
ming University of Science and Technology.
Plant culture and treatments
Seedlings were collected from the greenhouse. Before
hydroponic experiments, the seedlings were pre-washed
thoroughly with tap water to remove soil. Seedlings then
were sterilized with 2% sodium hypochlorite solution
for 2 h and washed thoroughly with distilled water. The
seedlings were cultured in a light-resistant box contain-
ing 250 ml of culture solution (0.5 mM CaCl2, pH 4.5)
in a growth chamber under a 14 h/25 °C day and 10 h/
22 °C night regime, with a light intensity of 100 μmol
photon m−2 s−1 (photosynthetic active radiation) at the
plant canopy level, and a relative air humidity of 80%.
After 24 h of pre-treatment, the plants were transferred
to 0.5 mM CaCl2 solution containing other chemicals:
(1) 0 (CK); (2) 400 μM AlCl3; (3) 50 μM SA; (4)
400 μM AlCl3 plus 50 μM SA or 100 μM PAC
(paclobutrazol, a synthetic inhibitor of SA). Each treat-
ment set 3 pots with 10 seedlings per repeat. The sam-
ples were harvested for analysis at 3 and 15 days of
treatment, respectively. The treatment solution was
changed once a day.
Analysis of dry matter partitioning
Dry matter partitioning was measured in terms of plant
dry weight after 15 days of stress imposition. Harvested
leaves were washed with deionized water for three
times, and then were blotted with absorbent paper. They
were separated into roots, stem and leaves with a scal-
pel. The dry weight data of roots, stem and leaves were
collected after detachment and drying to constant weight
at 80 °C for 96 h.
Analysis of Al in P. notoginseng leaves
The leaves of P. notoginseng, which are dried to con-
stant weight, are crushed. The total Al of leaves was
determined by microwave digestion (MDS-6G, China).
Pulverized tissue sample (0.2 g) was accurately
weighed, mixed with 10 ml HNO3 in a Teflon sample
cup and placed in a fume hood overnight at room
temperature. The 30% hydrogen peroxide of 2 ml was
added. Teflon PFA (perfluroalkosy) vessels were fitted
on a rotating table. The microwave power program was
set at 400 W, and temperature was held at a constant
Plant Soil (2019) 445:183–198
180 °C for 10 min (taking 25 min to reach 180 °C). After
cooling, the samples were filtered through acid-washed
papers into acid-washed 25 ml volumetric flasks (5%
HNO3 was used for acid washing). The Teflon PFA
vessels were then rinsed three times with distilled water
with washing being filtered into the flasks. The flasks
were filled to 25 ml with distilled water. In the meantime
the controls were carried out to eliminate inference. Al
was measures by inductively coupled plasma-atomic
emission spectrometry (ICP-AES) (Lee man-LABS,
Prodigy7, USA) (Gan et al. 2010).
Extract solution absorbance from dyed leaves and cell
viability
After different treatments, leaves were immersed in
0.25% aqueous Evans blue solutions for 5 h, then the
Chl in leaves were decolorized in boiling ethanol. The
blue precipitates in dead cells were solubilized with 25 ml
of 1% SDS-50% methanol and examined at 600 nm
using a UV spectrophotometer (UV-2600, SHIMADZU).
Absorbance at 600 nm in ethanol extract of untreated
control leaf was designated as 1 and the cell death in
treated samples was defined as ratios of absorbance in
treated leaf samples to control value (Liu and Lin 2011).
The cell viability was calculated following the formula as
[1-(a/ ad)] × 100%, where a is the extract solution absor-
bance from dyed leaves under Al stress and ad is the
absorbance of leaves cell death in treated samples.
Determination of total Chl
Total Chl were estimated by adopting the method de-
scribed by Lichtenthaler and Wellburn (1983) with some
modifications. Briefly, Chl was extracted from 0.1 g of
leaves with 8 ml of 100% acetone-alcohol (1:1). The
absorbance of the supernatant was measured at 645 nm,
652 nm and 663 nm. The total Chl was calculated using
the formula described by Lichtenthaler and Wellburn
(1983).
Measurements of photosynthetic characteristics and Chl
fluorescence
Pn, Gs, Tr, and Ci were measured of each plant with a
portable photosynthesis system (LCi, ADC Bioscientif-
ic Co., England) under the conditions of light intensity
of 100 μmol photon m−2 s−1, atmosphere temperature of
(25 ± 1) °C.
Plant Soil (2019) 445:183–198
Plants were dark-adapted for 30 min at 25 °C before
the Chl fluorescence parameters were measured. A total
of 24 leaf samples (3 independently grown sets of plants
provided 8 samples each) were measured using a pulse-
modulated fluorometer (Dual-PAM-100, WALZ, Ger-
many) for each treatment. Running the system accord-
ing to the manufacturer’s instructions, the following leaf
measurements were taken: Fm, dark-adapted minimum
fluorescence (F0), steady-state fluorescence (Fs), light-
adapted maximum fluorescence (Fm′), and minimal
fluorescence in the light-adapted state (F0′) (Zhang and
Liu 2018). The Fv/Fm is the maximum quantum effi-
ciency of PSII photochemistry. It can be calculated
based on the following equation: Fv/Fm = (Fm − F0)/
Fm = ΦPSII/qP. The effective quantum yield of PSII
(ΦPSII) was determined as (Fm′ − Fs) Fm′. The photo-
chemical quenching coefficient, qP = (Fm′ – Fs)/ (Fm′ –
F0′). NPQ was used as an indicator of the activity of
energy dissipation in the pigment bed of PSII, was
calculated as NPQ = (Fm − Fm′) Fm′.
Determination of non-structural carbohydrate
Sucrose, glucose, and fructose were extracted with 80%
ethanol at 80 °C three times (4 ml each for 30 min).
After centrifugation, the supernatant was pooled and
dried at 50 °C under vacuum. The residues were
redissolved in 1 ml distilled water and passed through
a 0.45 μm filter. Ten microliters of sample were then
injected into a HPLC (Agilent 1260, Agilent Technolo-
gies Inc., USA), containing an NH2P-50 4E column
(Shodex Asahipak, 4.6 mm × 250 mm) at 35 °C using
acetonitrile and water (70:30%) as eluent, with a flow
rate of 1.0 ml per min. The sucrose, fructose and glucose
were identified by the comparison of retention times to
known standards (Yang et al. 2019).
The pellet after extracting soluble sugars was
boiled in 0.2 M KOH for 30 min, adjusted to
pH of 5.5 with 1 M acetic acid, then digested
with 50 units of amyloglucosidase for 30 min at
55 °C. Starch was determined as glucose equiva-
lents (Chen and Cheng 2003). TNC was the sum
of starch, and TSC (sucrose + glucose + fructose).
Determination of hydrogen peroxide (H2O2)
concentration
H2O2 concentration was measured as described by
Velikova et al. (2000). Fresh leaf samples (0.5 g) were
187
homogenized in 5 ml of 0.1% (w/v) trichloroacetic acid
(TCA). The homogenate was centrifuged at 12,000 rpm
for 15 min and the supernatant was added to 10 mM
potassium phosphate buffer (pH 7.0) and 1 M potassium
iodide. The absorbance of the supernatant was recorded
at 390 nm. The concentration of H2O2 was calculated by
comparison with a standard calibration curve plotted
using known concentrations of H2O2.
Determination of Rubisco and RCA activity
The rubisco and RCA activity were determined using
the rubisco and rubisco activase assay kit (Genmed
Scientifics Inc., USA), respectively.
Total RNA extraction and gene expression analysis
Approximately 100 mg FW leaves were sampled
3 days after treated with different solutions, frozen
with liquid N2 and stored at −80 °C. Total RNA was
extracted using TRIzol Reagent (Invitrogen) accord-
ing to the manufacturer’s recommendations. A
Primer-Script First Strand cDNA synthesis kit
(TaKaRa, Japan) was used to synthesize the first-
strand cDNAs according to the manufacturer’s pro-
tocol in a total volume of 20 μl reaction mixture
containing 1 μg of total RNA, 4 μl 5 × PrimeScript
RT Master Mix and RNAase-free ddH2O. Gene-
specific primers were designed with primer 5.0 soft-
ware and synthesized by TsingKe Biological Tech-
nology (Beijing, China). The quantitative real-time
PCR program was conducted using a Light Cycler®
96 System. A 1 μl aliquot of cDNA was amplified in
the presence of 12.5 μl of 2 μl Power SYBR Green
PCR Master Mix (Applied Biosystems), 0.5 μl of
specific primers (final concentration, 0.2 μM) and
10.5 μl of sterilized water. PCR conditions were
95 °C for 2 min, 40 cycles of 95 °C for 15 s and
60 °C for 60 s. Expression of PnACT2 was used as
an internal control for target gene expression, and
every gene primers is synthesized according to the
primer sequence. Moreover, the Chl metabolism and
Calvin-Benson cycle key gene primers sequence of
P. notoginseng were shown in Supplemental Table1
(GenBank/NCBI). The relative gene expression
values were calculated with the 2-△△Ct method fol-
lowing Yang et al. (2018b).
188
Data analysis
Data were processed with Microsoft Excel software 2010
and means were compared by using Duncan’s multiple
comparison tests (P < 0.05). A one-way analyses of var-
iance (ANOVA) was performed to evaluate the level of
significance by using SPSS statistical package. Graphs
were plotted by using Prism 7.0 software. All the values
presented are means of replicates ± SE (standard error).
Results
Exogenous SA reduced the accumulation of Al
in P. notoginseng leaves
Compared to the control not treated with Al, the Al
concentration significantly increased by 24.40% in
leaves under Al treatment (Fig. 1). Compared to the
plants under Al treatment only, the Al concentration
decreased by 13.23% in plants under co-treatment with
SA and Al, whereas it increased by 33.99% under co-
treatment with PAC and Al.
Exogenous SA alleviated the inhibitory effects of Al
stress on P. notoginseng biomass accumulation
The dry matter quality of the roots, stems, and leaves of
P. notoginseng under different treatments showed
Fig. 1 Effect of different treatments on Al concentration in
P. notoginseng leaves. Two-year-old P. notoginseng seedlings
were exposed to 0.5 mM CaCl2 solution (pH 4.5) containing
0 μM AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA),
400 μM AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM
PAC (Al + PAC) for 15 days. Data are means ± SE (n = 3). Differ-
ent small letters mean significant differences at P < 0.05
Plant Soil (2019) 445:183–198
similar trends as that of the total biomass (Table 1).
Treatment with exogenous SA alleviated the inhibitory
effects of Al stress on the growth of P. notoginseng.
Compared with the plants only subjected to Al stress,
the biomass of the roots, stems, leaves, and the total dry
matter in P. notoginseng increased by 28.84%, 5.02%,
17.76%, and 21.78%, respectively, in plants treated with
exogenous SA and Al. However, PAC treatment further
inhibited the accumulation of dry matter in
P. notoginseng under Al toxicity (Table 1).
Exogenous SA reduced the Al-induced increase
of carbohydrate content in P. notoginseng leaves
The concentration of TNC in leaves followed the order
of sucrose > glucose > starch > fructose, while that in
roots were sucrose > starch > glucose > fructose. Under
the control conditions, the ratios of sucrose: starch:
glucose: fructose in leaves and root were 16:3:6:1 and
12:9:5:1, respectively. The concentration of TNC in
leaves and roots of P. notoginseng under Al stress in-
creased significantly by 29.19% and 40.20%, respec-
tively. SA significantly reduced the increase in TNC
concentration induced by Al, and the TNC concentra-
tion in leaves and roots decreased by 27.12% and
27.94%, respectively, compared with Al treatment. Al
stress decreased the starch concentration in the leaves
(52.37%) and roots (37.81%) with respect to the control
(Fig. 2). SA significantly lowered the Al-induced reduc-
tion in starch content in the roots and leaves by 36.26%
and 61.55%, respectively. Compared with the control,
the TSC content in leaves and roots were up-regulated
by 38.96% and 77.20% under Al treatment, respective-
ly, while they were down-regulated by 30.76% and
38.63% by the addition of SA. Sucrose, glucose and
fructose are the main components of TSC, among which
sucrose changes most significantly under different treat-
ments. As compared to the control, sucrose concentra-
tion significantly increased in the leaves and roots by
1.46 and 2.07 fold, respectively, in response to Al stress
(Fig. 2). The addition of SA significantly reduced the
Al-induced accumulation of sucrose by 36.17% and
44.35% in the leaves and roots, respectively. Glucose
had a similar changing trend with sucrose. The plants
exposed to Al stress had significant increases in glucose
as compared to the control, and the addition of SA
inhibited these increases in roots and leaves. Compared
with the control, Al treatment decreased the fructose
concentration, but there was no significant difference.
Plant Soil (2019) 445:183–198 189

Table 1 Effects of different treatment on dry matter production of root, stem and leaf of P. notoginseng. The data are given by mean values ±
SE (n = 25)

Treatments Roots dry weight Stems dry weight Leaves dry weight Total biomass Roots/total
(g/five plant) (g/five plant) (g/five plant) (g/five plant) biomass (%)

CK 2.949a 0.632a 1.701b 5.282a 55.825a

Al 1.627c 0.386d 1.247d 3.260c 49.912c
SA 3.109a 0.420b 2.153a 5.682a 54.708a
Al + SA 2.097b 0.405c 1.469c 3.970b 52.804b
Al + PAC 1.405d 0.391d 1.167e 2.963c 47.409c

The different letters mean separation within columns by Duncan’s multiple range test (P = 0.05)

Exogenous SA alleviated the inhibitory effects of Al
stress on the activity of TNC metabolism enzymes
of P. notoginseng leaves
Under Al stress, the activities of rubisco and RCA in the
leaves of P. notoginseng were significantly reduced by
40.00% and 22.71%, respectively. Compared with the
plants only subjected to Al stress, plants under Al stress
that were treated with exogenous SA exhibited
increased activities of rubisco (33.33%) and RCA
(13.34%). In contrast, PAC further decreased the rubisco
and RCA activities in plants under Al stress by 44.44%
and 26.78%, respectively (Fig. 3).
Exogenous SA alleviated the inhibitory effects of Al
stress on the expression of key genes of the Calvin cycle
Al stress significantly downregulated the expres-
sions of rbcL, rbcS, and RCA, which are key genes
of the Calvin cycle. Exogenous SA significantly
inhibited the downregulation of Al stress-induced
genes. Compared with the plants only exposed to
Al stress, the expression levels of rbcL, rbcS, and
RCA were upregulated 1.39-, 1.58-, and 1.56-fold,
respectively, in plants under Al stress treated with
exogenous SA. In contrast, PAC treatment had a
positive synergistic effect with Al on the downreg-
ulation of the expression of these Al stress-related
genes (Fig. 4).

Exogenous SA alleviated the inhibitory effects of Al

stress on photosynthetic parameters in P. notoginseng
leaves

It was found that the Pn, Tr, and Gs of

Fig. 2 Effects of different treatments on the TNC content in leaves
and roots of P. notoginseng. TNC refer to the sum of starch,
sucrose, glucose, fructose). Two-year-old P. notoginseng seedlings
were exposed to 0.5 mM CaCl2 solution (pH 4.5) containing 0 μM
AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA), 400 μM
AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. Data are means ± SE (n = 3). Different
small letters mean significant differences at P < 0.05
P. notoginseng leaves were reduced under Al stress
by 49.21%, 19.51%, and 16.79%, respectively
(Fig. 5). The co-treatment with SA and Al allevi-
ated the inhibitory effects of Al stress on the Pn,
Tr, and Gs. In particular, the Pn increased signif-
icantly by 52.16%. Treatment with PAC exacerbat-
ed the Al stress-induced decrease of the Pn in
P. notoginseng leaves. However, the Ci exhibited
an opposite trend compared with the other
190
Fig. 3 Rubisco and RCA activity of P. notoginseng leaves under
different treatments. Two-year-old P. notoginseng seedlings were
exposed to 0.5 mM CaCl2 solution (pH 4.5) containing 0 μM
AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA), 400 μM
photosynthetic parameters under the same treat-
ment (Fig. 5).
Exogenous SA alleviated the Al stress-induced toxicity
on cell viability in P. notoginseng leaves
While blue spots were not observed on the surfaces of
the leaves of plants not subjected to Al stress, a signif-
icant number of blue spots were seen in plants under Al
stress, implying that Al stress damages the integrity of
the plasma membranes of cells in the P. notoginseng
leaves. Treatment with exogenous SA reduced the area
and number of blue spots in plants under Al stress.
Treatment with PAC aggravated the damage to the in-
tegrity of the cell plasma membrane in the leaves of
Fig. 4 Relative expression of RCA, rbcL and rbcS of
P. notoginseng leaves in different treatments. RCA, rubisco
activase gene; rbcL, rubisco large subunit gene; rbcS, rubisco
small subunit gene. Two-year-old P. notoginseng seedlings were
exposed to 0.5 mM CaCl2 solution (pH 4.5) containing 0 μM
Plant Soil (2019) 445:183–198
AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. Data are means ± SE (n = 3). Different
small letters mean significant differences at P < 0.05
P. notoginseng plants under Al stress, which manifested
as enlarged blues spots (Fig. 6). The changes of the
absorbances of the extract solutions (Table 2) are con-
sistent with those depicted in Fig. 6, which also proves
that exogenous SA can alleviate the Al stress-induced
damage to the integrity of the cell plasma membrane in
P. notoginseng leaves.
Exogenous SA alleviated the reduction effects of Al
stress on the Chl in P. notoginseng leaves
Al stress reduced the concentrations of Chla, Chlb, and
Chla+b in P. notoginseng leaves. However, under Al
stress, treatment with exogenous SA increased the Chla,
Chlb, and Chla+b concentrations by 18.85%, 12.32%,
AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA), 400 μM
AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 3 days. Data are means ± SE (n = 3). Different
small letters mean significant differences at P < 0.05
Plant Soil (2019) 445:183–198 191

Fig. 5 Photosynthetic parameters

of P. notoginseng leaves under
different treatments. Two-year-
old P. notoginseng seedlings were
exposed to 0.5 mM CaCl2
solution (pH 4.5) containing
0 μM AlCl3 (CK), 400 μM AlCl3
(Al), 50 μM SA (SA), 400 μM
AlCl3 + 50 μM SA (Al + SA),
400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. Data are
means ± SE (n = 30). Different
small letters mean significant
differences at P < 0.05

and 14.89%, respectively (Fig. 7). In contrast, treatment
with PAC exacerbated the Al stress-induced decrease of
Chl concentration in P. notoginseng leaves.
Exogenous SA regulated the expression of Chl
synthesis-related genes
Al stress had a significant negative regulatory effect
on the expression of key enzymes involved in Chl
synthesis in P. notoginseng, such as members of the
PnCHL family (CHLM, CHLI, CHLG) and PnHEM
family (HEMA1, HEMB1, HEMC, HEME). Al
stress also significantly upregulated the expression
of the Chl degradation genes CLH2 and PPH1, but
had no significant effect on the expression of
PAO2. Compared with Al stress, under co-
treatment with SA and Al, the expressions of
CHLM, CHLG, HEMB1, HEMC and HEME were
upregulated 4.74-, 1.96-, 1.45-, 1.51-, and 2.01-
fold, respectively. The expressions of CLH2 and
PPH1 decreased by 48.49% and 46.19%, respec-
tively (Fig. 8a, b). In contrast, treatment with PAC
had a positive synergistic effect with Al on the
regulation of the expression of these genes.
Exogenous SA alleviated the reduction of Chl
fluorescence parameters induced by Al stress
in P. notoginseng leaves
Al stress significantly decreased the values of Fv/Fm,
Φ PSII , and qP, and enhanced the NPQ levels in
P. notoginseng leaves. Compared with the Al treatment

Fig. 6 Evans Blue dyeing pictures of P. notoginseng leaves after (pH 4.5) containing 0 μM AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM
15 days of exposure to different treatments. Two-year-old SA (SA), 400 μM AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 +
P. notoginseng seedlings were exposed to 0.5 mM CaCl2 solution 100 μM PAC (Al + PAC) for 15 days
192
Table 2 Extract solution absorbance from dyed leaves of
P. notoginseng seedling and their cell viability. The data are given
by mean values ± SE (n = 3)
Treatments Extract solution absorbance Cell viability (%)
CK 0.046 ± 0.004 d 97.39 ± 0.002 a
Al 0.072 ± 0.004 b 95.89 ± 0.002 c
SA 0.042 ± 0.004 d 97.89 ± 0.002 a
Al + SA 0.056 ± 0.006 c 96.78 ± 0.003 b
Al + PAC 0.093 ± 0.003 a 94.66 ± 0.002 d
Small letter means significant difference at P < 0.05
alone, under the co-treatment with Al and SA, the Fv/Fm,
ΦPSII, and qP values increased by 5.78%, 10.03%, and
4.19%, respectively, whereas under the co-treatment of
Al and PAC, they decreased by 19.80%, 22.88%, and
4.00%, respectively (Fig. 9). The NPQ under the co-
treatment of Al and SA was lower than that under the
Al treatment alone. However, Al + PAC treatment signif-
icantly increased the NPQ by 9.87% (Fig. 9).
Exogenous SA decreased the H2O2 concentration
in P. notoginseng leaves
Al stress significantly induced the increase of the H2O2
concentration in P. notoginseng leaves. Compared with
the plants only exposed to the Al treatment, the treat-
ment with exogenous SA significantly reduced the ac-
cumulation of H2O 2 in P. notoginseng leaves by
12.22%. In contrast, PAC promoted the increase of the
Fig. 7 Effect of different treatments on the total Chl concentration
of P. notoginseng leaves. Two-year-old P. notoginseng seedlings
were exposed to 0.5 mM CaCl2 solution (pH 4.5) containing 0 μM
AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA), 400 μM
Plant Soil (2019) 445:183–198
H2O2 concentration in P. notoginseng leaves by 26.85%
(Fig. 10).
Discussion
The accumulation of Al in plants can damage the cell
structure and cause metabolic disorders, thereby
inhibiting plant growth and development, and ultimately
reducing yield and quality. Accordingly, reducing the
accumulation of Al in plants is an urgent necessity to
alleviate crop yield reduction and quality degradation
caused by Al stress. Research by Zhu et al. (2019) sug-
gested that boron (B) promoted rice growth by reducing Al
in roots. Pandey et al. (2013) demonstrated that exogenous
SA significantly reduced Al uptake by rice, thereby in-
creasing dry matter accumulation. Our previous studies
have shown that treatment with exogenous SA significant-
ly reduced the accumulation of Al in P. notoginseng roots
(Yang et al. 2018b). In this study, exogenous SA also
lowered the Al concentration in the leaves of
P. notoginseng (Fig. 1). In addition, exogenous SA also
alleviated the inhibitory effect of Al on the accumulation of
dry matter in P. notoginseng (Table 1). The specific mech-
anism underlying this effect was analyzed in detail in the
present study.
Plant productivity is dependent upon the production,
translocation, storage, and utilization of carbohydrates,
which include TNC and SC. TNC accounts for about
10% of the dry biomass of the plant, and mainly includes
TSC and starch (Martnez-Vilalta et al. 2016). TNC is a
AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. The data are given by mean values ± SE
(n = 3). Different small letter means significant difference at
P < 0.05
Plant Soil (2019) 445:183–198 193

Fig. 8 Expression of Chl

synthesis (a) and degradation (b)
genes in leaves of P. notoginseng
under different treatments.
HEMA, glutamyl-tRNA
reductase; HEMB, 5-
aminolevulinate dehydrogenase;
HEMC, porphobilinogen
deaminase; HEME,
uroporphyrinogen III
decarboxylase; CHLM, Mg-
protoporphyrin IX
methyltransferase; CHLI, Mg-
chelatase; CHLG, chlorophyll
synthase; CLH, chlorophyllase;
PPH, pheophytinase; PAO,
pheophorbide a monooxygenase.
Two-year-old P. notoginseng
seedlings were exposed to
0.5 mM CaCl2 solution (pH 4.5)
containing 0 μM AlCl3 (CK),
400 μM AlCl3 (Al), 50 μM SA
(SA), 400 μM AlCl3 + 50 μM SA
(Al + SA), 400 μM AlCl3 +
100 μM PAC (Al + PAC) for
3 days. Data are means ± SE (n =
3). Different small letters mean
significant differences at P < 0.05

source of energy for plants and is closely related to the
growth and survival of plants under stress. Therefore, any
factors that affect the metabolism of TNC will affect the
growth of plants. The effects of heavy metal toxicity on
TNC metabolism are usually reflected in changes in the
concentrations of sucrose, glucose, fructose, and starch in
plant tissues (Shahid et al. 2019). For example, Li et al.
(2010) found that excessive manganese caused the accu-
mulation of glucose, fructose, and sucrose in Citrus
grandis seedlings. In the present study, TSC (glucose +
fructose + sucrose) was accumulated in P. notoginseng
under Al stress (Fig. 2). One possible reason for this was
the resistance of osmotic stress induced by Al. Kariya
et al. (2017) also concluded that the overexpression of
NtSUT1 (which primarily supports sucrose uptake via the
apoplastic pathway) at the root apex increases the soluble
sugar (TSC) content, resulting in greater cell survival and
root elongation under Al stress, leading to the Al-
tolerance phenotype. Starch can be decomposed into
TSC to enhance the ability of crops to resist stress. Al
stress caused significant decrease in starch concentration
in the leaves and roots of P. notoginseng were also found
in present study (Fig. 2). The decrease of starch concen-
tration under Al stress were mainly for the inhibition of
starch synthases activity (soluble starch synthase, ADP-
glucose pyrophosphorylase) and increased activity of
degradation enzymes (starch phosphorylase and β-amy-
lase) (Supplementary Figs.1 and 2). However, the TNC
concentration in plants under Al stress was increased,
which may be due to the limitation of the utilization of
TNC due to Al, resulting in a relative increase of the
TNC. El-Shazoly et al. (2019) reported that the reduction
of TNC utilization in wheat seedlings led to its increased
content under boron stress, which was significantly neg-
atively correlated with the accumulation of dry matter.
This explains why Al stress induced an increase in TNC
content but inhibited biomass accumulation.
Rubisco is the rate-limiting enzyme during the for-
mation of TNC. Although its catalytic activity is weak, it
can participate in the carbon assimilation process of
194 Plant Soil (2019) 445:183–198

Fig. 9 Effects of different

treatments on the maximum
quantum yield in the dark-adapted
state (Fv/Fm), the actual quantum
yield in the light-adapted steady
state (ΦPSII), the coefficients of
photochemical quenching (qP)
and the non-photochemical
quenching values (NPQ) of
P. notoginseng leaves. Two-year-
old P. notoginseng seedlings were
exposed to 0.5 mM CaCl2
solution (pH 4.5) containing
0 μM AlCl3 (CK), 400 μM AlCl3
(Al), 50 μM SA (SA), 400 μM
AlCl3 + 50 μM SA (Al + SA),
400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. The data
are given by mean values ± SE
(n = 24). Different small letter
means significant difference at
P < 0.05

plant photosynthesis after the activation of RCA, and its
activity is positively correlated with the Pn. Drought stress
was found to reduce rubisco activity, which could lead to a
decrease of photosynthesis in cotton (Zahoor et al. 2017).
Khairy et al. (2016) also suggested that Cd and Cu stress
reduced rubisco activity by inhibiting RCA activity in
tobacco, and ultimately inhibited photosynthesis. In the
Fig. 10 Concentration of H2O2 in leaves of P. notoginseng under
different treatment. Two-year-old P. notoginseng seedlings were
exposed to 0.5 mM CaCl2 solution (pH 4.5) containing 0 μM
AlCl3 (CK), 400 μM AlCl3 (Al), 50 μM SA (SA), 400 μM
AlCl3 + 50 μM SA (Al + SA), 400 μM AlCl3 + 100 μM PAC
(Al + PAC) for 15 days. The data are given by mean values ± SE
(n = 3). Different letters imply significant differences at P < 0.05
present work, the decrease of the Gs induced by Al stress
reduced the CO2 availability in the leaves and then
inhibited carbon fixation, thus leading to a decrease in
the Pn, as can be speculated from the lower rate of
rubisco-mediated carboxylation (Figs. 3, 4 and 5).
The increase of the biomass due to the weakening of the
inhibitory effects of Cd or Cu stress on tobacco growth via
the enhancement of rubisco and RCA activity by exoge-
nous sodium nitroprusside has also been confirmed
(Khairy et al. 2016). In this study, exogenous SA increased
the rubisco activity of P. notoginseng leaves under Al stress
by reducing the inhibitory effect of Al on RCA activity
(Fig. 3). In theory, this change should be beneficial to the
synthesis and accumulation of TNC, but it is notable that
the exogenous SA treatment significantly reduced the
increase in Al-induced TNC content (Fig. 2). This indi-
cates that SA promoted the synthesis of TNC under Al
stress by regulating the metabolism of starch and sugar,
and also caused a significant increase in the utilization of
TNC in P. notoginseng, resulting in a relative decrease in
the amount of TNC (Fig. 2). The increase in dry matter
accumulation provides strong evidence for this finding
(Table 1).
Chl participates in the processes of light energy
transmission, distribution, and transformation in photo-
synthesis, and affects the photosynthetic rate and the
Plant Soil (2019) 445:183–198
accumulation of organic matter. The dry matter in plants
mainly derives from photosynthesis, so the increase of
biomass is necessarily accompanied by an increase in
Chl. For instance, exogenous Mg was found to increase
the Chl of alfalfa under Al stress, which in turn in-
creased the photosynthetic rate and ultimately reduced
the inhibitory effect of Al on dry matter accumulation
(Yuan et al. 2018). Pandey et al. (2013) also found that
exogenous SA promoted dry matter accumulation in
rice by inhibiting Al absorption, thereby preventing the
loss of cell plasma membrane integrity in leaves. A
previous study suggested that exogenous SA alleviated
the Al-induced loss of plasma membrane integrity and
the decrease in cell viability by enhancing plant cell
growth via the activation of the antioxidant defense
system in tomato plants (Surapu et al. 2014). Under
Cd stress, treatment with exogenous SA alleviated the
decrease of Chl in Kentucky bluegrass and pea seedlings
(Guo et al. 2013; Popova et al. 2009). In the present
study, exogenous SA alleviated the loss of plasma mem-
brane integrity and the decrease of cell viability under
Al stress (Fig. 6, Table 2). These results suggest that
exogenous SA alleviated the damage of P. notoginseng
leaves by Al toxicity. In addition, exogenous SA signif-
icantly decreased the Al stress-induced reduction of Chl
by upregulating the Chl synthesis genes and downregu-
lating the expression of the degradation genes (Figs. 7,
and 8a, b). These studies indicate that exogenous SA
alleviated the decrease of photosynthetic pigments in-
duced by Al in P. notoginseng by maintaining the integ-
rity of leaf cell plasma membranes and regulating the
expression of Chl metabolism genes, which lays a ma-
terial foundation for improving photosynthesis efficien-
cy in P. notoginseng (Fig. 5).
Chl can be stimulated by light to produce fluores-
cence. The changes of fluorescence parameters can ac-
curately reflect the response characteristics of the plant
photosynthetic apparatus under environmental stress
(Yao et al. 2017). Fv/Fm, ΦPSII, qP, and NPQ are impor-
tant fluorescence parameters for evaluating photosyn-
thetic system damage in plants. The Fv/Fm value repre-
sents the maximum photochemical quantum yield of
PSII and reflects the maximum energy conversion effi-
ciency of PSII. It is frequently used as an indicator of
photoinhibition or stress damage to PSII. Under water
stress, the Fv/Fm value in soybean seedling leaves has
been found to significantly decrease, indicating that
stress can induce the photoinhibition of PSII (Tang
et al. 2017). In this study, exogenous SA significantly
195
alleviated the Al stress-induced reduction of Fv/Fm,
suggesting that exogenous SA can alleviate the Al
stress-induced damage to PSII and inhibit the decrease
of the potential maximum photosynthetic activity of
P. notoginseng. ΦPSII is the actual photochemical quan-
tum yield or efficiency of light utilization by photosyn-
thesis, which functions in the transfer of linear electrons.
Zhang et al. (2018) found that the ability of tomato
seedlings to convert light energy into chemical energy
was impaired by the decrease of ΦPSII under drought
stress. In this study, the ΦPSII values of P. notoginseng
leaves were decreased by Al stress, which indicates that
the electron transport chain was impaired and carbon
assimilation was reduced under Al stress. Exogenous
SA alleviated the inhibitory effects of Al on ΦPSII,
indicating that the light energy used for photochemical
reactions increased (Fig. 9). The qP value is the photo-
chemical quenching coefficient, which reflects the open
degree of the PSII center. The decrease of the qP value
in leaves of melon under Cd stress indicates a decrease
in the open degree of the PSII center under Cd stress
(Zhang et al. 2015b). The exogenous SA treatment in
this study reduced the Al-induced decrease in the qP
value (Fig. 9), indicating that treatment with exogenous
SA maintained more PSII reaction centers in an open
state so that more excitation energy was used for elec-
tron transport under Al stress. These photosynthetic
parameters in plants under Al stress were improved by
exogenous SA, suggesting that SA can alleviate the Al
stress-induced disruption of the structure and function of
the photosynthetic system; therefore, PSII was protected
from Al stress-induced damage and maintained high
photochemical activity (Ozfidankonakci et al. 2018).
The NPQ is a mechanism used by plants to protect
them from high light intensity, and represents the light
energy that is dissipated in the form of heat energy in the
PSII of plant leaves. An increase of the NPQ in
Xanthium strumarium under Mn stress has been previ-
ously observed, indicating that photosynthesis was
inhibited. In addition, the increase of the NPQ may
prevent the photo-damage caused by Mn stress and
protect the organs of the photosynthetic apparatus
against excess excitation energy (Pan et al. 2018). In
this study, Al stress induced the increase of the NPQ,
suggesting that Al stress resulted in the decrease of
electrons available for photosynthesis, dissipating ex-
cess energy through heat or other forms (Fig. 9). How-
ever, exogenous SA inhibited the increase of the NPQ
induced by Al stress, which was due to the SA-mediated
196
inhibition of the potential heat dissipation process of
PSII under Al stress, and thus avoided the generation
of oxidative stress (Moravcová et al. 2018). Zhang et al.
(2015b) also found that exogenous SA significantly
reversed the decrease of the Fv/Fm, qP, and ΦPSII values,
and the increase of the Cd stress-induced NPQ in melon
leaves. These results demonstrate that exogenous SA
reversed the damage caused by Al stress to the
P. notoginseng photosynthetic system and maintained
the maximum efficiency of the PSII, thereby promoting
the accumulation of dry matter under Al stress.
The decrease in PSII activity caused by abiotic stress
leads to a decrease in the utilization of light energy by
plants, which in turn leads to photoinhibition and the
accumulation of large amounts of H2O2; this ultimately
results in lipid peroxidation, photosynthetic organ damage,
and cell death (Ali et al. 2013; Sharma et al. 2012). H2O2 is
the receptor of the photosynthetic redox signal, and can
induce the change of the redox state (Gururani et al. 2015).
El-Shazoly et al. (2019) found that boron induced an
increase in the H2O2 concentration in rice leaves, which
then accumulated more TNC. This is due to the accumu-
lation of H2O2 as a result of the decrease of PSII activity
under stress. However, plants can protect themselves
against reactive oxygen species-induced damage by in-
creasing their TNC content, thereby reducing the damage
to the light system caused by heavy metals (Alho et al.
2019). It has been reported that the concentrations of Cd
and H2O2 in peas under Cd stress were reduced by treat-
ment with exogenous SA (Popova et al. 2009). Similarly,
in this study, the PSII activity was decreased and the H2O2
concentration was increased in leaves of P. notoginseng
under Al stress, both of which were significantly reversed
by exogenous SA. However, in contrast, PAC aggravated
the toxicity of Al stress (Fig. 10). These results indicate that
photoinhibition caused by the imbalance of the photosyn-
thetic redox signal transduction and PSII damage was
alleviated by the promotion of the SA signaling pathway.
Ultimately, the utilization rate of light energy was increased
in the leaves of P. notoginseng under Al stress.
Conclusion
Under Al stress, SA decreased the Al concentration in
P. notoginseng leaves, reduced the damage to the pho-
tosynthetic system, increased the utilization rate of light
energy, and then accelerated the process of carbon as-
similation. Eventually, the production of dry matter and
Plant Soil (2019) 445:183–198
its transfer to the roots were promoted, and the Al-
induced decrease of dry matter accumulation in
P. notoginseng was alleviated.
Acknowledgments This work was supported by the key project
at central government level: the ability establishment of sustain-
able use for valuable Chinese medicine resources (No. 2060302),
Natural Science Foundation of China (No. 81960690, 81891014
81460580), Ministry of Science and Technology of Yunnan Prov-
ince, China (No. 2017ZF014, 2017ZF001).
Compliance with ethical standards
Conflict of interest The authors declare no competing financial
interest.
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