Page 173 - AJWEP-22-5

Alleviating aluminum toxicity

indicates that Al stress induces an increase in flavonoid A B C
metabolites in wheat, which primarily assist in binding
Al ions and solubilizing P in the soil, thereby reducing
Al uptake. These findings underscore the importance
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of flavonoids in enhancing plant resilience to Al stress
and improving P availability in acidic soils.
Metabolomics can be used to identify metabolic
phenotypes, obtain qualitative and quantitative data
on many metabolites, construct metabolic networks,
and predict the functions of metabolites. Multivariate
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statistical analysis and liquid chromatography-tandem
mass spectrometry (LC-MS/MS) have revealed
significant differences in flavonoids in Coreopsis
tinctoria from different production areas. Different
geographical conditions and altitudes may lead to Figure 1. Camellia oleifera was treated with various
different metabolic mechanisms of flavonoids in phosphorus–aluminum solutions at different
Coreopsis tinctoria, which in turn affect its medicinal concentration ratios. (A) Group A4P_. (B) Group A4P5.
value. In this study, C. oleifera seedlings were (C) Group A_P5.
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utilized as experimental materials. The study entailed
administering varying concentrations of P and Al to A4P5-1, A4P5-2, and A4P5-3; and specimens of the
the seedlings, followed by metabolite analysis using A4P_ treatment group were labeled A4P_-1, A4P_-2,
ultrahigh-performance LC-MS/MS (UPLC-MS/MS). and A4P_-3. The samples were kept at −80°C in
This approach enabled the elucidation of the metabolic preparation for the downstream experiments.
regulatory network underlying the complex interaction
between P and Al. Furthermore, the research focused 2.2. Pretreatment and extraction of samples
on exploring the critical role of metabolites in the The leaf samples of C. oleifera were subjected to
mechanism by which P mitigates Al toxicity. vacuum freeze-drying treatment. Subsequently, a
grinderwas used to grind the samples into powder. The
2. Materials and methods powdered sample was weighed (50 mg) and added
to a specified internal calibration extraction solution,
2.1. Plant material and sample collection then vortexed once every 30 min for a total of 6 times.
C. oleifera seedlings were used as experimental subjects, Once the centrifugation process was completed,
and healthy plants at the same growth stage were selected the supernatant was filtered through a microporous
for experiments. The seedlings were treated with a membrane and stored in an injection bottle for
nutrient solution containing a combination of one of two UPLC-MS/MS analysis.
P concentrations (0 or 0.5 mM KH PO ) and one of two
2
4
Al concentrations (0 or 4 mM AlCl ·Al O) at pH 4.2. 2.3. Instrumentation
3
2
Seedlings were randomly divided into three treatment The instruments and equipment used in this study
groups: A_P5 (0 mmol/L Al, 0.5 mmol/L P), A4P5 included: a centrifuge (5424R, Eppendorf, Germany);
(4 mmol/L Al, 0.5 mmol/L P), and A4P_ (4 mmol/L constant-temperature metal mixer (MU-G02-0448,
Al, 0 mmol/L P) (Figure 1). C. oleifera seedlings were MIULab, China); grinder (MM 400, Retsch,Germany);
treated with nutrient solutions containing different one-ten-thousandth analytical balance (MS105DM,
combinations of P and Al concentrations. Treatments METTLER TOLEDO, Switzerland); centrifugal
were administered to each group at three-day intervals. concentrator (CentriVap, LABCONCO, USA);
After 2 months of treatment, the leaves of C. oleifera lyophilizer (Scientz-100F, SCIENTZ, China); vortex
were harvested, cleaned thoroughly using an ultrasonic mixer (VORTEX-5, Kyllin-Bell, Haimen, China);
cleaner, and placed in a freezer tube for snap freezing. ultrasonic cleaner (KQ5200E, SUPMILE, China);
Three biological replicates were prepared for each pipette (Research plus, Eppendorf, Germany);
treatment group. Specimens of the A_P5 treatment automation workstation (Biomek i5, Beckman
group were labeled A_P5-1, A_P5-2, and A_P5-3; Coulter, USA); film sealing instrument (Mini HES,
specimens of the A4P5 treatment group were labeled Monad, China).

Volume 22 Issue 5 (2025) 167 doi: 10.366922/AJWEP025150108