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Peer-reviewedCC BY-NC-ND 4.0BMC Plant Biology

Regulation of multi-metal stress responses in spinach by malic acid and Moringa leaf extract: implications for plant performance and phytoextraction

6 affiliations
  1. 1Department of Environmental Sciences, University of Gujrat, Hafiz Hayat Campus, 50700 Gujrat, Pakistan
  2. 2Department of Biology, College of Science, Qassim University, Burydah, 52571 Saudi Arabia
  3. 3Department of Biology, College of Science, Jouf University, Sakaka, 72341 Aljouf Saudi Arabia
  4. 4Department of Biology, Turabah University College, Taif University, Taif, 21995 Saudi Arabia
  5. 5Date Palm Research Centre of Excellence, King Faisal University, Al-Ahsa, 31982 Saudi Arabia
  6. 6Faculty of Agricultural Engineering, University of Aleppo, PO Box 8047, Aleppo, Syria

Abstract

Multi-metal contamination is a major limiting factor for phytoextraction with high biomass plants like spinach (Spinacia oleracea L.) from wastewater used in the surgery industry. This study investigated the integrated application of malic acid (10 mM) from rhizosphere and Moringa oleifera leaf extract (MLE) on the growth, antioxidant defense and multi-metal accumulation in spinach under 50% and 100% authentic surgical wastewater containing higher concentrations of Ni, Cu and Pb. Untreated 100% wastewater reduced plant height by 65.8%, it caused oxidative damage and suppressed biomass production. The dual treatment of 10 mM malic acid + MLE completely reversed these effects such as plant height which recovered by 42.8%; root SOD activity enhancement from 34.25 to 53.06 U mg-1 protein under 100% wastewater; ROS reduction and maintenance of photosynthetic pigments. At the same time, malic acid increased the bioavailability of the metals leading to significant increases in absolute accumulation by roots (under 100% wastewater, root Cu was 7046.67 µg and root Ni was 10510.69 µg). The combined rhizosphere–foliar application of malic acid and MLE against the surgical wastewater (SWW) containing several metals, which has been used in real conditions, has not yet been studied in spinach. This is the first report of the simultaneous rhizosphere application of malic acid and foliar application of Moringa oleifera leaf extract under authentic multi-metal surgical wastewater. The dual-route strategy not only enhanced absolute accumulation of Cu, Pb and Ni but also significantly alleviated oxidative stress and restored growth and photosynthetic performance, suggesting a low-cost approach for phytoextraction with a high-biomass crop.

Graphical abstract:

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BibTeX
@article{farid2026regulation,
  title = {{Regulation of multi-metal stress responses in spinach by malic acid and Moringa leaf extract: implications for plant performance and phytoextraction}},
  author = {Mujahid Farid and Fiza Farooq and Suliman Mohammed Suliman Alghanem and Haifa Abdul Aziz Sakit Alhaithloul and Mohammed S. Alotaibi and Muhammad Munir and Yousef Alhaj Hamoud and Hiba Shaghaleh},
  journal = {BMC Plant Biology},
  year = {2026},
  month = oct,
  doi = {10.1186/s12870-026-10087-7},
  url = {https://doi.org/10.1186/s12870-026-10087-7},
  note = {CC BY-NC-ND 4.0},
}
RIS
TY  - JOUR
TI  - Regulation of multi-metal stress responses in spinach by malic acid and Moringa leaf extract: implications for plant performance and phytoextraction
AU  - Mujahid Farid
AU  - Fiza Farooq
AU  - Suliman Mohammed Suliman Alghanem
AU  - Haifa Abdul Aziz Sakit Alhaithloul
AU  - Mohammed S. Alotaibi
AU  - Muhammad Munir
AU  - Yousef Alhaj Hamoud
AU  - Hiba Shaghaleh
PY  - 2026
DA  - 2026/10/10
JO  - BMC Plant Biology
DO  - 10.1186/s12870-026-10087-7
UR  - https://doi.org/10.1186/s12870-026-10087-7
AB  - Multi-metal contamination is a major limiting factor for phytoextraction with high biomass plants like spinach (Spinacia oleracea L.) from wastewater used in the surgery industry. This study investigated the integrated application of malic acid (10 mM) from rhizosphere and Moringa oleifera leaf extract (MLE) on the growth, antioxidant defense and multi-metal accumulation in spinach under 50% and 100% authentic surgical wastewater containing higher concentrations of Ni, Cu and Pb. Untreated 100% wastewater reduced plant height by 65.8%, it caused oxidative damage and suppressed biomass production. The dual treatment of 10 mM malic acid + MLE completely reversed these effects such as plant height which recovered by 42.8%; root SOD activity enhancement from 34.25 to 53.06 U mg-1 protein under 100% wastewater; ROS reduction and maintenance of photosynthetic pigments. At the same time, malic acid increased the bioavailability of the metals leading to significant increases in absolute accumulation by roots (under 100% wastewater, root Cu was 7046.67 µg and root Ni was 10510.69 µg). The combined rhizosphere–foliar application of malic acid and MLE against the surgical wastewater (SWW) containing several metals, which has been used in real conditions, has not yet been studied in spinach. This is the first report of the simultaneous rhizosphere application of malic acid and foliar application of Moringa oleifera leaf extract under authentic multi-metal surgical wastewater. The dual-route strategy not only enhanced absolute accumulation of Cu, Pb and Ni but also significantly alleviated oxidative stress and restored growth and photosynthetic performance, suggesting a low-cost approach for phytoextraction with a high-biomass crop.  Graphical abstract:
SN  - 1471-2229
ER  -