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<p>Heavy metal toxicity is a major abiotic stress affecting plant growth, productivity, and environmental sustainability. This study evaluated the in vitro responses of <i>Chenopodium ambrosioides</i> L. callus cultures to selected heavy metals, focusing on growth, tolerance, and metal accumulation. Callus induction was optimized using leaf, petiole, and stem explants on Murashige and Skoog (MS) medium supplemented with plant growth regulators. The highest induction rates (90% in stems and 100% in leaf and petiole explants) were achieved with 2&#xa0;mg/L dichlorophenoxyacetic acid (2,4-D) and 0.5&#xa0;mg/L 6-benzylaminopurine (BAP). Callus cultures were subsequently exposed to cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), and zinc (Zn) at concentrations of 0, 0.5, 1, and 1.5&#xa0;mg/L. Increasing metal concentrations caused a significant decline in callus dry weight (DW) and growth tolerance index (GTI), particularly under Cd and Co stress (r ≈ − 0.93 to − 0.96, p ≤ 0.01). In contrast, 0.5&#xa0;mg/L Cu enhanced growth (DW: 63.1&#xa0;mg; GTI: 105.16%), indicating a hormetic effect, while Cr and Zn at 0.5&#xa0;mg/L maintained near-control growth levels (Cu &gt; Zn ≥ Cr &gt; Co &gt; Cd at 0.5&#xa0;mg/L but Cr &gt; Cu &gt; Zn &gt; Co &gt; Cd at 1–1.5&#xa0;mg/L in medium). Metal accumulation increased significantly with concentration, reaching peak values of 622&#xa0;mg/L for Zn (1&#xa0;mg/L), 374.67&#xa0;mg/L for Cu (1&#xa0;mg/L), 323&#xa0;mg/L for Co (1&#xa0;mg/L), 242.67&#xa0;mg/L for Cd (1.5&#xa0;mg/L), and 152.67&#xa0;mg/L for Cr (0.5&#xa0;mg/L) (Zn &gt; Cu &gt; Cd &gt; Co &gt; Cr). Compared to controls, accumulation increased by up to 74–78 × for Cd, 48–54 × for Cr, 14.5–16.5 × for Co, 1.7–2.7 × for Zn, and 1.2–2.5 × for Cu (Cd &gt; Cr &gt; Co &gt; Zn &gt; Cu). Multivariate analyses (Pearson, Hierarchical Cluster, Multiple Regression) revealed strong negative correlations between growth and metal concentration, and positive correlations with accumulation. Principal component analysis (95.33% variance explained) clearly separated growth traits (DW, GTI) from accumulation. Bioaccumulation factor (BF) values were highest for Cd and Co [Cd (161.67–459.33) &gt; Co (99.33–197) &gt; Zn (2.95–4.29) ≥ Cu (1.12–1.41) ≥ Cr (0.56–0.66), while modified translocation factor (TFₚₛₑᵤ<sub>d</sub>ₒ) was greatest for Cd and Cr [Cd (78.35) &gt; Cr (48.72) &gt; Co (14.41) &gt; Zn (1.79) ≥ Cu (1.19)], indicating strong enrichment capacity. Bioaccumulation and translocation indices highlighted Cd and Co as strong accumulators through phytoextraction, Cr as showing stress-induced enrichment with limited uptake, and Cu and Zn as tightly regulated with minimal mobility and baseline response. Overall, <i>C. ambrosioides</i> callus cultures exhibit high metal uptake capacity and distinct tolerance strategies, supporting their use as a model for phytoremediation screening and stress physiology studies.</p> Graphical Abstract <p></p>

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High Phytoremediation of Chenopodium ambrosioides L. In vitro Callus Cultures to Cd, Co, Cr, Cu and Zn Stress Through Increase in Biomass, Growth Tolerance Index and Heavy Metal Accumulation After Optimizing Callusing Potential Using Different Explants and Plant Growth Regulators

  • 期刊论文

  • 发表日期：

    2026年7月8日

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  • Huda Enaya Mahood,
  • Virginia Sarropoulou,
  • Theocharis Chatzistathis

摘要

Heavy metal toxicity is a major abiotic stress affecting plant growth, productivity, and environmental sustainability. This study evaluated the in vitro responses of Chenopodium ambrosioides L. callus cultures to selected heavy metals, focusing on growth, tolerance, and metal accumulation. Callus induction was optimized using leaf, petiole, and stem explants on Murashige and Skoog (MS) medium supplemented with plant growth regulators. The highest induction rates (90% in stems and 100% in leaf and petiole explants) were achieved with 2 mg/L dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L 6-benzylaminopurine (BAP). Callus cultures were subsequently exposed to cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), and zinc (Zn) at concentrations of 0, 0.5, 1, and 1.5 mg/L. Increasing metal concentrations caused a significant decline in callus dry weight (DW) and growth tolerance index (GTI), particularly under Cd and Co stress (r ≈ − 0.93 to − 0.96, p ≤ 0.01). In contrast, 0.5 mg/L Cu enhanced growth (DW: 63.1 mg; GTI: 105.16%), indicating a hormetic effect, while Cr and Zn at 0.5 mg/L maintained near-control growth levels (Cu > Zn ≥ Cr > Co > Cd at 0.5 mg/L but Cr > Cu > Zn > Co > Cd at 1–1.5 mg/L in medium). Metal accumulation increased significantly with concentration, reaching peak values of 622 mg/L for Zn (1 mg/L), 374.67 mg/L for Cu (1 mg/L), 323 mg/L for Co (1 mg/L), 242.67 mg/L for Cd (1.5 mg/L), and 152.67 mg/L for Cr (0.5 mg/L) (Zn > Cu > Cd > Co > Cr). Compared to controls, accumulation increased by up to 74–78 × for Cd, 48–54 × for Cr, 14.5–16.5 × for Co, 1.7–2.7 × for Zn, and 1.2–2.5 × for Cu (Cd > Cr > Co > Zn > Cu). Multivariate analyses (Pearson, Hierarchical Cluster, Multiple Regression) revealed strong negative correlations between growth and metal concentration, and positive correlations with accumulation. Principal component analysis (95.33% variance explained) clearly separated growth traits (DW, GTI) from accumulation. Bioaccumulation factor (BF) values were highest for Cd and Co [Cd (161.67–459.33) > Co (99.33–197) > Zn (2.95–4.29) ≥ Cu (1.12–1.41) ≥ Cr (0.56–0.66), while modified translocation factor (TFₚₛₑᵤdₒ) was greatest for Cd and Cr [Cd (78.35) > Cr (48.72) > Co (14.41) > Zn (1.79) ≥ Cu (1.19)], indicating strong enrichment capacity. Bioaccumulation and translocation indices highlighted Cd and Co as strong accumulators through phytoextraction, Cr as showing stress-induced enrichment with limited uptake, and Cu and Zn as tightly regulated with minimal mobility and baseline response. Overall, C. ambrosioides callus cultures exhibit high metal uptake capacity and distinct tolerance strategies, supporting their use as a model for phytoremediation screening and stress physiology studies.

Graphical Abstract