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Effects of nickel sulphate and lead acetate trihydrate on heavy metal stress-related gene activities in forage pea (Pisum sativum ssp. arvense L.) in Turkiye

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Overview

This study measured the expression of four heavy metal stress-related genes (APX, CAT, MT, PCS) by qRT-PCR in three forage pea cultivars (Kurtbey, Pembe Incisi, Kirazli) of Pisum sativum ssp. arvense exposed to nickel sulphate (20 and 40 mg/L) and lead acetate trihydrate (20 and 40 mg/L) in a greenhouse pot experiment. It is a gene-expression and morphological study; it does not report tissue metal concentrations in food units. The study documents that pea plants respond to nickel and lead through antioxidant and chelation pathways (APX, CAT, MT, PCS), and that the magnitude of the stress response depends on cultivar genetics and metal dose.

Key numbers

The study reports fold-change in mRNA expression (relative to control), not tissue metal concentrations:

  • APX fold-change in Kirazli, lead acetate trihydrate 40 mg/L: approximately 43-fold above control (Figure 1, p.4-5).
  • MT fold-change in Kurtbey, lead acetate trihydrate 20 mg/L: approximately 8.7-fold above control (Figure 3, p.5).
  • PCS fold-change in Kurtbey, lead acetate trihydrate 20 mg/L: approximately 8.7-fold above control (Figure 4, p.5).
  • One-way ANOVA (Dunnett’s test) found significant differences among means for APX gene expression across all 15 samples (p = 0.0472) (Figure 5, p.6).

Morphological data (Table 3, p.7-8): control plants had a mean plant height of 46.88 cm. Lead acetate trihydrate at 20 mg/L reduced mean plant height to 39.66 cm (p < 0.05). Nickel sulphate at 20 mg/L raised plant height to 47.33 cm (not significant vs control). Stem diameter, node number, internode length, stipule width, and stipule length were also measured; most treatment effects on morphology were non-significant at p < 0.05 except for treatment effects on plant height and stipule length.

Methods (brief)

Pot experiment, greenhouse, Pazar district, Rize, Turkiye, 2024. Split-plot design, 3 replications, 3 cultivars x 5 treatments (control, NiSO4 20 mg/L, NiSO4 40 mg/L, lead acetate trihydrate 20 mg/L, lead acetate trihydrate 40 mg/L). Plants treated 4 weeks after sowing; treated by mixing equal amounts of solution into each pot. RNA isolation using Ecotech Plant Total RNA Kit. cDNA synthesis using Thermo Scientific PCR protocol. qRT-PCR using Ecotech SYBR Green qCR Assays kit, Roche Lightcycler 96. Beta-Actin used as housekeeping control. 2-delta-delta CT method (Livak and Schmittgen 2001). ANOVA in IBM SPSS 29.0. Tukey’s HSD test at p < 0.05. No tissue metal concentrations measured; study design is gene expression only.

Implications

This study is context for the uptake pathway section of the frozen peas and pea-protein pages: it documents that peas activate antioxidant defense (APX, CAT) and metal-chelation genes (MT, PCS) under nickel and lead stress. These responses indicate root-to-shoot metal translocation is occurring but do not provide food-concentration data. The 20-40 mg/L solution concentrations used are experimentally elevated stress levels, not typical agricultural soil concentrations.

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