The Role of Lignin in the Compartmentalization of Cadmium in Maize Roots Is Enhanced by Mycorrhiza

Abstract

In nature, arbuscular mycorrhizal fungi (AMF) play a crucial role in the root systems of plants. They can help enhance the resistance of host plants by improving the compartmentalization of toxic metal contaminants in the cell walls (CWs). However, the functions and responses of various CW subfractions to mycorrhizal colonization under Cd exposure remain unknown. Here we conducted a study to investigate how Cd is stored in the cell walls of maize roots colonized by Funneliformis mosseae. Our findings indicate that inoculating the roots with AMF significantly lowers the amount of Cd in the maize shoots (63.6 ± 6.54 mg kg^-1 vs. 45.3 ± 2.19 mg kg^-1, p < 0.05) by retaining more Cd in the mycorrhized roots (224.0 ± 17.13 mg kg^-1 vs. 289.5 ± 8.75 mg kg^-1, p < 0.01). This reduces the adverse effects of excessive Cd on the maize plant. Additional research on the subcellular distribution of Cd showed that AMF colonization significantly improves the compartmentalization of 88.2% of Cd in the cell walls of maize roots, compared to the 80.8% of Cd associated with cell walls in the non-mycorrhizal controls. We observed that the presence of AMF did not increase the amount of Cd in pectin, a primary binding site for cell walls; however, it significantly enhanced the content of lignin and the proportion of Cd in the total root cell walls. This finding is consistent with the increased activity of lignin-related enzymes, such as PAL, 4CL, and laccase, which were also positively impacted by mycorrhizal colonization. Fourier transform infrared spectroscopy (FTIR) results revealed that AMF increased the number and types of functional groups, including -OH/-NH and carboxylate, which chelate Cd in the lignin. Our research shows that AMF can improve the ability of maize plants to tolerate Cd by reducing the amount of Cd transferred from the roots to the shoots. This is achieved by increasing the amount of lignin in the cell walls, which binds with Cd and prevents it from moving through the plant. This is accomplished by activating enzymes related to lignin synthesis and increasing the exposure of Cd-binding functional groups of lignin. However, more direct evidence on the immobilization of Cd in the mycorrhiza-altered cell wall subfractions is needed.

Keywords: Cd tolerance; arbuscular mycorrhizal fungus (AMF); cell walls (CWs); lignin; maize (Zea mays).

Figure 1

The growth (A,B), biomass (C), plant height (D), basal perimeter (E), and chlorophyll content (F) of maize plants evaluated based on two conditions: those inoculated with F. mosseae (M+) and those not inoculated (M−), and under two Cd supplementation levels: 0 mg kg⁻¹ (Cd0) and 25 mg kg⁻¹ (Cd25) for a period of 50 days. The data presented in the results represent the mean with standard error (n = 8), and statistically significant differences are indicated with asterisks (*). The level of significance is denoted as follows: (*, p < 0.05; **, p < 0.01; ***, p < 0.001, t-test). NS indicates no statistically significant difference (p > 0.05, two-way ANOVA).

Figure 2

This data shows the concentration (A) and translocation factor of Cd (B) that accumulated in maize dry weight (DW), as well as the Cd concentration in maize fresh weight (FW) (C) and the Cd proportion (D) of various root subcellular fractions. The maize was either inoculated (M+) or not inoculated (M−) with F. mosseae and treated with Cd0 or Cd25. The data presented in the results represent the mean with standard error (n = 8), and statistically significant differences are indicated with asterisks (*). The level of significance is denoted as follows: (*, p < 0.05; **, p < 0.01; ***, p < 0.001, t-test). The subcellular fractions were divided into CW (cell walls) and nCW (remaining fractions excluding cell wall components).

Figure 3

Cd concentration (A), Cd proportion (B), lignin content (C), and pectin content (D) in the cell wall (measured in fresh weight, FW) of roots that were either inoculated (M+) or not inoculated (M−) with F. mosseae, and exposed to either 0 mg kg⁻¹ (Cd0) or 25 mg kg⁻¹ Cd (Cd25) supplementation. The data presented are the means ± SE ((A,B), n = 4; (C,D), n = 8), and statistical significance is indicated by asterisks (* for p < 0.05, ** for p < 0.01, t-test). NS indicates no statistically significant difference (p > 0.05, two-way ANOVA).

Figure 4

FTIR characteristics of functional groups with Cd binding potential from total cell wall (A), lignin (B) and pectin fractions (C) of un-inoculated (M−) or inoculated (M+) maize roots under 0 mg kg⁻¹ and 25 mg kg⁻¹ Cd supplementation.

Figure 5

Activities of cell wall-related enzymes of maize roots inoculated (M+) or not inoculated (M−) with F. mosseae under 0 (Cd0) and 25 mg kg⁻¹ Cd supplementation (Cd25). Phenylalanine ammonialyase (PAL) (A), 4-Coumarate:CoA ligase activity (4CL) (B), laccase activity (C), pectin methylase (PME) (D) and α1,4 galacto syltransferase (α1,4 GalT) (E). The results are presented as means ± SE (n = 4), with statistically significant differences indicated by * (p < 0.05), ** (p < 0.01) or *** (p < 0.001) using a t-test. NS indicates no statistically significant difference (p > 0.05, two-way ANOVA).