Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection

Drought stress greatly affects the growth and development of plants in coal mine spoils located in the Inner Mongolia grassland ecosystem. Arbuscular mycorrhizal fungi (AMF) can increase plant tolerance to drought. However, little is known regarding the contribution of AMF to plants that are grown in different types of coal mine spoils under drought stress. To evaluate the mycorrhizal effects on the drought tolerance of maize (Zea mays L.) grown in weathered (S1) and spontaneously combusted (S2) coal mine spoils, a greenhouse pot experiment was conducted to investigate the effects of inoculation with Rhizophagus intraradices on the growth, nutrient uptake, carbon:nitrogen:phosphorus (C:N:P) stoichiometry and water status of maize under well-watered, moderate and severe drought stress conditions. The results indicated that drought stress increased mycorrhizal colonization and decreased plant dry weights, nutrient contents, leaf moisture percentage of fresh weight (LMP), water use efficiency (WUE) and rehydration rate. A high level of AMF colonization ranging from 65 to 90% was observed, and the mean root colonization rates in S1 were lower than those in S2. In both substrates, inoculation with R. intraradices significantly improved the plant growth, P contents, LMP and WUE and decreased the C:P and N:P ratios of plants under drought stress. In addition, maize grown in S1 and S2 exhibited different wilting properties in response to AMF inoculation, and plant rehydration after drought stress occurred faster in mycorrhizal plants. The results suggested that inoculation with R. intraradices played a more positive role in improving the drought stress resistance of plants grown in S2 than those grown in S1. AMF inoculation has a beneficial effect on plant tolerance to drought and effectively facilitates the development of plants in different coal mine spoils.