Jazbia Shirin ¹ Yongjing Chen ¹ Azhar Hussain Shah ² Yanmei Da ¹ Guowei Zhou ^1* Qingye Sun ^1*

• ¹ Anhui University, Hefei, China
• ² Department of Biotechnology and Genetics Engineering, Hazara University, Mansehra, Pakistan

Microplastics (MPs) have garnered global attention as emerging contaminants due to their adaptability, durability, and robustness in various ecosystems. Still, studies concerning their combination with heavy metals (HMs), their interactions with soil biota, and how they affect soil physiochemical properties and terrestrial plant systems are limited. Our study was set to investigate the combined effect of HMs (cadmium, arsenic, copper, zinc and lead) contaminated soil of Tongling and different sizes (T1 = 106 µm, T2 = 50 µm, and T3 = 13 µm) of polystyrene microplastics on the soil physiochemical attributes, both bacterial and fungal diversity, compositions, AMF (arbuscular mycorrhizal fungi), plant pathogens in the soil, and their effect on Lactuca sativa by conducting a greenhouse experiment. According to our results, the combination of HMs and polystyrene microplastic (PS-MPs), especially the smaller PS-MPs (T3), was more lethal for the lettuce growth, microbes and soil. The toxicity of combined contaminants directly reduced the physio-biochemical attributes of lettuce, altered the lettuce's antioxidant activity and soil health. T3 at the final point led to a significant increase in bacterial and fungal diversity. In contrast, overall bacterial diversity was higher in the rhizosphere, and fungal diversity was higher in the bulk soil. Moreover, the decrease in MPs size played an important role in decreasing AMF and increasing both bacterial and fungal pathogens, especially in the rhizosphere soil. Functional prediction was found to be significantly different in the control treatment, with larger MPs compared to smaller PS-MPs. Environmental factors also played an important role in the alteration of the microbial community. This study also demonstrated that the varied distribution of microbial populations could be an ecological indicator for tracking the environmental health of soil.Overall, our work showed that the combination of HMs and smaller sizes of MPs was more lethal for the soil biota and lettuce and also raised many questions for further studying the ecological risk of PS-MPs and HMs.