3种纳米材料对水稻幼苗生理及根际细菌群落结构的影响
Effects of Three Nanomaterials on Physiology and Rhizosphere Bacterial Community Structure of Rice Seedlings
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摘要: 随着纳米技术的快速发展，评估人工纳米材料(ENMs)对植物-微生物系统的潜在危害至关重要。本研究通过盆栽试验，分析不同浓度(0、0.50、1.00和2.00 mg·g-1)的纳米材料即纳米二氧化硅(nSiO2)、纳米二氧化钛(nTiO2)和纳米氧化锌(nZnO)对水稻幼苗生理和根际细菌群落结构的影响。研究结果显示，3种纳米材料处理后，水稻幼苗的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均显著增加(P<0.05，下同)，可溶性蛋白(SP)含量仅在2.00 mg·g-1 nTiO2和0.50 mg·g-1 nZnO处理时显著降低；2.00 mg·g-1 nSiO2处理及1.00 mg·g-1和2.00 mg·g-1 nZnO处理均可显著降低水稻幼苗的株高(PH)、鲜质量(FW)和干质量(DW)，nTiO2处理则对其没有显著性影响。高通量测序结果表明，与不加纳米材料的对照(CK)处理比，3种纳米材料处理的根际土壤优势菌门为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)和拟杆菌门(Bacteroidetes)，根际促生及反硝化细菌如芽孢杆菌属(Bacillus)、黄色土源菌(Flavisolibacter)、Kaistobacter、红游动菌属(Rhodoplanes)和Candidatus_Solibacter菌属的丰度均有显著提升。Pearson相关分析表明，Catellatospora等属的相对丰度与水稻的抗氧化酶活性和生物量呈显著正相关，而Candidatus_Koribacter等属的相对丰度与其呈显著负相关。同时，nSiO2和nZnO处理浓度分别为1.00 mg·g-1和2.00 mg·g-1的水稻根际土壤细菌多样性均降低，群落结构变化明显，而nTiO2处理对其影响不显著；nSiO2和nTiO2处理均诱导水稻根际细菌中编码氨基酸代谢等基因丰度显著升高，nZnO处理则降低与细胞运动等相关功能基因丰度。综上所述，3种尺度相同的纳米材料可直接对水稻幼苗产生生理毒性，但因nZnO和nTiO2的水力直径分别为最小和最大而相应表现出最强和最弱的毒性效应。此外，3种纳米材料尤其是nSiO2和nZnO还可通过改变根际土壤细菌基因功能，降低水稻根际土壤细菌多样性并改变群落组成及结构，间接造成水稻幼苗氧化应激和渗透胁迫，进而不同程度地影响幼苗生长和发育，其中nZnO的抑制效果最显著。3种纳米材料对水稻幼苗生理和根际细菌群落表现出的毒性大小顺序：nZnO > nSiO2 > nTiO2。本研究结果为纳米材料对水稻及根际土壤微生物的潜在危害及农田生态系统的环境保护与资源利用提供了科学依据。Abstract: With the rapid development of nanotechnology, it is critical to assess the potential harm of artificial nanomaterials (ENMs) on plant-microbial systems. In this study, the effects of different concentrations (0, 0.50, 1.00 and 2.00 mg·g-1) of nanomaterials such as nanometer silica, nanometer titanium peroxide and nanometer zinc oxide (nSiO2, nTiO2 and nZnO) on the physiological characteristics and rhizosphere bacterial community structure of rice seedlings were analyzed by pot experiments. The results showed that the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in rice seedlings increased significantly after the treatment of three nanomaterials (P<0.05, the same below), and the soluble protein (SP) content significantly decreased only when treated with 2.00 mg·g-1 nTiO2 and 0.50 mg·g-1 nZnO. The treatment of 2.00 mg·g-1 nSiO2 and 1.00 mg·g-1 and 2.00 mg·g-1 nZnO significantly reduced plant height (PH), fresh weight (FW) and dry weight (DW) of rice seedlings, while nTiO2 treatment had no significant effects on PH, FW and DW of rice seedlings. The results of high-throughput sequencing showed that compared with the control (CK), the dominant phyla were Proteobacteria, Acidobacteria, Chloroflexi and Bacteroidetes, and the abundances of plant growth promoting bacteria and denitrifying bacteria such as Bacillus, Flavisolibacter, Kaistobacter, Rhodoplanes and Candidatus_Solibacter significantly increased in the rhizosphere soil treated with three kinds of nanomaterials. Pearson correlation analysis showed that the relative abundances of Catellatospora and other genera were significantly positively correlated with the antioxidant enzyme activity and the biomass of rice seedlings, while the relative abundances of Candidatus_Koribacter and other genera were significantly negatively correlated with it. At the same time, the bacterial diversity decreased and the community structure changed significantly in rice rhizosphere soil treated with 1.00 mg·g-1 nSiO2 or 2.00 mg·g-1 nZnO, but nTiO2 treatment had no significant effect on it. The treatment with nSiO2 or nTiO2 significantly increased the abundance of genes related to amino acid metabolism in rice rhizosphere bacteria, while nZnO treatment decreased the abundance of functional genes related to cell movement. In conclusion, three nanomaterials with the same size can directly produce physiological toxicity to rice seedlings. However, nZnO and nTiO2 showed the strongest and the weakest toxic effects due to the smallest and the largest hydraulic diameters respectively. Three kinds of nanomaterials, especially nSiO2 and nZnO, indirectly caused oxidative stress and osmotic stress, and affected the growth and development of rice seedlings by changing gene function, reducing the diversity and changing the community structure in bacteria from rice rhizosphere soil. Among them, nZnO had the strongest inhibitory effect. The order of the toxicity of three nanomaterials to physiological functions of rice seedlings and bacterial community in the rhizosphere was as follow: nZnO > nSiO2 > nTiO2. The results of this study provided a scientific basis to reveal the potential damage of nanomaterials to rice and microorganisms in rhizosphere soil, and conduct the environmental protection and resource utilization of farmland ecosystem.
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