RESPONSE CHARACTERISTICS OF VALLISNERIA NATANS AND RHIZOSPHERE MICROBIAL COMMUNITY UNDER SEDIMENT-CADMIUM POLLUTION
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摘要:
研究以典型沉水植物苦草(Vallisneria natans)为研究对象, 通过在0、1、5、10、20和50 mg/kg沉积物镉(Cd)污染环境下苦草鲜重、株高和叶绿素(Chlorophyll)及丙二醛(Malondialdehyde, MDA)、超氧化物歧化酶(Superoxide Dismutase, SOD)和过氧化物酶(Peroxidase, POD)活性测定来分析苦草对镉的耐受效应, 并借助生物/底泥富集因子(BSAF)和植物转运因子(TF)来了解苦草的富集能力, 同时利用16S rRNA基因扩增子测序技术研究苦草根际细菌的群落结构对沉积物镉污染环境下的响应特征。结果表明: (1) 当Cd≥20 mg/kg处理时, 苦草植株鲜重、株高、叶绿素含量和抗逆能力下降, SOD和POD活性先上升后下降, MDA含量上升; (2) BSAF均超过1表明苦草能有效地富集污染底泥中的镉, 转运系数(TF)均小于1, 说明苦草依靠根滤机制来富集镉, 建议在收获苦草时需要定期连根去除; (3) 测序结果表明苦草根际由放线菌门、厚壁菌门和变形菌门等37个门细菌组成, 包含鞘氨醇单胞菌属、黄杆菌属、马赛菌属、芽孢杆菌属和节杆菌属等植物促生细菌, 这些细菌可能在促进苦草生长、耐受和富集重金属镉起到重要作用。研究分析了苦草在镉胁迫环境下形态学和生理生化等相关指标的变化和苦草根际细菌群落组成及其对镉胁迫的响应特征, 为后续提高苦草修复效果提供了基础。
Abstract:In this study, the typical submerged macrophyte Vallisneria natans was selscted, and the fresh weight, plant height, chlorophyll, malondialdehyde (MDA), superoxide dismutase (SOD) and peroxidase (POD) activities of Vallisneria natans were measured at 0, 1, 5, 10, 20 and 50 mg/kg of sediment cadmium contamination to analyze the cadmium tolerance effect of Vallisneria natans, and the enrichment capacities were indicated by the values of Biota-sediment accumulation factor (BSAF) and translocation factor (TF), while the 16S rRNA gene amplicon sequencing technique was used to study the community structure of Vallisneria natans rhizosphere bacteria response to the sediment cadmium contamination. The results showed that: (1) When Cd≥20 mg/kg treatment, the fresh weight, plant height, chlorophyll content and the stress resistance decreased of Vallisneria natans. The activities of SOD and POD increased firstly and then reduced, MDA content increased with increasing concentration; (2) in all the treatments, BSAF exceeded 1, indicating that Vallisneria natans could effectively enrich cadmium in the contaminated sediment, and TF was less than 1, indicating that Vallisneria natans relied on root filtration mechanism to enrich cadmium, and needed to be harvested regularly; (3) the sequencing results showed that the rhizosphere bacteria of Vallisneria natans were mainly composed of 37 phylum, such as Actinobacteria, Firmicutes and Proteobacteria, including plant growth-promoting bacteria such as Sphingomonas, Flavisobacter, Massilia, Bacillus, and Arthrobacter, which may play important roles in promoting the growth, tolerance, and enrichment of cadmium in Vallisneria natans. In this study, we preliminarily analyzed the changes of morphological, physiological and biochemical indicators of Vallisneria natans under varied cadmium stress environments, and the composition of Vallisneria natans rhizosphere bacterial community and its response characteristics to cadmium stress, which provide guidance for the subsequent improvement of Vallisneria natans restoration efficiency.
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Keywords:
- Sediment /
- Cadmium /
- Restoration /
- Microorganisms community /
- Vallisneria natans
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图 1 不同浓度的镉处理对苦草叶片叶绿素、MDA、SOD和POD含量的变化
Figure 1. Changes of contents of Chlorophyll, MDA, SOD and POD in leaves of Vallisneria natans treat with different concentrations of Cd (mean±SD, n=3)
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图 2 不同浓度的镉处理苦草叶片ROS分布
Figure 2. The distribution of ROS in the leaves of Vallisneria natans treat with different concentrations of Cd
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图 3 不同镉浓度处理下沉积物微生物物种分类热图(属水平)
Figure 3. Heatmap of sediment microbial species under different cadmium concentrations treatments (genus level)
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图 4 镉浓度与沉积物、苦草指标相关性分析
Figure 4. Correlation analysis of cadmium concentration with indexes of sediment and Vallisneria natans (* P≤0.05, ** P≤0.01)
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图 5 属分类水平下沉积物微生物群落与环境因子、苦草生理生化相关性分析
Figure 5. Correlation analysis of sediment microbial community with environmental factors and Vallisneria natans physiology and biochemistry indexes at genus taxonomic level (* P≤0.05, ** P≤0.01)
表 1 不同浓度的镉处理对苦草生长的影响
Table 1 Effects of different Cd concentrations on growth of Vallisneria natans (mean±SD, n=3)
处理组Treatment
(mg/kg)整株生物量
Whole plant (g)株高
Plant height (cm)叶片
Leaf (mg/kg)根
Root (mg/kg)BSAF TF 0 3.123±0.534a 31.7±1.700a — — — — 1 2.712±0.157ab 27.4±0.432b 7.53±0.17e 15.33±0.78e 13.01±0.25a 0.49±0.01b 5 2.517±0.449abc 26.3±1.700b 14.73±0.37d 63.96±0.70d 6.84±0.31b 0.23±0.00d 10 2.429±0.393abc 24.9±0.660bc 32.30±0.13c 134.44±0.37c 6.01±0.16c 0.24±0.00d 20 2.092±0.209bc 23.7±1.247c 61.25±1.46b 183.52±1.66b 4.69±0.04d 0.33±0.00c 50 1.942±0.170b 18.5±0.508d 221.67±6.72a 365.84±4.75a 5.08±0.09d 0.61±0.01a 注: 表中数据为3个重复的平均值; 同一列相同右上角含有不同英文上标字母表示有显著差异(P<0.05); 下同Note: Data are means of triplicates. Means in each line sharing the different superscript are significantly different determined by Least test (P<0.05), the same applies below 
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表 2 沉积物pH、氮磷和酶活性
Table 2 pH, nitrogen and phosphorus, and enzyme activities of sediments (mean±SD, n=3)
处理组Treatment (mg/kg) 镉Cd (mg/kg) pH 总氮 TN (mg/kg) 总磷 TP (mg/kg) 脲酶 S-UE [μg/(d·g)] 磷酸酶 S-NP [μmol/(d·g)] 0 0.01±0.00e 5.46±0.02f 492.93±48.05a 134.58±10.01ab 97.76±4.09a 5.09±0.04a 1 0.72±0.01e 6.25±0.08e 324.07±52.56b 156.46±19.55a 93.86±3.92ab 4.52±0.15b 5 4.53±0.22d 6.37±0.01d 390.71±6.90b 127.90±11.36ab 80.96±12.05abc 4.50±0.22b 10 8.39±0.11c 6.62±0.02c 334.82±16.78b 139.77±12.49ab 66.56±5.71c 2.52±0.09d 20 17.58±0.46b 6.74±0.04b 350.79±50.02b 123.45±7.73b 76.87±13.01bc 2.93±0.08c 50 46.12±0.74a 7.04±0.01a 537.36±34.83a 147.19±18.38ab 88.30±3.64ab 2.77±0.04cd
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