新疆地震断裂带次生植物对土壤微生物群落结构及代谢的影响
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摘要
为了解新疆地震断裂带次生植物对土壤微生物群落结构及代谢的影响,本文利用末端限制性片段长度多态性(terminal restriction fragment length polymorphism,T-RLFP)与BIOLOG技术,结合测定土壤主要化学性质,以同土层裸地为对照,对13种生长于地震断裂带的植物根际土壤微生物群落结构及代谢特征进行研究。主要研究内容与试验结果如下:
1.依据土壤分析方法标准(DB/6500 B11)测定对照和不同植物根际土壤的pH、有机质、全氮、速效氮、速效磷、速效钾。结果表明,样地土壤呈碱性(pH 8.04-8.51),由方差分析(ANOVA)可知不同植物根际土壤化学性质差异均显著(p<0.05)。9种植物根际土壤pH高于对照;土壤养分在不同植物间变化范围较大,总体趋势表现为植物根际土壤养分多高于裸地。其中蓍、东方旱麦草的根际土壤各项养分含量普遍较高,而新疆方枝柏较低。由此可知,植被的恢复在不同程度上改善了土壤的养分。
2.利用T-RLFP技术研究次生植物根际土壤细菌的群落结构,选用细菌16S rDNA通用引物8F/1492R进行扩增,HhaⅠ、RsaⅠ进行酶切。结果显示,不同植物改变了土壤微生物优势类群组成,增加了优势类群种类,尤其东方旱麦草、蓍对根际微生物作用最明显。变形菌门(Proteobacteria)细菌在植物根际土壤优势类群中占最大比例(49.55%)。典范对应分析(CCA)表明,细菌优势类群的分布与pH呈显著负相关(HhaⅠ酶切:r=-0.82,p<0.001; RsaⅠ酶切:r=-0.76,p<0.01)。由此推定植被的恢复增加了土壤细菌多样性,改变了土壤细菌群落结构,同时优势类群的分布还与土壤pH有关。
3.利用T-RLFP技术研究次生植物根际土壤古菌的群落结构,选用古菌16S rDNA通用引物21F/958R进行扩增,HhaⅠ、RsaⅠ进行酶切。结果显示,在皱纹柳、直穗柳、东方旱麦草、西北绢蒿、鼠掌老鹳草、欧亚绣线菊、蓬子菜、密刺蔷薇8种植物根际土壤扩增出古菌,各植物根际土壤古菌群落间相似性较低,泉古菌门(Crenarchaeota)和广古菌门(Euryarchaeota)为主要类群。典范对应分析(CCA)表明,优势类群的分布与土壤有机质呈显著正相关(HhaⅠ酶切:r=0.94,p<0.001; RsaⅠ酶切:r=0.74,p<0.05),由相关分析可知土壤速效磷含量与古菌群落各多样性指数呈显著正相关(p<0.05),土壤总氮含量与均匀度指数E呈显著正相关(p<0.05)。总体而言,部分植被可在土壤0～20cm深度富集古菌,且古菌群落因植物种类不同呈较大差异,同时土壤有机质、速效磷也对古菌类群分布产生影响。
4.选取次生植物中草本、灌木优势植物各3种,利用BIOLOG技术研究微生物群落代谢功能。结果显示:6种植物根际土壤的平均颜色变化率(AWCD)差异显著(p<0.05),且均高于裸地。次生植物不影响碳源利用的丰富度(p>0.05),但改变优势度和均匀度(p<0.05)。主成分分析(PCA)表明,不同植物根际土壤微生物群落利用碳源存在差异,其对糖类、氨基酸类、羧酸类碳源利用不同是造成差异的主要因素。随着植物的出现,土壤微生物利用碳源的类型由酚类向糖类、羧酸类转变。相关分析发现,土壤速效钾含量与聚合物类碳源(r=-0.84)、胺类(r=-0.83)的利用呈负相关。由此推断次生植物能显著增强土壤微生物碳源利用能力,改变碳源利用类型,且部分类型碳源的利用受速效钾含量的影响。
The aim of this study was to investigate the effects of secondary plants on Soil microbial community structure and metabolism in Xinjiang earthquake fault zone. The rhizosphere soils collected from 13 different secondary plants were studied by testing soil chemical properties and employing terminal restriction fragment length polymorphism (T-RFLP) and BIOLOG techniques. Unplanted soil in the same depth served as control. The investigated contents and results showed that:
1. Soil pH, organic matter, total nitrogen, available nitrogen, available phosphorus and available potassium were measured according to the standard soil analysis methods (DB/6500 B11). The results showed that the soils were alkaline (pH=8.04-8.51), Analysis of variance(ANOVA) indicated that chemical properties of 13 rhizosphere soils were significantly different (p<0.05). The pH in most (9) rhizosphere soils were higher than control; soil nutrients among the different plants varied widely, the overall trend showed rhizosphere soil nutrients were higher than control, and soil nutrients contents in the rhizosphere of Achillea millefolium and Eremopyrum orientale were relatively high, while the one in Sabina pseudosabina were low. The results presumed that the vegetation restoration improved the soil nutrients in varying degrees.
2. T-RLFP technique were used to study bacterial community structures in rhizosphere soil of secondary plants, we choosed universal primers 8F/1492R to amplify bacterial 16SrDNA, and HhaⅠ, RsaⅠenzymes to digest. As a result, the different plants changed the composition of dominant bacterial groups in soil and increased the sorts of dominant bacterial groups. Achillea millefolium and Eremopyrum orientale had the most significant effects. Proteobacteria accounted for the largest proportion (49.55%) in the soil. Canonical correspondence analysis (CCA) showed that the distribution of dominant bacterial groups was negatively correlated with pH significantly (HhaⅠdigestion: r =- 0.82, p <0.001; RsaⅠdigestion: r =- 0.76, p <0.05). These results suggested that vegetation restoration increased soil bacterial diversity, and had influence on the structures of soil bacterial communities. The distribution of dominant bacterial groups was also related to soil pH.
3. T-RLFP technique were used to study archaeal community structures in rhizosphere soil of secondary plants, we choosed universal primers 21F/958R to amplify archaeal 16SrDNA, and HhaⅠ, RsaⅠe nzymes to digest. The results showed that archaea were found only in thoes rhizosphere soils of Salix vistita, Salix rectijulis, Eremopyrum orientale, Seriphidium nitrosum, Geranium sibiricum, Spiraea media, Galium verum and Rosa spinosissima. Results in the detection of archaea showed that the similarities of archaeal communities among different rhizosphere soils were low. Crenarchaeota and Euryarchaeota were the main groups. Canonical correspondence analysis (CCA) showed that the distribution of dominant archaeal groups was positively correlated with soil organic matter (HhaⅠdigestion: r = 0.94, p<0.001; RsaⅠdigestion: r = 0.74, p<0.05). Correlation analysis showed that soil available phosphorus content was positively correlated with all diversity indices of archaeal communities (p<0.05), and total nitrogen content showed a significant positive correlation (p<0.05) to the evenness index in HhaⅠDigestion results. The results showed that the vegetation restoration can enrich archaea at 0-20 cm soil. Because of the differences of plants, archaeal community presented very differently. Soil organic matter and available phosphorus also affected the distribution of archaea.
4. We selected 3 dominant secondary plants from each herbaceous and shrubs, and used BIOLOG technique to study microbial communities metabolic function. As a result, the average well color development(AWCD)of 6 rhizosphere samples were distinctly(p<0.05) different, and higher than control. Secondary plants had no significant(p>0.05) impact on richness of carbon source utilization, but changed the dominance and evenness (p<0.05). The principal component analysis (PCA) showed that the utilization of carbon sources were different among samples. Those differences were mainly attributed to the use of carbohydrates, amino acids and carboxylic acids. As appearance of plants, the types of carbon sources utilized by microorganisms were changed from phenol to carbohydrates and carboxylic acid. In addition, available potassium content in soil was negatively correlated with the use of polymers (-0.84) and amines(-0.83). These results suggested that the secondary plants can significantly enhance the ability of microorganisms to carbon source utilization, and change the types of carbon sources utilized by microorganisms. The utilization of some carbon sources were affected by available potassium content.
1.依据土壤分析方法标准(DB/6500 B11)测定对照和不同植物根际土壤的pH、有机质、全氮、速效氮、速效磷、速效钾。结果表明,样地土壤呈碱性(pH 8.04-8.51),由方差分析(ANOVA)可知不同植物根际土壤化学性质差异均显著(p<0.05)。9种植物根际土壤pH高于对照;土壤养分在不同植物间变化范围较大,总体趋势表现为植物根际土壤养分多高于裸地。其中蓍、东方旱麦草的根际土壤各项养分含量普遍较高,而新疆方枝柏较低。由此可知,植被的恢复在不同程度上改善了土壤的养分。
2.利用T-RLFP技术研究次生植物根际土壤细菌的群落结构,选用细菌16S rDNA通用引物8F/1492R进行扩增,HhaⅠ、RsaⅠ进行酶切。结果显示,不同植物改变了土壤微生物优势类群组成,增加了优势类群种类,尤其东方旱麦草、蓍对根际微生物作用最明显。变形菌门(Proteobacteria)细菌在植物根际土壤优势类群中占最大比例(49.55%)。典范对应分析(CCA)表明,细菌优势类群的分布与pH呈显著负相关(HhaⅠ酶切:r=-0.82,p<0.001; RsaⅠ酶切:r=-0.76,p<0.01)。由此推定植被的恢复增加了土壤细菌多样性,改变了土壤细菌群落结构,同时优势类群的分布还与土壤pH有关。
3.利用T-RLFP技术研究次生植物根际土壤古菌的群落结构,选用古菌16S rDNA通用引物21F/958R进行扩增,HhaⅠ、RsaⅠ进行酶切。结果显示,在皱纹柳、直穗柳、东方旱麦草、西北绢蒿、鼠掌老鹳草、欧亚绣线菊、蓬子菜、密刺蔷薇8种植物根际土壤扩增出古菌,各植物根际土壤古菌群落间相似性较低,泉古菌门(Crenarchaeota)和广古菌门(Euryarchaeota)为主要类群。典范对应分析(CCA)表明,优势类群的分布与土壤有机质呈显著正相关(HhaⅠ酶切:r=0.94,p<0.001; RsaⅠ酶切:r=0.74,p<0.05),由相关分析可知土壤速效磷含量与古菌群落各多样性指数呈显著正相关(p<0.05),土壤总氮含量与均匀度指数E呈显著正相关(p<0.05)。总体而言,部分植被可在土壤0～20cm深度富集古菌,且古菌群落因植物种类不同呈较大差异,同时土壤有机质、速效磷也对古菌类群分布产生影响。
4.选取次生植物中草本、灌木优势植物各3种,利用BIOLOG技术研究微生物群落代谢功能。结果显示:6种植物根际土壤的平均颜色变化率(AWCD)差异显著(p<0.05),且均高于裸地。次生植物不影响碳源利用的丰富度(p>0.05),但改变优势度和均匀度(p<0.05)。主成分分析(PCA)表明,不同植物根际土壤微生物群落利用碳源存在差异,其对糖类、氨基酸类、羧酸类碳源利用不同是造成差异的主要因素。随着植物的出现,土壤微生物利用碳源的类型由酚类向糖类、羧酸类转变。相关分析发现,土壤速效钾含量与聚合物类碳源(r=-0.84)、胺类(r=-0.83)的利用呈负相关。由此推断次生植物能显著增强土壤微生物碳源利用能力,改变碳源利用类型,且部分类型碳源的利用受速效钾含量的影响。
The aim of this study was to investigate the effects of secondary plants on Soil microbial community structure and metabolism in Xinjiang earthquake fault zone. The rhizosphere soils collected from 13 different secondary plants were studied by testing soil chemical properties and employing terminal restriction fragment length polymorphism (T-RFLP) and BIOLOG techniques. Unplanted soil in the same depth served as control. The investigated contents and results showed that:
1. Soil pH, organic matter, total nitrogen, available nitrogen, available phosphorus and available potassium were measured according to the standard soil analysis methods (DB/6500 B11). The results showed that the soils were alkaline (pH=8.04-8.51), Analysis of variance(ANOVA) indicated that chemical properties of 13 rhizosphere soils were significantly different (p<0.05). The pH in most (9) rhizosphere soils were higher than control; soil nutrients among the different plants varied widely, the overall trend showed rhizosphere soil nutrients were higher than control, and soil nutrients contents in the rhizosphere of Achillea millefolium and Eremopyrum orientale were relatively high, while the one in Sabina pseudosabina were low. The results presumed that the vegetation restoration improved the soil nutrients in varying degrees.
2. T-RLFP technique were used to study bacterial community structures in rhizosphere soil of secondary plants, we choosed universal primers 8F/1492R to amplify bacterial 16SrDNA, and HhaⅠ, RsaⅠenzymes to digest. As a result, the different plants changed the composition of dominant bacterial groups in soil and increased the sorts of dominant bacterial groups. Achillea millefolium and Eremopyrum orientale had the most significant effects. Proteobacteria accounted for the largest proportion (49.55%) in the soil. Canonical correspondence analysis (CCA) showed that the distribution of dominant bacterial groups was negatively correlated with pH significantly (HhaⅠdigestion: r =- 0.82, p <0.001; RsaⅠdigestion: r =- 0.76, p <0.05). These results suggested that vegetation restoration increased soil bacterial diversity, and had influence on the structures of soil bacterial communities. The distribution of dominant bacterial groups was also related to soil pH.
3. T-RLFP technique were used to study archaeal community structures in rhizosphere soil of secondary plants, we choosed universal primers 21F/958R to amplify archaeal 16SrDNA, and HhaⅠ, RsaⅠe nzymes to digest. The results showed that archaea were found only in thoes rhizosphere soils of Salix vistita, Salix rectijulis, Eremopyrum orientale, Seriphidium nitrosum, Geranium sibiricum, Spiraea media, Galium verum and Rosa spinosissima. Results in the detection of archaea showed that the similarities of archaeal communities among different rhizosphere soils were low. Crenarchaeota and Euryarchaeota were the main groups. Canonical correspondence analysis (CCA) showed that the distribution of dominant archaeal groups was positively correlated with soil organic matter (HhaⅠdigestion: r = 0.94, p<0.001; RsaⅠdigestion: r = 0.74, p<0.05). Correlation analysis showed that soil available phosphorus content was positively correlated with all diversity indices of archaeal communities (p<0.05), and total nitrogen content showed a significant positive correlation (p<0.05) to the evenness index in HhaⅠDigestion results. The results showed that the vegetation restoration can enrich archaea at 0-20 cm soil. Because of the differences of plants, archaeal community presented very differently. Soil organic matter and available phosphorus also affected the distribution of archaea.
4. We selected 3 dominant secondary plants from each herbaceous and shrubs, and used BIOLOG technique to study microbial communities metabolic function. As a result, the average well color development(AWCD)of 6 rhizosphere samples were distinctly(p<0.05) different, and higher than control. Secondary plants had no significant(p>0.05) impact on richness of carbon source utilization, but changed the dominance and evenness (p<0.05). The principal component analysis (PCA) showed that the utilization of carbon sources were different among samples. Those differences were mainly attributed to the use of carbohydrates, amino acids and carboxylic acids. As appearance of plants, the types of carbon sources utilized by microorganisms were changed from phenol to carbohydrates and carboxylic acid. In addition, available potassium content in soil was negatively correlated with the use of polymers (-0.84) and amines(-0.83). These results suggested that the secondary plants can significantly enhance the ability of microorganisms to carbon source utilization, and change the types of carbon sources utilized by microorganisms. The utilization of some carbon sources were affected by available potassium content.
引文
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