摘要
2006年7月至2007年12月,采用“非浸没培养皿法”(non-flooded Petri dish method)和活体观察法对小陇山自然保护区麻沿林区的土壤纤毛虫群落进行了定性、定量研究。利用花盆实验,通过添加二萜类化合物对该地区土壤纤毛虫群落的毒性进行了研究。并从实验土壤中分离出膨胀肾形虫、僧帽肾形虫和粘游仆虫,通过急性和慢性毒性试验,研究了二萜类化合物对土壤纤毛虫种群的毒性。主要结果如下:
1、麻沿林区土壤纤毛虫群落结构的原位研究共鉴定到土壤纤毛虫91种,其中包括7个未定名种和14个中国土壤纤毛虫新纪录种,隶属于3纲、11目、29科、42属。其中,下毛目(Hypotrichida)为优势类群,肾形目(Colpodida)和前口目(Prostomatida)为次优势类群,侧口目(Pleurostomatida)、缘毛目(Peritrichida)和寡毛目(Oligotrichida)为偶见类群,膨胀肾形虫(Colpoda inflata)、长篮环虫(Cyrtolophosis elongata)、大口薄咽虫(Leptopharynx eurystoma)和长圆膜袋虫(Cyclidium oblongum)为优势种。原位研究结果表明,小陇山麻沿林区土壤纤毛虫群落物种丰富,特有和稀有物种繁多,群落结构复杂,林区土壤纤毛虫群落与我国已有研究报道的其他地区的均不相似。同时麻沿林区土壤纤毛虫的C/P系数为0.710,反映了麻沿林区比较适合土壤纤毛虫生长繁殖。
2、模拟生态实验中,定性研究共鉴定到纤毛虫88种,其中对照组75种,实验组随添加的二萜类化合物浓度升高,纤毛虫物种数减少,群落结构简单化;回归分析表明纤毛虫物种数和二萜类化合物浓度成极显著负相关关系。当施药浓度超过32.5 mg·kg~(-1)时土壤纤毛虫优势种和C/P系数发生变化,此时优势种为膨胀肾形虫(Colpoda inflata)、长篮环虫(Cyrtolophosis elongata)、苔藓膜袋虫(Cyclidium muscicola)、水藓薄咽虫(Leptopharynx sphagnetorum)、大弹跳虫(Halteria grandinella)、小尖毛虫(Oxytricha minor)、有肋薄咽虫(Leptopharynx costatus)和前口虫属一种(Frontonia sp.);C/P系数大于1,表明二萜类化合物使土壤纤毛虫群落结构遭到了相当大的破坏。
定量研究发现,在相同曝露时间,不同施药浓度下土壤纤毛虫丰度与对照组相比均有极明显的降低,而同一施药浓度,随着曝露时间逐渐延长,纤毛虫的数量逐渐回升。二萜类化合物残留浓度测定结果表明,即使土壤中二萜类化合物残留浓度很低,也对纤毛虫群落有显著的抑制作用。非参数多个独立样本检验分析结果表明,各浓度组间纤毛虫的丰度存在极显著差异。
3、种群毒性试验中,12 h和24 h的急性毒性试验结果显示,浓度对数与纤毛虫死亡概率单位之间存在极显著的正相关关系。即随二萜类化合物浓度增大,对纤毛虫种群的毒性增大。膨胀肾形虫、僧帽肾形虫和粘游仆虫12 h-LC50分别为161.4 mg·L~(-1),94.8 mg·L~(-1),83.7 mg·L~(-1); 24 h-LC50依次为114.9 mg·L~(-1),92.3 mg·L~(-1),65.8 mg·L~(-1)。单因子方差分析(ANOVA)显示:与对照组相比,不同浓度的二萜类化合物对三种纤毛虫的致死率存在极显著差异(P﹤0.01);各浓度组之间也存在极显著差异(P﹤0.01)。以上结果表明二萜类化合物对纤毛虫种群的急性毒性作用显著。
慢性毒性试验结果显示,不同浓度二萜类化合物培养液中三种纤毛虫的种群密度和种群增长率明显低于对照组,而世代时间明显高于对照组。三种纤毛虫在不同浓度二萜类化合物培养液中的种群密度和种群增长率与对照之间存在着极显著的差异(P﹤0.01)。综合以上结果:在试验浓度范围内,二萜类化合物对膨胀肾形虫、僧帽肾形虫和粘游仆虫的种群生长有明显的抑制效应。
对纤毛虫个体形态的影响方面,二萜类化合物使三种纤毛虫个体形态发生显著变化。
The community characteristic of soil ciliates in the Mayan Forest Region of the National Nature Reserve of Xiaolong Mountains was studied quantitativly and qualitativly by“non-flooded petri dish method”and observation in vivo.Toxicity of Diterpenoids on community of soil ciliates in this area was studied by pot test,and that on population of Colpoda inflata,Colpoda cucullus and Euplotes muscicola was studied by acute and chronic toxicity test.The study was carried out from July of 2006 to December of 2007.The main results are as follows:
1.In the study in situ,91 species,including 7 unnamed species and 14 new records of soil ciliates in China,belonging to 3 classes,11 orders,29 families and 42 genera,were identified.Hypotrichida was the dominant group,Colpodida and Prostomatida were the subdominant groups,Pleurostomatida,Peritrichida and Oligotrichida were the incidental groups,and Colpoda inflata,Cyrtolophosis elongata, Leptopharynx eurystoma and Cyclidium oblongum were the dominant species.The results showed that there were rich species and endemic and rare species in the community of soil ciliates,the community structure of soil ciliates in the Mayan Forest Region of Xiaolong Mountains was complex and was quite dissimilar to that in other areas with reported results of study in China.The C/P coefficient of soil ciliates was 0.710,which showed that the natural environments of Mayan Forest Region of Xiaolong Mountains was suitable for growth and reproduction of soil ciliates.
2.In the simulation test of soil ecosystem,totally 88 species were identified in qualitative study,75 species in which were in control groups.The species numbers decreased with increasing of centration of Diterpenoids in test groups,the community structure of soil ciliates was simplified. The results of regression analysis showed that there were significant negative correlations between the species numbers and the concentrations of Diterpenoids. When the concentrations were more than 32.5 g·kg~(-1),the dominant species and C/P coefficient changed, and the dominant species were Colpoda inflata,Cyrtolophosis elongata, Cyclidium muscicola,Leptopharynx sphagnetorum, Halteria grandinella, Oxytricha minor,Leptopharynx costatus and Frontonia sp.,the C/P coefficient were more than one,which showed that the community structure of soil ciliates were greatly disturbed.
Quantitative study showed that the abundance of ciliates decreased markedly compared to control groups with different concentrations at the same exposed time,but that increased gradually with extending exposed time at the same concentrations. The results of determined residual concentrations showed that the community structure of soil ciliates was significantly restrained even in lower concentrations. The result of K-independent sample of nonparametric tests showed that there were significant difference on the abundance of ciliates among different concentrations groups.
3.In toxicity test on population, 12 h and 24 h acute toxicity test showed that there were significant positive correlations between the concentration logarithm and the mortality probability units, i.e. the toxicity of Diterpenoids on populations of soil ciliates increased with the concentrations of Diterpenoids increasing. 12 h-LC50 of Colpoda inflate,Colpoda cucullus and Euplotes muscicola were 161.4 mg·L~(-1),94.8 mg·L~(-1),83.7 mg·L~(-1) respectively,and 24 h-LC50 was in sequence of 114.9 mg·L~(-1),92.3 mg·L~(-1),65.8 mg·L~(-1). ANOVA showed that,compared with the control groups,a significant difference (P<0.01) were found between the mortality rate of ciliates and the concentrations of Diterpenoids and it occured also among the groups in terms of different concentration. Considering all the above,the acute toxicity of Diterpenoids on populations were significant.
The chronic toxicity test showed that population densities and population growth rates of test groups were obviously lower than ones in control groups while the generation times were obviously longer than ones in control groups.The analysis indicated that there were a significant difference (P<0.01) between the population densities and the growth rates of test groups and those of control groups.According to above results,within the test concentration ranges,the population growth of Colpoda inflata,Colpoda cucullus and Euplotes muscicola were restrained significantly by Diterpenoids.
The results of effects on body shape indicated that the body shape of Colpoda inflata and Colpoda cucullus changed significantly.
引文
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