云南元谋干热河谷深色有隔内生真菌（DSE）的研究
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摘要
金沙江位于长江中上游,金沙江干热河谷是在地势、地貌、自然和人类活动等综合因素作用条件下形成的,主要分布在云南和四川境内。这一特殊生态系统的突出特点就是全年气温高,蒸发量大,年蒸发量大于年降水量。云南干热河谷地区由于人口密度大、开发时间早、强度大,因此也是水土流失最严重的地区,对长江流域生态系统造成严重影响。国家已把长江中上游地区的生态治理列为国家重点环境治理工程,其中干热河谷地区的植被恢复是这一工程中的重中之重。深色有隔内生菌(Dark Septate Endophytes,DSE)是土壤生态系统中的一类广泛分布并具有重要的生态功能的微生物,在陆地生态系统和水生生态系统均有发现,并在陆地生态系统的植被恢复和土壤修复过程中发挥重要的作用。
本研究以金沙江干热河谷(云南元谋段,简称元谋干热河谷)3种生境——原生地、耕地和休耕地为研究对象,对各生境中自然生长的植物根中DSE的定殖情况进行了全面调查,从中分离并得到深色有隔内生真菌(DSE)菌株;通过形态学观察和对核糖体小亚基ITS1-5.8S rDNA-ITS2序列分析,对这些菌株的系统发育地位进行初步鉴定;从分离到的DSE菌株中,选择形态特征典型的菌株以玉米为宿主植物,进行回接试验;在不同干旱胁迫水平处理下,以玉米为宿主植物,接种典型菌株,考查接种真菌对玉米的生长及其抗旱性影响。研究取得的主要结果和结论如下:
用碱解离、酸性品红染色法对元谋干热河谷耕地、休耕地和原生地三种生境中30科64种植物(土著植物、农作物和引种植物)根样的深色有隔内生真菌(DSE)及丛枝菌根(Arbuscular Mycorrhizas, AM)状况进行了调查,发现所有被调查的植物均被深色有隔内生真菌(DSE)感染,AMF的感染率分别为原生地86%(24种),耕地78%(21种)和休耕地96%(25种)。
从三种生境中自然生长的植物根内分离并得到的不同DSE菌株用PDA培养基培养,根据其形态学特征和ITS 1-5.8S rDNA-ITS 2序列分析,将其中的77-2、105-2、108-1等8株菌鉴定为枝孢菌属,其它菌株由于鉴定手段的欠缺,仅根据其DNA序列比对还不能将其归为相关的属、种。
将枝孢菌属真菌菌株与宿主植物玉米进行回接实验说明,在轻度(10%)、中度(20%)、重度(30%)干旱胁迫条件下,枝孢菌属的部分种类对宿主植物玉米的抗旱性具有有益作用。
Jinsha River is located in the middle and upper reaches of Yangtze River. Hot-dry valley of Jinsha River, which is distributed mainly in Yunnan and Sichuan province, is formed under the comprehensive actions of geological, landscape, natural and human activities. This special ecosystem is charactered by its high annual temperature and heavy evaporation and its annual evaporation is higher than its annual precipitation. High population density and intensive exploitation in Hot-dry valley of Yunnan have been resulting in most serious soil and water loss, which have already threatened the ecosystems of drainage area of Yangtze River. So, the ecological improvement projects of the middle and upper reaches of Yangtze River have been listed as the key environmental improvement projects by Chinese government, in which the vegetation restoration of hot-dry valley is the most important core project. Dark septate endophytes are a assembly of fungi which widely distribute in soil and may have important ecological functions in natural ecosystems. Their existence has been found in terrestrial ecosystems and aquatic ecosystems. Moreover, they may play an important role in the restoration and biomediation of terrestrial ecosystems.
Three habitats, representing cultivated lands (CL), old fields (OF) and never-cultivated fields (NCF), which located in hot-dry valley of Yuanmou, southwest China, were chosen as study sites. Based on the investigation of colonization in plants of three different ecosystems, 39 fungal isolates were obtained from surface sterilized root segments. Taxonomic affinity of the isolates was studied according to morphological observation and phylogenetic analysis of ITS1-5.8S-ITS2 region of rDNA. Inoculation experiments with maize were done with the isolates which are typical strains. Under different drought stress conditions, there are the effects of inoculation of strains on the tolerance of maize. The main results and conclusions of the thesis are:
The DSE and AMF colonization of 64 plant species representing native plants, crops and introduced plants in CL, OF and NCF of hot-dry valley of Yuanmou was investigated by alkaline lysising and acid fuchsin staining. It was found that all plants surveyed in the three habitats were infected by DSE. The frequency of AMF in NCF, AL, OF are 86%, 78%, 96%. The results indicate that the plants are highly dependent on DSE and AMF in this hot and arid ecosystem.
According to the morphological characters of the isolates on PDA and phylogenetic analysis of ITS regions, 8 isolates, including 77-2, 105-2,108-1 were identified to Cladosporium spp..
After maize were inoculated with Cladosporium spp. and, DSE structures were observed in their roots, and no pathogenic symptom appeared in the maize. It has a positive effects to maize on different drought stress conditions. These results suggest that some species of Cladosporium are candidates of DSE.
本研究以金沙江干热河谷(云南元谋段,简称元谋干热河谷)3种生境——原生地、耕地和休耕地为研究对象,对各生境中自然生长的植物根中DSE的定殖情况进行了全面调查,从中分离并得到深色有隔内生真菌(DSE)菌株;通过形态学观察和对核糖体小亚基ITS1-5.8S rDNA-ITS2序列分析,对这些菌株的系统发育地位进行初步鉴定;从分离到的DSE菌株中,选择形态特征典型的菌株以玉米为宿主植物,进行回接试验;在不同干旱胁迫水平处理下,以玉米为宿主植物,接种典型菌株,考查接种真菌对玉米的生长及其抗旱性影响。研究取得的主要结果和结论如下:
用碱解离、酸性品红染色法对元谋干热河谷耕地、休耕地和原生地三种生境中30科64种植物(土著植物、农作物和引种植物)根样的深色有隔内生真菌(DSE)及丛枝菌根(Arbuscular Mycorrhizas, AM)状况进行了调查,发现所有被调查的植物均被深色有隔内生真菌(DSE)感染,AMF的感染率分别为原生地86%(24种),耕地78%(21种)和休耕地96%(25种)。
从三种生境中自然生长的植物根内分离并得到的不同DSE菌株用PDA培养基培养,根据其形态学特征和ITS 1-5.8S rDNA-ITS 2序列分析,将其中的77-2、105-2、108-1等8株菌鉴定为枝孢菌属,其它菌株由于鉴定手段的欠缺,仅根据其DNA序列比对还不能将其归为相关的属、种。
将枝孢菌属真菌菌株与宿主植物玉米进行回接实验说明,在轻度(10%)、中度(20%)、重度(30%)干旱胁迫条件下,枝孢菌属的部分种类对宿主植物玉米的抗旱性具有有益作用。
Jinsha River is located in the middle and upper reaches of Yangtze River. Hot-dry valley of Jinsha River, which is distributed mainly in Yunnan and Sichuan province, is formed under the comprehensive actions of geological, landscape, natural and human activities. This special ecosystem is charactered by its high annual temperature and heavy evaporation and its annual evaporation is higher than its annual precipitation. High population density and intensive exploitation in Hot-dry valley of Yunnan have been resulting in most serious soil and water loss, which have already threatened the ecosystems of drainage area of Yangtze River. So, the ecological improvement projects of the middle and upper reaches of Yangtze River have been listed as the key environmental improvement projects by Chinese government, in which the vegetation restoration of hot-dry valley is the most important core project. Dark septate endophytes are a assembly of fungi which widely distribute in soil and may have important ecological functions in natural ecosystems. Their existence has been found in terrestrial ecosystems and aquatic ecosystems. Moreover, they may play an important role in the restoration and biomediation of terrestrial ecosystems.
Three habitats, representing cultivated lands (CL), old fields (OF) and never-cultivated fields (NCF), which located in hot-dry valley of Yuanmou, southwest China, were chosen as study sites. Based on the investigation of colonization in plants of three different ecosystems, 39 fungal isolates were obtained from surface sterilized root segments. Taxonomic affinity of the isolates was studied according to morphological observation and phylogenetic analysis of ITS1-5.8S-ITS2 region of rDNA. Inoculation experiments with maize were done with the isolates which are typical strains. Under different drought stress conditions, there are the effects of inoculation of strains on the tolerance of maize. The main results and conclusions of the thesis are:
The DSE and AMF colonization of 64 plant species representing native plants, crops and introduced plants in CL, OF and NCF of hot-dry valley of Yuanmou was investigated by alkaline lysising and acid fuchsin staining. It was found that all plants surveyed in the three habitats were infected by DSE. The frequency of AMF in NCF, AL, OF are 86%, 78%, 96%. The results indicate that the plants are highly dependent on DSE and AMF in this hot and arid ecosystem.
According to the morphological characters of the isolates on PDA and phylogenetic analysis of ITS regions, 8 isolates, including 77-2, 105-2,108-1 were identified to Cladosporium spp..
After maize were inoculated with Cladosporium spp. and, DSE structures were observed in their roots, and no pathogenic symptom appeared in the maize. It has a positive effects to maize on different drought stress conditions. These results suggest that some species of Cladosporium are candidates of DSE.
引文
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