Knowledge Resource Center for Ecological Environment in Arid Area
| 中国西北三省区微藻多样性调查及产油微藻的分离筛选 | |
| 其他题名 | Microalgal diversity, separation and screening of oleaginous microalgae in 3 Northwestern regions of China |
| 徐亮亮 | |
| 出版年 | 2014 |
| 学位类型 | 硕士 |
| 导师 | 胡春香 |
| 学位授予单位 | 中国科学院大学 |
| 中文摘要 | 微藻被认为是最具有前途替代化石能源的生物质原料。但目前可以商业利用的微藻种质资源还非常贫乏。因此，获得优良产油藻株不仅是建立新型的微藻能源产业的首要前提，更是确立国家能源战略资源的制高点。本文在对我国宁夏回族自治区、甘肃省、青海省三个地区的土壤(荒漠、草甸、农田、滩涂)和淡水(湖泊、水库、河流、池塘)生境中的藻类资源进行普查的基础上，首先对各种生境中的藻类资源尽可能地进行了分离、纯化、培养及油脂含量的分析，并尝试分析了藻类含油量与自然生境间的可能关系；然后，对油含量高于30%的藻株进一步进行了18S rDNA鉴定，并对5株油脂产率高的优良藻株进行了营养等生长条件的摸索，主要结果如下： \n地处我国季风气候与大陆气候过渡带的宁夏回族自治区，发现了最丰富的藻类资源，其中陆生生境中有334种，主要为席藻、微鞘藻、小球藻和绿球藻种类；淡水生境中有527种，主要为衣藻、小球藻、舟形藻种类。地处大陆干旱气候带的甘肃地区，藻类资源较少，其中陆生中75种，主要为色球藻、席藻、微鞘藻、绿球藻、小球藻；淡水中336种，主要为衣藻、小球藻、脆杆藻、针杆藻、舟形藻、桥弯藻、角甲藻等。地处内陆高寒气候带的青海地区，藻类资源也较少，其中陆生藻有90种，主要为席藻、微鞘藻、绿球藻、小球藻；水生藻有360种，主要为小球藻、脆杆藻、针杆藻、舟形藻。 \n对调查地区微藻种类多样性的分布格局进一步分析发现，在不同气候区的同种土壤藻类多样性普遍差异较小，其中宁夏地区干旱和半湿润滩涂种类多样性较高；淡水生境中，不同气候区的湖泊、河流、池塘藻类多样性差异较大，其中宁夏地区干旱区的河流、池塘，甘肃地区干旱区的湖泊、池塘及青海地区的半干旱区河流微藻种类多样性较高。 \n经过对362个水样、243个土样的稀释分离和平板分离，在所调查地区中共分离出微藻1934株，其中陆生生境中的培养分离效率平均在46.5%以上，水体中在26.7%以上。 \n纯化藻株含油量的检测发现，低含油量(<20%)藻株比例普遍最大(50%)，有973 株；中含油量(20-40%)的有660株(34%)，主要是小球藻属；高含油量(>40%)的有300株(16%)，主要是小球藻属、月牙藻属。 \n各地带间比较发现，甘肃水体中含油量藻株所占比例较高(31.33%)；青海水体中，高含油量藻株所占比例较高(30.17%)。 \n在采样较为详细的宁夏回族自治区，高海拔地区藻株的生物量和油脂生产率普遍低于低海拔地区；水生微藻的生物量低于陆生微藻，但油含量和油脂生产率高于陆生微藻。陆生生境中，农田微藻的生物量和油含量平均值较高；水生生境中，河流微藻的生物量和油含量平均值较高。含油量高于30%和40%的藻株在草甸和水库中比例较高。 \n在含油量较高的10属 556 株微藻的比较研究中发现，盘星藻属和绿球藻属的生物量普遍较高，月牙藻属和空星藻属的油含量普遍较高，纤维藻属和盘星藻属油脂产率普遍较高。 \n比较BG-11、CSI、f/2三种培养基发现，BG-11中微藻的生物量普遍较高，CSI中的油含量较高。 \n对5株(Ankistrodesmus fusiformis NG-264，Chlorella sorokiniana NG-323，Chlorella sorokiniana NT-337，Monoraphidium dybowskii NG-417，Desmodesmus intermedius NT-536)高产油微藻的生长特性进行了比较分析发现，它们在各自适宜的营养条件下，油脂产率分别达到81.3mg/L.d，157.8mg/L.d，109.5mg/L.d，127.6mg/L.d，201.3mg/L.d，充分显示它们都是非常有应用潜力的优良藻株。 |
| 英文摘要 | As bio-feedstock, microalgae are recognized as the most promising alternatives to fossil energy. But by now, very few oleaginous microalgae have been really applied at commercial scale. Therefore, to obtain the higher oleaginous microalgae is the first premise to establish microalgal energy industry. This study investigated microalgal resources in the different soil (desert, meadow, farmland and shallow) and aquatic (lake, reservoir, river and pond) habitats in Ningxia Hui Autonomous Region, Gansu and Qinghai provinces. Based on the above investigation, the microalgae were firstly isolated, purified and cultured as many as possible. Then the lipid contents of all cultured microalgae were analyzed, and the possible relationships between the lipid content and natural habitats were discussed. Finaly, the all strains that lipid content are higher than 30% were further identified via 18S rDNA sequencing, and 5 strains of high lipid productity were optimized for nutrition and other growth conditions. The main results are as follows: \nNingxia Hui Autonomous Region, locating in the monsoon and continental climate transition zone, has the most abundant microalgal resources among our investigated three regions. In this region, as much as 334 species of microalgae were identified in the terrestrial environment, mainly belonging to Phormidium, Microcoleus, Chlorella and Chlorococcum; 527 species of microalgae were found in freshwater habitats, mainly belonging to Chlamydomonas, Chlorella and Navicula. Gansu province, locating in the continental arid climatic zone, has the least algal resources among the three regions. In this area, only 75 species of microalgae were identified in the terrestrial habitats, mainly belonging to Chroococcus, Phormidium, Microcoleus, Chlorococcum and Chlorella; 336 species in the freshwater environments, mainly belonging to Chlamydomonas, Chlorella, Fragilaria. Synedra, Navicula, Cymbella and Ceratium. Qinghai province locates in Alpine climate zones in the inland, also with less microalgal species. There are 334 species mainly Phormidium, Microcoleus, Chlorococcum and Chlorella in the terrestrial habitats, and 527 species mainly Chlorella, Fragilaria. Synedra and Navicula in aquatic habitats. \nThe distribution pattern showed that microalgal diversity in the same types of terrestrial habitats of different climate zones had only little variation. And the arid and semi-humid areas locating in Ningxia had the relatively higher diversity. On the contrary, the variations in the same types of aquatic habitats of different climatic zones were much greater. Among the freshwater habitats, the rivers and ponds in the arid area of Ningxia, the lakes and ponds in the arid region of Gansu, and the rivers in the semi-arid area of Qinghai all had relatively higher species diversity. \nAfter dilution and agar plate isolation for 362 aquatic samples and 243 soil samples, total 1934 strains of microalgae were isolated and purified. The averaged isolation efficiency in terrestrial habitats was higher than 46.5%, while about 26.7% in water habitats. \nThe lipid content of all purified strains showed, 50% strains had low lipid content (<20%), 34% strains (mainly Chlorella) had 20%-40% lipid content; 16% strains (mainly Chlorella and Selenastrum) had much higher lipid content (>40%). \nAmong the climatic zones, the proportion of oil-rich strains in Gansu aquatic habitats was relatively higher (31.33%), also in Qinghai (30.17%). \nIn detailed survey of Ningxia area, biomass and lipid productivity of all strains from higher altitudes were generally less than those from lower altitudes; biomass of aquatic strains was generally lower than that of terrestrial ones, while lipid content was higher in the aquatic strains. In terrestrial habitats, the mean biomass and lipid content were higher in farmlands; while in aquatic habitats, were that in rivers. In addition, the relative larger proportion of strains possessing higher lipid content (>30% and 40%) occurred in meadow and reservoir. \nAfter comparison of 556 oil-rich strains from 10 genera, it was found biomass of Pediastrum and Chlorococcum were generally higher, while lipid content of Selenastrum and Coelastrum was higher, lipid productivity of Ankistrodesmus and Pediastrum was higher. \nWhen the isolated strains were respectively cultured in BG-11, CSI, F/2, it was found their biomasses in BG-11 were generally highest, while lipid contents were highest in CSI. \nFurthermore, 5 strains candidates , namely Ankistrodesmus fusiformis NG-264，Chlorella sorokiniana NG-323，Chlorella sorokiniana NT-337，Monoraphidium dybowskii NG-417 and Desmodesmus intermedius NT-536, their nutrient conditions were optimized, and the lipid productivities were respectively carried out 81.3mg/L.d, 157.8mg/L.d, 109.5mg/L.d, 127.6mg/L.d and 201.3mg/L.d, which showed they were all very potential strains. |
| 中文关键词 | 西北地区 ; 多样性 ; 产油微藻 ; 分离 ; 筛选 |
| 英文关键词 | Northwestern China Diversity Oleaginous microalgae Isolation Screen |
| 语种 | 中文 |
| 国家 | 中国 |
| 来源学科分类 | 生物工程 |
| 来源机构 | 中国科学院水生生物研究所 |
| 资源类型 | 学位论文 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/287418 |
| 推荐引用方式 GB/T 7714 | 徐亮亮. 中国西北三省区微藻多样性调查及产油微藻的分离筛选[D]. 中国科学院大学,2014. |
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