共生细菌对盐生小球藻亚砷酸盐代谢的影响

为探讨共生细菌对盐生小球藻（Chlorella salina）亚砷酸盐（As（III））富集和形态转化的影响,从C.salina培养液中分离培养出一株共生细菌,并采用抗生素处理C.salina获得无菌藻,设置0,75,150,300,750 μg/LAs（III）溶液处理带菌和无菌的C.salina,7d后测定C.salina对As（III）的吸收、吸附和富集量,以及培养液和藻细胞内As的形态,计算带菌和无菌C.salina对As（III）的氧化率和去除率;使用不同浓度的As（III）处理共生细菌7d,并以300μg/L的As（III）处理共生细菌不同时间,计算两种情况下共生细菌对培养液中As（III）的氧化率和去除率.结果表明：C.salina的共生细菌为根癌农杆菌（Agrobacterium tumefaciens WB-1）;与无菌C.salina相比,带菌C.salina生长更快、对As（III）的耐性更强.带菌C.salina对As（III）的富集量为103.10~448.12mg/kg、氧化率为78.93%~96.88%、去除率为18.92%~55.21%,显著高于无菌C.salina的富集量（11.68~91.39mg/kg）、氧化率（14.46%~26.39%）和去除率（12.82%~29.15%）,也高于A.tumefaciens WB-1对As（III）的氧化率（4.51%~30.61%）和去除率（1.86%~16.19%）.此外,带菌C.salina胞内还检测到少量的As（III）和甲基砷,而在无菌C.salina胞内没有甲基砷存在.共生细菌促进了盐生小球藻对As（III）的富集和形态转化.     

微藻无机氮利用过程中的稳定氮同位素分馏

选用两种δ15N差异显著的铵态氮和硝态氮,分别设置不同浓度的铵态氮和硝态氮来处理莱茵衣藻和蛋白核小球藻,通过分析微藻的稳定氮同位素组成变化,来研究微藻利用不同浓度、不同形态无机氮过程中的稳定氮同位素分馏特征。结果显示,在未添加无机氮的条件下,微藻利用有机氮时,生长缓慢,稳定氮同位素基本上不存在分馏;在添加低浓度无机氮(≤20 mmol/L)时,微藻的生长和稳定氮同位素分馏都随着无机氮浓度的增加而增加;而添加高浓度无机氮(20 mmol/L)时,微藻的生长趋于稳定,铵态氮条件下的微藻稳定氮同位素分馏继续加大,而硝态氮条件下的微藻稳定氮同位素分馏反而减小,可能与此时微藻硝酸还原酶的活力减小有关。     

不同碳源及光照对小球藻生长和产油脂的影响

考察了3种碳源(葡萄糖、NaHCO3和乙酸钠)在不同初始质量浓度和光照条件下对小球藻(Chlorella vulgaris)生长和产油脂的影响。采用OD680和生物量来评价小球藻的生长情况;以溶剂浸提法提取生物油脂,并以油脂质量分数和油脂产量来描述产油脂特性。结果表明,经过9 d的培养,3种碳源中葡萄糖是最佳有机碳源。由其培养的小球藻生长速率最快,由NaHCO3培养的小球藻的生长效果不如葡萄糖,而乙酸钠不利于小球藻的生长。随着光照的增强,小球藻光合效率提高,生物量逐渐提高,5 000 lx最利于小球藻的生长,而1 600 lx最利于小球藻油脂的积累。研究表明,光照5 000 lx下,初始质量浓度为15 g/L的葡萄糖作为碳源是小球藻适宜的生长和产油条件,获得了3.17 g/L的最大生物量和1.025 g/L的最大油脂产量。     

有机磷农药对蛋白核小球藻的毒性相互作用研究

水体中农药复合污染产生的毒性效应具有潜在风险。为系统考察有机磷农药(OPs)混合物对淡水生态系统中绿藻的联合毒性效应,以马拉硫磷(MIT)、敌敌畏(DDVP)、敌百虫(TRC)、乐果(DIT)和氧乐果(OMT)等5种OPs作为混合物组分,运用直接均分射线法设计9组二元混合物体系共45条混合物射线。利用96孔微板测定5种OPs及其二元混合物对蛋白核小球藻(C. pyrenoidosa)的生长抑制毒性,通过基于置信区间的组合指数法分析混合物的联合毒性及毒性相互作用。结果表明,以p EC50为毒性指标,5种OPs对C. pyrenoidosa的毒性大小顺序为:TRCMITDDVPOMTDIT,OPs对C. pyrenoidosa的毒性大小受其中心磷原子的电正性影响;因混合组分的不同,部分OPs混合物对C. pyrenoidosa的联合毒性依赖于组分浓度比; OPs混合物对C. pyrenoidosa的毒性相互作用以加和为主,部分发生拮抗作用,发生拮抗作用的混合体系具有低效应区域呈加和作用,高效应区域呈拮抗作用的规律;与MIT混合的体系均有发生拮抗作用,且依赖于MIT浓度,MIT浓度比例越高,拮抗作用越强,OPs混合物的毒性相互作用与组分浓度比相关; OPs混合物的毒性相互作用组分浓度比依赖性与其联合毒性的组分浓度比依赖性规律不相关。     

乙草胺对蛋白核小球藻的毒性效应研究

乙草胺是我国使用量最大的除草剂之一,在水体中广泛存在。已有研究证明,乙草胺对人类、老鼠和鱼类具有毒害效应,而关于其对浮游植物影响的研究较少。以蛋白核小球藻(Chlorella pyrenoidosa)为模型,使用1～10 000μg·L~(-1)的乙草胺对其进行7 d的暴露实验,考察小球藻生长性能、叶绿素含量、光合作用产氧量以及光合作用相关基因(pbsA、rbcL和rbcS)表达的变化。结果表明,较低浓度的乙草胺可刺激蛋白核小球藻生长,而较高浓度乙草胺则会抑制其生长;并且乙草胺会通过影响小球藻叶绿素的含量而影响光合作用产氧量;小球藻光合作用相关基因pbsA、rbcL和rbcS表达大多显著上升,这可能是对乙草胺胁迫响应的反馈调节。研究表明,乙草胺会对蛋白核小球藻的生长及光合作用产生影响。     

Effects of cultivation strategies on the cultivation of Chlorella sp. HQ in photoreactors

Heterotrophic cultivation caused high level of ROS and high lipids accumulation. HMTC is the best culture strategy for improving the microalgal biomass. Chlorella sp. HQ had great nutrient removal capacity under five culture strategies. The effects of cultivation strategies (including autotrophic cultivation (AC), heterotrophic cultivation (HC), fed-batch cultivation (FC), heterotrophic+ autotrophic two-stage cultivation (HATC), and heterotrophic+ mixotrophic two-stage cultivation (HMTC)) on the growth and lipid accumulation of Chlorella sp. HQ and its total nitrogen (TN) and total phosphorus (TP) removal in secondary effluent were investigated in column photoreactors. The results showed that the TN and TP removal rates ranged between 93.72%–95.82% and 92.73%–100%, respectively, under the five different strategies. The microalgal growth potential evaluated by the maximal growth rate (R_max) was in the order of HMTC>HC>FC>AC>HATC. The values of biomass, total lipid yield, triacylglycerols (TAGs) yield, and total lipid content of the microalga cultivated in the last 5 d increased significantly, but the TAGs productivities of the five strategies were lower than those in the first 7 d. Compared with all the other cultivation strategies, the TAGs productivity and yield after 12 d of cultivation under the heterotrophic condition reached the highest values accompanying the highest level of intracellular reactive oxygen species (ROS), in which the TAGs yield reached 40.81 mg/L at the end of the cultivation period. The peaks in TAGs yield and ROS level suggested that HC was beneficial for lipids accumulation via regulating the cellular redox status and exerting ROS stress on microalgal cells. In summary, HMTC was the best cultivation strategy for improving the microalgal biomass and HC was the best strategy for microalgal TAGs accumulation to produce biodiesel.     

光照条件对跑道池光生物反应器内蛋白核小球藻生长特性的影响

本文采用多孔曝气管的曝气方式,以蠕动泵为藻液循环的动力,研究了气体流量、光照强度、光照周期等因素对跑道池光生物反应器内蛋白核小球藻生长与代谢特性的影响,并利用傅里叶红外光谱半定量法对不同光照条件下藻细胞内蛋白质、油脂以及多糖含量变化进行了分析。研究结果表明,适当的气体流量、光照强度和光照周期有利于跑道池内蛋白核小球藻细胞的生长和代谢。在本实验条件下,在最佳气体流量2 000 m L/min条件下,光照强度为3 500μW/cm2,光照周期为16∶8时最有利于蛋白核小球藻的生长;光照强度为2 000μW/cm2和光照周期为16∶8的条件均有利于藻细胞内油脂的积累。     

改进的硫酸盐-聚乙烯醇法包埋藻菌脱氮除磷研究

采用改进的硫酸盐-PVA固定化法将藻与菌混合固定。在三种不同藻菌比的情况下，固定化系统对氮去除均可达100％，但是去除速度与固定的细菌量有关，细菌量越大，对氮的去除速度越快；对磷的去除随实验进行，最大去除率逐步下降，其下降速度与固定藻的量有关，藻量越大，下降速度越慢。由此说明脱氮的主要贡献者是细菌，而藻对除磷起了主要作用。为达到有效的脱氮除磷，应适当提高固定化藻的浓度，藻菌比应大于2：1。透射电镜照片显示，在聚乙烯醇载体上，藻类的生长没有受到限制。     

Biodegradation of 2,4,6-trichlorophenol in the presence of primary substrate by immobilized pure culture bacteria

In this study, pure strains that are capable of utilizing 2,4,6-trichlorophenol have been isolated from the mixed culture grown on substrates containing chlorophenolic compounds. Studies have been carried out on the capability of these isolated pure strains in suspended and immobilized forms to decompose 2,4,6-trichlorophenol. Additionally, the influence of primary substrates (e.g., phenol, 2-chlorophenol, 3-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol) on the decomposition of 2,4,6-trichlorophenol by the isolated pure strains grown in immobilized form is also investigated. The results are: Through bacterial isolation and identification, three pure strains have been obtained: Pseudomonas spp. strain 01, Pseudomonas spp. strain 02 and Agrobacterium spp. Whether in suspended or immobilized forms, all strains have poor removal efficiencies of 2,4,6-trichlorophenol. However, addition of 200 mg/l phenol will enable the immobilized Pseudomonas spp. strain 01, and Pseudomonas spp. strain 02 to achieve 65% and 48% removal of 2,4,6-trichlorophenol, respectively. Addition of phenol will assist the immobilized Pseudomonas spp. strain 02 in achieving removal of 2,4,6-trichlorophenol but the removal efficiency is not good if the phenol concentration is too low. The optimum phenol concentration should be between 200 and 400 mg/l.     

Early stage toxicity of excess copper to photosystem II of Chlorella pyrenoidosa–OJIP chlorophyll a fluorescence analysis

Acute toxicity of excess Cu on the photosynthetic performance of Chlorella pyrenoidosa was examined by using chlorophyll a fluorescence transients and JIP-test after exposure to elevated Cu concentrations for a short time period. High Cu concentration resulted in a significant suppression in photosynthesis and respiration. The absorption flux (ABS/RC) per PSII reaction center increased with increasing Cu concentration, but the electron transport flux (ET₀/RC) decreased. Excess Cu had an insignificant effect on the trapping flux (TR₀/RC). The decline in the efficiency, with which a trapped exciton can move an electron into the electron transport chain further than Q_A−(Ψ₀), the maximal quantum yield of primary photochemistry (?_P0), and the quantum yield of electron transport (?_E0) were also observed. The amount of active PSII reaction centers per excited cross section (RC/CS) was also in consistency with the change of photosynthesis when cells were exposed to excess Cu concentration. JIP-test parameters had a good linear relationship with photosynthetic O₂ evolution. These results suggested that the decrease of photosynthesis in exposure to excess Cu may be a result of the inactivation of PSII reaction centers and the inhibition of electron transport in the acceptor side.