水稻RBX1基因cDNA的分离及其过量表达载体对水稻的转化

RBX1蛋白是基于Cullin蛋白的E3复合体中的一个亚基,对于基于Cullin蛋白的E3复合体结合E2及Cullin蛋白的活化具有重要作用.利用RT-PCR分离到水稻RBX1cDNA,序列比对分析显示,水稻RBX1与拟南芥、人、果蝇及酵母中该同源蛋白的氨基酸序列一致性分别为80%、74.4%、72%和51.2%.酵母双杂交实验显示,水稻RBX1与水稻Cullin4蛋白有着相互作用.将水稻RBX1cDNA插入植物表达质粒pHB,利用农杆菌介导法将构建好的植物表达载体导入水稻,获得植株38株.以基因组DNA为模板对潮霉素抗性基因片段进行扩增,初步确定其中32株为转基因植株.图5表1参17     

利用硫杆菌淋滤电镀污泥中的重金属(英文)

从广东云浮硫铁矿矿山酸性废水中分离得一株氧化亚铁硫杆菌(Thiobacillus ferrooxdans),从新兴县含硫化氢温泉中分离得一株氧化硫硫杆菌 (Thiobacillus thiooxidans), 并将其应用于电镀污泥的生物淋滤的研究,该污泥含铬、镍及铜分别为80.74 g/kg自然风干污泥、37.34g/kg自然风干污泥和13.42 g/kg自然风干污泥.利用未经驯化的氧化亚铁硫杆菌和氧化硫硫杆菌,以底物类型、菌液比、初始pH值和污泥浓度为四个因素,设置3个水平进行L9(34)正交实验,得出电镀污泥的最适淋滤条件:污泥浓度0.1%、硫酸亚铁和硫代硫酸钠作底物、菌液比T.f:T.t=:1:1、初始pH值2.0.为了将分离得的两种硫杆菌应用于更高浓度的电镀污泥,对所得菌进行驯化,以提高该菌种的重金属耐性,并应用于高浓度污泥的生物淋滤.经过驯化后的细菌,成功地适应了电镀污泥环境.结合上述最适条件,加入了合适的营养底物及还原剂之后,电镀污泥中的重金属,如铜、镍等,在30℃酸性好氧的环境中,能够在7天内被有效地淋滤出来,混合菌对污泥浓度为0.5%的污泥中铜的滤出率达90%以上,镍的滤出率40%以上;对1%的污泥的淋滤效果较差;对铬的淋滤效果不明显.本文还讨论了两株硫杆菌对高毒性电镀污泥的适应性.即在一定的污泥浓度范围内,氧化亚铁硫杆菌和氧化硫硫杆菌能够氧化底物,并产生硫酸将污泥中的重金属溶解,两株硫杆菌能够加快镍和铜的滤出效率.     

利用城市污泥生产苏云金杆菌生物农药

用城市污水污泥和经酸水解后的城市污泥作为培养基发酵苏云金杆菌Bacillusthringiensis(B.t),与常规的黄豆粉培养基作比较.检测B.t发酵液的pH值变化,总细胞数及活孢子数以及在发酵48h后,用2龄家蚕作试虫比较3种发酵液的生物毒性.结果表明,用污泥培养的B.t总细胞数及活孢子数略低于黄豆粉培养基生产的B.t,但两者的细菌总数都在同一数量级,差别不显著.用污泥培养B.t,芽孢大量形成109个/mL的时间,比用黄豆粉培养基生产的B.t约提前10h,发酵周期明显缩短.生物毒性实验表明用污泥培养的B.t孢晶混合液比用黄豆粉生产的B.t孢晶混合液毒性高,在相同的稀释倍数下,家蚕的死亡率提高了10%,酸水解污泥毒性略差.     

两种硫杆菌对河涌底泥重金属的生物沥滤

自矿山酸性废水中分离纯化得到氧化亚铁硫杆菌(Thiobacillus ferrooxidans , T.f)与氧化硫硫杆菌(Thiobacillus thiooxidans, T.t),利用这2 种硫杆菌对广州郭村河涌底泥中Cd、Cr、Pb、Ni、Cu 和Zn 6 种重金属进行序批式生物沥滤试验.结果表明,T.f 菌对重金属的浸出能力高于T.t 菌.对Cd、Cr、Ni、Cu 和Zn 的浸出率T.f 菌最高可达60%、45%、70%、90%和75%,T.t 菌则分别为45%、34%、50%、65%和55%.二者对Pb 的去除率相当,均在25%左右.向T.f 菌分别提供S 粉与Fe²⁺,2 种底物对Cd、Ni、Cu 和Zn 的浸出效果无明显影响,而对Cr和Pb 的浸出,投加S 粉的浸出效果更好.同时向T.f 菌投加S 粉与Fe²⁺ 2 种底物,其浸出效果反而下降.可能是T.f 菌代谢生成的SO₄^2-与Fe³⁺易生成黄铁矾等沉淀物质,与溶出的重金属发生共沉淀,从而降低其浸出效果.     

Biodegradation of 2-naphthol and its metabolites by coupling Aspergillus niger with Bacillus subtilis

To explore biodegradation of 2-naphthol and its metabolites accumulated in wastewater treatment, a series of bio-degradation experiments were conducted. Two main metabolites of 2-naphthol, 1,2-naphthalene-diol and 1,2-naphthoquinone, were identified by high-performance liquid chromatography with standards. Combining fungus Aspergillus niger with bacterium Bacillus subtilis in the treatment enhanced 2-naphthol degradation e ciency, lowered the accumulation of the two toxic metabolites. There were two main phases during the degradation process by the kinetic analysis: 2-naphthol was first partly degraded by the fungus, producing labile and easily accumulated metabolites, and then the metabolites were mainly degraded by the bacterium, attested by the degradation processes of 1,2-naphthalene-diol and 1,2-naphthoquinone as sole source of carbon and energy. Sodium succinate, as a co-metabolic substrate, was the most suitable compound for the continuous degradation. The optimum concentration of 2-naphthol was 50 mg/L. The overall 2-naphthol degradation rate was 92%, and the CODCr removal rate was 80% on day 10. These results indicated that high degradation rate of 2-naphthol should not be considered as the sole desirable criterion for the bioremediation of 2-naphtholcontaminated soils/wastewater.     

Anaerobic benzene biodegradation by a pure bacterial culture of Bacillus cereus under nitrate reducing conditions

A pure culture using benzene as sole carbon and energy sources was isolated by screening procedure from gasoline contaminated soil. The analysis of the 16S rDNA gene sequence, morphological and physiological characteristics showed that the isolated strain was a member of genus Bacillus cereus. The biodegradation performance of benzene by B. cereus was evaluated, and the results showed that benzene could be e ciently biodegraded when the initial benzene concentration was below 150 mg/L. The metabolites of anaerobic nitrate-dependent benzene oxidation by strain B. cereus were identified as phenol and benzoate. The results of substrate interaction between binary combinations for benzene, phenol and benzoate showed that the simultaneous presence of benzene stimulated the degradation of benzoate, whereas the addition of benzene inhibited the degradation of phenol. Benzene degradation by B. cereus was enhanced by the addition of phenol and benzoate, the enhanced e ects were more pronounced at higher concentration. To our knowledge, this is the first report that the isolated bacterial culture of B. cereus can e ciently degraded benzene under nitrate reducing conditions.     

细菌氧化法制取黄钾铁矾的研究

黄钾铁矾是金属硫化物在酸性条件下氧化形成的主要次生矿物,嗜酸氧化亚铁硫杆菌在9K液体培养基中能迅速把Fe2+氧化成Fe3+,而Fe3+易水解,最终产生黄钾铁矾类沉淀。文章为确定其最佳生成条件,在初始pH值分别为1.40、2.10、2.46、3.03、3.52、4.10、4.90、5.90,温度分别为25℃、30℃、35℃、40℃条件下,考察了体系中Fe2+和总铁浓度的变化及黄钾铁矾产量的大小,通过XRD、FTIR、XRF等方法对其化学组成和结构做了分析鉴定。结果表明:初始pH值为4.1、温度在30℃时是细菌氧化法制取黄钾铁矾的最佳条件,经鉴定,此最佳条件下制得的样品89%是黄钾铁矾,含少量氢黄钾铁矾和黄铵铁矾。     

1株嗜酸硫杆菌的分离鉴定及其对污染底泥中重金属的去除效果

采用常规细菌分离方法,从河流底泥中筛选到1株细菌FD97,通过序批式摇瓶培养考察了不同温度下(22~40℃)该菌株对污染底泥中Zn、Cu和Cr的去除效果.结果表明,形态学和16S rDNA序列同源性分析鉴定菌株FD97为嗜酸硫杆菌属(Acidithiobacillussp.).在22~40℃条件下,以Acidithiobacillussp.FD97为主要菌株的生物沥浸作用可有效去除污染底泥中Zn、Cu和Cr.沥浸处理16 d,Zn、Cu和Cr的去除率可分别达70%、90%和25%.从底泥pH值变化、SO42-产生以及重金属去除率角度衡量不同温度的底泥沥浸处理效果依次为:34℃28℃≈40℃22℃.控温28℃较适宜今后的实际应用.底泥重金属生物沥浸去除率的大小与底泥pH值变化密切相关,与温度无直接关系.当底泥pH值降至5.0、3.5和2.5时,底泥中Zn、Cu和Cr分别开始溶出,pH值降为2.0,重金属去除率达最大.     

Alcaligenes sp. XJ-T-1利用废弃油脂生产破乳剂研究

生物破乳剂是一种应用于油水乳状液分离的新型破乳剂.本研究对筛选得到的1株高效破乳剂产生菌XJ-T-1（鉴定结果为Alcaligenes sp.）代谢产生的生物破乳剂进行了理化性质、破乳能力和废弃油脂利用能力分析.采用废弃油脂Ⅱ为碳源,可提高生物破乳剂产量4.6倍,XJ-T-1在以液体石蜡和废弃油脂培养8　d后该菌能使水的表面张力从72 mN/m下降到32 mN/m, CMC-1分别为10、 20;以液体石蜡为碳源培养得到的生物破乳剂在油包水型、水包油型模型乳状液的破乳率分别达到了96%、 50%;而以废弃油脂Ⅱ为碳源培养得到的生物破乳剂的破乳率分别为97.8%、 65％;采用煤油脱出率、乳状液脱除率和水脱出率3种评价方式对破乳效果评价发现生物破乳剂最先作用于乳状液的连续相;采用TLC和IR对废弃油脂生产破乳剂得到的破乳剂有效成分进行鉴定,结果为脂肽与石蜡培养产物相同.     

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene (BaP) was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons (PAHs) (such as phenanthrene, pyrene and fluoranthene) as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.