非光合固碳微生物的碳源组合优化

二氧化碳(CO_2)过量排放而引起的全球变暖是目前全球面临的重大环境问题。微生物固碳是实现二氧化碳资源利用的一种重要方式。探索无需光照的高效固碳微生物对于更广泛环境条件下的微生物固碳具有重要意义。在从全球各大海域筛选富集出非光合微生物茵群的基础上,本文构建了在不同的电子供体条件下(铵和氢气)促进非光合微生物茵群生长的最佳混合碳源组合,得到如下结果:在以NH_4~+为电子供体的条件下,优化后的碳源组合为374.24 mgC/L碳酸钠(Na_2CO_3),54.76 mgC/L碳酸氢钠(NaHCO_3)和0 mgC/LCO_2时,最佳响应值TOC为3.06 mg/L。最佳响应值TOC低于以Na_2CO_3为单一碳源时,但高于以CO_2或NaHCO_3为单一碳源时;在以H_2为电子供体条件下,使用优化后的混合碳源为0.26 mg/L Na_2CO_3、0.59mg/L NaHCO_3和71.48mL/L CO_2时,非光合微生物菌群的固碳效率可达27.62 mg/L,较以CO_2为单一碳源提高35%左右。这可能意味着有H_2条件下非光合微生物菌群中的微生物可能以羟基丙酸固碳途径为主,而且多条固碳途径均能被混合菌群利用。     

阿尔山不同时期落叶松根际土壤固氮菌的多样性研究

利用传统的细菌分离培养,结合16S rDNA序列分析等方法,对阿尔山地区不同时期落叶松根际可培养固氮菌群落的多样性进行分析,以揭示落叶松根际固氮菌的多样性及群落结构的季节变化规律,为森林生态系统的可持续发展提供理论依据。结果显示:(1)从阿尔山落叶松根际土壤中共计分离纯化细菌菌株112株,分属于14属41种,包括假单胞菌属、伯克氏菌属、根瘤菌属、叶杆菌属、节杆菌属、类芽孢杆菌属、沙雷菌属、欧文菌属、短小杆菌属、芽孢杆菌属、肠杆菌属、不动杆菌属、柄杆菌属、红球菌属;其中优势菌群为假单胞菌属,次优势菌群为叶杆菌属、伯克氏菌属和节杆菌属。(2)季节变化对落叶松固氮菌群的变化有显著影响,表现为4月份和10月份最优势类群为γ-变形菌纲(γ-proteobacteria)中的假单胞菌属,6月份和8月份的最优势类群相同,但组成有所差别,其中6月份优势菌群包括假单胞菌属、短小杆菌属、红球菌属、节杆菌属,8月份的优势菌群为假单胞菌属、不动杆菌属、肠杆菌属、短小杆菌属、红球菌属和节杆菌属。(3)不同时期的物种均匀度指数(McIntosh index)差异显著,8月份最大,4月份最小,变化范围在0.83～1.164之间;物种丰富度指数(Shannon-Wiener index) 6月份和8月份显著高于4月份和10月份;优势度指数(Simpson index) 4月份和10月份显著高于6月份和8月份。研究表明,阿尔山地区落叶松根际微生物的多样性较高,群落相对复杂,分离的14个菌属多为根际促生菌,且不同时期固氮菌的群落组成受季节的影响明显。     

Ammoniacal silver staining of proteins: mechanism of glutaraldehyde enhancement

When the conditions for detecting proteins by ammoniacal silver staining (B. R. Oakley, D. R. Kirsch, and N. R. Morris (1980) Anal. Biochem. 105, 361-363.) following gel electrophoresis were varied, it was noted that glutaraldehyde pretreatment was necessary for maximal staining, which could not be explained simply as the result of "fixation." Further studies indicated that glutaraldehyde enhancement of protein staining with this silver reagent was probably due to oxidation of the aldehyde groups by silver ions, resulting in metallic silver depositions within the gel which act as nucleation sites for additional metallic silver localization in the protein bands upon the addition of formaldehyde developer. This proposed mechanism is consistent with the Tollen's reaction, as well as some aspects of the photographic process. Consistent with this notion, silver-staining intensities are directly related to mole percentage lysine of various standard proteins.     

A new, sensitive method for enzyme kinetic studies of scarce glucosides

The maize β-glucosidase Zm-p60.1 is important for the regulation of plant development through its role in the targeted release of free cytokinins from cytokinin-O-glucosides, their inactive storage forms. Enzyme kinetics studies using these scarce substrates close to physiological concentrations are difficult due to two reasons: (a) Available methods are mainly suited for end-point kinetics. (b) These methods are not sufficiently sensitive when using scarce glucoside substrates.We developed a glucose assay using a system comprising three enzymes β-glucosidase, glucose oxidase and horseradish peroxidase, with the new substrate N-acetyl-3,7-dihydroxyphenoxazine-Amplex Ultra Red reagent (Molecular Probes). A calibration curve was constructed for resorufin and validation was carried out by comparing our method with the standard spectrophotometric method using p-nitrophenyl-β-d-glucopyranoside. In comparison with the other methods, this method is more sensitive, precise and accurate. The assay is rapid and hence suited for continuous kinetics, it is readily adapted to suit automated procedures, and potential applications include its use in studying the physiological role(s) of enzymes that cleave scarce glucoside substrates.     

长喙田菁-Azorhizobium caulinodans共生固氮体系在华南地区的生长、结瘤、固氮和种子生产

研究了长喙田菁－Ａｚｏｒｈｉｚｏｂｉｕｍｃａｕｌｉｎｏｄａｎｓ共生固氮体系在华南地区的生长、结瘤、固氮和种子生产．结果表明,长喙田菁－Ａ．ｃａｕｌｉｎｏｄａｎｓ共生固氮体系在华南地区生长正常,并具旺盛的茎根瘤结瘤作用．经６５天的生长,其在湛江地区的单位面积生物量（干物质）和单位面积Ｎ产量分别为２８７５２和６８１ｋｇ·ｈｍ－２,远优于普通田菁的１６５２０和３５２ｋｇ·ｈｍ－２;茎瘤菌Ａ．ｃａｕｌｉｎｏｄａｎｓ品系ＡＲ１１１和ＡＲ５６在华南地区混合接种效果良好,植株茎瘤结瘤率达到１００％,平均单株茎瘤个数为１８２个,单株茎瘤鲜重约为１．２ｇ,茎瘤生物量在茎根瘤总生物量中所占比重为７０％,而其根瘤生物量略高于普通田菁;长喙田菁在华南地区能够正常开花结实,在栽植密度为４×１０４株·ｈｍ－２的条件下,其种子产量达３２００ｋｇ·ｈｍ－２．     

Effect of taxol and okadaic acid on microtubule dynamics in thimerosal-arrested primary mouse oocytes: a confocal study

A pulse of thimerosal (TMS), a sulfhydryl reagent, induces an instantaneous, complete and long-lasting microtubule interphasic network disassembly in mouse primary oocytes, correlated with the irreversible inhibition of meiosis reinitiation This inhibition is bypassed by dithiothreitol (DTT) while thiosalicylic acid, an analog of TMS, does induce neither microtubules depolymerisation nor inhibition of reinitiation and resumption of meiosis. This strongly suggests that the dramatic and pleiotropic inhibitory effect of TMS is specifically related to its sulfhydryl group oxidising activity of critical molecules among which tubulin. In contrast to DTT, okadaic acid (OA), known to bypass the inhibitory effect of drugs interfering with protein kinase activities, induces a late chromatin condensation and GVBD in TMS-pulsed oocytes as compared to the control situation, with no significant concomitant microtubule assembly. These cytological features are suggested to be indirectly induced by a late MAPK activation and confirm that a very early thiol oxidation induced by TMS exerts a much more dramatic effect on resumption of meiosis than any pharmacological manipulation of protein kinase activities leading to activation of MPF. Finally, taxol was shown to promote tubulin polymerisation even when microtubules were irreversibly disassembled by thiol oxidation but fails to restore the ability to undergo maturation.     

Diazocyclopentadiene (DACP), a light sensitive reagent for the ethylene receptor in plants

Diazocyclopentadiene (DACP) has been shown to be an effective reagent for the ethylene receptor. Treatment of mung bean sprouts or tobacco leaves with DACP in the light or in the dark inactivates much of the ethylene binding. In the light, inactivation seems to be permanent, while in the dark, the site becomes active again after the DACP diffuses away. The compound is 10 times more effective in the light than in the dark. DACP inhibits banana ripening indicating the physiological receptor is involved. It also overcomes the inhibitory effect of ethylene on mung bean seedling growth (Km = 0.09 µl/1 E) at low ethylene levels. At high ethylene levels, an apparent high ethylene level site becomes apparent (Km = 50 µl/1 E) and growth is inhibited.     

Solid-phase distribution of heavy metals as affected by single reagent extraction in dredged sediment derived surface soils

ABSTRACT

EDTA is useful to assess mobile metal pools in polluted soils and sediments. There is a need to enhance our understanding of the significance of metal fractions released. The impact of single reagent extraction with 0.05 mol L^?1 EDTA on the solid phase distribution of trace metals in surface soils sampled from confined dredged sediment disposal sites was investigated. Not extracted and EDTA extracted soils were subjected to sequential extraction to fractionate the total contents into: (1) easily exchangeable and carbonate bound fraction; (2) reducible fraction; (3) oxidisable fraction; and (4) residual fraction. With EDTA, significant portions of metals associated with the acid extractable and reducible fractions were released. The oxidisable and residual fractions remained unaffected for most of the investigated metals except for the organic matter associated metals (Cu and Pb). A decrease in the residual fraction after EDTA-extraction for Cu and Pb was attributed to artifacts of the sequential extraction procedure.     

Membrane Topology and Functional Importance of the Periplasmic Region of ABC Transporter LolCDE

The LolCDE complex is an ATP-binding cassette transporter that mediates the release of newly synthesized lipoproteins from the cytoplasmic membrane of gram-negative bacteria, which results in the initiation of outer-membrane sorting of lipoproteins through the Lol pathway. LolCDE is composed of one copy each of membrane subunits LolC and LolE, and two copies of nucleotide-binding subunit LolD. In this study, we examined the membrane topology of LolC and LolE by PhoA fusion analysis. Both LolC and LolE were found to have four transmembrane segments with a large periplasmic loop exposed to the periplasm. Despite similarities in sequence and topology, the accessibility of a sulfhydryl reagent to Cys introduced into the periplasmic loop suggested that the structure of the periplasmic region differs between LolC and LolE. Inhibition of the release of lipoproteins by the sulfhydryl reagent supported a previous proposal that LolC and LolE have distinct functions.     

Real-time background suppression during frequency domain lifetime measurements

We describe real time background suppression of autofluorescence from biological samples during frequency domain or phase modulation measurements of intensity decays. For these measurements the samples were excited with a train of light pulses with widths below 1 ps. The detector was gated off for a short time period of 10 to 40 ns during and shortly after the excitation pulse. The reference signal needed for the frequency domain measurement was provided by a long-lifetime reference fluorophore which continues to emit following the off-gating pulse. Both the sample and the reference were measured under identical optical and electronic conditions avoiding the need for correction of the photomultiplier tube signal for the gating sequence. We demonstrate frequency domain background suppression using a mixture of short- and long-lifetime probes and for a long-lifetime probe in human plasma with significant autofluorescence.