5,6,7,8-Tetrahydromethanopterin-dependent enzymes involved in methanogenesis

Abstract In the process of methanogenesis, 5,6,7,8-tetrahydromethanopterin (H₄MPT) is the carrier of the C₁ unit at the formyl through methyl state of reduction. By the transfer of a formyl group from formylmethanofuran, 5-formyl- and 10-formyl-H₄MPT are formed in hydrogenotrophic and methylotrophic organisms, respectively. Cyclohydrolysis of the 5- and 10-formyl derivatives then yields 5,10-methenyl-H₄MPT, which is reduced in two subsequent coenzyme F₄₂₀-dependent reactions to 5-methyl-H₄MPT. Following the transfer of the methyl group to coenzyme M, the substrate of the terminal step in methanogenesis, methylcoenzyme M, is produced. In this paper properties of the enzymes catalyzing the individual H₄MPT-dependent reactions are discussed.     

Crystal structures and enzymatic properties of three formyltransferases from archaea: environmental adaptation and evolutionary relationship

Formyltransferase catalyzes the reversible formation of formylmethanofuran from N(5)-formyltetrahydromethanopterin and methanofuran, a reaction involved in the C1 metabolism of methanogenic and sulfate-reducing archaea. The crystal structure of the homotetrameric enzyme from Methanopyrus kandleri (growth temperature optimum 98 degrees C) has recently been solved at 1.65 A resolution. We report here the crystal structures of the formyltransferase from Methanosarcina barkeri (growth temperature optimum 37 degrees C) and from Archaeoglobus fulgidus (growth temperature optimum 83 degrees C) at 1.9 A and 2.0 A resolution, respectively. Comparison of the structures of the three enzymes revealed very similar folds. The most striking difference found was the negative surface charge, which was -32 for the M. kandleri enzyme, only -8 for the M. barkeri enzyme, and -11 for the A. fulgidus enzyme. The hydrophobic surface fraction was 50% for the M. kandleri enzyme, 56% for the M. barkeri enzyme, and 57% for the A. fulgidus enzyme. These differences most likely reflect the adaptation of the enzyme to different cytoplasmic concentrations of potassium cyclic 2,3-diphosphoglycerate, which are very high in M. kandleri (>1 M) and relatively low in M. barkeri and A. fulgidus. Formyltransferase is in a monomer/dimer/tetramer equilibrium that is dependent on the salt concentration. Only the dimers and tetramers are active, and only the tetramers are thermostable. The enzyme from M. kandleri is a tetramer, which is active and thermostable only at high concentrations of potassium phosphate (>1 M) or potassium cyclic 2,3-diphosphoglycerate. Conversely, the enzyme from M. barkeri and A. fulgidus already showed these properties, activity and stability, at much lower concentrations of these strong salting-out salts.     

巴氏新小绥螨对柑桔全爪螨处理的枳橙叶片挥发物的行为反应

【目的】研究巴氏新小绥螨Neoseiulus barkeri对柑桔全爪螨Panonychus citri及刺吸式昆虫为害柑桔叶片释放的挥发物的行为反应,揭示巴氏新小绥螨的嗅觉反应特点。【方法】采用GC-MS顶空进样法对枳橙叶片常见挥发性化合物、针刺枳橙叶片及其柑桔全爪螨雌成螨取食枳橙叶片挥发物进行定性分析,确定每类化合物的相对保留指数,构建枳橙叶片常见及受害挥发性化合物特征谱库。利用嗅觉测定技术分析巴氏新小绥螨对枳橙叶片挥发物的行为反应。【结果】针刺处理和柑桔全爪螨取食影响枳橙叶片挥发物的组成和含量。两种方法处理时枳橙叶片释放的主要物质为α-蒎烯、水芹烯、4-异丙基甲苯。随着处理加重,增量释放的物质为:cis-π-罗勒烯、月桂烯、柠檬烯、异松油烯,减量表达的物质为2-乙基-1-乙醇和十一烷。在10~(-2)、10~(-4)、10~(-6)和10~(-8)g/m L浓度下,正己醛、正壬醛、乙酸辛酯和正庚醛对巴氏新小绥螨有强烈的引诱作用(P>0.05);月桂烯在10~(-2)、10~(-4)和10~(-6)g/m L浓度下对巴氏新小绥螨有强烈的引诱作用(P>0.05);正壬醇和正辛醇随着浓度增加,对巴氏新小绥螨的引诱作用降低;苯甲醛对巴氏新小绥螨的引诱作用较弱。【结论】巴氏新小绥螨对柑桔全爪螨及刺吸式口器昆虫为害柑桔叶片释放出的挥发物各组分具有不同的行为反应,柑桔及其刺吸式害虫生境中的嗅觉线索在巴氏新小绥螨的寄主定位和生境选择中起着重要作用。     

Inhibition of Methanosarcina barkeri strain 227 by cadmium

Abstract The effect of cadmium (Cd) on methane formation from methanol and/or H₂–CO₂ by Methanosarcina barkeri was examined in a defined growth medium and in a simplified buffer system containing 50 mM Tes with or without 2 mM dithiothreitol (DTT). No inhibition of methanogenesis by high concentrations of cadmium was observed in growth medium. Similarly, little inhibition of methanogenesis by whole cells in the Tes buffer system was observed in the presence of 430 μM Cd or 370 μM mercury (Hg) with 2 mM DTT. When the concentration of DTT was reduced to 0.4 mM, almost complete inhibition of methanogenesis from H₂–CO₂ and methanol by 600 μM Cd was observed. In the absence of DTT, 150 μM Cd inhibited methanogenesis from H₂–CO₂ completely and from methanol by 97%. Methanogenesis from H₂–CO₂ was more sensitive to Cd than that from methanol.     

Hydrogen metabolism during methanogenesis from acetate by Methanosarcina barkeri

A tritium exchange assay and a sensitive gas chromatographic technique were used to demonstrate that hydrogenase was active and that hydrogen was produced by Methanosarcina barkeri strain MS grown on acetate. Both methane and hydrogen production rates were dependent on the concentration of acetate in the medium. H₂ was produced at 0.5–2% of the rate of CH₄ formation. Chloroform and potassium cyanide, inhibitors of methanogenesis from acetate, inhibited H₂ production but not hydrogenase activity. The addition of hydrogen gas to cell suspensions did not inhibit CH₄ or carbon dioxide production from the methyl group of acetate. H₂ production appears to be linked to several intracellular redox processes which follow the cleavage of acetate.     

How larvae of Thrips tabaci reduce the attack success of phytoseiid predators

Neoseiulus barkeri (= Amblyseius mckenziei) and Amblyseius cucumeris (Acari:Phytoseiidae) are used as control agents of Thrips tabaci (Insecta:Thripidae) in greenhouse crops. Their success in capturing prey larval stages is related to both the feeding state of the predators and to the size of the larvae. When starved, predators are more successful in seizing larvae. Upon contact with a starved predator second stage prey larvae incur a lower death risk than first stage larvae. The larvae of T. tabaci reduce the attack success of their predators by jerking the abdomen and by producing a drop of rectal fluid. When this defensive behaviour is prevented by anaesthetising the larvae with CO₂, predator attack success increases. Anaesthesia does not, however, level out the difference in death risk of the two larval stages. Conceivable causes for this discrepancy are discussed.Availability of suitable prey is dependent on the dynamics of the age structure of the prey population and, hence, may be lower than total thrips density suggests. If so, alternative food sources may be important to maintain the predator population.

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Zusammenfassung Neoseiulus barkeri (= Amblyseius mckenziei) und Amblyseius cucumeris (Acari: Phytoseiidae) werden zur Bekämpfung von Thrips tabaci (Insecta: Thripidae) in Gewächshauskulturen eingesetzt. Sowohl der Ernährungszustand der räuberischen Milben als auch die Grösse der Thripslarven haben Einfluss auf das Ausmass der Beutenahme. Die Prädatoren sind erfolgreicher, wenn sie eine Zeitlang ohne Nahrung gehalten wurden. Beim Zusammentreffen mit einer ausgehungerten Raubmilbe besteht für Thripslarven des zweiten Stadiums ein geringeres Risiko erbeutet und gefressen zu werden als für Larven des ersten Stadiums. T. tabaci Larven mindern den Angriffserfolg der Prädatoren durch kräftiges Hin- und Herschlagen des Abdomens und durch Abgabe eines Tropfens Rektalflüssigkeit. Wird dieses Abwehrverhalten der Larven durch Anaästhesie mit CO₂ verhindert, erhöht sich der Angriffserfolg der Prädatoren. Anästhesie nivelliert jedoch nicht das für beide Larvenstadien unterschiedlich hohe Risiko erbeutet zu werden. Mögliche Ursachen für diesen Unterschied werden diskutiert.Die Verfügbarkeit geeigneter Beutetiere hängt ab von der zeitlichen Entwicklung der Altersstruktur ihrer Population. Das Angebot an wirklich geeigneten Beutetieren kan also unter Umständen geringer sein, als dies die Gesamtthripsdichte zunächst vermuten lässt. Ist das der Fall, dürften alternative Nahrungsquellen für die Ernährung der Prädatorenpopulation wichtig sein.

     

巴氏钝绥螨rDNA的ITS基因片段序列分析

采用酚-氯仿抽提法提取了采自江西赣南的巴氏钝绥螨Amblyseiusbarkeri Hughes,1948基因组DNA。以相应引物对巴氏钝绥螨核糖体ITS基因进行PCR扩增,直接测序,得到了652bp的碱基片段（国际基因库索引号FJ392365）,其碱基序列A、C、G、T含量分别为193bp（29．60％）、114bp（17．48％）、144bp（22．09％）、201bp（30．83％）,并对其与其他植绥螨5．8SrDNA及两侧ITS序列进行了分析。     

Retrospective spatial analysis of the pollination of two miniature epiphytic orchids with different pollination strategies in a coffee plantation in Soconusco,Chiapas, Mexico

Quantitative and spatial data for orchid pollination are scarce and may be important tools for reintroduction and conservation; however, conclusions cannot be drawn on the basis of the typically infrequent and unpredictable pollination events. We carried out a novel, retrospective, spatial analysis of the pollination of the entire population of two miniature orchids, Erycina crista‐galli and Notylia barkeri, on coffee bushes in plantations at 900 m in Soconusco, south‐eastern Mexico. The numbers of mature flowering plants of both species in the experimental site were similar. Notylia barkeri produced nearly four times as many flowers, but a similar proportion of the total number of flowers produced was pollinated (1.23% and 1.48% for N. barkeri and E. crista‐galli, respectively). An estimated 29 919 977 (±4 983 995) seeds were produced by N. barkeri, nearly 12 times more than E. crista‐galli at 1 009 414 (±147 000). The pollinators of N. barkeri chose flower clusters at random and pollinated various flowers within a patch, whereas the pollinators of E. crista‐galli chose patches of flowers slightly more systematically, with less dependence on flower density, and appeared to dedicate less attention to each patch. For both species, pollinators slightly favoured larger clusters of flowers and left many individual and groups of flowers unvisited. To restore populations of these orchids in coffee plantations as a replacement habitat, N. barkeri should be planted in small, separate groups and E. crista‐galli in larger groups of individuals, dispersed regularly throughout the selected site to maximize the possibility that the flowers will be discovered by pollinators. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 448–459.