生物固碳途径研究进展

生物固碳是地球碳循环过程的重要组成部分。自然界已经发现了六条天然生物固碳途径,但自然途径不仅能量利用效率低下,而且人工改造提升固碳效率难度大。随着合成生物学的发展,新的人工固碳途径不断涌现。相对于天然途径,人工固碳途径具有路线短、耗能少、原子经济性高等优点,有望在不久的将来能够替代天然固碳途径,实现固碳效率的大幅提高,是解决人类能源与环境问题的有效途径之一。主要总结了天然固碳途径和人工固碳途径的代谢原理和关键固碳酶的酶学特征,并对未来发展趋势进行展望。     

毛乌素沙地固碳功能菌群落随生物结皮发育阶段的演变特征

生物结皮是干旱区生态系统光合固碳的重要贡献者,固碳功能菌是生物结皮碳固定过程中的关键功能群,而毛乌素沙地生物结皮有关固碳功能菌群落多样性的研究未见详细报道。通过qPCR和高通量测序,研究了毛乌素沙地固碳功能菌基因丰度、群落多样性在生物结皮发育过程中的演变规律及其关键环境因子。结果表明,form IAB、IC和ID的基因丰度均随生物结皮发育呈升高趋势,在地衣结皮最高,在藓结皮又明显降低;form IAB的Chao1和form IC的Shannon指数随生物结皮演替呈显著上升趋势,而 form IC和ID的Chao1则在藻结皮达到峰值。物种组成上,IAB型固碳功能菌以蓝藻门的颤藻目占优势,其相对丰度在藻结皮最高,随生物结皮演替逐渐降低;IC型固碳功能菌在裸沙中以土壤红杆菌目为主, 而在三种生物结皮中以生丝微菌目、亚硝化单胞菌目和红螺菌目占优势; ID型固碳菌则以硅藻门的舟形藻目占优势,其丰度在裸沙中最高。IAB和IC型固碳功能菌群落结构在藻结皮和地衣结皮类似,而与裸沙和藓结皮差异显著,裸沙与藓结皮亦具显著差异。生物结皮演替过程中生物类群和土壤理化特征的改变为固碳功能菌提供了不同的生态位,土壤总有机碳、全氮、铵态氮、全磷、速效磷和pH对固碳功能菌群落进行综合调控,通过对固碳功能菌的筛选作用,最终改变固碳功能菌群落组成和结构。为深入理解荒漠生态系统生物结皮的光合固碳功能的微生物学机制提供了理论支撑。     

黄土高原人工苜蓿草地固碳效应评估

苜蓿草地作为黄土高原草地的重要组成部分,在黄土高原草地生态系统碳循环占有重要地位。同时由于苜蓿具有固氮和固碳的双重作用,使它在低碳农业的发展中具有重要意义。本文以一年生（建植初期）、三年生（盛产期）、五年生（稳产期）和十年生（衰败期）的人工苜蓿草地为研究对象,利用地上地下生物量和土壤碳贮量,定量分析了人工苜蓿草地不同发育阶段的固碳能力;结合种植面积,对黄土高原区人工苜蓿草地的总固碳效应进行了评价。研究结果表明：苜蓿草地地上生物固碳能力以三年生和五年生最强,并显著高于其他年限;地下生物固碳和土壤固碳量均以十年苜蓿草地最高,且显著高于其他年限。总体而言,人工苜蓿草地以土壤固碳为主,并集中在0~80cm土壤,各年限0-80cm土层土壤固碳量分别占到总固碳量的41.02%、39.43%、41.56%、39.59%;而地下生物固碳主要发生在0~20cm土层,各年限该层生物固碳量分别占到地下生物总固碳量的45.74%、 55.68%、53.12%和 43.28%。苜蓿草地在整个生命周期中总固碳量随生长年限逐年增长,但增长速度不明显,各年限单位面积总固碳量分别为12.69kg/m2、13.49 kg/m2、13.31 kg/m2和14.83 kg/m2。估算得到黄土高原的人工苜蓿草地年总固碳量为1.430?1011 Kg（143.0 Tg）,其中地上、地下、土壤分别为5.43Tg、1.15Tg和136.4Tg。本研究结果为正确分析区域碳循环和发展低碳大农业提供了重要理论参考。     

多肽固相合成的研究进展

多肽固相化学合成法是蛋白质研究领域非常重要的研究方法之一,主要可以分为Boc方法和Fmoc方法,在生物药物、蛋白质工程、免疫学等研究中得到了广泛的应用。该文论述了多肽固相合成法的原理,比较了两种典型合成方法的优缺点,介绍了适用于该方法的多肽种类,提出了多肽合成过程中存在的问题与解决策略,最后展望了多肽合成法的应用前景,以供相关领域的研究人员参考。     

毛萼田菁茎瘤豆血红蛋白含量与固氮活性的关系

在个体瘤发育的不同阶段和植株生长的不同时期,以及在不同的光照条件下,分别测定茎瘤的固氨活性和豆血红蛋白含量。结果表明,毛萼田菁茎瘤的固氨活性和豆血红蛋白含量呈正相关,豆血红蛋白在茎瘤的共生固氮中起着重要的作用。     

氮输入对植物光合固碳的影响研究进展

植物光合固碳(C)是生物固C的重要途径和生态系统C循环中的重要环节。在全球环境变化背景下,研究氮(N)输入对植物光合固C的影响,对于更好的认识C、N循环过程及生态系统对全球变化的响应过程等具有重要意义。N输入是否能够增加植物固C取决于生态系统类型以及生态系统的N饱和度;草原和湿地生态系统N输入的临界负荷值较高,干旱、半干旱荒漠地区较低;N输入可能改变植物光合固C在各器官的分配,主要由植物生理、自身生长节律和环境养分等决定。由于物种和生态系统类型的差异,N输入对植物固C的影响仍具有很大的不确定性,目前缺乏准确、定量表达N输入对生态系统光合和C同化物分配影响的数学表达方法和过程算法。未来应着重加强N输入下C同化物分配的生物地球化学模型和N、P富集下植物光合固C耦合模型研究,并应用同位素标记和分子生物学技术,从生态系统角度综合探讨N输入下植物光合固C的分配和转化特征。     

生物炭对土壤有机碳矿化的激发效应及其机理研究进展

近年来由于生物炭具有碳素稳定性强和孔隙结构发达等特性,其在土壤固碳减排方面的作用研究受到广泛关注.然而当生物炭进入土壤环境后最终是增加土壤碳的储存还是促进土壤碳的排放?目前学术界对该问题仍存在争议.生物炭对土壤有机碳的激发效应及其机理研究有待进一步深入开展.本文在分析生物炭自身碳素组分和稳定性、孔隙结构及表面形态特征的基础上,综述了添加生物炭对土壤本底有机碳矿化产生激发效应的研究进展,分别阐述了产生正激发和负激发效应(即促进和抑制矿化)的机制机理,认为正激发效应主要是基于生物炭促进土壤微生物活性增强、生物炭中易分解组分的优先矿化以及由此引发的土壤微生物的共代谢作用,而负激发效应主要是基于生物炭内部孔隙结构和外表面对土壤有机质的包封作用和吸附保护作用、生物炭促进土壤有机-无机复合体形成的稳定化作用、生物炭对土壤微生物及其酶活性的抑制作用.最后对今后相关研究方向进行了展望,以期为生物炭在土壤固碳减排方面的应用提供理论依据.     

不同耕作处理下大豆生物固N能力及对系统N素的贡献

2002年至2003年在黄土高原研究了4个耕作处理,传统耕作（t）、传统耕作＋秸秆覆盖（ts）、免耕（nt）和免耕＋秸秆覆盖（nts）下大豆的生物固N百分率（％Ndfa）、固N数量及其对春玉米-冬小麦-夏大豆轮作系统中N素的贡献。结果表明,在t、ts、nt和nts处理下2002年的生物固N百分率为17．6％、34．3％、22．4％和19．3％,2003年则为58．5％、62．4％、54．9％和43．8％,其中2003年的生物固N百分率比2002年分别高出69．9％、45．O％、59．3％和56．1％,固N数量高出56．2％、33．8％、49．5％和43．1％。大豆生物固N百分率、生物固N数量与生物量呈正相关关系,在ts处理下显著相关。土壤NO3-N含量和大豆固N数量呈负相关,大豆植株吸N量占土壤NO3-N的百分比在2002年为：t（88．1）〉ts（84．6）〉nts（78．7）〉nt（63．6）,2003年为：t（115．5）〉ts（104．2）〉nts（99．8）〉nt（95．8）。2002年大豆对该轮作系统的N素贡献分别为6．6（t）、11．6（ts）、6．5（nt）和6．1（nts）kg／hm^2,生物固N量占总N输入量的百分比为14．6（t）、21．5（ts）、14．9（nt）和12．9（nts）;2003年大豆对系统的N素贡献分别为14．9（t）、17．6（ts）、12．9（nt）和10．7（nts）kg／hm2,生物固N量占总N输入的百分比为63．2％（t）、58．5％（nt）、47．7％（ts）和39．9％（nts）。年际变异造成了大豆生物固N的年际差异,秸杆覆盖＋耕作因改善水分状况,而促进了大豆的生物固N作用。     

生物炭施用的固碳减排潜力及农田效应

气候变暖及粮食安全是保证人类可持续发展的重要课题。生物炭具有较高的稳定性、较高碳含量等特点,能增加土壤碳储量,提高土壤物理及化学性质,提高农田产出,能应对高温胁迫及土壤退化双重压力,具有一举多赢的生态环境效益,在缓解温室效应及粮食危机方面展现出巨大的潜力。综合前人研究成果,分析了生物炭固碳减排潜力及农田效应影响因素(包括:生物炭原料、制备温度、施用量、土壤类型等)。综合固碳减排及提高产出两方面因素,提出了较合适的生物炭施用标准,即300—700℃制备的农林废弃物生物炭,且施用量不超过5%。对生物炭固碳减排及田间效应领域未来的研究方向进行了展望。     

岩溶区水生生态系统微藻的生物碳泵效应

微藻在水生生态系统的碳固定中扮演重要角色。本文综述了岩溶区水生生态系统生物碳泵的提出、岩溶区微藻生物碳泵作用、影响微藻固碳的主要环境因素以及岩溶区微藻固碳的研究进展,并提出了亟待解决的关键科学问题,为深入研究岩溶区水生生态系统微藻固碳能力及生物碳泵机制、科学认识岩溶生态系统的碳汇潜力、丰富和完善岩溶碳循环理论提供参考。     

斯氏假单胞菌A1501固氮新基因PST1305的功能分析

摘要：【目的】研究斯氏假单胞菌A1501基因组“固氮岛”中PST1305基因在A1501生物固氮过程中所起的作用。【方法】利用同源重组与三亲接合的方法构建PST1305的非极性突变株。乙炔还原法测定固氮酶活。RT-PCR分析PST1305基因与其周围基因转录单元的关系,Real-Time PCR比较PST1305在最佳固氮与非固氮条件下表达水平的差异。【结果】突变株np1305的固氮酶活显著降低,功能互补菌株np1305Comp能基本恢复细胞的固氮作用。PST1305与其上游的nifB、fdxN、下游的nifQ等基因位于同一个转录单元,组成一个操纵子。基因芯片表明,PST1305基因在固氮比非固氮条件下表达量显著上调（约38.7倍）,Real-Time PCR验证支持这一结果。【结论】PST1305基因参与固氮过程,其突变会影响固氮酶的活性,该基因可能通过参与A1501固氮酶电子传递或者固氮酶的氧保护过程影响固氮效率。     

Nitrogenase gene diversity and microbial community structure: a cross-system comparison

Biological nitrogen fixation is an important source of fixed nitrogen for the biosphere. Microorganisms catalyse biological nitrogen fixation with the enzyme nitrogenase, which has been highly conserved through evolution. Cloning and sequencing of one of the nitrogenase structural genes, nifH, has provided a large, rapidly expanding database of sequences from diverse terrestrial and aquatic environments. Comparison of nifH phylogenies to ribosomal RNA phylogenies from cultivated microorganisms shows little conclusive evidence of lateral gene transfer. Sequence diversity far outstrips representation by cultivated representatives. The phylogeny of nitrogenase includes branches that represent phylotypic groupings based on ribosomal RNA phylogeny, but also includes paralogous clades including the alternative, non-molybdenum, non-vanadium containing nitrogenases. Only a few alternative or archaeal nitrogenase sequences have as yet been obtained from the environment. Extensive analysis of the distribution of nifH phylotypes among habitats indicates that there are characteristic patterns of nitrogen fixing microorganisms in termite guts, sediment and soil environments, estuaries and salt marshes, and oligotrophic oceans. The distribution of nitrogen-fixing microorganisms, although not entirely dictated by the nitrogen availability in the environment, is non-random and can be predicted on the basis of habitat characteristics. The ability to assay for gene expression and investigate genome arrangements provides the promise of new tools for interrogating natural populations of diazotrophs. The broad analysis of nitrogenase genes provides a basis for developing molecular assays and bioinformatics approaches for the study of nitrogen fixation in the environment.     

固氮酶的研究进展

固氮酶将N2还原为NH3的过程是自然界实现氮循环的重要环节。固氮酶是由γ2型二聚体组成的Fe蛋白和α2β2异四聚体组成的MoFe蛋白组成。固氮酶催化的机制包括铁蛋白的氧还循环和钼铁蛋白的氧还循环两部分。Klebsiella pneumoniae的nif基因簇由20个基因组成,构成了8个转录单位,总长度24206bp,其操控机制是多水平、多层次的调控过程。同时综述了固氮酶的多样性,目前已经发现的有钼铁固氮酶、钒铁固氮酶、铁铁固氮酶以及在Strpomyces thermoauophicus内存在的与已知的三种固氮酶体系明显不同的固氮体系。     

Effect of dissolved oxygen on nitrogen fixation by A. vinelandii. I. Free cell cultures

As part of an investigation into the use of biological nitrogen fixation for fertilizer ammonia production, continuous culture studies of respiration and nitrogen fixation in the aerobic bacteria Azotobacter vinelandii under oxygen-limited conditions were conducted. Respiration and growth rates followed Monod forms with respect to dissolved oxygen concentration. However, specific nitrogen fixation rate and nitrogenase activity exhibited maximum values at dissolved oxygen concentrations of ca. 0.02 mM (10% of air saturation). These results suggest careful control of oxygen in the environment is necessary to optimize fixed nitrogen production by this organism.     

Hydrogen metabolism and energy costs of nitrogen fixation

Abstract The high energy costs of biological nitrogen fixation are partly caused by hydrogen production during the reduction of dinitrogen to ammonia. Some nitrogen-fixing organisms can recycle the evolved hydrogen via a membrane-bound uptake hydrogenase. The energetic aspects of hydrogen metabolism and nitrogen fixation are discussed.
Studies on both isolated nitrogenase proteins and nitrogen-fixing chemostat cultures show that energy limitation will result in a high hydrogen production by nitrogenase. In plant- Rhizobium symbiosis, the supply of oxygen or photosynthetate is the limiting factor for nitrogen fixation. In both cases, nitrogen fixation is energy-limited, and it is concluded that a large amount of hydrogen is produced during nitrogen fixation in these symbioses.
Hydrogen reoxidation yields less energy than the oxidation of endogenous substrates, and therefore expression of hydrogenase under oxygen-limited conditions is energetically unfavourable. Moreover, hydrogen reoxidation can never completely regain the energy invested during hydrogen production. The controversial reports of the effect of hydrogen reoxidation on the efficiency of nitrogen fixation are being discussed.
The determination of the energy costs of nitrogen fixation (expressed as the amount of ATP needed to fix 1 mol of N₂) using chemostat cultures is described. Calculations show that the nitrogenase-catalysed hydrogen production has more influence on the efficiency of nitrogen fixation than the absence or presence of a hydrogen uptake system.     

The natural history of nitrogen fixation

In recent years, our understanding of biological nitrogen fixation has been bolstered by a diverse array of scientific techniques. Still, the origin and extant distribution of nitrogen fixation has been perplexing from a phylogenetic perspective, largely because of factors that confound molecular phylogeny such as sequence divergence, paralogy, and horizontal gene transfer. Here, we make use of 110 publicly available complete genome sequences to understand how the core components of nitrogenase, including NifH, NifD, NifK, NifE, and NifN proteins, have evolved. These genes are universal in nitrogen fixing organisms-typically found within highly conserved operons-and, overall, have remarkably congruent phylogenetic histories. Additional clues to the early origins of this system are available from two distinct clades of nitrogenase paralogs: a group composed of genes essential to photosynthetic pigment biosynthesis and a group of uncharacterized genes present in methanogens and in some photosynthetic bacteria. We explore the complex genetic history of the nitrogenase family, which is replete with gene duplication, recruitment, fusion, and horizontal gene transfer and discuss these events in light of the hypothesized presence of nitrogenase in the last common ancestor of modern organisms, as well as the additional possibility that nitrogen fixation might have evolved later, perhaps in methanogenic archaea, and was subsequently transferred into the bacterial domain.     

固氮蓝细菌束毛藻生物固氮策略研究进展

固氮蓝细菌束毛藻(Tricodesmium)是海洋中丰度最高的固氮微生物,贡献了约42%的海洋生物固氮,为海洋生态系统提供了新的氮源,驱动海洋初级生产力和食物网,在海洋生物地球化学循环中发挥重要作用。作为海洋中“新氮”主要贡献者,束毛藻是一种不产生异形胞的丝状固氮蓝细菌。因为生物固氮的关键酶固氮酶对氧气十分敏感,一般固氮蓝细菌通常产生异形胞或采用夜间固氮的方式进行生物固氮,避免氧气对固氮酶的抑制作用。近年来研究发现,束毛藻具有一套独特的生物固氮体系,能够使同一藻丝在白天同时完成光合作用和生物固氮,并具有复杂的调控机制。本文综述了近年来束毛藻生物固氮策略的最新研究进展,介绍了其生物固氮和光合作用之间的精密调控机制,对拓展固氮微生物尤其是海洋蓝细菌固氮机制的认识具有借鉴意义。     

Variation in rates of nitrogen fixation in termites: response to dietary nitrogen in the field and laboratory

Abstract. Termites contribute nitrogen to their habitat through the nitrogenase activity of their bacterial symbionts. Previous studies indicate that high levels of dietary nitrogen suppress nitrogen fixation in termites. We examined the effects of dietary nitrogen on fixation rates in termites in both field and laboratory experiments. Ten field cplonies of Reticulitermes were collected and assayed for nitrogenase activity in July 1993, October 1993, January 1994, and April 1994. The nitrogen content of the wood collected with each colony was determined. There was no correlation between termite nitrogen fixation rates and the amount of nitrogen in their food for any of the four collection periods. In laboratory experiments, nitrogen fixation rates decreased when termites were fed filter paper treated with 2% and 5% ammonium nitrate or a 5% mixture of the amino acids proline, tryptophan and leucine, compared to water-treated controls. By contrast, the nitrogenase activity of termites fed filter paper treated with 2% and 5% ammonium phosphate, a mixture of the amino acids histidine, serine and aspartic acid, or 2% and 5% urea did not differ from the controls. However, nitrogenase activity increased when termites were fed with 2% uric acid. No clear association exists between termite nitrogen fixation and the nitrogen content of their food.     

以茎尖组培苗检测甘蔗体内内生固氮菌的固氮活性

以巴西引进的甘蔗品种‘B1’、‘B3’、‘B5’、‘B8’以及广西地区栽培品种‘新台糖16号’和‘桂糖11号’的茎尖进行组织培养,对其继代4次的茎尖组培苗进行有氮与无氮培养比较以及于温室中无氮盆栽品种比较试验的结果表明,经茎尖培养的甘蔗品种其原有的固氮能力不丧失,4个巴西甘蔗品种的固氮能力和在无氮下的生长能力强于广西主栽品种‘新台糖16号’和‘桂糖11号’,无氮培养的巴西甘蔗品种氮含量较高,固氮活性较强,缺氮症状出现时间较迟;无氮盆栽的巴西甘蔗品种株高、分蘖、叶片的固氮活性等均优于‘新台糖16号’和‘桂糖11号’。