大气CO2浓度升高对春玉米土壤呼吸的影响

为探讨春玉米不同生育期土壤呼吸速率对大气CO₂浓度升高的响应,以黄土高原旱作春玉米为研究对象,通过改进的开顶式气室（OTC）模拟大气CO₂浓度升高的环境,在田间条件下设置自然大气CO₂浓度（CK）、OTC对照（OTC,CO₂浓度同CK）与CO₂浓度升高（OTC+CO₂,OTC系统自动控制CO₂浓度700 μmol/mol）3种处理。研究了旱区覆膜高产栽培春玉米播前（V0）、六叶期（V6）、九叶期（V9）、吐丝期（R1）、乳熟期（R3）、蜡熟期（R5）及完熟期（R6）土壤呼吸速率对大气CO₂浓度升高的响应特征,以及大气CO₂浓度升高对土壤呼吸速率的温度与水分效应的影响。研究发现,OTC+CO₂处理土壤呼吸速率,与CK相比,在R3和R5期分别增加43%、104%（P<0.05）,与OTC相比,R3和R5期分别提升了63%、109%（P<0.05）;OTC处理与CK相比,在整个生育期对土壤呼吸影响不显著;3种处理条件下,土壤温度和水分随生育期变化趋势基本一致,土壤呼吸速率与土壤温度和水分分别呈指数相关和抛物线型相关;结果表明：大气CO₂浓度升高对土壤呼吸的影响因生育期而异,土壤温度和土壤水分是影响旱地农田土壤呼吸的重要因素,CO₂浓度升高会使土壤呼吸温度效应值（Q₁₀）降低,土壤呼吸对土壤水分响应的阈值提高。     

杉木人工林不同深度土壤CO2通量

土壤CO₂通量具有明显的时间和空间变异性。土壤温度和含水量是影响土壤CO₂通量的重要因素,同时,不同深度的土壤CO₂通量对温度和含水量变化的响应差异较大,因此,研究土壤CO₂通量和影响因素随土壤深度的变化,对于准确评估土壤碳排放具有重要意义。选择福建三明杉木人工林(Cunninghamia lanceolata)作为研究对象,利用非散射红外CO₂浓度探头和Li-8100开路式土壤碳通量系统,并使用Fick扩散法计算了0-60cm深度土壤CO₂的通量,结果表明:(1)5种扩散模型计算的表层(5cm)CO₂通量与Li-8100测量结果均具有显著相关性(P<0.01),Moldrup气体扩散模型计算结果较好。(2)土壤CO₂浓度随深度的增加而升高,但60cm深度以下土壤CO₂浓度开始降低;不同深度土壤CO₂浓度的日变化均呈现单峰型;0-60cm土壤CO₂通量日通量均值变化范围为0.54-2.17μmol m^-2 s^-1;(3)指数拟合分析显示,5、10cm和60cm深度处土壤CO₂通量与温度具有显著相关性,Q₁₀值分别为1.35、2.01和4.95。不同深度土壤含水量与CO₂通量的相关性不显著。     

甘肃马先蒿不同寄生密度对紫花针茅内生真菌共生体生理特性的影响

【目的】根寄生植物持续掠夺禾草体内营养物质成为禾草生长过程中的生物逆境,禾草内生真菌提高冷季型禾草对生物和非生物逆境耐受能力。然而,有关禾草内生真菌对根寄生逆境下禾草生理过程调控作用的研究鲜有报道。【方法】开展温室盆栽试验,以带菌(E+)和不带菌(E-)紫花针茅为研究对象,研究甘肃马先蒿不同寄生密度对紫花针茅抗氧化酶活性、渗透调节物质和根系活力影响的动态变化规律。【结果】甘肃马先蒿寄生显著增加紫花针茅抗氧化酶活性、丙二醛和脯氨酸含量,而根系活力却快速降低;高密度寄生紫花针茅植株生理特性指标显著高于低密度寄生或自然生长植株;同时,E+紫花针茅抗氧化酶活性、脯氨酸含量和根系活力显著高于E-植株,而E-植株丙二醛含量显著高于E+植株。【结论】禾草内生真菌通过增强抗氧化酶活性、调节细胞膜透性和增强根系生长能力的途径提高紫花针茅对根寄生逆境的耐受能力,利用植物替代方法带菌紫花针茅可以作为一种生物防治手段用于防控根寄生杂草。     

色季拉山急尖长苞冷杉林不同层次土壤CO2浓度对短时降雨的响应

为阐明不同层次土壤CO₂浓度日变化特征及对短时降雨的响应,以西藏东南部色季拉山急尖长苞冷杉（Abies georgei var. smithii）林为研究对象,在自然降雨条件下,分析短时降雨及水分再分布过程中各层次土壤CO₂浓度变化特征。结果表明：在0-60 cm层次内,土壤CO₂浓度随土壤层次的加深而显著增加（P < 0.01）,二者之间呈显著对数函数关系（R=0.9764,P < 0.01）;短时降雨脉冲使表层5 cm土壤CO₂浓度显著下降,而10 cm层次土壤CO₂浓度显著增加;在降雨和水分再分布阶段,5 cm与10 cm层次土壤CO₂浓度之间极显著负相关,10、20、40 cm和60 cm之间均呈极显著正相关（P < 0.01）;5 cm层次土壤含水量显著影响0-60 cm剖面CO₂浓度,降雨阶段,二者之间极显著线性正相关（P < 0.001）,而水分再分布阶段,二者之间符合极显著幂函数负相关（P < 0.001）。即降雨引起表层土壤含水量的快速增加,显著提高土壤剖面CO₂浓度,而降雨停止后,有利于土壤CO₂向土表的释放;土壤温度和含水量对CO₂浓度的影响效应在各层次之间表现不一致,除40 cm均为正效应外,其他各层均表现为相反的影响效应。这些结果表明,短时降雨使各层次土壤含水量增加,减少土壤表面CO₂释放量,使下层土壤体系中CO₂浓度升高,在分析土壤CO₂通量时间变化时,应考虑短时降雨对不同层次土壤CO₂的影响。     

甘肃马先蒿不同寄生密度对紫花针茅内生真菌共生体生理特性的影响北大核心CSCD

【目的】根寄生植物持续掠夺禾草体内营养物质成为禾草生长过程中的生物逆境,禾草内生真菌提高冷季型禾草对生物和非生物逆境耐受能力。然而,有关禾草内生真菌对根寄生逆境下禾草生理过程调控作用的研究鲜有报道。【方法】开展温室盆栽试验,以带菌(E+)和不带菌(E-)紫花针茅为研究对象,研究甘肃马先蒿不同寄生密度对紫花针茅抗氧化酶活性、渗透调节物质和根系活力影响的动态变化规律。【结果】甘肃马先蒿寄生显著增加紫花针茅抗氧化酶活性、丙二醛和脯氨酸含量,而根系活力却快速降低;高密度寄生紫花针茅植株生理特性指标显著高于低密度寄生或自然生长植株;同时,E+紫花针茅抗氧化酶活性、脯氨酸含量和根系活力显著高于E-植株,而E-植株丙二醛含量显著高于E+植株。【结论】禾草内生真菌通过增强抗氧化酶活性、调节细胞膜透性和增强根系生长能力的途径提高紫花针茅对根寄生逆境的耐受能力,利用植物替代方法带菌紫花针茅可以作为一种生物防治手段用于防控根寄生杂草。     

CO2浓度升高对龙血树和春羽生长及光合生理的影响

采用开顶式气室（OTC）控制模拟环境,研究了二氧化碳（CO₂）浓度升高对龙血树和春羽叶片光合生理及叶绿素荧光参数的影响。结果表明:（1）随着CO₂浓度增加,龙血树和春羽幼苗叶面积和株高均呈显著增加趋势。（2）龙血树和春羽幼苗叶片净光合速率随着CO₂浓度的升高均呈先增加后降低的趋势,而同期蒸腾速率和气孔导度显著降低。（3）随着CO₂浓度的增加,龙血树和春羽幼苗叶片最大光量子产量、实际光能转化效率均呈先升后降的趋势,而同期非光化学猝灭系数呈先降低后升高的趋势;光化学猝灭系数总体上也呈先升高后降低的趋势,但差异不显著。研究表明,一定时间的高浓度CO₂处理提高了龙血树和春羽的净光合速率,促进了植物的生长,但随着处理时间的延长,这种促进作用逐渐降低进而消失,并以春羽幼苗表现得更突出;即高浓度CO₂可能破坏了龙血树及春羽生长后期光系统Ⅱ反应中心结构,导致叶片光合能力降低。     

地质封存CO2泄漏对玉米根系形态的影响

碳捕集与封存（Carbon Capture and Storage,CCS）是应对全球气候变化、实现煤炭清洁利用的有效手段之一,但是地质封存的CO₂存在泄漏的风险,可能对农田生态系统产生重大威胁,影响我国粮食安全。根系生长是地上部和地下部相互作用、相互促进的统一过程,其形态特征对作物生产力有显著影响,但CCS泄漏对植物根系的影响评估尚不多见。本文以玉米为研究对象,采用盆栽底部通入CO₂的方法模拟不同CO₂泄漏情景,研究CK（0 g m^-2 d^-1）和G1000（1000 g m^-2 d^-1）和G2000（2000 g m^-2 d^-1）三种泄漏情景下CO₂对玉米根系形态的影响。结果表明：CO₂泄漏对玉米根系形态有明显的影响,随着泄漏量的增大总根长从40290.81 cm减少至21448.18 cm,减少46.77%,其中细根大幅减少;CO₂泄漏造成玉米明显减产,最大减产率达26.64%;玉米的地上部生物量较地下部生物量对CO₂泄漏更加敏感。综合来看,随着CO₂泄漏量增大,对玉米根的生长、地上部生物量、地下部生物量以及产量有显著的抑制作用。作物根系形态对封存CO₂泄漏的响应可为CCS泄漏监测和生态修复提供系统科学依据。     

大气CO2浓度升高对B型烟粉虱大小、酶活及其寄主的选择性影响

以CO₂浓度升高为作用因子,观测了连续3代饲养在转Bt基因棉(GK-12)和亲本棉泗棉3号(S3)上B型烟粉虱的个体大小、解毒酶活性及第3代烟粉虱的选择性和产卵量。结果表明,CO₂浓度和棉花品种处理对第2、3代烟粉虱卵长、伪蛹大小、雌雄成虫大小的影响均不显著,但CO₂浓度升高使第1代烟粉虱的伪蛹长、雌虫翅展和雌、雄虫长显著低于对照CO₂浓度下生长的烟粉虱;取食S3的伪蛹长、雌虫长和雌虫翅展分别比相应的对照浓度下的低2.81%、2.95%、0.94%,取食GK-12的雌虫翅展和雄虫长均比相应的对照浓度下的低2.08%和2.58%。3种解毒酶的活性测定显示,取食高CO₂浓度处理的转基因棉GK-12上的F3代烟粉虱的谷胱甘肽S转移酶(Glutathione Stransferase, 简称GSTs)活性和取食亲本棉S3的F1代的乙酰胆碱酯酶(Acetylcholinesterase;简称AchE)的活性分别比对照处理的增加45.73%和27.68%;而羧酸酯酶(Carboxylic Ester hydrolase;简称 CarE)在取食4个处理的棉花上烟粉虱的活性均无显著的差异;而取食对照CO₂条件下GK-12棉花的F1代烟粉虱GSTs活性比取食S3棉花低35.12%。对3个因子的交互分析结果显示,C0₂浓度对3种酶活影响均不显著,品种对GSTs的影响显著,世代、CO₂×品种及CO₂×品种×世代间的交互作用对AchE酶活影响显著,各因子对CarE的活性影响均不显著。烟粉虱的选择实验结果表明,4种处理的F3代B型烟粉虱均喜好选择在高CO₂处理的棉花,而且选择高CO₂处理的GK-12的数量多于其他3个处理,其中对照浓度下生长的烟粉虱选择高CO₂的GK-12比高CO₂处理的S3、对照浓度处理的GK-12 和S3分别增加9.175%, 19.89%、27.93%; 而高浓度下生长的烟粉虱选择高CO₂的GK-12比高CO₂处理的S3、对照浓度处理的GK-12 和S3分别增加12.56%,21.05%,28.73%。72h的产卵量检测发现,烟粉虱在高CO₂处理的2种棉花上的产卵量也是多于对照条件下的棉花,对照条件下的棉花相比,高CO₂和对照浓度下生长的烟粉虱的在高CO₂处理的2种棉花上的产卵量分别增加24.55%和19.03%;在高CO₂浓度下生长的F3代烟粉虱更喜好在高CO₂浓度处理的GK-12上产卵,与对照浓度下的GK-12相比增加26.93%,差异达到显著水平。     

水葱和香蒲叶经济性状对模拟增温和CO2浓度倍增的响应

气候变化是国际社会共同关注的环境问题,植物对气候变化的响应反映了植物应对气候变化的生长和生存策略。叶经济性状与植物对资源的获取、利用和储存直接相关,并且受到温度条件和CO₂浓度的显著影响。该文采用人工环境控制系统封顶式生长室研究广布湿地植物水葱(Scirpus validus)和香蒲(Typha orientalis)的叶经济性状对模拟增温(现行环境温度+2 ℃)和CO₂浓度倍增(增至850 μmol·mol^-1)的响应。结果表明:(1)增温处理下,水葱净光合速率、氮含量和磷含量显著降低,但其胞间CO₂浓度和比叶重显著增加; CO₂浓度倍增处理下,水葱胞间CO₂浓度和净光合速率均显著降低,但比叶重显著增加。(2)增温处理下香蒲的比叶重显著增加,而氮含量和磷含量显著降低; 香蒲的光合参数、氮含量和磷含量在CO₂浓度倍增处理下均显著降低,而比叶重显著增加。(3)水葱的比叶重、氮含量、磷含量、净光合速率、气孔导度、胞间CO₂浓度与主成分分析的两个环境变量相关; 而香蒲的经济性状均与两个环境变量相关,表明这些经济性状在香蒲响应增温和CO₂浓度变化过程中发挥重要作用。(4)除碳含量外,水葱和香蒲的其他经济性状参数包括净光合速率、气孔导度、蒸腾速率、胞间CO₂浓度、氮含量、磷含量和比叶重均在响应增温和CO₂浓度倍增过程中发挥重要作用。总体而言,该研究结果反映了水葱和香蒲在功能性状上对增温和CO₂浓度倍增的响应策略。两种植物的光合能力和养分含量在两种处理下虽然均受到显著的抑制作用,但是其抗逆能力升高,表明增温和CO₂浓度升高不利于水葱和香蒲的生长。     

预处理方式对香蒲和芦苇种子萌发的影响

有性繁殖是植物种群形成与维持的主要方式。为探索退化湿地的快速恢复方法,为松花江下游退化湿地恢复提供科学依据,本研究开展了香蒲和芦苇种子快速发芽的有性繁殖实验。研究采用滤纸为发芽基质,通过变温培养试验,以未浸种处理为参照,分析了蒸馏水、双氧水(H₂O₂)、硝酸钾(KNO₃)和高锰酸钾(KMnO₄)溶液浸种的预处理方式对香蒲、芦苇种子发芽率和发芽速率的影响。结果表明:不同预处理方式对香蒲、芦苇种子的发芽率和发芽速率均具有显著影响。KMnO₄溶液浸种再清洗处理条件下,香蒲种子发芽率和发芽速率均显著高于其他处理,平均发芽率可达未浸种处理条件下的3.1倍,发芽速率为16.17±0.80。芦苇种子的发芽率和发芽速率在经KNO₃溶液浸泡再清洗处理后效果最佳,种子发芽率达96%-99%,发芽速率达28.43±0.71。因此,分别对香蒲、芦苇种子采用KMnO₄和KNO₃溶液浸泡再清洗的预处理方式可以缩短出苗时间,提高发芽率,从而加速湿地植被恢复进程。     

Interconversions of different forms of vitamin B6 in tobacco plants

There are six different vitamin B₆ (VB₆) forms, pyridoxal (PL), pyridoxamine (PM), pyridoxine (PN), pyridoxal 5′-phosphate (PLP), pyridoxamine 5′-phosphate (PMP), and pyridoxine 5′-phosphate (PNP), of which PLP is the active form. Although plants are a major source of VB₆ in the human diet, and VB₆ plays an important role in plants, the mechanisms underlying the interconversions of different VB₆ forms are not well understood. In this study, in vitro tobacco plants were grown on Murashige and Skoog (MS) basal media supplemented with 100 mg/L of PM, PL or PN and the abundance of the different B₆ vitamers in leaf tissue was quantified by high performance liquid chromatography (HPLC). The total amount of VB₆ was about 3.9 μg/g fresh weight of which PL, PM, PN, PLP and PMP accounted for 23%, 14%, 37%, 20% and 6%, respectively. Tobacco plants contained a trace amount of PNP. Supplementation of the culture medium with any of the non-phosphorylated vitamers resulted in an increase in total VB₆ by about 10-fold, but had very little impact on the concentrations of the endogenous phosphorylated vitamers. Administration of either PM or PN increased their endogenous levels more than the levels of any other endogenous B₆ vitamers. PL supplementation increased the levels of plant PN and PM significantly, but not that of PL, suggesting that efficient conversion pathways from PL to PN and PM are present in tobacco. Additionally, maintenance of a stable level of PLP in the plant is not well-correlated to changes in levels of non-phosphorylated forms.     

Light- and seasonal-induced plasticity in leaf morphology, N partitioning and photosynthetic capacity of two temperate deciduous species

Processes involved in leaf photosynthetic acclimation to light and throughout the growing season were investigated in two hardwood species (Acer saccharum and Betula alleghaniensis), which differed in their level of shade-tolerance. For both species, variation in traits related to (i) leaf morphology (LMA, leaf mass:area ratio), (ii) leaf N content (N_A, leaf nitrogen content on an area basis and N_M, N concentration in leaf dry mass), (iii) leaf N partitioning among photosynthetic functions (P_r, N allocated to Rubisco, and P_b, N allocated to bioenergetics), and (iv) leaf photosynthetic capacity (V_cmax, maximal carboxylation rates, and J_max, maximal light-driven electron flow) were assessed at three different times during the growing season (early, mid- and late summer) and under four contrasting light regimes (40, 17, 6 and 2% of full sunlight). For both species, light-driven variation in most traits was greater than their seasonally driven variation. Furthermore, results showed for both species the pre-eminence of LMA changes in the light-driven acclimation of N_A. Importance of N_M to variation in N_A was restricted to seasonal acclimation, especially for the less shade-tolerant species, B. alleghaniensis. Similarly, for both species, light-driven acclimation of leaf photosynthetic capacities was tightly related to variation in N_A, which was related to LMA changes. However, variation in P_r and P_b better explained seasonally driven variation in V_cmax and J_max, specifically under lower light levels, where N_A was low. Thus, the great variability observed for leaf activity in response to contrasting light environments was related to efficient morphological adjustments, regardless of species level of shade-tolerance. Finally, physiological adjustments were mainly involved in fine-scale changes observed during seasonally driven acclimation of leaves, when LMA was constrained to a slight range of variation.     

Selection of a highly virulent fungal isolate, Metarhizium anisopliae CLO 53, for controlling Hoplia philanthus

The June beetle, Hoplia philanthus Füessly (Coleoptera: Scarabaeidae), has become a widespread and destructive insect pest of lawns, sport turf, pastures, and horticultural crops in Belgium. The virulence of 34 entomopathogenic fungal isolates from the genera Metarhizium, Beauveria, and Paecilomyces to third-instar H. philanthus was tested in bioassays by dipping larvae in 10(7)conidia/ml suspensions. Two isolates of Metarhizium anisopliae (CLO 53 and CLO 54) caused maximally 90% mortality 10 weeks post-inoculation while other isolates only caused mortalities between 10 and 62%. The virulence of M. anisopliae CLO 53 was further tested by exposing H. philanthus larvae to conidial serial concentrations of 10(4)-10(9)conidia/g sandy soil for up to 11 weeks at 15, 20 or 25 degrees C. Mortality was dependant on the fungal concentration, exposure time, and temperature. Eleven weeks after inoculation, the LC50 values for this isolate ranged from 1.3 to 4.0 x 10(6), 1.0 to 3.2 x 10(5), and 2.5 x 10(4) to 10(5)conidia/g soil at 15, 20, and 25 degrees C, respectively. The LT50 values for this isolate ranged from 3.5 to 21.7, 2.4 to 18.7, and 2.9 to 16.1 weeks at concentrations of 10(9) and 10(4)conidia/g soil at 15, 20, and 25 degrees C, respectively. In glasshouse pot experiment with perennial ryegrass (Lolium perenne L.), the isolate CLO 53 caused mortalities of 50 and 88% of H. philanthus larvae 10 weeks after application of 10(4) and 10(6)conidia/cm(2) soil surface, respectively. The present results suggest that the Belgian isolate CLO 53 has excellent potential for biological control of H. philanthus.     

Two groups of entomopathogenic bacteria, Photorhabdus and Xenorhabdus, share an inhibitory action against phospholipase A2 to induce host immunodepression

Photorhabdus and Xenorhabdus are two genera of entomopathogenic bacteria having a mutualistic relationship with their respective nematode hosts, Heterorhabditis and Steinernema. One of the pathogenic mechanisms of these bacteria includes host immunodepression, which leads to lethal septicemia. It has been known that X. nematophila inhibits phospholipase A2 (PLA2) to induce host immunodepression. Here, we tested the hypothesis of PLA2 inhibition using another bacterial species involved in other genera. P. temperata subsp. temperata is the intestinal symbiont of an entomopathogenic nematode, H. megidis. The bacteria caused potent pathogenicity in a dose-dependent manner against the fifth instar larvae of a test target insect, Spodoptera exigua, as early as 24 h after the intra-hemocoelic injection. In response to the live bacterial injection, hemocyte nodulation (a cellular immune response) and prophenoloxidase (pPO) activation were inhibited, while the injection of heat-killed bacteria significantly induced both immune reactions. The immunodepression induced by the live bacteria was reversed by the addition of arachidonic acid, the catalytic product of phospholipase A2. In contrast, the addition of dexamethasone, a specific PLA2 inhibitor to the heat-killed bacterial treatment, inhibited both immune capacities. In addition to a previously known PLA2 inhibitory action of X. nematophila, the inhibition of P. temperata temperata on PLA2 suggests that bacteria symbiotic to entomopathogenic nematodes share a common pathogenic target to result in an immunodepressive state of the infected insects. To prove this generalized hypothesis, we used other bacterial species (X. bovienni, X. poinarii, and P. luminescens) involved in these two genera. All our experiments clearly showed that these other bacteria also share their inhibitory action against PLA2 to induce host immunodepression.     

The lethal and sub-lethal consequences of entomopathogenic nematode infestation and exposure for adult pine weevils, Hylobius abietis (Coleoptera: Curculionidae)

Entomopathogenic nematodes (EPN) frequently kill their host within 1-2 days, and interest in EPN focuses mainly on their lethality. However, insects may take longer to die, or may fail to die despite being infected, but little is known about the effects of EPN infection on insects, other than death. Here we investigate both lethal and sub-lethal effects of infection by two EPN species, Steinernema carpocapsae and Heterorhabditis downesi, on adults of the large pine weevil, Hylobius abietis. Following 12 h nematode-weevil contact in peat, S. carpocapsae killed a significantly higher proportion of weevils (87-93%) than H. downesi (43-57%) at all concentrations tested. Less than 10% of weevils were dead within 2 days, and weevils continued to die for up to 10 days after exposure (LT₅₀ of 3 days or more). In a separate experiment, live weevils dissected 6 days after a 24 h exposure to nematodes on filter paper harbored encapsulated and dead nematodes, showing that weevils could defend themselves against infection. Some live weevils also harbored live nematodes 6 days after they had been removed from the nematode infested medium. Feeding by weevils was not affected by infection with, or exposure to, either species of EPN. We discuss these results in relation to the use of EPN in biological control against H. abietis.     

Mechanisms of enhanced cellulosic bioethanol fermentation by co-cultivation of Clostridium and Thermoanaerobacter spp

Engineering microbial consortia capable of efficient ethanolic fermentation of cellulose is a strategy for the development of consolidated bioprocessing for bioethanol production. Co-cultures of cellulolytic Clostridium thermocellum with non-cellulolytic Thermoanaerobacter strains (X514 and 39E) significantly improved ethanol production by 194-440%. Strain X514 enhanced ethanolic fermentation much more effectively than strain 39E in co-cultivation, with ethanol production in X514 co-cultures at least 62% higher than that of 39E co-cultures. Comparative genome sequence analysis revealed that the higher ethanolic fermentation efficiency in strain X514 was associated with the presence of a complete vitamin B(12) biosynthesis pathway, which is incomplete in strain 39E. The significance of the vitamin B(12)de novo biosynthesis capacity was further supported by the observation of improved ethanol production in strain 39E by 203% following the addition of exogenous vitamin B(12). The vitamin B(12) biosynthesis pathway provides a valuable biomarker for selecting metabolically robust strains for bioethanol production.     

Thermodynamic and kinetic characterisation of individual haems in multicentre cytochromes c3

The characterisation of individual centres in multihaem proteins is difficult due to the similarities in the redox and spectroscopic properties of the centres. NMR has been used successfully to distinguish redox centres and allow the determination of the microscopic thermodynamic parameters in several multihaem cytochromes c₃ isolated from different sulphate-reducing bacteria. In this article we show that it is also possible to discriminate the kinetic properties of individual centres in multihaem proteins, if the complete microscopic thermodynamic characterisation is available and the system displays fast intramolecular equilibration in the time scale of the kinetic experiment. The deconvolution of the kinetic traces using a model of thermodynamic control provides a reference rate constant for each haem that does not depend on driving force and can be related to structural factors. The thermodynamic characterisation of three tetrahaem cytochromes and their kinetics of reduction by sodium dithionite are reported in this paper. Thermodynamic and kinetic data were fitted simultaneously to a model to obtain microscopic reduction potentials, haem-haem and haem-proton interacting potentials, and reference rate constants for the haems. The kinetic information obtained for these cytochromes and recently published data for other multihaem cytochromes is discussed with respect to the structural factors that determine the reference rates. The accessibility for the reducing agent seems to play an important role in controlling the kinetic rates, although is clearly not the only factor.     

Structure/function studies on a S-adenosyl-L-methionine-dependent uroporphyrinogen III C methyltransferase (SUMT), a key regulatory enzyme of tetrapyrrole biosynthesis

The crystallographic structure of the Pseudomonas denitrificans S-adenosyl-L-methionine-dependent uroporphyrinogen III methyltransferase (SUMT), which is encoded by the cobA gene, has been solved by molecular replacement to 2.7A resolution. SUMT is a branchpoint enzyme that plays a key role in the biosynthesis of modified tetrapyrroles by controlling flux to compounds such as vitamin B(12) and sirohaem, and catalysing the transformation of uroporphyrinogen III into precorrin-2. The overall topology of the enzyme is similar to that of the SUMT module of sirohaem synthase (CysG) and the cobalt-precorrin-4 methyltransferase CbiF and, as with the latter structures, SUMT has the product S-adenosyl-L-homocysteine bound in the crystal. The roles of a number of residues within the SUMT structure are discussed with respect to their conservation either across the broader family of cobalamin biosynthetic methyltransferases or within the sub-group of SUMT members. The D47N, L49A, F106A, T130A, Y183A and M184A variants of SUMT were generated by mutagenesis of the cobA gene, and tested for SAM binding and enzymatic activity. Of these variants, only D47N and L49A bound the co-substrate S-adenosyl-L-methionine. Consequently, all the mutants were severely restricted in their capacity to synthesise precorrin-2, although both the D47N and L49A variants produced significant quantities of precorrin-1, the monomethylated derivative of uroporphyrinogen III. The activity of these variants is interpreted with respect to the structure of the enzyme.     

Detection of metallo-β-lactamase genes in clinical specimens by a commercial multiplex PCR system

hyplex®-MBL ID Multiplex PCR-ELISA, a novel method for identifying metallo-β-lactamase genes directly in clinical specimens, was evaluated using a consecutive collection of 326 samples from three hospitals in Greece characterized by high prevalence of VIM producers. The method exhibited high sensitivity (98.0%) and specificity (98.6%) and was proven reliable in detecting bla_VIM genes in blood, urine, pus, and sputum samples that, as confirmed by conventional methods, contained various VIM-producing species. Future multicenter studies should be considered for the thorough evaluation of this method and its potential diagnostic utility.     

Tuning of functional heme reduction potentials in Shewanella fumarate reductases

The fumarate reductases from S. frigidimarina NCIMB400 and S. oneidensis MR-1 are soluble and monomeric enzymes located in the periplasm of these bacteria. These proteins display two redox active domains, one containing four c-type hemes and another containing FAD at the catalytic site. This arrangement of single-electron redox co-factors leading to multiple-electron active sites is widespread in respiratory enzymes. To investigate the properties that allow a chain of single-electron co-factors to sustain the activity of a multi-electron catalytic site, redox titrations followed by NMR and visible spectroscopies were applied to determine the microscopic thermodynamic parameters of the hemes. The results show that the redox behaviour of these fumarate reductases is similar and dominated by a strong interaction between hemes II and III. This interaction facilitates a sequential transfer of two electrons from the heme domain to FAD via heme IV.