I. Pathogenic variation in fungi and bacteria and mycorrhizal compatibility: Pathogenic variation in Verticillium albo-atrum isolates from hop (Humulus lupulus)

Non-lethal (M strain) ‘fluctuating’ wilt was first identified in hops in 1924. Lethal (V strain) ‘progressive’ wilt appeared in the 1930s and spread rapidly through Kent, destroying all commercial varieties. By 1965 the disease was largely controlled by use of new resistant cultivars, accompanied by restricted nitrogen fertilisers, non-cultivation (with weed control by overall herbicides), and high standards of field hygiene. Between 1968 and 1971 severe wilt developed in previously highly resistant clones, used as ‘controls’ in routine field resistance tests. Verticillium isolates from the two farms supplying the natural inoculum for these tests during 1968–71 were more virulent than previously encountered V strains, some causing severe wilt in all resistant cultivars. Subsequent surveys supported by pathogenicity tests against resistant cultivars indicated the existence of two ‘super virulent’ strains on eight farms in Kent. The appearance of mild (M) strains in the 1920s, virulent (VI) strains in the 1930s, and ‘super-virulent’ (V2 and V3) strains in the 1960s follow 400 years of apparent freedom from wilt. This suggests that the initial breakage of resistance barriers was critical and subsequent variations in an apparently complex, polygenically controlled resistance system are rapidly being matched by the pathogen.     

Detection of chitinase activity after polyacrylamide gel electrophoresis

Commercial Streptomyces griseus and Serratia marcescens chitinases and purified wheat germ W1A and hen egg white lysozymes were subjected to polyacrylamide gel electrophoresis under native conditions at pH 4.3. After electrophoresis, an overlay gel containing 0.01% (W/V) glycol chitin as substrate was incubated in contact with the separation gel. Lytic zones were revealed by uv illumination with a transilluminator after staining for 5 min with 0.01% (W/V) Calcofluor white M2R. As low as 500 ng of purified hen egg lysozyme could be detected after 1 h incubation at 37 degrees C. One band was observed with W1A lysozyme and several bands with the commercial microbial chitinases. The same system was also used with native polyacrylamide gel electrophoresis at pH 8.9. Several bands were detected with the microbial chitinases. The same enzymes were also subjected to denaturing polyacrylamide gel electrophoresis in gradient gels containing 0.01% (W/V) glycol chitin. After electrophoresis, enzymes were renatured in buffered 1% (V/V) purified Triton X-100. Lytic zones were revealed by uv after staining with Calcofluor white M2R as for native gels. The molecular weights of chitinolytic enzymes could thus be directly estimated. In denaturing gels, as low as 10 ng of purified hen egg white lysozyme could be detected after 2 h incubation at 37 degrees C. Estimated molecular weights of St. griseus and Se. marcescens were between 24,000 and 72,000 and between 40,500 and 73,000, respectively. Some microbial chitinases were only resistant to denaturation with sodium dodecyl sulfate while others were resistant to sodium dodecyl sulfate and beta-mercaptoethanol.     

Inhibition of microbial arsenate reduction by phosphate

The ratio of arsenite (As(III)) to arsenate (As(V)) in soils and natural waters is often controlled by the activity of As-transforming microorganisms. Phosphate is a chemical analog to As(V) and, consequently, may competitively inhibit microbial uptake and enzymatic binding of As(V), thus preventing its reduction to the more toxic, mobile, and bioavailable form - As(III). Five As-transforming bacteria isolated either from As-treated soil columns or from As-impacted soils were used to evaluate the effects of phosphate on As(V) reduction and As(III) oxidation. Cultures were initially spiked with various P:As ratios, incubated for approximately 48 h, and analyzed periodically for As(V) and As(III) concentration. Arsenate reduction was inhibited at high P:As ratios and completely suppressed at elevated levels of phosphate (500 and 1,000 μM; P inhibition constant (K(i))～20-100 μM). While high P:As ratios effectively shut down microbial As(V) reduction, the expression of the arsenate reductase gene (arsC) was not inhibited under these conditions in the As(V)-reducing isolate, Agrobacterium tumefaciens str. 5B. Further, high phosphate ameliorated As(V)-induced cell growth inhibition caused by high (1mM) As pressure. These results indicate that phosphate may inhibit As(V) reduction by impeding As(V) uptake by the cell via phosphate transport systems or by competitively binding to the active site of ArsC.     

Grass invasion of a hardwood forest is associated with declines in belowground carbon pools

Invasive plant species affect a range of ecosystem processes but their impact on belowground carbon (C) pools is relatively unexplored. This is particularly true for grass invasions of forested ecosystems. Such invasions may alter both the quantity and quality of forest floor inputs. Dependent on both, two theories, ‘priming’ and ‘preferential substrate utilization’, suggest these changes may decrease, increase, or leave unchanged native plant‐derived soil C. Decreases are expected under ‘priming’ theory due to increased soil microbial activity. Under ‘preferential substrate utilization’, either an increase or no change is expected because the invasive plant's inputs are used by the microbial community instead of soil C. Here, we examine how Microstegium vimineum affects belowground C‐cycling in a southeastern US forest. Following predictions of priming theory, M. vimineum's presence is associated with decreases in native‐derived, C pools. For example, in September 2006 M. vimineum is associated with 24%, 34%, 36%, and 72% declines in total organic, particulate organic matter, mineralizable (a measure of microbially‐available C), and microbial biomass C, respectively. Soil C derived from M. vimineum does not compensate for these decreases, meaning that the sum of native‐ plus invasive‐derived C pools is smaller than native‐derived pools in uninvaded plots. Supporting our inferences that C‐cycling accelerates under invasion, the microbial community is more active per unit biomass: added ¹³C‐glucose is respired more rapidly in invaded plots. Our work suggests that this invader may accelerate C‐cycling in forest soils and deplete C stocks. The paucity of studies investigating impacts of grass invasion on C‐cycling in forests highlights the need to study further M. vimineum and other invasive grasses to assess their impacts on C sink strength and forest fertility.     

Culturable Heterotrophic Bacteria Associated with Healthy and Bleached Scleractinian Madracis decactis and the Fireworm Hermodice carunculata from the Remote St. Peter and St. Paul Archipelago, Brazil

We report on the first characterization of the culturable heterotrophic bacteria of the scleractinian Madracis decactis. In addition, we characterized the culturable bacteria associated with the fireworm Hermodice carunculata, observed predating partially bleached coral colonies. Our study was carried out in the remote St. Peter and St. Paul Archipelago (SPSPA), Mid-Atlantic Ridge, Brazil. We constituted a 403 isolates collection and subsequently characterized it by means of pyrH and 16S rRNA partial sequences. We identified Photobacterium, Bacillus, and Vibrio species as members of the culturable microbiota of healthy M. decactis. V. campbellii, V. harveyi, V. communis, and V. maritimus were the most commonly found Vibrio species in healthy corals, representing more than 60 % of all vibrio isolates. Most of the vibrios isolated from the fireworm’s tissues (n = 143; >90 %) were identified as V. shiloi. However, we did not recover V. shiloi from bleached M. decactis. Instead, we isolated V. communis, a novel Photobacterium species, Bacillus, Kocuria, and Pseudovibrio, suggesting a possible role of other facultative anaerobic bacteria and/or environmental features (such as water quality) in the onset of bleaching in SPSPA’s M. decactis.     

The frequency of Familial Mediterranean fever gene mutations and genotypes at Kirikkale and comparison with the mean of regional MEFV mutation frequency of Turkey

In this study we have retrospectively analysed the mutation spectrum of the 351 Familial Mediterranean fever patients referred to K?r?kkale University Faculty of Medicine, Department of Medical Genetics Laboratory over a period of 5 years and compared them with Turkey’s mean. We have found 11 different mutations, including rare mutations such as F479L, K695R, M680I(G/A) and 45 different genotypes showing the heterogeneity of MEFV mutations in Central Anatolia. The most three prevalent mutations were M694V (14.8 %), E148Q (7.1 %) and M680I(G/C) (4.1 %) in accordance with the literature. We have also investigated R202Q in our routine molecular diagnosis. Mutation causing R202Q (c.605G > A) change was described as a frequent polymorphism and G allele was found in linkage disequilibrium (LD) with M694V. There are limited number of studies investigating R202Q, some of them implicate that its homozygote state is disease causing. We showed the high frequency of R202Q (23.7 %) with and without M694V in all the groups analysed and its high LD rate with M694V in the diagnosed group. Our study is reflecting the mutational heterogeneity of MEFV and summarize mutational spectrum of Turkey’s geographical regions and overall Turkey.     

Vetch aphid, <Emphasis Type="Italic">Megoura crassicauda</Emphasis> (Hemiptera: Aphididae), parasitism does not reduce the bean production of narrow-leaved vetch, <Emphasis Type="Italic">Vicia sativa</Emphasis> subsp. <Emphasis Type="Italic">nigra</Emphasis> (Fabaceae)

Megoura crassicauda Mordvilko (Hemiptera: Aphididae) is a dominant aphid species found on Vicia sativa subsp. nigra (L.) Ehrh. (Fabaceae) in the spring. Worker ants of Formica japonica, the dominant ant species attracted to the extrafloral nectaries of V. s. nigra, often attack ladybirds (Coccinella septempunctata), which are aphid enemies. However, the workers of F. japonica do not attack or exclude M. crassicauda, the non-myrmecophilous aphid. It appears that the “bodyguard” retained by the plant guards the plant’s herbivore by attacking the herbivores’ enemies, rather than guarding the plant itself. The relationship between V. s. nigra and M. crassicauda was observed in the field to examine and evaluate the cost of parasitism. Parasitism by M. crassicauda delayed flower bud formation markedly in V. s. nigra but did not kill the plants. V. s. nigra plants that were parasitized showed a net bean production similar to that of the non-parasitized controls. The parasitism rate of M. crassicauda increased when extrafloral nectaries were used by F. japonica. These results may indicate that M. crassicauda provides V. s. nigra with benefits by preventing other serious disadvantages.     

川西亚高山不同林龄云杉人工林土壤微生物群落结构

以川西亚高山云杉人工林林地土壤为对象,采用磷脂脂肪酸(PLFA)法研究了4种不同林龄(50、38、27和20年)的人工林土壤微生物多样性和群落结构特征.结果表明: 随着林龄的增加,土壤有机碳和全氮含量逐步增加;土壤微生物Shannon多样性和Pielou均匀度指数则呈现先增后减的趋势.土壤微生物总PLFAs量、细菌PLFAs量、真菌PLFAs量、放线菌PLFAs量以及丛枝菌根真菌PLFAs量均表现为随林龄的增加而增加.主成分分析(PCA)表明,不同林龄人工林的土壤微生物群落结构之间存在显著差异,其中,第1主成分(PC1)和第2主成分(PC2)共同解释了土壤微生物群落结构总变异的66.8%.冗余分析(RDA)表明,对土壤微生物群落结构产生显著影响的环境因子分别为土壤有机碳、全氮、全钾以及细根生物量.随着人工造林时间的延长,土壤肥力和微生物生物量增加,森林生态系统的恢复进程稳定.     

On the movement and alignment of DNA during 120 degrees pulsed-field gel electrophoresis.

The displacement per pulse of lambda, T4, and G DNA during pulsed-field agarose gel electrophoresis has been measured for a fine mesh of pulse durations T between 0.02 and 120 s. The slopes of these curves show that the DNA moves by two distinct processes, designated 1 and 2, depending upon the pulse duration T. Process 1 operates at short T and causes dx/dT to decrease gradually with increasing T. This process is independent of molecular weight M. Process 2 is effective at longer T and causes dx/dT to rise sharply in sigmoidal fashion at a value of T which increases as M1.2, finally reaching a plateau of 1.4 microns/s for E = 4 V/cm. The shape of the dx/dT curve and its dependence on M lead directly to 4 zones of separation in plots of mobility vs M for different T. The alignment of the 3 DNAs during PFGE was measured by fluorescence-detected linear dichroism for E between 4 and 10 V/cm. These results are used in developing a molecular understanding of the mobility data.     

Allelic exclusion of a T cell receptor-beta minilocus.

A TCR-beta minilocus in germline configuration (beta M) has previously been shown to undergo rearrangement and expression in transgenic mice. To study allelic exclusion of TCR miniloci, several beta M transgenic mouse lines were generated and crossed with mice transgenic for a functionally rearranged TCR V beta 2 gene (beta R). PCR analysis of beta M beta R double transgenic mice revealed almost complete suppression of endogenous TCR V beta gene rearrangements, but blockage of minilocus V beta rearrangements was achieved with only one of five minilocus transgenic lines. This result cannot be explained by copy number or arrangement of the multiple miniloci integrated. It appears that the minilocus is not autonomously regulated which suggests that sequences flanking the integration sites influence accessibility of the minilocus for rearrangement and allelic exclusion. However, although productively rearranged genes were formed in double transgenic mice, surface expression of minilocus-encoded beta chains was not detected. This indicates that allelic exclusion may operate at a level after gene rearrangement.     

不同施肥黑土微生物量氮变化特征及相关因素

研究长期施用两种不同量有机肥(M2、M4)和化肥(NPK)的黑土微生物量N在作物生长季的变化特征．结果表明,施用有机肥黑土微生物量N显著高于施用化肥(NPK)和不施肥(CK),微生物量N季节波动小．微生物量N为M4 25．52～239．12mg·kg^-1,M2 10．40～94．31mg·kg^-1．NPK6．27～87．04mg·kg^-1,CK9．15～69．81mg·kg^-1,同一处理最大值与最小值相差7～14倍．M2、NPK处理微生物量N最大值出现在抽雄吐丝期,M4处理最大值出现在拔节期,CK处理最大值出现在播种期;不同处理微生物量N的差异并未因季节变化及玉米生育时期影响而明显改变．微生物量N的动态变化与极少数黑土生物、理化特性指标动态变化显著相关;微生物量N与黑土生物、理化特性,植物氮、磷、钾有极显著的正相关关系,与土壤含水量、籽粒粗蛋白含量呈显著正相关关系．     

Professor Natal’ya N. Traugott’s Contribution to Studies on the Development of Central Speech Mechanisms

The name of Natal’ya Nikolaevna Traugott is closely associated with the history of human higher nervous activity studies in Russia. Destiny gifted to N.N. Traugott the opportunity to communicate with many outstanding scientists; she regularly attended the “Pavlov Wednesdays”, took part in V.M. Bechterev’s clinical discussions, visited L.S. Vygotskii’s lectures on the problems of children higher psychic functions development. For many years N.N. Traugott had a professional relationship with A.R. Luriya and his colleagues, at the suggestion of L.A. Orbeli she had headed the laboratory of pathology of higher nervous function from the beginning of 1956. The paramount importance for the development of new directions in evolutionary physiology had N.N. Traugott’s works on the principles of cortical dysfunction in different psychopathological conditions and her recommendations aimed at a strategy for individual corrections of speech in children.     

五节芒定居对尾矿砂重金属形态、微生物群落功能及多样性的影响

选择尾矿砂裸地和4个五节芒(Miscanthus floridulus)定居地(RI、RII、RIII和RIV)为样地, 分别研究了五节芒定居对尾矿砂重金属形态转化和微生物功能参数的影响。结果表明: 五节芒自然定居显著地提高了尾矿砂碳酸盐结合态和硫化物-有机物结合态重金属比例(p＜0.05), 降低了尾矿砂残渣态重金属的比例(p＜0.05)。土壤微生物群落的纤维素分解作用、酚转化作用、固氮作用、氨化作用、硝化作用、有机磷转化作用、功能多样性、4类不同碳源(碳水化合(CH)、聚合物(PL)、胺类化合物(AM)和杂合物(ML))均随着五节芒自然定居显著提高(p＜0.05)。而氨基酸(AA)的利用强度却显著下降(p＜0.05)。典范相关分析(CCA)表明: 土壤微生物功能参数的总体变化与土壤碳酸盐结合态和硫化物-有机物结合态重金属的含量呈显著正相关, 与残渣态重金属含量呈显著负相关。该研究结果表明, 五节芒定居不仅促进了尾矿砂重金属朝着沉淀态和螯合态方面转化, 而且还显著地改善了尾矿砂微生物群落的功能发挥。因而, 五节芒在重金属矿业废弃地恢复实践中具有较大的应用潜力。     

穗状狐尾藻与不同生长期苦草种间竞争研究

水生植物之间的竞争作用在水体生态系统恢复过程中具有重要作用。本研究采用取代系列实验方法,对穗状狐尾藻(Myriophyllum spicatum L.)与不同生长期(幼苗期和成株期)苦草(Vallisneria natans(Lour.)Hara)的种间竞争进行了研究。结果显示,对于不同生长期的苦草,穗状狐尾藻均具有较强的竞争优势,成株期苦草的竞争能力强于其幼苗期;苦草成株组中的穗状狐尾藻平均株高比苦草幼苗组中的穗状狐尾藻平均株高增加了128.6%(P0.05),但苦草成株组中的穗状狐尾藻植株平均干重比苦草幼苗组中的穗状狐尾藻植株平均干重降低了62.8%(P0.05);苦草幼苗组中的穗状狐尾藻分枝较多、株丛大,对苦草幼苗形成了较大的遮光作用;苦草成株组中的穗状狐尾藻分枝少、植株较长,顶端漂浮于水面生长,表明受苦草的种间竞争压力增大,但仍小于穗状狐尾藻的种内竞争;另外,苦草幼苗组中的苦草根长比苦草成株组中的苦草根长平均长28.6%(P0.05),表明在与穗状狐尾藻竞争过程中,苦草幼苗更趋于竞争地下资源。本研究结果说明穗状狐尾藻竞争能力强于苦草,苦草幼苗受穗状狐尾藻的竞争影响较大。     

Evolutionary significance of morphogenetic mechanisms

Different approaches to interpretation of ontogenesis are compared. The importance is emphasized of Ber’s theory of developmental patterns and the law of embryonic similarity. The significance of evolutionary rationalization of morphogenetic mechanisms is discussed (V.N. Beklemishev, V.A. Dogel’ and O.M. Ivanova-Kazas). There are two main pathways of rationalization of morphogenetic mechanisms: by means of epithelial morphogenesis and at the expense of oligocellular primordia (V.N. Beklemishev). When the developmental plans of the taxonomical types are determined, it is important to take into consideration the diversity of morphogenetic mechanisms within a given group in order to reveal morphological spectrums of morphogenetic mechanisms. A comparison of morphogenetic spectrums in different groups provides a wide evolutionary picture and helps us to reconstruct the phylogeny. Influence of morphogenetic mechanisms on the structure of definitive forms is demonstrated: construction technologies determine types of organs, anatomical and histological systems, and even symmetrical features of organisms and their complex parts.     

Preparation and properties of stretched polytene chromosomes. 2. Mechanism of stretching]

Isolated polytene chromosomes were stretched in a 0.125 M NaCl solution with constant speed, by constant force and by cyclically changing force. For each regime, the dependence of chromosome length on the time and force magnitude were recorded. From this it may be concluded that three processes are involved in chromosome stretching: viscoelastic deformation, viscous flow of DNP segments, and cristallization, i.e. intermolecular cross-linking of neighbour segments. At a high rate stretching (V greater than Vo) chromosome may be torn like at small deformation; when rate is V greater than Vo chromosome deformation is mostly viscoelastic; at rates V approximately Vo viscous flow of DNP segments if predominant. We estimate Vo approximately less than 3--6 mum/s. Electron microscopy shows that during chromosome stretching its DNP fibers are oriented along chromosome axis without detectable breaks.     

Transmembrane region of wild-type and mutant M13 coat proteins. Conformational role of beta-branched residues.

Although transmembrane (TM) segments of integral membrane proteins are putatively alpha-helical in conformation, beta-sheet promoters (Val, Ile, Thr) often account for approximately 40% of TM residue composition. We are examining the conformational role(s) of these residues, using as a model system the major coat protein of the filamentous bacteriophage M13. This 50-residue protein, which is located at the Escherichia coli host membrane during phage reproduction, contains a prototypic 19-residue hydrophobic midregion (residues 21-39: YIGYAWAMVVVIVGATIGI). Using "Eckstein" site-directed mutagenesis, we have generated several viable M13 coat protein mutants with beta-branched amino acid substitutions within their TM region. Mutant coat proteins, including Ile32----Val (I32V) and Ala27----Thr (A27T), were obtained in milligram quantities by growing M13 mutant phages in liter preparations, confirming that these coat proteins are capable of assuming their normal biological function(s) in phage reproduction. Circular dichroism spectroscopy performed in the membrane-mimetic medium of deoxycholate micelles indicated comparable alpha-helical contents of mutants I32V and A27T to wild-type protein. 13C nuclear magnetic resonance experiments with mutant A27T demonstrated that the combination of additional beta-branched content and introduction of an -OH substituent induced chemical shift and temperature-dependent changes and influenced the local protein environment at sites up to 12 residues remote from the mutation site. In contrast, mutant I32V (of which a salient feature is a mid-TM pentavaline segment) behaved very similarly to wild-type coat. These findings are interpreted in terms of the range of TM secondary structure and stability which can be accommodated by viable M13 coat protein mutants.     

Adaptation to temperature stress by Vibrio fischeri facilitates this microbe's symbiosis with the Hawaiian bobtail squid (Euprymna scolopes)

For microorganisms cycling between free‐living and host‐associated stages, where reproduction occurs in both of these lifestyles, an interesting inquiry is whether adaptation to stress during the free‐living stage can impact microbial fitness in the host. To address this topic, the mutualism between the Hawaiian bobtail squid (Euprymna scolopes) and the marine bioluminescent bacterium Vibrio fischeri was utilized. Using microbial experimental evolution, V. fischeri was selected to low (8°C), high (34°C), and fluctuating temperature stress (8°C/34°C) for 2000 generations. The temperatures 8°C and 34°C were the lower and upper growth limits, respectively. V. fischeri was also selected to benign temperatures (21°C and 28°C) for 2000 generations, which served as controls. V. fischeri demonstrated significant adaptation to low, high, and fluctuating temperature stress. V. fischeri did not display significant adaptation to the benign temperatures. Adaptation to stressful temperatures facilitated V. fischeri’s ability to colonize the squid host relative to the ancestral lines. Bioluminescence levels also increased. Evolution to benign temperatures did not manifest these results. In summary, microbial adaptation to stress during the free‐living stage can promote coevolution between hosts and microorganisms.     

种植香根草对铜尾矿废弃地基质化学和生物学性质的影响

通过实地调查取样和室内分析,研究铜陵水木冲铜尾矿废弃地不同时期种植香根草(Vetiveria zizanioides L.)群落(近期种植香根草群落(V.zizanioides communities were established in the recent stage,JX),中期种植香根草群落(V.zizanioides communities were established in the middle stage,ZX)和早期种植香根草群落(V.zizanioides communities were established in the early stage,OX))对尾矿基质化学性质、微生物量和土壤酶活性的影响,探讨人工植被恢复对铜尾矿废弃地基质系统的修复作用。结果表明:香根草的定植能延缓铜尾矿的酸化过程,且随着香根草定植时间的延长,0—5 cm和5—20 cm层尾矿基质中总氮和速效磷含量提高(其中,0—5 cm层总氮积累更加显著),OX下0—5 cm表层基质总氮和速效磷的平均值分别是JX下的4.64倍和22.44倍。基质微生物量C、N含量和脱氢酶、过氧化氢酶、脲酶活性也随香根草种植时间的延长而有不同程度的升高,且基质化学性质对微生物量和酶活性有影响,其中基质微生物量C、N含量、脱氢酶和过氧化氢酶活性均与电导率呈显著或极显著负相关性;而基质微生物量N和4种酶活性均与总氮含量呈显著或极显著正相关性,表明总氮含量是影响基质微生物量N和酶活性的主要因子;基质微生物量N、脱氢酶和过氧化氢酶活性还与速效磷含量呈极显著正相关性。基质中Cu、Pb含量对脱氢酶、过氧化氢酶活性和微生物量均有显著抑制作用,而Zn对基质微生物活性有一定的激活作用。生长在尾矿废弃地上的香根草不仅显著地改善了铜尾矿废弃地的基质化学性质,且有利于基质微生物量和酶活性的增加,是一种良好的矿业废弃地生态修复物种。