Activités enzymatiques digestives chez Palaemon serratus: Variations annuelles de l''acrophase des rythmes circadiens

Evidence has been obtained for a circadian rhythm of digestive enzymatic activities in Palaemon serratus. This rhythm does not show the same characteristics during the course of a whole year. The number and intensity of activity maxima are not the same for both amylases and the proteases. On the other hand, the photophase plays a role in the temporal regulation of these rhythms but modifies its principal characteristics. Eyestalk ablation does not abolish these rhythms.     

Cartographie de restriction comparée de la séquence d''ADN hautement répétée coupée par Bam HI chez Papio papio et P. cynocephalus

Highly repeated DNA sequences from two baboon species (Papio papio and P. cynocephalus) have been compared using restriction endonucleases. The two species share a 343 base pairs tandemly repeated DNA, that is cut once by Bam HI. Papio cynocephalus differs from P. papio by loss of an EcoRI star site in the repeated sequence.     

毕赤酵母底盘芳香族氨基酸合成途径改造生产肉桂酸及对香豆酸*

目的:改造毕赤酵母使其异源合成类黄酮生物合成途径的重要中间体肉桂酸、对香豆酸,并优化前体芳香族氨基酸生物合成途径以提高毕赤酵母的生产能力。方法:在毕赤酵母GS115中利用乙醇诱导型人工转录系统表达Rhodotorula glutinis来源的苯丙氨酸解氨酶,并在该重组菌株中分别过表达胞内芳香族氨基酸生物合成途径中的关键酶或其突变体以进行优化。结果:异源表达苯丙氨酸解氨酶可使毕赤酵母将自身产生的L-苯丙氨酸、L-酪氨酸转化为肉桂酸(38.8 mg/L)、对香豆酸(34.2 mg/L),而通过过表达相关酶进行优化,最终肉桂酸和对香豆酸的产量分别达到124.1 mg/L和302.0 mg/L。结论:利用新的异源宿主毕赤酵母成功合成了肉桂酸、对香豆酸,并对胞内的芳香族氨基酸生物合成途径进行了优化,表明毕赤酵母具有生产黄酮类化合物的应用潜力,也为其他芳香族氨基酸衍生物或植物化合物在毕赤酵母中的异源合成奠定了基础。     

Dosages protéiques et profils d''activités enzymatiques comparés chez les deux espèces de babouins Papio anubis et P. cynocephalus

Biochemical levels of serum proteins and enzymes were investigated in two populations of the baboons Papio anubis and P. cynocephalus. The mean average levels of proteins or enzyme activity are the same in the two species. The only difference concerns the level of an inhibitor of pseudocholinesterase in a few individuals of Papio anubis.     

Motility and chemotactic response of Pseudomonas fluorescens toward chemoattractants present in the exudate of Macrophomina phaseolina

Pseudomonas fluorescens strains (LAM1-hydrophilic) and (LAM2-hydrophobic) showed positive chemotaxis towards attractants (sugars, amino acids, polyols and organic acids) present in the exudate of Macrophomina phaseolina (a soilborne plant pathogenic fungus). The varied response of motility traits such as speed, rate of change in direction (RCDI) and net to gross displacement ratio (NGDR) was observed for different chemoattractants. Swimming speed of the strains was highest in 10-fold diluted exudate or 100–1000 μM strength of different attractants, but further dilutions significantly decreased the swimming speed (P = 0.05). Chemotactic response of P. fluorescens was positively correlated with swimming speed (P = 0.05; r = 0.76). Relative to control, the RCDI values decreased 1.5-fold in amino acids or sugars, and 1.2-fold in polyols or organic acids. With increase in swimming speed, the NGDR of both strains also increased, but the RCDI decreased. Both hydrophilic and hydrophobic strains did not show significant differences in their motility traits. The results demonstrate that M. phaseolina exudate contains chemical attractants that serve as signal for flagellar motility of P. fluorescens. Motile P. fluorescens strains thus may consume fungal exudate as nutrients, and thus spores could offer a niche for these bacteria in soil.     

Variations des zymogrammes estérasiques chez les larves de Palaemon serratus soumises à différentes durées de choc thermique

Palaemon serratus at larval stage 2, acclimatized at 16°, were subjected to a temperature increase of 15° for periods of 5, 20 or 40 min with or without return to the initial temperature for 12 h. The soluble protein concentration of larvae subjected to a thermal shock of 20 min duration is lower than in control larvae. Total esterase-2C activity per mg of soluble protein after a thermal shock of 20 min duration is less than in control larvae but, in contrast with larvae subjected to a shock of 40 min duration, the initial activity is restored after 12 hours. Esterase-2C activity zymograms, after polyacrylamide gradient gel electrophoresis, show twelve isozymes. The activity of each isozyme examined varies according to the duration of the thermal shock.     

Stearidonic acid (18:4ω3) in Primula florindae

Stearidonic acid (18:4ω3), which is reported to be of rare occurrence in the plant kingdom and which is of considerable dietary and pharmaceutical interest has been found in three closely related Primula species. It occurs, together with γ-linolenic acid (3–4% of the seed oil total fatty acids), in significant percentages in Primula florindae (11%), P. sikkimensis (14%) and P. alpicola (14%). 18:4(ω3 may also be of chemotaxonomic interest in the genus Primula, as high levels may be typical for section Sikkimensis. The only commercial plant source of stearidonic acid known so far is the seed oil of Ribes nigrum.     

七星瓢虫成虫对枸杞木虱的捕食作用

为了测定七星瓢虫成虫对枸杞木虱4种虫态的捕食作用,分别在室内测定七星瓢虫的捕食功能反应、种内干扰、自身密度干扰、捕食偏好性以及在田间七星瓢虫对枸杞木虱的捕食效果.结果表明: 七星瓢虫对枸杞木虱的捕食功能反应符合Holling Ⅱ型方程,其中对卵的最大捕食量为112.6粒,对1～2龄若虫、3～5龄若虫、成虫的最大捕食量分别为536、415和113.9头;田间罩笼试验结果证明,七星瓢虫成虫在其生长周期30 d内能使枸杞木虱总虫口密度下降80.1%;七星瓢虫对1～2龄枸杞木虱若虫的搜寻效率参数a=0.9451,处理时间参数T_h=0.001865,整体优于卵、3～5龄若虫与成虫,且在每皿100头的猎物密度下七星瓢虫的最大捕食率能达80.2%,益害比参考值为1∶100.七星瓢虫对枸杞木虱的捕食作用受自身密度的影响显著大于种内干扰.在混合猎物密度为每皿100头下,七星瓢虫更偏好木虱成虫,在密度为每皿300头下,七星瓢虫更偏向于木虱若虫.表明七星瓢虫是很有控制潜力的捕食性天敌,人工释放七星瓢虫成虫可有效取食枸杞木虱初孵若虫,降低木虱为害.     

青杄中sPPa1的cDNA序列克隆及其生物信息学分析

以青杄(Picea wilsonii)均一化cDNA文库为模板,通过RACE方法克隆得到青杄PPa1基因cDNA全长,对该cDNA序列、核苷酸序列的相似性、理化性质、疏水性、二级结构、三级结构及是否跨膜进行了分析预测;进行了多序列比对并构建了系统树,同时对PPa1在青杄各组织中的表达量进行了检测。结果表明:青杄PPa1基因共由216个氨基酸组成,分子量为24.55 kD,理论PI为5.83,属可溶性蛋白;二级结构主要由α-螺旋、不规则卷曲和β-折叠构成;PPa1在青杄花粉中表达量最高。研究为进一步研究青杄PPa1的功能奠定了基础。     

The Peniophorella praetermissa species complex (Basidiomycota)

The corticioid basidiomycete Peniophorella praetermissa has long been regarded as a morphologically variable species complex. An ITS-based phylogenetic study based on a worldwide sampling was carried out using parsimony and Bayesian inference. The resulting trees feature three major clades, further divided into well-supported subclades. These could be considered as distinct species, a contention that is further supported by crossing test data. Only two out of the eight phylogenetic lineages identified can be distinctly morphologically characterized: P. odontiaeformis and P. subpraetermissa. P. odontiaeformis is an odontioid species with a paleotropical distribution whereas the taxa in the remaining subclades have smooth basidiomata and are distributed in temperate areas. P. subpraetermissa is known only from the type collection and is distinguished microscopically by its reddish brown apically encrustated cystidia. Taxa in the remaining subclades are impossible to distinguish from each other morphologically, and therefore, are viewed as a species complex, P. praetermissa s. lat. One of the subclades, which is widely distributed but restricted to the Northern hemisphere, is proposed to represent P. praetermissa s. str. An epitype is selected from the same area as the holotype, among the specimens studied here. However, the geographically most widespread clade with many representatives from both hemispheres is here referred to as P. pertenuis, a taxon that was previously considered a synonym to P. praetermissa.     

Expression of a Bacillus subtilis pectate lyase gene in Pichia pastoris

The methylotrophic yeast Pichia pastoris is an attractive heterologous protein expression host, mainly for genes from higher eukaryotes. However, no successful examples for the expression of bacterial gene encoding pectate lyase in P. pastoris have been reported. The present study reports for the first time the cloning and functional expression of the bacterial Bacillus subtilis gene encoding alkaline pectate lyase in P. pastoris. A molecular weight of 43,644 Da was calculated from the deduced amino acid sequence. A pectate lyase activity as high as 100 U/ml was attained in the fermentation broth of P. pastoris GS 115, which was about 10 times higher than when the gene is expressed in Escherichia coli. The recombinant pectate lyase was purified to homogeneity and maximal activity of the enzyme was observed at 65 °C, and pH 9.4. The recombinant enzyme showed a wider pH and thermal stability spectrum than the purified pectate lyase from B. subtilis WSHB04-02. Pectate lyase activity slightly increased in the presence of Mg²⁺ (ion) but decreased in the presence of other metal ions. Analysis of polygalacturonic acid degradation products by electrospray ionization-mass spectrometry revealed that the degradation products were unsaturated trigalacturonic acid and unsaturated bigalacturonic acid, which confirms that the enzyme catalyzes a trans-elimination reaction.     

胡杨WINDs基因克隆及功能初步分析*

目的: WIND(WOUND INDUCED DEDIFFERENTIATION),是属于ERF/AP2 (ETHYLENE RESPONSE FACTOR/ APETALA 2)家族的一种重要转录因子,该类基因最早被发现在拟南芥中可以与乙烯响应元件GCC-BOX和脱水响应元件DRE结合,响应干旱信号和调节乙烯水平。最近的研究发现WIND基因在植物伤口信号回应、愈伤组织形成及不定芽的产生过程中也发挥了关键作用。已有的研究阐述了WIND基因在拟南芥中控制愈伤组织形成及不定芽再生的机制,但其在木本植物中的功能尚不明确,将探究WIND基因在胡杨中与伤口信号响应及不定芽再生相关的功能,同时为在分子水平上解决胡杨再生问题提供理论依据。方法: 采用基因克隆、qRT-PCR、转基因表型分析等方法研究WIND基因在胡杨外植体伤口响应和再生不定芽过程中的作用。结果: 克隆胡杨WIND家族中的基因PeWIND1和PeWIND2,发现其编码区序列长度分别为1 050 bp和1 032 bp,编码349个和343个氨基酸,亚细胞定位均在细胞核中。组织特异性分析显示PeWIND1和PeWIND2在胡杨根、茎、叶、愈伤组织中均有表达,且在愈伤组织中表达量最高。时间表达特异性显示,在经伤口刺激后的24 h内,PeWIND1和PeWIND2基因均呈现先升高后降低的表达趋势,且均在伤口刺激后1 h达到表达量峰值。转基因植株表型统计发现,过表达PeWIND1和PeWIND2基因后转基因植株不定芽再生能力增强。结论: 在胡杨叶片有伤口刺激后,PeWIND1和PeWIND2响应伤口信号,表达量先升高后降低,PeWIND1和PeWIND2能够促进杨树茎段再生不定芽。     

Evidence for conspecificity of Piper methysticum forst. f. and Piper wichmannii C. DC

Morphological, chemical, cytological and genetic evidence demonstrating the absence of taxonomic distinction between Piper methysticum and Piper wichmannii are reviewed. Piper methysticum is not a separate species, but rather a group of sterile cultivars selected from somatic mutants of P. wichmannii. As P. methysticum was described first (1786), it has priority and P. wichmannii (1910) is superfluous. A new subspecific classification is suggested that makes a distinction between the sterile cultivars (P. methysticum var. methysticum) and the wild populations (P. methysticum var. Wichmannii).     

Growth of the sponge Pseudosuberites (aff.) andrewsi in a closed system

Explants of the Indo-Pacific sponge Pseudosuberites aff. andrewsi were fed with the microalgae Chlorella sorokiniana and Rhodomonas sp. It was microscopically observed that these algae were ingested and digested by the sponge cells, suggesting that they were consumed by the sponges. The algae were further used for two growth experiments with five explants of P. aff. andrewsi and four explants of P. andrewsi. Growth was measured as the increase in projected body area. The explants showed considerable growth (up to 730% in 54 days for P. aff. andrewsi and up to 680% in 22 days for P. andrewsi), which is much higher than previously reported growth rates for sponges. Growth started after a stationary phase of 5–20 days in which the projected body area did not increase. The growth of P. aff. andrewsi appeared to be linear and was inhibited at the end of the experiment. Two explants of P. andrewsi showed exponential growth instead of linear growth. Hence, no general statements about the growth kinetics of these sponges can be made at this time. However, the high growth rates found in this study suggest a promising future for cultivation of sponges in closed systems.     

农杆菌介导海洋草酸青霉转化体系及聚酮合酶Pks生物学功能*

为研究南海柳珊瑚共附生草酸青霉SCSGAF0023的聚酮合酶(PKS)生物学功能,采用农杆菌介导法构建草酸青霉SCSGAF0023的Pks敲除株ΔPks,比较野生菌株及ΔPks的生长发育及环境适应性差异。以草酸青霉SCSGAF0023分生孢子为受体,p0380-hygB为双元载体,成功实现草酸青霉SCSGAF0023的遗传转化。结果表明:农杆菌浓度为OD₆₀₀=0.5,在200μmol/L 乙酰丁香酮(AS)诱导下与10⁷个/ml草酸青霉SCSGAF0023孢子于25℃共孵育时转化效率最高。基于上述转化体系,成功获得Pks敲除株ΔPks,并首次证实Pks正向调控草酸青霉SCSGAF0023产孢,但不影响其对环境的适应性。这为进一步系统研究真菌PKSs及聚酮化合物对真菌生长发育与环境适应性的影响提供素材。     

Evidence of natural reciprocal hybridisation between Pinus uliginosa and P. sylvestris in the sympatric population of the species

Peat-bog pine Pinus uliginosa Neumann has become extinct or rare in many parts of Europe. It is supposed to hybridize with P. sylvestris but the influence of hybridization to genetic erosion of the P. uliginosa gene pool is unknown. In the presented study, the crossability between P. uliginosa and P. sylvestris was analyzed in a sympatric population at Węgliniec reserve in Poland. The aim of the study was to prove natural hybridisation and to estimate the influence of this phenomenon on possible natural gene pool erosion of peat-bog pine. A sequence polymorphism in the trnF–trnL cpDNA region of P. uliginosa and P. sylvestris was used to develop the species diagnostic PCR-RFLP marker. The marker of paternally transmitted cpDNA was applied in haplotype analyses of the progeny from open-pollinated P. uliginosa seeds (collected in 2000–2002) and from P. sylvestris ones (in 2002). An inconsistency in species diagnostic cpDNA haplotypes of seedlings and parental trees was observable both for P. uliginosa and P. sylvestris (about 1% and 2% of hybrids seeds, respectively). The results prove the occurrence of reciprocal hybridisation between the species. The influence of hybridisation on the natural gene pool protection and measures to reintroduce peat-bog pine is discussed.     

Composition en lipides et acides gras de deux esèces de Taphrina parasites de Prunus domestica

Four strains of both Taphrina pruni and T. institiae were cultivated under identical conditions and and lipids and fatty acids were quantitatively analysed at two stages of their development. Tri- and diglycerides are the major neutral lipids in both species. Phosphatidylcholine and phosphatidylethanolamine are the most abundant polar lipids. Qualitatively, the two species show identical fatty acid contents, except for margaric acid (17:0) which was only found in Taphrine pruni. Quantitatively there are several differences: palmitoleic acid (16:1) occurs in reasonable amounts regularly and only in Taphrina pruni. The ratios 16:0/18:0 and 18:1/18:2 are generally higher for T. insititiae whereas the degree of unsaturation of fatty acids is higher in the former. The results are discussed with regard to data on other fungal species.     

不同改良措施对太子参根际土壤酚酸含量及特异菌群的影响

太子参为石竹科多年生草本植物,以根部入药,药用价值高,市场需求大,但其栽培过程中存在严重的连作障碍问题,连作导致产量品质严重下降,病虫害猖獗,探索太子参连作障碍形成的机制及其消减措施意义重大.本研究以广泛栽培的“柘参2号”为试验材料,采用土壤农化分析、高效液相色谱(HPLC)分析、荧光定量PCR(qPCR)等技术对不同连作年限或不同消减措施下太子参根际土壤的主要养分、酚酸含量动态及特异菌群变化进行分析.结果显示: 太子参连作导致产量显著下降,与正茬相比产量下降达43.5%,而水旱轮作和微生物菌肥处理可不同程度缓解其障碍效应.太子参连作土壤主要养分(如速效氮、速效磷、速效钾、全钾)未降反升,但导致土壤酸化.HPLC分析表明,太子参连作下或者随生长时期的推进,土壤酚酸类物质并未表现出明显的积累效应,膨大中期和收获期重茬土壤中大部分酚酸含量甚至低于正茬土壤.进一步荧光定量PCR分析发现,太子参连作导致前期分离筛选到的几类病原菌(如尖孢镰刀菌、踝节霉菌、Kosakonia sacchari)的绝对含量显著上升,而水旱轮作和微生物菌肥处理可有效降低3类病原菌的含量,改善微生态结构.综上认为,太子参连作障碍的发生并非由土壤养分匮乏或者酚酸持续积累直接造成,可能主要与连作下土传病原微生物的大量繁殖爆发有关,本研究结果为深入揭示太子参连作障碍的形成机制及消减修复机制提供了理论依据.     

Pseudo-nitzschia species population dynamics in the Quoddy Region, Bay of Fundy

The population dynamics of Pseudo-nitzschia species from the Quoddy Region of the Bay of Fundy was examined in a field-based study. Five stations were sampled over a period of 11 weeks during the course of one discrete bloom episode in 2003 with seven species of Pseudo-nitzschia found and enumerated: P. americana, P. delicatissima, P. pseudodelicatissima, P. fraudulenta, P. pungens, P. seriata and P. subpacifica. We related species abundance to physical and chemical properties of seawater (transparency, fluorescence, silicate, phosphate, nitrite + nitrate, ammonia, nitrite, oxygen, sigma-t, tidal level, tidal state and total depth of water column) using canonical correspondence analysis (CCA) to identify factors explaining the greatest amount of variance in their temporal and spatial distribution patterns. Our results indicate that abundance of species and groups of species correlated well with certain specific chemical and physical properties of seawater. P. pseudodelicatissima and P. delicatissima abundance was positively correlated with nitrates and P. americana with depth of the water column. P. pungens was more abundant in samples with higher concentrations of phosphates and lower concentrations of nitrates. P. seriata abundance was negatively associated with total fluorescence. P. subpacifica and P. fraudulenta abundances were not statistically related to any of the variables examined. Our data therefore provides direction for testable, hypothesis driven experiments that could provide predictive insights into the occurrence of certain harmful algal bloom (HAB) species should specific environmental variables be further affected along the gradients extracted in this study.