Phylogenetic and populational study of the Tuber indicum complex

When examined using SEM, Chinese samples of Tuber indicum and T. sinense displayed the same ascospore ornamentation as that of T. pseudohimalayense, T. indicum, collected in India by Duthie in 1899, and samples renamed T. himalayense in 1988. The different authors who named the four taxa (T. indicum, T. himalayense, T. sinense, T. pseudohimalyense) described differences in the surface of the peridium which could be considered as usual variations within a single species. We consider T. indicum, T. himalayense, T. sinense and T. pseudohimalayense as one species, T. indicum. Within this T. indicum complex, according to ITS and β-tubulin sequences, there are two groups in China, which could be considered as geographical ecotypes. This study is the first to identify a genetic and phylogeographical structure within the Chinese Tuber species.     

Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria

Efficient protein secretion is very important in biotechnology as it provides active and stable enzymes, which are an essential prerequisite for successful biocatalysis. Therefore, optimizing enzyme-producing bacterial strains is a major challenge in the field of biotechnology and protein production. In this study, the Gram-positive model bacterium Bacillus subtilis was optimized for heterologous protein secretion using a novel approach. Two lipolytic enzymes, cutinase from Fusarium solani pisi and a cytoplasmatic esterase of metagenomic origin, were chosen as reporters for heterologous protein secretion. In a systematic screening approach, all naturally occurring (non-lipoprotein) Sec-type signal peptides (SPs) from B. subtilis were characterized for their potential in heterologous protein secretion. Surprisingly, optimal SPs in cutinase secretion were inefficient in esterase secretion and vice versa, indicating the importance of an optimal fit between the SP and the respective mature part of the desired secretion target proteins. These results highlight the need for individually optimal signal peptides for every heterologous secretion target. Therefore, the SP library generated in this study represents a powerful tool for secretion optimization in Gram-positive expression hosts.     

Origin and taxonomic status of the Palearctic population of the stem borer Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae)

The major pest of maize in Mediterranean Europe, the stem borer Sesamia nonagrioides (Lefèbvre) (Lepidoptera: Noctuidae), has a fragmented distribution, north and south of the Sahara. The present study aimed: (1) to clarify the uncertain taxonomic status of the Palearctic and sub‐Saharan populations which were first considered as different species and later on as subspecies (Sesamia nonagrioides nonagrioides and Sesamia nonagrioides botanephaga) and (2) to investigate the origin of the Palearctic population which extends from Spain to Iran, outside what is considered typical for this mainly tropical genus. We reconstructed the evolutionary history of both populations using one nuclear and two mitochondrial genes. The sub‐Saharan taxon was fragmented in two isolated populations (West and East) whose mitochondrial genes were distant by 2.3%. The Palearctic population was included in the East African clade and its genes were close or identical to those of a population from Central Ethiopia, where the species was discovered for the first time. Similarly, in Africa, the alleles of the nuclear gene were distributed mainly in two West and East clades, whereas some Palearctic alleles belonged to the West clade. The Palearctic population originated therefore from East and West Africa and is the progeny of the cross between these two African populations. The main species concepts were in agreement, leading to the conclusion that the three populations are still conspecific. In the surveyed regions, the species therefore does not include two subspecies but three isolated populations. The Palearctic population suffered from severe bottlenecks that resulted in the fixation of one East African mitochondrial genome and the large reduction in its genetic diversity compared to the African populations. The data suggest that natural colonization of the Palearctic region was more plausible than human introduction. The allelic distribution of the Palearctic population was similar to that of species that survived the last glaciation. It is concluded that the African populations expanded during the last interglacial, crossed the Sahara and mixed in North Africa where fixation of the East mitochondrial genome occurred. The species then colonized Europe westward through only one eastern entrance. The coalescent‐based estimate of the time to the ancestor of the Palearctic population was 108 000 years, which is consistent with this scenario. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 904–922.     

Thermal change alters the outcome of behavioural interactions between antagonistic partners

1. Concerns about climate change often trigger the question whether physiological and behavioural responses of species will enable them to persist. However, species do not exist alone and are largely dependent on interactions with others within communities. 2. In the present study, a mechanistic approach is used to test the hypothesis that inter‐specific differences in metabolic response to unpredictable short‐term thermal changes can change the outcome of host–parasitoid behavioural interactions. 3. The effect of a drop or a rise of 5 °C on resting metabolic rates (RMR) of the main aphid pest of cereal crops in Western Europe, the host Sitobion avenae Fabricius and its main natural enemy, the parasitoid Aphidius rhopalosiphi De Stefani‐Perez was measured. Also, defence and attack behaviours were measured for host and parasitoid separately as well as in interaction, since behavioural strategies of both species largely determine parasitism success. 4. The results showed that, when no change in temperature occurred, parasitoids had the highest oviposition rate. However, only with a rise of temperature behavioural interactions were disrupted: the parasitoid attack rate decreased whereas the aphid defence rate increased. This alteration in behaviour was associated with a stronger thermal response of RMR in hosts than in parasitoids, suggesting that species‐specific thermal responses of RMR could give valuable information on changes in the outcome of species interactions under warm spells but not under cold ones. 5. It was shown that relatively modest thermal changes with non‐lethal effects can have profound consequences for interacting co‐evolved species which may affect ecosystem services, such as biological control of pest populations.     

Lipids in cell biology: how can we understand them better?

Lipids are a major class of biological molecules and play many key roles in different processes. The diversity of lipids is on the same order of magnitude as that of proteins: cells express tens of thousands of different lipids and hundreds of proteins to regulate their metabolism and transport. Despite their clear importance and essential functions, lipids have not been as well studied as proteins. We discuss here some of the reasons why it has been challenging to study lipids and outline technological developments that are allowing us to begin lifting lipids out of their “Cinderella” status. We focus on recent advances in lipid identification, visualization, and investigation of their biophysics and perturbations and suggest that the field has sufficiently advanced to encourage broader investigation into these intriguing molecules.Lipids are fundamental building blocks of all cells and play many important and varied roles. They are key components of the plasma membrane and other cellular compartments, including the nuclear membrane, the endoplasmic reticulum, the Golgi apparatus, and trafficking vesicles such as endosomes and lysosomes. The lipid composition of different organelles, cell types, and ultimately tissues can vary substantially, suggesting that different lipids are required for different functions (Saghatelian et al., 2006 ; Klose et al., 2013 ). Mammalian cells express tens of thousands of different lipid species and use hundreds of proteins to synthesize, metabolize, and transport them. Although the complexity and diversity of lipids approach those of proteins, we have a much poorer understanding of their functions, making lipids in many ways the “Cinderellas” of cell biology. We discuss here recent advances, and challenges, in investigating how these molecules contribute to the many biological processes in which they participate.Like proteins, lipids can have structural (e.g., by stabilizing different membrane curvatures) or signaling roles. Posttranslational lipidation of proteins (e.g., palmitoylation or farnesylation) and carbohydrate-linked lipids (glycolipids) are also important examples of cellular lipid pools. It is clear that higher-order organization is key to most lipid functions, and they are believed to assemble into signaling platforms that contain both lipids and proteins (Kusumi et al., 2012 ). Some lipid microdomains are termed lipid rafts, and much ongoing effort is being focused on defining their parameters (Simons and Sampaio, 2011 ; Suzuki et al., 2012 ; Klotzsch and Schutz, 2013 ). Because of this focus on lipid rafts, we have a better understanding of the properties of proposed raft lipids (e.g., sphingomyelins and cholesterol) than of many other lipid species. Much of what we know about lipids has come from studying synthetic membranes with specific lipid compositions. Although model membranes usually consist of very few (<10) lipid species, they have been useful for understanding the biophysical properties of lipids. At the other end of the spectrum, lipids have been implicated in various diseases, with cholesterol metabolism being a prominent example. We are now getting to a point at which we can address the roles of lipids in the middle of the spectrum—in cells.     

荧光标记复合PCR扩增微卫星位点的构建与优化

本研究采用毛细管电泳技术,构建并优化了荧光标记复合PCR同时扩增多个微卫星位点。主要过程为：首先根据设计所扩增微卫星位点的期望长度,将9个微卫星位点分成两组,5个位点用FAM（蓝色）标记,4个位点用HEX（绿色）标记;两种荧光类型分组优化,用琼脂糖胶电泳检测。其次,荧光标记的复合PCR扩增8个中华绒螯蟹样品的9个微卫星位点,采用ABI3730xl毛细管电泳检测,以ROX500（红色）为长度标准物,结果经Genemapper3.5软件 分析,检测结果表明毛细管电泳检测荧光标记复合PCR产物不仅精确读取微卫星位点的长度（分辨率高达1bp）,还能区分微卫星位点复制时滑链所引起的“回声斑”;调整各微卫星位点引物比列使所有位点扩增强弱均匀。最后,逐一检测复合PCR基本参数（dNTP浓度、 PCR程序和模版DNA用量）对复合PCR产物的影响,优化PCR。结果表明通过毛细管电泳检测荧光标记复合PCR产物来读取微卫星位点的基因型具有精确性、高效性和稳定性。     

盐度对互花米草枯落物分解释放硅、碳、氮元素的影响

为了研究潮汐湿地盐度对枯落物分解过程中硅、碳、氮元素释放的影响,通过室内模拟不同盐度(0、5、15和30)对互花米草枯落物茎和叶分解释放过程中硅、碳、氮元素的动态变化进行测定。结果表明:(1)互花米草茎和叶枯落物失重率和分解速率均随盐度增加而降低。(2)互花米草茎和叶枯落物分解水体中硅含量均随着盐度升高而增加,并且盐度30处理下,枯落物分解硅释放量显著高于盐度0和5(P0.05)。而分解末期生物硅残留量则随盐度升高而降低。(3)不同盐度处理茎枯落物分解碳释放量无显著差异,但叶枯落物分解碳释放量在盐度5、15和30处理中显著高于淡水(P0.05)。(4)互花米草茎枯落物分解释放到水中的NH_4~+-N含量随着盐度的升高而减少,NO_3~--N含量与之相反。研究单因素盐度对枯落物分解及元素释放的影响,可以为预测潮汐湿地枯落物分解对盐水入侵的响应机制提供参考,为湿地生源要素生物地球化学循环过程研究提供基础依据。     

南海北部浮游植物生长对营养盐的响应

2004年夏季作者在南海北部海域研究了浮游植物生长的营养动力学,结合物理-化学过程对浮游植物生物量分布的影响与机制进行了研究,阐明了水平对流和中尺度涡对营养盐分布的影响及浮游植物生长和现存生物量对其的响应。受西南季风和东向沿岸流作用所形成的Ekman输送的影响,南海北部海岸带表层海水作离岸运动,使深层富含营养盐的冷水爬坡涌升到表层来补充,激发浮游植物生物量迅速增长。海区反气旋涡使海水辐聚下沉,造成水体具高温、低盐、高溶解氧浓度、低营养盐浓度和低浮游植物生物量。同时通过现场营养盐加富试验,发现该海域营养盐是浮游植物生长的主要限制因子,而且是多种营养元素共同限制了浮游植物的生长,添加单一的营养盐并不能促进浮游植物的生长。在生物量出现增长的试验组中,营养盐添加不仅促使浮游植物生物量的增长,而且也改变了浮游植物的粒级结构和群落结构。例如,在站S1008,培养前叶绿素a浓度为0.28 mg.m-3,加富培养60 h后浮游植物生物量在NP和NPSi的试验组中有显著的增加,叶绿素a浓度分别达1.07 mg.m-3和1.19 mg.m-3;培养前粒度分级叶绿素a主要以Pico级份占优势,而加富试验结束后,在NP和NPSi的试验组以Nano级份占优势,其它试验组仍以Pico级份占优势;同时,在培养后生物量出现增长的试验组,浮游植物群落的优势类群从甲藻向硅藻演替。