鉴别桐花树毛颚小卷蛾幼虫、蛹及成虫雌雄的方法

报道如何在林间迅速、准确地根据外部形态特征区分桐花树毛颚小卷蛾Lasiognatha cellifera(Meyrick)幼虫、蛹及成虫雌雄的方法。这对于了解其在林间的性比,观察其生物学特性、种群动态及开展预测预报等十分重要。     

利用末龄幼虫蜕和蛹壳对苹果蠹蛾基因的无创伤检测方法

苹果蠹蛾Cydia pomonella（L.）是我国重大农业入侵害虫,对我国果业健康发展造成严重威胁。在利用基因编辑等技术对相关基因进行功能研究时,通常采用单对交配策略对纯合突变体进行筛选。因此,在配对前明确个体基因型,避免对虫体造成损伤尤为重要。本研究分别收集末龄幼虫蜕（10个蜕）和蛹壳（1个、5个、10个、15个蛹壳）,通过基因组DNA提取、目的基因PCR扩增、琼脂糖凝胶电泳检测和PCR产物测序,以评估该方法作为苹果蠹蛾基因检测的可行性。结果显示,以苹果蠹蛾末龄幼虫10个蜕及5个以上蛹壳提取的基因组DNA作为模板,扩增精巢特异性丝氨酸/苏氨酸蛋白激酶（Testis specific serine/threonine protein kinase,TSSK1）基因,PCR产物经琼脂糖凝胶电泳检测得到单一、明亮的条带,经测序证实该产物是目的基因序列。基于高效及节约成本的原则,拟推荐利用10个末龄幼虫蜕或10个蛹壳提取基因组DNA、通过PCR扩增和测序进行苹果蠹蛾的无损伤基因型检测。本研究建立了一种利用幼虫蜕和蛹壳进行苹果蠹蛾无损伤、高效的基因检测方法,为提高苹果蠹蛾基因功能研究的效率奠定了基础。     

A non-destructive screenable marker,OsFAST, for identifying transgenic rice seeds

The production of transgenic plants has contributed greatly to plant research. Previously, an improved method for screening transgenic Arabidopsis thaliana seeds using the FAST (Fluorescence-Accumulating-Seed Technology) method and FAST marker was reported. Arabidopsis seeds containing the FAST marker may be visually screened using a fluorescence stereomicroscope or blue LED handy-type instrument. Although the FAST method was originally designed for Arabidopsis screens, this study endeavors to adapt this method for the screening of other plants. Here, an optimized technology, designated the OsFAST method, is presented as a useful tool for screening transgenic rice seeds. The OsFAST method is based on the expression of the OsFAST-G marker under the control of a seed-embryo-specific promoter, similar to the Arabidopsis FAST-G marker. The OsFAST method provides a simple and non-destructive method for identifying transgenic rice seeds. It is proposed that the FAST method is adaptable to various plant species and will enable a deeper analysis of the floral-dip method.Key words: Oryza sativa, oleosin, seed, green fluorescent protein, transformation, screenable markerThe production of transgenic plants has significantly enhanced many areas of plant science research. Antibiotic/herbicide-resistance genes are traditionally used as screenable markers for the selection of transgenic plants. However, this approach does have disadvantages. First, antibiotics or herbicides occasionally inhibit the growth of transgenic plants, regardless of the incorporation of antibiotic- or herbicide-resistance genes¹ into the transgenic plants. Second, the identification of resistant transgenic plants requires that the seed population be sown onto plates containing antibiotics or herbicides. Third, the selection process is slow and labor intensive, often involving the screening of vast numbers of potentially transgenic seeds on selective plates.To overcome these disadvantages, an improved approach for selecting transgenic Arabidopsis thaliana, designated the FAST (Fluorescence-Accumulating-Seed Technology) method, was developed. This method employs the use of a fluorescent protein that is expressed in seeds and used as a visual screenable marker for the identification of transgenic seeds. The seed-specific protein oleosin, a family of oil-body-membrane proteins,² has an important role as a size regulator of oil bodies.³ AtOLE1, the most abundant oleosin, functions in the freezing tolerance of Arabidopsis seeds.⁴ A plasmid containing an AtOLE1-GFP fusion gene controlled by the AtOLE1 promoter was constructed and designated the FAST-G (Fluorescence-Accumulating-Seed Technology with OLE1-GFP) marker. Interestingly, Arabidopsis seeds containing the FAST-G marker emitted clear fluorescence under a fluorescence stereomicroscope or blue LED handy-type instrument. The transgenic seeds were visually identified by the seed fluorescence without the use of antibiotics or herbicides, thus indicating that the FAST method offers a nondestructive approach. The FAST marker permits the identification of homozygous seeds among the T2 population with a false discovery rate of less than 1% as a co-dominant screenable marker. In contrast to conventional methods using antibiotics or herbicides, the FAST method reduces the amount of time required to acquire homozygous transgenic plants from 7.5 months to 4 months. The fluorescence of the FAST-G marker was limited to a specific organ (i.e., in seeds) and a specific time (i.e., during dormancy), desirable characteristics of selectable and/or screenable markers. Furthermore, the FAST marker does not require sterile seeding and the handling of large numbers of plants.     

基于外部形态特征的红火蚁野外快速识别方法研究

红火蚁Solenopsis invicta Buren是一种严重威胁公共设施、人类安全、农林生产和生物多样性的重大入侵害虫。自2004年在我国大陆广东省首次发现以来,其发生范围急剧扩大,目前已扩散至南方11个省(市),而在2016年底红火蚁已突破封锁防线侵入浙江省。本文通过对浙江省仙居县23个乡镇(街道)中的草皮、苗圃、废品站、塑料加工厂等高风险传播红火蚁的生境进行火腿肠诱饵诱集调查,共获得10种蚂蚁,且隶属于4亚科7属,并以其身体颜色、体长、头形及与身体的协调性、腹柄结共4个指标特征分别与红火蚁进行比对、分析,建立一种直观易懂的野外快速识别红火蚁的方法,以期提高判定红火蚁的准确性及疫情普查效率。     

快速鉴别菱角水螟雌雄蛹的方法

【目的】本研究拟建立一种快捷、准确地鉴别菱角水螟Parapoynx crisonalis(Walker)蛹性别的方法。【方法】基于蛹的腹部末端第8、9两腹节的腹面外部形态学特征进行判别。【结果】雌雄蛹的腹面外部形态学特征主要区别在于雄蛹第8腹节平滑无裂缝,第9腹节中央具一较短纵裂缝,其两侧具明显的半圆形瘤状突起,而雌蛹第8腹节中央具一长纵裂缝,其两侧较为平坦、无明显凸起。依据该特征鉴别菱角水螟蛹雌雄的准确率为100%。【结论】该方法在田间进行雌雄蛹快速鉴别行之有效,对提前掌握菱角水螟田间性比、下一代种群动态的预测预报工作具有重要意义。     

ＤＮＡ粗提取与鉴定的简易方法

介绍一种在中学可行的ＤＮＡ提取与鉴定的简易方法,该法可以不使用 离心机和一些特殊的昂贵的药品。     

A rapid non-destructive DNA extraction method for insects and other arthropods

Preparation of arthropods for morphological identification often damages or destroys DNA within the specimen. Conversely, DNA extraction methods often destroy the external physical characteristics essential for morphological identification. We have developed a rapid, simple and non-destructive DNA extraction technique for arthropod specimens. This technique was tested on four arthropod orders, using specimens that were fresh, preserved by air drying, stored in ethanol, or collected with sticky or propylene glycol traps. The technique could be completed in 20 min for Coleoptera, Diptera and Hemiptera, and 2 min for the subclass Acarina, without significant distortion, discolouration, or other damage to the specimens.     

A non-destructive selection method for resistance to fusicoccin in Arabidopsis thaliana

A mutagenised population of seeds of Arabidopsis thaliana was allowed to germinate in the presence of the positively charged aminoglycoside hygromycin (4 μg/ml) and the fungal toxin fusicoccin (5×10^–6 m). This hygromycin concentration, which is non-toxic by itself, becomes toxic when used together with fusicoccin, which stimulates cation uptake. Seeds that had germinated after 3–5 days and produced seedlings with green cotyledons were potentially resistant to fusicoccin and were therefore transferred into sterile Magenta vessels containing 1/2-strength Murashige and Skoog medium. This selection procedure is non-destructive, i.e. it allows the recovery of viable seedlings and their growth into adult plants thus permitting direct physiological characterisation. Received: 16 February 1998 / Revision received: 11 August 1998 / Accepted: 13 August 1998     

A simple method for the collection of Necator americanus larvae

A simple method for the collection of third-stage larvae of Necator americanus has been described. This technique provides repeated recovery of very clean larvae from cultures in moderate numbers.     

A method for measuring the size of early euphausiid larvae

A simple method to measure early euphausiid larvae is tested on 35 specimens of Euphausia superba obtained in the Weddell-Scotia Confluence region (ranging from Calyptopes I to Furcilia III) against the measures obtained on the same specimens with a graduated eyepiece. The arrangement of the test includes two observers, four magnifications (from 7.5× to 62.5×) whenever feasible and two replications of each measurement. A total of 953 measurements were analyzed in an incomplete random blocks ANOVA design not finding significant differences between magnifications, methods, observers and their interactions. It was found that the relative differences between methods were of the same magnitude as the differences between replications (approximately 5%). The proposed method is less demanding on laboratory work, thus allowing the measurement of the large number of specimens needed to estimate size frequency distributions.     

A novel method for identifying hydrophobicity on fungal surfaces

Fungal surface hydrophobicity has many ecological functions and water contact angles measurement is a direct and simple approach for its characterization. The objective of this study was to evaluate if in-vitro growth conditions coupled with versatile image analysis allows for more accurate fungal contact angle measurements. Fungal cultures were grown on agar slide media and contact angles were measured utilizing a modified microscope and digital camera setup. Advanced imaging software was adopted for contact angle determination. Contact angles were observed in hydrophobic, hydrophilic and a newly created chronoamphiphilic class containing fungi taxa with changing surface hydrophobicity. Previous methods are unable to detect slight changes in hydrophobicity, which provide vital information of hydrophobicity expression patterns. Our method allows for easy and efficient characterization of hydrophobicity, minimizing disturbance to cultures and quantifying subtle variation in hydrophobicity.     

A statistical method for identifying differential gene-gene co-expression patterns

MOTIVATION: To understand cancer etiology, it is important to explore molecular changes in cellular processes from normal state to cancerous state. Because genes interact with each other during cellular processes, carcinogenesis related genes may form differential co-expression patterns with other genes in different cell states. In this study, we develop a statistical method for identifying differential gene-gene co-expression patterns in different cell states. RESULTS: For efficient pattern recognition, we extend the traditional F-statistic and obtain an Expected Conditional F-statistic (ECF-statistic), which incorporates statistical information of location and correlation. We also propose a statistical method for data transformation. Our approach is applied to a microarray gene expression dataset for prostate cancer study. For a gene of interest, our method can select other genes that have differential gene-gene co-expression patterns with this gene in different cell states. The 10 most frequently selected genes, include hepsin, GSTP1 and AMACR, which have recently been proposed to be associated with prostate carcinogenesis. However, genes GSTP1 and AMACR cannot be identified by studying differential gene expression alone. By using tumor suppressor genes TP53, PTEN and RB1, we identify seven genes that also include hepsin, GSTP1 and AMACR. We show that genes associated with cancer may have differential gene-gene expression patterns with many other genes in different cell states. By discovering such patterns, we may be able to identify carcinogenesis related genes.     

TIM-Finder: A new method for identifying TIM-barrel proteins

Background  

The triosephosphate isomerase (TIM)-barrel fold occurs frequently in the proteomes of different organisms, and the known TIM-barrel proteins have been found to play diverse functional roles. To accelerate the exploration of the sequence-structure protein landscape in the TIM-barrel fold, a computational tool that allows sensitive detection of TIM-barrel proteins is required.     

A network module-based method for identifying cancer prognostic signatures

Discovering robust prognostic gene signatures as biomarkers using genomics data can be challenging. We have developed a simple but efficient method for discovering prognostic biomarkers in cancer gene expression data sets using modules derived from a highly reliable gene functional interaction network. When applied to breast cancer, we discover a novel 31-gene signature associated with patient survival. The signature replicates across 5 independent gene expression studies, and outperforms 48 published gene signatures. When applied to ovarian cancer, the algorithm identifies a 75-gene signature associated with patient survival. A Cytoscape plugin implementation of the signature discovery method is available at http://wiki.reactome.org/index.php/Reactome_FI_Cytoscape_Plugin     

A new phylogenetic method for identifying exceptional phenotypic diversification

Currently available phylogenetic methods for studying the rate of evolution in a continuously valued character assume that the rate is constant throughout the tree or that it changes along specific branches according to an a priori hypothesis of rate variation provided by the user. Herein, we describe a new method for studying evolutionary rate variation in continuously valued characters given an estimate of the phylogenetic history of the species in our study. According to this method, we propose no specific prior hypothesis for how the variation in evolutionary rate is structured throughout the history of the species in our study. Instead, we use a Bayesian Markov Chain Monte Carlo approach to estimate evolutionary rates and the shift point between rates on the tree. We do this by simultaneously sampling rates and shift points in proportion to their posterior probability, and then collapsing the posterior sample into an estimate of the parameters of interest. We use simulation to show that the method is quite successful at identifying the phylogenetic position of a shift in the rate of evolution, and that estimated rates are asymptotically unbiased. We also provide an empirical example of the method using data for Anolis lizards. [This article was published online on September 20, 2011. An error in a co‐author's name was subsequently identified. This notice is included in the online and print versions to indicate that both have been corrected September 21, 2011.]     

A method for identifying sounds used in the classification of alarm calls

In this study, we present a methodology that identifies acoustic units in Gunnison's prairie dog alarm calls and then uses those units to classify the alarm calls and bouts according to the species of predator that was present when the calls were vocalized. While traditional methods measure specific acoustic parameters in order to describe a vocalization, our method uses the variation in the internal structure of a vocalization to define possible information structures. Using a simple representation similar to that used in human speech to identify vowel sounds, a software system was developed that uses this representation to recognize acoustic units in prairie dog alarm calls. These acoustic units are then used to classify alarm calls and their associated bouts according to the species of predator that was present when the alarm calls were vocalized. Identification of bouts with up to 100% accuracy was obtained. This work represents a first step toward revealing the details of how information is encoded in a complex nonhuman communication system. Furthermore, the techniques discussed in this paper are not restricted to a database of prairie dog alarm calls. They could be applied to any animal whose vocalizations include multiple simultaneous frequencies.     

A primer on the use of mouse models for identifying direct sex chromosome effects that cause sex differences in non-gonadal tissues

In animals with heteromorphic sex chromosomes, all sex differences originate from the sex chromosomes, which are the only factors that are consistently different in male and female zygotes. In mammals, the imbalance in Y gene expression, specifically the presence vs. absence of Sry, initiates the differentiation of testes in males, setting up lifelong sex differences in the level of gonadal hormones, which in turn cause many sex differences in the phenotype of non-gonadal tissues. The inherent imbalance in the expression of X and Y genes, or in the epigenetic impact of X and Y chromosomes, also has the potential to contribute directly to the sexual differentiation of non-gonadal cells. Here, we review the research strategies to identify the X and Y genes or chromosomal regions that cause direct, sexually differentiating effects on non-gonadal cells. Some mouse models are useful for separating the effects of sex chromosomes from those of gonadal hormones. Once direct “sex chromosome effects” are detected in these models, further studies are required to narrow down the list of candidate X and/or Y genes and then to identify the sexually differentiating genes themselves. Logical approaches to the search for these genes are reviewed here.