牛GDF-9基因的克隆与组织表达的分析

为了检测牛GDF-9基因mRNA在怀孕期牛生殖系统的表达情况,为GDF-9基因在雌性动物繁殖上的研究提供一定的理论依据。本试验根据GenBank中报道的牛GDF-9基因序列设计引物,从牛卵巢、输卵管、子宫、肾脏中提取RNA,采用RT-PCR技术进行cDNA扩增,扩增产物与载体pGM-T连接,转化大肠杆菌DH5α,筛选阳性克隆,测序。结果表明,已成功克隆的阳性重组子经测序鉴定其片段大小与预期大小完全一致,为264 bp。序列同源性分析比较表明,该cDNA及其推导的氨基酸序列与绵羊GDF-9cDNA和氨基酸序列同源性分别为97%和94%。以-βactin为内参照物,采用RT-PCR技术进行半定量分析,通过检测比较牛GDF-9基因在卵巢、输卵管、子宫、肾脏组织中mRNA的表达情况。结果表明,牛GDF-9基因在卵巢中表达量最高,在输卵管、子宫、肾脏中均有表达,但表达量不高。     

Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Impact of lakeshore development on green frog abundance

Many amphibian species exhibit metapopulation spatial dynamics and temporally are faced with local population extinction and re-colonization. These natural population fluctuations can exhibit stochastic effects when human-caused alteration and fragmentation of habitats occur during sensitive life-cycle events. In this study, we explored the effects of shoreline development on adult green frogs Rana clamitans melanota on lakes (n=24) of northern Wisconsin. We estimated green frog abundance using both auditory and direct observation surveys. The immediate shoreline habitat was mapped and placed into a Geographical Information System (GIS) for analysis. Adult green frog populations were significantly lower on lakes with varying degrees of shoreline house and cottage development than lakes with little or no development. A negative linear relationship existed between shoreline development densities and the number of adult green frogs. However, house and cottage densities alone did not directly explain this reduction. Analysis of variance (ANOVA) identified that the amount of suitable habitat, not development density, significantly affected green frog abundance. Therefore, greater development densities significantly decrease breeding habitat quality, resulting in lower adult frog abundance. These and other findings suggest that lakeshore development regulations are not protecting sensitive amphibian species.     

Numerical taxonomic studies in the generaTriticum L. andPisum L.

Summary Up till now numerical taxonomy has been applied to cultivated plants only to a limited extent though this method seems to be especially suitable for handling the great diversity of these plants. Moreover the theory of numerical taxonomy has not been studied sufficiently.Therefore studies with cultivated plants were begun with the aim to investigate the fitness of different numerical taxonomic techniques for their classification. Materials used: 1) 24 species ofTriticum and additionally 24 species ofAegilops 2) 1225Triticum taxa from Iran (mostlyT. aestivum) 3) 312 taxa ofPisum (mostlyP. sativum).Using 10 coefficients and 7 clustering methods inTriticum andAegilops in most cases a clear separation of both genera resulted as well as a species arrangement withinTriticum according to the genomes.The main aim when dealing with the wheat from Iran was the investigation of the influence of different character sets. At this the coefficient ofGower and the unweighted pair-group method using arithmetic averages (UPGMA) were applied. The separation of the different species is satisfying clear while the representatives of the different varieties ofT. aestivum could not always be found in one cluster. This effect increased when adding further characters sets.InPisum the influence of 11 similarity coefficients to the dendrogram formation has been investigated. With two-state characters the simple matching coefficient and with mainly multistate characters the Canberra metric gave best results (using UPGMA and information measures SUMRAT and SAMRAT).The hitherto existing results show the complicatedness of the evaluation of numerical taxonomic methods in their application to cultivated plants. Therefore generalizations are not yet possible.

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Numerisch-taxonomische Studien in den GattungenTriticum L. undPisum L.

     

Wasseraufnahme und Wasserverbrauch von Ackerbohne und Hafer auf einer Löß-Parabraunerde

Water uptake and water use of field beans and oats grown on a loess-derived grey-brown podzolic soil (Eutroboralf) The terms of the water-balance equation were determined, when field beans (Vicia faba L.) and oats (Avena sativa L.) were grown on a loess-derived soil during two seasons (1982 and 1983). A specific objective of this investigation was to quantify the water uptake from different layers and the total transpiration of both crops, as field beans are known to be susceptible to water shortage. Beside soil physical measurements climatological data for calculation of potential evapotranspiration were recorded. Plants were analyzed due to leaf area and root length density once a week. Actual evapotranspiration including interception, as determined by the soil physical approach, was split up by calculation procedures into actual evaporation, interception and actual transpiration. Total root length and root length density of field beans were much smaller and the rooting system was shallower as compared with oats. Development of leaf area and of roots was slower with beans than oats and was retarted by 2 to 3 weeks. Accordingly the time of maximum transpiration was found begin of June with oats and at begin of July with field beans. Despite reduced root growth Vicia faba transpired 250 mm in total, that is 86 % of what was found for the cereal crop (290 mm). Water uptake field beans however, was restricted to the upper 80-cm profile with 90 % of total uptake. The water uptake per unit length of root was substantial higher with beans than with oats. Due to the delayed development of the bean crop the losses by evaporation and seepage exceeded those from the soil grown to oats by 40mm (64 %). These investigations support the conclusion that yield stability of field beans may be substantially improved by selection of new varieties with increased rooting depth.     

Occurrence of zearalenone-4-beta-D-glucopyranoside in wheat

An LC-MS method was developed for the analysis of zearalenone-4-beta-D-glucopyranoside and zearalenone in wheat (Triticum aestivum). The limit of determination for zearalenone-4-beta-D-glucopyranoside and zearalenone was 10 microg/kg. The recovery rates were calculated to be 69% and 89% at a concentration of 100 microg/kg for zearalenone-4-beta-D-glucopyranoside and zearalenone, respectively. Twenty-four Bavarian wheat samples from a 1999 harvest were analyzed. Zearalenone was present in 22 of 24 field samples, the levels ranged from 11 to 860 microg/kg. Zearalenone-4-beta-D-glucopyranoside was found in 10 of the zearalenone positive samples (42%) at levels ranging from 17 to 104 microg/kg. The amounts of zearalenone-4-beta-D-glucopyranoside were correlated to those of zearalenone (r2 = 0.86, b = 0.10). After gastrointestinal hydrolyzation, zearalenone-4-beta-glucopyranoside might be implicated in the development of a zearalenone-syndrome. Therefore, more attention should be focused on conjugated mycotoxins in food and feed.     

Interlinked fast and slow positive feedback loops drive reliable cell decisions

Positive feedback is a ubiquitous signal transduction motif that allows systems to convert graded inputs into decisive, all-or-none outputs. Here we investigate why the positive feedback switches that regulate polarization of budding yeast, calcium signaling, Xenopus oocyte maturation, and various other processes use multiple interlinked loops rather than single positive feedback loops. Mathematical simulations revealed that linking fast and slow positive feedback loops creates a "dual-time" switch that is both rapidly inducible and resistant to noise in the upstream signaling system.