Restriction pattern analysis of deoxyribonucleic acid isolated from callus and cell suspension of actinorhizal and non-actinorhizal Betulaceae

Using cell suspensions, a method was elaborated to isolate high-molecular-weight genomic deoxyribonucleic acid (DNA; 65 MDa or more) from members of the Betulaceae: Alnus incana (L.) Moench, Alnus glutinosa (L.) Gaertn. and Betula papyrifera Marsh. The method was also effective for isolation of DNA from callus cells. Based on the chemical lysis of protoplasts, this procedure yielded 130 μg (callus) to 250 μg (cell suspension) of DNA (g fresh cells)⁻¹, with a ratio A₂₀₀/A₂₈ of 1.7–2.0. The purified DNA obtained, formed distinct bands when restricted fragments were electrophoresed. Among the 10 endonucleases used for restriction analysis of Alnus glutinosa, Alnus incana and Betula papyrifera genomes, PvuI1 (EC 3.1.23.33) was unique in giving identical patterns for the two Ainus species. An unusual pattern occurred when Al-2 DNA was restricted with Ava II (EC 3.1.23.4). It formed a ladder with a repeating fragment unit of 181 base pairs long. With the enzymes tested, no differences in restriction patterns were observed among clones of Alnus incana (AI-2 vs AI-2), Betula papyrifera (BP-4 vs BP-8) and subclones of Ainus glutinosa AG-1 (PLFJ709 vs LF1709), suggesting genetic stability of the Betulaceae cultures.     

Occurrence of hemoglobin in the nitrogen-fixing root nodules of Alnus glutinosa

A true hemoglobin (Hb) was shown to be present in the root nodules of Alnus glutinosa L. After purification by gel filtration and ion exchange, the Hb formed a stable complex with oxygen. This oxygen complex could then be converted to carboxyhemoglobin by treatment with CO. Optical absorption spectra typical of Hb were observed. The molecular weight was estimated to be 15 100 by gel filtration, and 18 300 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Hb was largely insoluble when the initial homogenization was done in the absence of a detergent. Under these conditions much of the Hb appears to be associated with clusters of Frankia , the nitrogen-fixing actinomycete that infects plant cells within the nodules. The exact localization of the Hb in vivo is uncertain. The relatively low average concentration of Hb in Alnus nodules suggests that it is either confined to a relatively small fraction of total nodule volume, or has a function other than facilitation of O₂ transport.     

Relationship between the chromatoid body and the acrosomal system in early spermatids of Myxine glutinosa L.

Summary A transient close relationship between the chromatoid body and the developing acrosome is demonstrated in early spermatids of Myxine glutinosa.This work was supported by the Norwegian Research Council for Science and Humanities (NAVF, Grant Nr. D 61.44) and the Austrian Fonds zur Förderung der wissenschaftlichen Forschung, Projekt 2183     

Expression of Nicotiana glutinosa Ribonucleases in Escherichia coli

We previously isolated from Nicotiana glutinosa leaves three distinct cDNA clones, NGR1, NGR2, and NGR3, encoding a wound-inducible RNase NW, and putative RNases NGR2 and NGR3, respectively. In this study, we produced RNases NW and NGR3 in Escherichia coli and purified them to homogeneity. RNase NGR3 had non-absolute specificity toward polynucleotides, although RNase NW preferentially cleaved polyinosinic acid (Poly I). Both RNases NW and NGR3 were more active toward diribonucleoside monophosphates ApG, CpU, and GpU. Furthermore, kinetic parameters for RNase NW (K _m, 0.778 mM and k _cat, 1938 min^?1) and RNase NGR3 (K _m, 0.548 mM and k _cat, 408 min^?1) were calculated using GpU as a substrate.     

地黄GGPPS1基因克隆及表达分析

以地黄为材料,通过分析地黄转录组数据,设计特异性引物,克隆了地黄牻牛儿基牻牛儿基焦磷酸合酶（geranylgeranyl pyrophosphate synthase,GGPPS）基因的cDNA序列,命名为RgGGPPS1,GenBank登录号为KU258808。同时在生物信息学分析的基础上,进行原核表达、纯化以及组织特异性表达分析。结果显示：（1）RgGGPPS1基因开放阅读框为987 bp,编码328个氨基酸。（2）生物信息学分析结果显示,RgGGPPS1蛋白含有2个富含天冬氨酸的基序(DDXXXXDD和DDXXD),与芝麻等双子叶植物中的GGPPS蛋白相似性较高。（3）利用构建的原核表达载体pET 32a RgGGPPS1在大肠杆菌BL21(DE3)菌株中成功表达RgGGPPS1重组蛋白,采用Ni²⁺亲和层析得到了纯化的RgGGPPS1重组蛋白。（4）荧光定量PCR结果显示,RgGGPPS1基因在根中表达量最高,叶、茎中表达量较低。研究结果为进一步研究RgGGPPS1基因在地黄环烯醚萜苷生物合成途径中的功能奠定了基础。     

地黄育种研究进展

地黄是一种玄参科药用植物,是我国著名的"四大怀药"之一,具有重要的药用价值和经济价值。该文综述了地黄的引种、选择育种、杂交育种、组织培养辅助育种、转基因育种和DNA分子标记辅助育种等育种研究的成果,以期为地黄育种工作者提供参考。     

正交试验优化梓醇的微波辅助提取工艺

目的:通过正交实验优选了地黄中梓醇的微波提取工艺。方法:以地黄粗提液中梓醇含量为指标,HPLC为含量测定方法,采用正交实验法,选取乙醇浓度(A)、料液比(B)、提取时间(C)和提取次数(D)4个因素,每个因素选取3个水平进行实验,确定了最佳提取工艺。结果:研究结果表明乙醇浓度为60%,料液比为4,微波提取3次,每次3 min为梓醇的最佳提取工艺。结论:微波辅助提取地黄中梓醇效率高,提取完全,方法可行。     

The ecological effect of phenotypic plasticity — Analyzing complex interaction networks (COIN) with agent-based models

Analyzing complex dynamics of ecological systems is complicated by two important facts: First, phenotypic plasticity allows individual organisms to adapt their reaction norms in terms of morphology, anatomy, physiology and behavior to changing local environmental conditions and trophic relationships. Secondly, individual reactions and ecological dynamics are often determined by indirect interactions through reaction chains and networks involving feedback processes.

We present an agent-based modeling framework which allows to represent and analyze ecological systems that include phenotypic changes in individual performances and indirect interactions within heterogeneous and temporal changing environments. We denote this structure of interacting components as COmplex Interaction Network (COIN).

Three examples illustrate the potential of the system to analyze complex ecological processes that incorporate changing phenotypes on the individual level:

• A model on fish population dynamics of roach (Rutilus rutilus) leads to a differentiation in fish length resulting in a conspicuous distribution that influences reproduction capability and thus indirectly the fitness.

• Modeling the reproduction phase of the passerine bird Erithacus rubecula (European Robin) illustrates variation in the behavior of higher organisms in dependence of environmental factors. Changes in reproduction success and in the proportion of different activities are the results.

• The morphological reaction of plants to changes in fundamental environmental parameters is illustrated by the black alder (Alnus glutinosa) model. Specification of physiological processes and the interaction structure on the level of modules allow to represent the reaction to changes in irradiance and temperature accurately.

Applying the COIN-approach, individual plasticity emerges as a structural and functional implication in a self-organized manner. The examples illustrate the potential to integrate existing approaches to represent detailed and complex traits for higher order organisms and to combine ecological and evolutionary aspects.