Inheritance and reliability of random amplified polymorphic DNA-markers in two consecutive generations of common carp (Cyprinus carpio L.)

Random amplified polymorphic DNA (RAPD) markers have been used in a variety of genetic studies in fisheries and aquaculture. Most population studies are performed without preliminary data demonstrating the Mendelian inheritance and reproducibility of RAPD markers. In this study, the inheritance and reproducibility of RAPD markers was examined in two consecutive generations of common carp, Cyprinus carpio L. Variability and segregation of RAPD markers were investigated in one F₁ progeny and three F₂ progenies. Seventy-four RAPD markers were generated by five primers using DNA extracted from the initial ornamental (koi) common carp female and wild-type colour common carp male. Fifty-five of these RAPD markers were transmitted to the F₁ progeny and the inheritance patterns were analysed. Twenty RAPD markers were fully reproducible and demonstrated dominant simple Mendelian inheritance patterns in two consecutive generations. Twenty-four RAPD markers were not reproducible in all progenies. Thirteen markers displayed inheritance ratios in the progenies that did not fit simple Mendelian inheritance patterns. Non-reproducibility of RAPD markers and distorted ratios may be caused by the absence of amplification, poor amplification or by the appearance of artefact bands. Random amplified polymorphic DNA markers with poor reproducibility and non-Mendelian inheritance can lead to misinterpretations of data in population studies, resulting in errors in the estimation of genetic diversity within and between individual populations. Therefore, it is recommended to first identify the set of reproducible RAPD markers that demonstrate Mendelian inheritance before application of the RAPD technique in population studies.     

Diatomaceous earth foliar spraying along with adjuvants in pistachio orchards associated with the common pistachio psylla,Agonoscena pistaciae

The common pistachio psylla, Agonoscena pistaciae, is a serious global pest menacing pistachio orchards. Considering the dangers of using excessive chemical pesticides, it seems that using natural insecticides such as diatomaceous earth is a suitable way to lower the residual amount of highly hazardous pesticides. In this study, the effects of diatomaceous earth with different additives, including dipotassium hydrogen phosphate, polyurethane glue as a wood adhesive, and potassium silicate, were investigated in several concentrations over two years in orchard conditions. Although all treatments showed significant effects, the most effective treatments were (diatomaceous earth+dipotassium hydrogen phosphate) and (diatomaceous earth+polyurethane glue). Therefore, the use of diatomaceous earth combined with the additive materials mentioned can potentially be a safe method for the integrated management of the common pistachio psylla.     

Genetic interpretations of the effects of bulk breeding on four populations of beans (Phaseolus vulgaris L.)

Summary The effects of bulk breeding on yield and its components in two high-yeilding and two low-yielding bean populations when grown at crop densities are interpreted in terms of gene action and selection. There was considerable additive variation in the crosses, dominance effects were variable but tended to be negative. The results were compared with results obtained at low density by another worker with the same populations, and this showed that interpretations of gene action in hybrid vigour depend on the test environment used. The interpretations were also compared with plant breeding interpretations made on the same material (Hamblin, 1977), and it was found that a single pattern of response to natural selection, in plant breeding terms, may be explained by more than one genetic interpretation and, conversely, a single genetic interpretation may result in different patterns of response in plant breeding terms. The value of composite crosses in plant breeding are discussed and it is concluded that there is a greater chance of producing improved lines from a large number of simple crosses than from a smaller number of composite crosses.     

Identification of a novel filovirus in a common lancehead (Bothrops atrox (Linnaeus, 1758))

I performed metaviromic analysis of publicly available RNA-seq data from reptiles to understand the diversity of filoviruses (family Filoviridae). I identified a coding-complete sequence of a filovirus from the common lancehead (Bothrops atrox (Linnaeus, 1758)), tentatively named Tapajós virus (TAPV). Although the genome organization of TAPV is similar to mammalian filoviruses, our phylogenetic analysis showed that TAPV forms a cluster with a fish filovirus. However, TAPV is still distantly related to all the known filoviruses, suggesting that TAPV can be assigned as a species of a novel genus in Filoviridae. To our knowledge, this is the first report identifying a filovirus in reptiles, and thus contributes to a deeper understanding of the diversity and evolution of filoviruses.     

"兰箭3号"春箭筈豌豆叶绿体全基因组草图及特征分析

箭筈豌豆(Vicia sativa)是自花授粉的二倍体一年生豆科牧草,可为我国高海拔地区的反刍动物提供优质蛋白粗饲料。以"兰箭3号"春箭筈豌豆为研究对象,采用DNase I法纯化叶绿体,利用第二代高通量测序平台Illumina Hiseq2000进行测序,并对"兰箭3号"叶绿体全基因组序列的结构特征进行分析。结果表明,"兰箭3号"叶绿体基因组仅包括一个单拷贝的反向重复序列,其叶绿体全基因组大小为121 883bp,共编码了109个基因,包括4个核糖体RNA(rRNA)基因,29个转运RNA(tRNA)基因,75个蛋白质编码基因和1个假基因。"兰箭3号"在叶绿体基因组结构、基因种类、排列顺序上与其它豆科植物基本一致。其叶绿体全基因组序列已在GenBank注册,序列号为KU053796。"兰箭3号"叶绿体全基因组测序的成功为叶绿体分子生物学研究奠定了基础,并可有效地促进箭筈豌豆遗传育种和分子进化研究。     

空肠弯曲菌共同抗原单克隆抗体的研制及其在细菌检验中的应用 Ⅱ.杂交瘤细胞株的建立及特性鉴定

应用B-淋巴细胞杂交瘤技术,建立了5株稳定分泌抗空肠弯曲菌共同抗原(CA)单克隆抗体(McAb)的杂交瘤细胞株(4A7,9D6、9C2、11A1、11D4)。在5株杂交瘤分泌的McAb中,有4株(4A7、9D6、11A1、11D4)针对CA中的耐热抗原成分,对其中4A7、9D6两株分泌的McAb进行免疫印迹分析表明,其与CA中大约为7.7×104u的蛋白移行带出现特异反应;一株(9C2)针对CA中的不耐热抗原成分。对杂交瘤细胞的核型分析表明,其染色体数在93-108之间。体外传代4个月,仍保持稳定分泌抗体的特性。这种McAb具有较强的特异性,除与幽门弯曲菌出现弱阳性反应外,与参试的其它31种非弯曲菌属的细菌均为阴性反应。对不同地区、不同来源的516株空肠弯曲菌的鉴定表明,制备的McAb与现有空肠弯曲菌均出现阳性反应,与常规鉴定法的符合率为100％。     

燕麦与箭薚豌豆不同混作模式对根际土壤微生物数量的影响

 本试验以燕麦与箭薚豌豆不同种植模式的根际土壤为对象,对其土壤微生物数量进行了初步研究。结果表明,根际土壤细菌、真菌和放线菌数量在燕麦拔节孕穗期较低,开花期较高,随着生育进程的推进至成熟收获期数量呈逐渐减少的趋势。在不同生育期,间混作处理比单作燕麦显著提高了根际土壤细菌数量（P＜０．０５）。收获期土壤微生物以细菌占绝大多数,其中间作混播根际土壤细菌数量是单作燕麦的１．１倍左右,单作箭薚碗豆是单作燕麦根际土壤细菌数量的１．３倍;土壤微生物总数量以燕麦与箭薚豌豆间作（１∶２）和燕麦与箭薚豌豆混播（１∶２）处理较高,单作燕麦最少。说明间作混播种植方式能起到提高根际土壤微生物数量的作用。     

Characterization of acetylcholinesterase and its molecular forms in organs of five freshwater teleosts

The activity and molecular forms of acetylcholinesterase (AChE, EC 3.1.1.7) were characterized in the brain, heart, white skeletal muscle and liver of the Siberian sturgeon (Acipenser baeri), European eel (Anguilla anguilla), wels (Silurus glanis), common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix), belonging in four families. The brain had the highest (between 183.2±10.6 and 1361.2±189.7 mU mg^-1 protein and between 4.7±0.4 and 23.5±2.7 U g^-1 wet tissue) and the liver the lowest (between 25.3±1.9 and 126.5±22.7 mU mg^-1 protein and between 1.0±0.1 and 4.5±0.5 U g^-1 wet tissue) activity in all these fish, with the exception of the wels, where the heart contained the smallest amount of AChE (26.2±7.5 mU mg^-1 protein and 1.2±0.3 U g^-1 wet tissue). The highest tissue AChE activity was found in the carp and lowest in the sturgeon. The solubility properties and molecular forms of AChE in the four tissues were studied by extraction in high-salt medium (1.0 M NaCl) with and without the detergent Triton X-100 (0.5%, v/v). The proportions of detergent-soluble (DS) and salt-soluble (SS) forms of AChE varied considerably from one species to another, but a general tendency could be observed: the proportion of DS AChE was generally higher in the brain (between 70.4±4.1 and 82.5±1.2%) and lower in the heart (between 12.0±2.0 and 51.3±2.3%) and skeletal muscle (between 16.0±2.0 and 64.4±0.7%). Velocity sedimentation centrifugation revealed that most tissues contained the G₄ or A₄ form. Three tissues (brain, heart, and liver) of the Siberian sturgeon contained a relatively high proportion of the G1 form (between 24.3±2.4 and 28.6±4.9%), while the skeletal muscle of the wels contained only the most complex type of AChE, i.e., A₁₂. Amongst the tissues studied, the brain and liver mostly contained amphiphilic globular forms of the G₄ type, while the heart and skeletal muscle were rich in asymmetric forms (A₄ and A₁₂). It was concluded that the differences revealed in the activities of AChE and in the distribution of its molecular forms may be connected with the variation in behavioral habits (feeding and swimming patterns), and the developmental stage of the nervous system.     

Growth testing of two breeds of common carp, Cyprinus carpio L. (Hungarian mirror and Třeboň scaly carp), in ponds with low and high stocking density

The growth of two breeds of common carp, Cyprinus carpio L., was tested in ponds under the climatic conditions of South Bohemia. T?eboň scaly carp (TR) and Hungarian mirror carp (M2) were kept in both low and high stocking densities during the second growing season and then stocked together for communal testing during the third growing season. Before the communal testing, the mean initial weights of fish from low‐ and high‐density stocks differed significantly (374.1 vs. 227.7 g for the TR breed and 766.7 vs. 317.3 g for the M2 breed respectively, P<0.01). After communal testing, mean weights of fish from low‐ and high‐density stocks gained 761.8 vs. 543.8 g for the TR breed and 1339.7 vs. 706.7 g for the M2 breed respectively. These observed weights were also significantly different (P<0.01). However, the test of corrected weight gain, i.e. gain not related to the initial weight of fish, revealed insignificant differences (P>0.01) between the weight gains after correction, i.e. the effect of different initial weights was successfully eliminated. These results seem to confirm the applicability of this method for the assessment of growth of purebred common carp under the climatic conditions of Central European fish farms.     

Ionoregulatory responses to temperature change in two species of freshwater fish

We examined the ionoregulatory responses to temperature changes in two species of freshwater fish that differ in thermal preferences; the stenothermal, cold-water rainbow trout (Oncorhynchus mykiss) and the more eurythermal, warm-water common shiner (Notropis cornutus). We found that rainbow trout maintained constant plasma Na⁺ levels over the entire temperature regime (5–20 °C). Upon transfer from 15 °C (holding temperature) to 5 and 10 °C, rainbow trout experienced a significant drop in Na⁺ uptake (J_in ^Na), but after two weeks J_in ^Na had upregulated to warm temperature levels. Further, Na⁺ efflux (J_out ^Na) fell significantly at the colder temperatures. As a result, trout at the lowest temperatures were still in ion balance. When trout were exercised to exhaustion both O₂ consumption (MO₂) and J_out ^Na rose significantly at all temperatures, but while MO₂ continued to be dependent upon temperature, J_out ^Na was high and constant. In contrast to the trout, common shiners experienced a 20% drop in plasma Na⁺ at 5 °C. Upon exposure to cold temperatures they experienced a reduced J_in ^Na, and showed no signs of acclimation during the subsequent two weeks. Likewise J_out ^Na was constant at all temperatures. These findings raise questions regarding the degree to which fish employ homeostatic mechanisms designed to defend a set- point (i.e., steady-state) osmolarity and ionic composition.