Nuclear transfer in loach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the loach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 °C in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days. It was found that gastrula cells incubated at 4 °C had the same totipotence as blastula cells. The optimal UV dosage for inactivation of the oocyte chromatin was 180–240 mJ cm⁻². Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.     

奶牛冷冻胚胎规模化移植效果研究

为了提高奶牛良种化程度,对奶牛冷冻胚胎的规模化移植效果进行了研究。结果表明:PG二次肌注法的发情率显著高于(P<0.05)PG一次肌注法,两种方法的发情同期率均超过85%,受体牛发情时间多集中在肌注PG后48～72h;胚胎解冻后的复苏率达到82.58%,且早期囊胚与囊胚的复苏率要高于桑椹胚(P>0.05);共移植受体牛218头,受胎97头,受胎率为44.5%,且春、秋季的移植效果差异不显著(P>0.05)。为奶牛冻胚的规模化移植研究提供了参考依据。     

波尔山羊胚胎移植影响因素的分析

波尔山羊胚胎移植效果受诸多因素的影响，供体状况、药物、季节等因素使供体超排效果产生较大的差异，而受体状况、胚胎质量等对受体的妊娠率产生较大的影响，这些因素阻碍了波尔山羊胚胎移植的产业化应用。本文对以上影响波尔山羊胚胎移植效果的因素进行了分析，以期为提高胚胎移植效率提供依据，从而促进波尔山羊胚胎移植产业化进程。     

家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

A grower-friendly method to transfer predacious mites to commercial orchards

The establishment of predacious mites in commercial orchards may be accelerated by the transfer of pruned wood in winter and summer from donor orchards to release orchards. Following winter pruning, 3-year-old and older wood is collected and transported as soon as possible in bundles to a release orchard for distribution. If the release orchard is composed of dwarf trees, then one or two bundles of 5 kg each are placed vertically at the base of the trunk of every tree in the block (0.5 to 1 ha); if the trees are of standard size, then four or five bundles used. Following summer pruning, annual shoots and suckers are distributed immediately in a release orchard composed of dwarf trees by placing 12–15 branches on the foliage of fruit-bearing branches; if the release orchard is composed of standard trees, then 50 branches are used. The pruned wood should have 20–25 leaves and not less than one predator per leaf. The release orchard should have a light infestation (two or three mites per leaf) of pest tetranychids. These phytophagous mites would serve as food and help establish the predators. The release orchard grower should develop a pest management program based on the same groups of pesticides used in the donor orchard. http://www.phytoparasitica.org posting Aug. 31, 2005.     

Meiotic competence of mouse oocytes reconstructed by replacing the germinal vesicle with a male pronucleus and somatic nucleus

The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes.     

中国东北和华北地区紫花苜蓿根瘤菌遗传多样性研究

【目的】紫花苜蓿(Medicago sativa L.)被誉为牧草之王,近年来,中国东北和华北地区种植面积不断扩大,但紫花苜蓿质量与产量仍不足以满足中国畜牧业发展的需求。本研究旨在分析中国东北和华北地区紫花苜蓿根瘤菌遗传多样性,为紫花苜蓿高效固氮根瘤菌的筛选与应用提供参考。【方法】采用表面消毒和平板划线法从根瘤中分离纯化根瘤菌单菌落;使用BOX-PCR方法对供试菌株进行基因型划分;选取代表菌株进行持家基因(atpD、glnII和rpoB)和共生基因(nifH和nodC)的系统发育分析。【结果】从中国东北和华北19个采样地共分离纯化了499株根瘤菌,BOX-PCR可将供试菌株分为37种BOX型,BOX型存在显著的地理分布现象,同时寄主品种对根瘤菌基因型具有一定的选择作用。97.60%(487/499)的根瘤菌为Sinorhizobium meliloti。其余12株分别为S. adhaerens、Mesorhizobium huakuii、Rhizobium loessense、R.mesosinicum、R. vignae、Mesorhizobium sp.和Phyllobacteri...     

Human serum albumin interaction with paraquat studied using spectroscopic methods

The interaction between herbicide paraquat and human serum albumin (HSA) was investigated by fluorescence and UV/Vis absorption spectroscopy. Paraquat can strongly quench the intrinsic fluorescence of HSA by static quenching and nonradiative energy transferring; The hydrophobic and electrostatic interactions play a major role in stabilizing the complex. The binding site number n and apparent binding constant K_A, corresponding thermodynamic parameters ΔG, ΔH, ΔS at different temperatures, were calculated. The distance r between donor (HSA) and acceptor (paraquat) was obtained according to fluorescence resonance energy transfer. The effect of paraquat on the conformation of HSA was analyzed using synchronous fluorescence spectroscopy.     

基于迁移学习和双线性CNN的细粒度菌菇表型识别

为了对细粒度菌菇进行表型识别,在双线性卷积神经网络细粒度图像识别框架基础上,提出了一种基于迁移学习和双线性Inception-ResNet-v2网络的菌菇识别方法。利用Inception-ResNet-v2网络的特征提取能力,结合双线性汇合操作,提取菌菇图像数据的细粒度特征,采用迁移学习将ImageNet数据集上预训练的模型参数迁移到细粒度菌类表型数据集上。试验表明,在开源数据集和个人数据集上,识别精度分别达到87.15%和93.94%。开发了基于Flask框架的在线菌类表型识别系统,实现了细粒度菌菇表型的在线识别与分析。