Variation among stallions in sperm quality after single layer centrifugation

Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p < .05–p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples.     

AANAT基因在滩羊不同繁殖时期卵巢中的转录差异及DNA甲基化程度分析

试验旨在检测5-羟色胺-N-乙酰基转移酶（AANAT）基因在绵羊休情季节和繁殖季节（卵泡期和黄体期）卵巢组织中的转录差异,并分析转录差异是否由DNA甲基化修饰程度改变所导致。试验采用自然环境条件和饲养管理一致,且体重差异在0.5 kg范围内的空怀母滩羊作为试验动物,采集其休情期、卵泡期和黄体期（每个时期3只）的卵巢组织,采用SYBR染料法进行实时荧光定量PCR检测AANAT基因在滩羊不同繁殖时期卵巢组织中的转录水平。随后针对转录水平有差异的两个时期（休情期和卵泡期）的样本,利用MethPrimer 2.0在线软件预测AANAT基因启动子区和第一外显子区的CpG岛;用重亚硫酸盐测序法（BSP法）检测AANAT基因启动子区及第一外显子区的甲基化程度。试验结果显示,滩羊休情期卵巢组织中AANAT基因转录水平显著低于卵泡期的AANAT基因转录水平（P<0.05）,休情期与黄体期滩羊卵巢组织中AANAT基因的转录水平差异不显著（P>0.05）。滩羊卵巢组织中AANAT基因启动子区上存在着一个长度为173 bp的CpG岛,第一外显子区存在着一个长度为118 bp的CG岛。然而,两个甲基化岛区内的单个CpG位点甲基化程度在滩羊休情期和卵泡期之间均不存在显著差异,暗示AANAT基因的表达受甲基化修饰外的因素调控。本研究结果可为进一步探讨AANAT基因在季节性发情和卵泡成熟中的功能提供参考资料。     

DNA barcoding for identification of cryptic species in the field and existing museum collections: a case study of Aethomys and Micaelamys (Rodentia: Muridae)

DNA barcoding has been proposed as a method for species identification. However, this method has been criticised for its over-reliance on a single mitochondrial gene. In this study, four mitochondrial gene regions and one nuclear gene region were used to investigate their different abilities to identify tissue associated with museum specimens of Aethomys chrysophilus, Aethomys ineptus and Micaelamys namaquensis. Aethomys chrysophilus and the more recently elevated A. ineptus are indistinguishable on morphological grounds; however, their ranges are largely parapatric with only one syntopic locality currently known. All of the mitochondrial gene regions were able to separate M. namaquensis from A. chrysophilus and A. ineptus, but they varied in their abilities to resolve differences between A. chrysophilus and A. ineptus. The sequence results identified a specimen from KwaZulu-Natal that was misclassified and should have been identified as A. ineptus. Seven specimens that had not been reclassified following the elevation of A. ineptus to species level were identified as A. ineptus. Individuals of A. chrysophilus from Malawi could not be classified as either A. chrysophilus or A. ineptus, and may be a hybrid or a new, distinct species. This study indicates that DNA barcoding may be used to separate M. namaquensis from A. chrysophilus and A. ineptus, and although it was not able to separate A. chrysophilus and A. ineptus, it did indicate specimens from Malawi may be a new cryptic species.     

棉花分子育种研究进展

本文主要综述了我国近２０年来棉花分子育种在理论基础、技术方法、育种机理以及新品种培育等方面研究取得的成就及新进展，并讨论了棉花分子育种的发展方向     

Developing a Recognition System of Gun-code

Intelligent recognition system of gun code is designed , which achieves online recognition of gun code. We applied video camera to get image, and some special technique in this deformed character recognition. As well, we applied dynamic template matching to get a better result.     

Avoiding Obstacle Planning for Robots on Molecular Optimization Algorithm

Interest in DNA computing has increased overwhelmingly since Adleman successfully demonstrated its capability to solve Hamiltonian Path Problem. This article introduces the improving method in virtue of the biological thery of DNA technology, a new molecular algorithm is advanced. After a numerical simulation, the result shows that it avoids the prematurely and lower convergent speed of the classic genetic algorithm, and inherits global search capability, the validity and the speed of the genetic algorithm have been increased. The best result can be obtained in few iterative times. It is fit for solving path planning problem.     

以冻融法快速纯化高活力基因工程Taq DNA聚合酶

用含有TaqDNA聚合酶基因的pTaq表达质粒转化E.coli DH5α菌株，IPTG诱导表达Taq DNA聚合酶，利用该酶的热稳定性，反复2次－70℃，75℃交替冻融以裂解细胞释放胞浆；以高速离心除去冻融变性的细胞碎片及核酸蛋白的复合物以达到快速纯化Taq DNA聚合酶的目的。PCR扩增反应和SDS－PAGE分析表明所制备的Tap DNA聚合酶的纯度，活力，敏感性，特异性均达到试验要求。该方法具有快速简便的优点。     

变铅青链霉菌DNA的研究——Ⅱ.位点特异性降解与DNA修饰的关系

变铅青链霉菌的DNA上存在着一种异常的修饰,使其在含有微量Fe~(++)的缓冲液中电泳时,双链DNA遭到降解。DNA的切割是位点特异性的。与已知修饰特征的DNA进行同步试验发现,变铅青链霉菌的这种特异性修饰与目前所发现的修饰系统(如DNA甲基化)均不相同,很可能是一种新的修饰系统。     

Mutation Techniques for Gene Discovery and Crop Improvement

One of the most important breakthroughs in the history of genetics ）was the discovery that mutations can be artificially induced in organisms （van Harten, 1998）. Artificially induced mutations, by physical and chemical mutagens, have greatly advanced the understanding of genetics of higher organisms. Starting in the late 1960＇s, the International Atomic Energy Agency （IAEA） and the Food and Agriculture Organization （FAO） of the United Nations sponsored extensive research on mutation induction and their application to breeding of food and indnstrial crops that resulted in the introduction of new varieties of rice, wheat, barley, apples, citrus, sugar cane, banana, and others （more than 2 500 officially released new varieties are to be found in the FAO/IAEA Mutant Varieties Database） （hitp：//www-mvd.iaea.org/MVD/default.htm）. However, the usefulness of mutation techniques has been underappreciated in research communities, particularly during the last decade, when more and more researchers and breeders were rushing into molecular marker techniques and transgenic plants. In this paper, after a brief review of the past accomplishments of mutation induction and its application, we discuss the uniqueness of induced mutations in gene discovery and how to integrate induced mutants into functional genomics programs;