Extensive dispersal of Roanoke logperch (Percina rex) inferred from genetic marker data

The dispersal ecology of most stream fishes is poorly characterised, complicating conservation efforts for these species. We used microsatellite DNA marker data to characterise dispersal patterns and effective population size (N_e) for a population of Roanoke logperch Percina rex, an endangered darter (Percidae). Juveniles and candidate parents were sampled for 2 years at sites throughout the Roanoke River watershed. Dispersal was inferred via genetic assignment tests (ATs), pedigree reconstruction (PR) and estimation of lifetime dispersal distance under a genetic isolation‐by‐distance model. Estimates of N_e varied from 105 to 1218 individuals, depending on the estimation method. Based on PR, polygamy was frequent in parents of both sexes, with individuals spawning with an average of 2.4 mates. The sample contained 61 half‐sibling pairs, but only one parent–offspring pair and no full‐sib pairs, which limited our ability to discriminate natal dispersal of juveniles from breeding dispersal of their parents between spawning events. Nonetheless, all methods indicated extensive dispersal. The AT indicated unrestricted dispersal among sites ≤15 km apart, while siblings inferred by the PR were captured an average of 14 km and up to 55 km apart. Model‐based estimates of median lifetime dispersal distance (6–24 km, depending on assumptions) bracketed AT and PR estimates, indicating that widely dispersed individuals do, on average, contribute to gene flow. Extensive dispersal of P. rex suggests that darters and other small benthic stream fishes may be unexpectedly mobile. Monitoring and management activities for such populations should encompass entire watersheds to fully capture population dynamics.     

泥鳅线粒体DNA控制区结构分析及遗传多样性研究

采用PCR和DNA测序技术对4个泥鳅（Misgurnus anguillicaudatus）群体的线粒体DNA控制区序列进行比较，研究其遗传多样性和控制区的结构。结果显示，用于分析的线粒体DNA控制区片段序列为918～920bp。32个序列中共发现了56个多态位点，其中32个转换位点，19个颠换位点，5个转换与颠换同时存在的位点，定义了32种单倍型。同时对控制区结构进行分析，识别了其终止序列区（ETAS）、中央保守区（CD）和保守序列区（CSB）的关键序列。4个地理群体的单倍型多样性（Hd）、核苷酸多样性（Pi）和平均核苷酸差异数（K）分别为0．992、0．012和10．698。群体间的平均Kimura双参数遗传距离（Kimura 2-parameter distance，K2-P）、遗传分化指数（Fst）、基因交流值（Nm）和分子方差分析（AMOVA）均表明4个泥鳅群体具有较高的遗传分化，基因交流贫乏。利用核苷酸序列构建的NJ分子系统树揭示4个群体分为2个谱系，除范县群体外，其余各群体间相互交叉聚集。     

虹鳟杂交胚胎(2n♀×3n♂)的发育状况及其DNA倍性分析

雄性三倍体虹鳟(Oncorhyncus mykiss)可以发育至生理成熟并表现出较强的雄性副性征,性腺发育基本正常；雌性三倍体虹鳟性腺几乎不发育,并且存在着类雄性化的发育趋势.本研究拟通过对虹鳟杂交胚胎(2n♀×3n♂)发育状况及其DNA倍性的分析,进而确认三倍体虹鳟是否具有与二倍体虹鳟类似的生育能力并分析其原因.选取经两个世代家系选育的虹鳟优良品系二倍体雌性虹鳟作为母本,以热休克方法人工诱导的优质雄性三倍体虹鳟为父本,常规人工采集精卵授精,获得三倍体雄性与二倍体雌性杂交胚胎,观察杂交胚胎的发育进程,通过流式细胞仪检测杂交胚胎的DNA含量,进而与二倍体雌雄交配获得的胚胎对照确定杂交胚胎的倍性.结果表明,杂交胚胎的受精率、发眼率和孵化率均显著地低于对照组,杂交胚胎多数在发眼期之前死亡,破膜的胚胎在仔鱼期上浮前全部死亡.杂交胚胎的DNA含量分布范围较大,大致可以分为两个水平,接近84％的胚胎的DNA含量介于二倍体与三倍体之间,约为对照组DNA含量的2.5倍；约有16％的胚胎的DNA含量在三倍体和四倍体之间.本研究认为,染色体非整倍体化可能会引起杂交胚胎细胞核质不相容,进而导致基因表达调控紊乱,从而使杂交胚胎不能正常发育,无法完成由母体卵黄供给的内源营养向外源营养的转换,成为导致杂交胚胎发育过程中大量死亡,以及杂交仔鱼在破膜之后开始进食之前全部死亡的一个重要原因.     

产蛋前期和产蛋高峰期鸡全基因组甲基化差异及其对肝脏转录组影响的研究

旨在通过对产蛋前期和产蛋高峰期鸡肝全基因组甲基化差异进行分析,解析基因组甲基化对不同发育阶段肝中基因表达差异的影响。本研究采用全基因组重亚硫酸盐测序（WGBS）技术对产蛋前期（20周龄）和产蛋高峰期（30周龄,各3只DNA混池）卢氏绿壳蛋鸡肝全基因组的甲基化水平进行检测,并与已有的肝mRNA转录组数据进行整合分析,探讨基因组甲基化对不同生理阶段基因表达差异的影响。结果表明,全基因组范围约有4%的胞嘧啶（C）发生了甲基化（mC）;两个生理阶段的总体甲基化水平基本一致。共检测到670个差异甲基化区域（DMRs）和356个差异甲基化基因（DMGs）。基因本体（GO）和相关信号通路（KEGG）分析发现,超甲基化DMGs显著富集在发育的正向调控、细胞形态改变的调控、VEGF信号通路、肌动蛋白细胞骨架的调控、粘着斑及间隙连接等相关过程,低甲基化DMGs显著富集在胚胎消化道形态的发生、间充质细胞增殖的正向调控、淀粉和蔗糖代谢及Wnt信号通路等相关过程。基因不同功能区域甲基化水平与基因表达水平有关,启动子（promoter）及基因体（gene body）区域甲基化水平与基因表达水平呈显著负相关,其他区域（内含子、3'UTR）的甲基化水平与基因的表达水平无明显关系。其中,与肝脂质代谢相关的候选基因RASD1、HAO1、UBE2O、MSRB3受甲基化调控。本研究绘制了不同生理时期卢氏绿壳蛋鸡全基因组甲基化图谱,结合mRNA转录组数据阐述了DNA甲基化在基因表达方面的调控作用,并鉴定出了不同生理时期受甲基化调控的基因,为深入研究表观遗传调控在不同生理时期蛋鸡肝代谢中的作用机制提供参考。     

割手密线粒体DNA的简易快速提取方法

高效快速地分离提取高质量的线粒体DNA(mtDNA)是研究植物线粒体基因及其起源进化的重要前提。为获得高质量的mtDNA进行割手密资源的线粒体基因序列扩增及测序分析,本研究以割手密黄化苗为材料,通过简单差速离心分离得到线粒体,经DNaseⅠ消化处理,去除核DNA杂质得到较纯的线粒体,然后经过5%SDS和蛋白酶K充分裂解线粒体,利用饱和的苯酚/氯仿(1:1)和氯仿两次抽提除去蛋白质,并经过RNase A的消化处理除去RNA,无水乙醇沉淀等一系列操作得到mtDNA。所提取的mtDNA样品经紫外吸收光度测定,琼脂糖凝胶电泳和PCR扩增分析其浓度和纯度,结果表明利用该方法提取的mtDNA纯度高,完全能满足后续PCR分析及分子克隆测序的要求。该方法不仅DNA提取纯度高,操作简单、快速经济,可为今后开展甘蔗及其各野生种的研究提供技术支持。     

外源DNA导入小麦后的变异系生物学特性及胚乳蛋白的研究

应用受粉后的花粉管能通道Ｃ４作物高梁ＤＮＡ和抗逆性强的长穗偃麦草ＤＮＡ导入小麦，结果在不同组合中出现了不同类型的广泛变异，在三个组合中已选育出几个稳定遗传的优良变异系，其生物学性状大多是介于原受体和供体之间。主要特性是叶功能期延长，籽粒千粒重和单株粒重增加，增产显著，抗锈病能力增强等，其胚乳蛋白电泳图谱发季了明显变化，在变异系中产生了新的蛋白质组分，而且Ａ区和Ｂ区增加的是蛋白质群，相反原受体６８－     

Sex-linked SSR markers in hemp

Hemp is a dioecious plant with sex chromosomes X and Y, the male sex being heterogametic. The quality of the fibre depends on the sex type. The sex chromosomes can be characterized by molecular markers. In this report, sex‐linked simple sequence repeat (SSR) markers are described. One SSR marker was polymorphic in both the populations derived from single crosses, two other markers in but one of the two populations. Three alleles were detected for two SSR markers indicating polymorphism not only between X and Y, but also between different X chromosomes. In addition, several sex‐linked RAPD markers were detected in one population. Recombination within the sex chromosomes was observed for nearly all markers.     

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;     

Characterization of the A4 cytoplasmic male-sterile lines of sorghum using RFLP of mtDNA

Three sorghum cytoplasmic male sterile lines CSV4 A(V), CSV4 A(G₁) and CSV4 A(M), grouped as A₄, were compared with a milo (A₁) and two other non-milo (A₂ and A₃) cytoplasms for their RFLP patterns of mitochondrial DNA (mtDNA). A 9.7 kb clone from pearl millet mtDNA discriminated each of the three A₄ entries whereas other maize and pearl millet mtDNA clones used could not distinguish this group completely. The molecular differences within the A₄ cytoplasmic group offer some explanation for the inconsistency in the fertility restoration behaviour of these A₄ lines obtained with a definite set of testers in the field. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of 'quick and dirty' DNA extraction methods for marker-assisted selection in rice (Oryza sativa L.)

Six simple methods for extracting DNA from rice seedlings were evaluated for marker‐assisted selection (MAS). The assessment of each method was based on PCR amplification of SSR markers, DNA yield and purity, time and cost. Based on these criteria, two methods were selected as being superior to other methods. The best two methods included the standard method developed at the International Rice Research Institute (IRRI), which utilizes a sodium dodecyl sulfate extraction buffer followed by chloroform/isoamyl alcohol extraction and a previously published method using sodium hydroxide and Tris. These two methods produced nearly identical PCR amplification results. The sodium hydroxide method is considerably simpler, quicker and cheaper than the standard IRRI method, and may be particularly useful for many applications of MAS or high‐resolution mapping. This method was also adapted into an effective high throughput method utilizing 96‐well plates emphasizing its versatility.