孑遗植物银杏群体遗传多样性的ISSR分析

采用ISSR分子标记技术,对江苏泰兴、美国纽约的栽培银杏(Ginkgo biloba)群体和中国3个可能为野生的银杏自然群体(浙江西天目山、贵州务川、湖北大洪山区)的遗传多样性水平和群体遗传结构进行了研究。用13个引物对5个群体共66个样品进行扩增,共得到88个清晰的扩增位点,其中多态性位点62个,多态位点百分率(PPB)为70．45％。POPGENE分析结果表明：与其他裸子植物相比,银杏具有丰富的遗传变异(He=0．2408;Ho=0．3599)。贵州务川群体的遗传多样性水平最高(PPB=56．82％,He=0．2089,Ho=0．3087),江苏泰兴栽培群体(PPB=34．09％,,Ho=0．1269,Ho=0．1858)和美国纽约的栽培群体(PPB=23．86％,,He=0．0884,Ho=0．1312)的遗传多样性水平较低。Nei′s遗传多样性分析和AMOVA分析表明,3个可能的自然群体间出现了一定程度的遗传分化(Gst=0．1476,Φst=14．26％)。群体间一定程度的遗传分化可能是人为选择压力和基因流障碍引起的。根据Nei′s遗传距离矩阵分别构建了群体间和个体间的遗传关系树状图。由UPGMA聚类分析可知,贵州务川群体与浙江西天目山群体优先聚类;美国纽约群体与湖北大洪山群体具有较近的亲缘关系,它们可能为同一野生群体的后裔。通过对银杏群体遗传结构的分析并结合群落学调查研究,结果表明：贵州务川银杏群体很可能为野生自然群体。基于银杏群体遗传学和生态学的研究结果,建议在自然银杏群体最适生境和遗传多样性最高的贵州务川建立银杏保护区。由于银杏群体间出现了一定程度的分化,建议3个自然群体间可进行植株和幼苗相互移栽,以提高群体间的基因交流,以最大限度地保护银杏的遗传多样性。     

银杏ISSR-PCR扩增反应体系的建立与优化

目的:为了对银杏进行分子鉴定和遗传关系的分析,建立银杏ISSR-PCR的最佳扩增反应体系。方法:采用正交设计和单因素梯度实验,对影响ISSR-PCR反应体系的5个主要因素(Mg2+、dNTP、引物、模板DNA及Taq DNA聚合酶)进行筛选及优化。结果:银杏25μL ISSR最佳扩增反应体系包含10×Taq反应缓冲液、2.5 mmol/L MgCl2、0.45 mmol/L dNTP、1.2μmol/L引物(UBC861)、10 ng模板DNA及0.9 U Taq DNA聚合酶,使用此ISSR扩增反应体系,获得了10株不同性别银杏DNA的清晰条带,验证了该体系的稳定性。结论:优化的反应体系为采用ISSR分子标记技术对银杏进行遗传多样性分析、遗传育种和转基因等研究奠定了一定的理论基础。     

银杏观赏品种遗传关系的AFLP分析

从64对EcoRⅠ／MseⅠ引物（其中MseⅠ引物为荧光标记物）中筛选出8对扩增产物多态性高、谱带清晰的引物,对来自美国、荷兰、日本、法国和我国的21个银杏观赏品种的遗传关系进行了研究。结果表明：8对引物共产生1117条谱带,229个特异位点（其中缺失带54条、单态带175条）,多态带983条,多态带的比例为88％;每对引物鉴别效率为100％。14个国外银杏观赏品种平均多态带的比例35．86％,7个国内品种平均多态带的比例31．51％。21个观赏品种之间相似系数为0．4899-0．8499。当相似系数为0．7300时,供试观赏品种可分为四类,来源地相同的品种并不单独聚成一类,中国和法国的品种分别属于其中的三类。根据对观赏品种的特异位点、相似系数、聚类结果等进行综合分析表明,‘塔形银杏’、‘垂乳银杏’、‘筒叶银杏’、‘大耳银杏’、‘斑叶银杏’、‘展冠银杏’、‘垂枝银杏’、‘沂源叶籽银杏’这8个品种是银杏观赏品种中的重要特异种质。     

天目山银杏群体遗传变异的同工酶分析

本文利用同工酶电泳方法,研究了天目山银杏群体的遗传变异性,计算了4种同工酶(GDH,G-6PDH,PRX,SDH)8个位点上的等位基因频率和每个位点的平均杂合率。结果表明:天目山银杏群体的遗传变异性较小,各位点的平均杂合率仅为0.150±0.004,呈现较大程度的遗传同一性,因此认为天目山银杏很可能是僧人在寺庙旁栽植的银杏留下的后代。     

不同地理种群银杏大蚕蛾COI基因序列变异与遗传分化

银杏大蚕蛾Caligula japonica是亚洲东部的特有种, 既是一种重要的林业害虫, 也是一种珍贵的野生蚕类资源。为了揭示银杏大蚕蛾地理种群间的内在联系, 测定了我国分布的12个地理种群的线粒体细胞色素氧化酶C亚基I（COI）基因部分序列（GenBank登录号: FJ358506-FJ358517）, 对地理种群间的序列变异和遗传分化进行了分析。结果表明: 银杏大蚕蛾地理种群间的COI序列同源性高达99%~100%, 显示出比较小的遗传差异。序列对准后从供试COI序列中仅鉴定出9个变异位点和6个单元型, 其中3种是共享单元型。系统发育分析结果表明种群间已经按地理位置形成了一定的地理格局, AMOVA分析显示北方组和南方组之间已经具有明显的遗传分化（FST=0.478, P<0.001）。综合分析, 我们认为北方组和南方组之间的遗传分化可能与差异巨大的生态条件有关。研究结果为银杏大蚕蛾的种群遗传学和生态学研究提供了一个基本的分子生物学线索。     

不同地域银杏大蚕蛾的遗传多样性

银杏大蚕蛾Dictyoploca japonica Moore是一种重要的林业害虫,近年来在我国局部地区暴发且有逐步蔓延的趋势。利用ISSR技术对10个不同地域的银杏大蚕蛾的基因组DNA进行遗传多样性分析。结果表明:所采用的35个随机引物中,有20个引物扩增出清晰且重复性好的条带,扩增总片断数为299条,其中有250个位点具有多态性,多态性位点比率为83·61%,不同地域银杏大蚕蛾间的遗传距离(D)在0·2315~0·4147之间。根据D值,由UPGMA聚类分析软件绘制了分子进化树,发现不同地域银杏大蚕蛾已发生种群分化。     

分子遗传保护研究进展

介绍了遗传保护的内涵和遗传保护的途径及存在的问题,从遗传多样性检测、种群内遗传变异及进化分化的鉴别、就地或移地保护物种的繁殖与遗传保护、遗传结构地理差异、种群大小的遗传基础与保护等方面综述了遗传保护研究的进展。     

DNA遗传标记技术在林业生产中的应用

林木种子及苗木鉴别技术的准确性和可靠性是提高营林造林质量的关键环节。传统的种苗鉴别技术的局限性往往使之不能有效鉴别林木种子的品种及种源。ＤＮＡ遗传标记技术则能提供稳定、准确、可靠的种及品种特异性标记,因而成为林业生产上极具潜力的品种鉴定手段。本文综述了目前国际上ＤＮＡ遗传标记技术的研究进展,并讨论了开展我国林木种苗ＤＮＡ鉴定技术研究的若干设想。     

SSR标记技术在葡萄品种鉴别及遗传育种上的应用

SSR标记因其数量多、重复性好、出现频率高和共显性遗传等优点在植物的种质资源品种鉴别、遗传育种等方面得到广泛应用。简要介绍了SSR标记在葡萄上的发展和葡萄遗传多样性分析、品种鉴定、DNA指纹图谱构建和遗传育种等方面的应用,并对SSR标记未来在葡萄育种应用的研究方向作出了展望。     

银杏DNA提取及RAPD分析

采用SDS裂解液和苯酚/氯仿/异戊醇提取液从银杏叶中提取银杏总DNA,并进行DNA样品分光光度测定和琼脂糖凝胶电泳分析。通过引物筛选和反应参数优化,选用3个随机引物对DNA样品进行RAPD扩增,获得较为清晰并有一定多态性差异的扩增谱带,初步摸索出适合于以银杏叶为材料的DNA提取方法和RAPD扩增程序,为研究银杏遗传多态性及种质资源研究提供一种实用的分析方法。     

Genetic discrimination and the law.

The use of genetic tests can lead to genetic discrimination, discrimination based solely on the nature of an individual's genotype. Instances of the discriminatory uses of genetic tests by employers and insurance companies have already been reported. The recently enacted Americans with Disabilities Act of 1990 (ADA), together with other federal and state laws, can be used to combat some forms of this discrimination. In this article we define and characterize genetic discrimination, discuss the applicability of the various relevant federal and state laws, including the ADA, in the areas of employment and insurance discrimination, explore the limitations of these laws, and, finally, suggest some means of overcoming these limitations.     

Genetic discrimination and the public entities and public accommodations Titles of the Americans with Disabilities Act.

The introduction of newly developed medical genetic diagnostic tests has been accompanied by social problems involving privacy issues and genetic discrimination. Previous studies of genetic discrimination have focused on the areas of employment and insurance. In this paper, we provide six hypothetical illustrative cases of genetic discrimination involving access to public entities and to private entities considered to be public accommodations. We argue that many of these forms of genetic discrimination that arise in both the public and private sectors should be prohibited by Titles II and III, respectively, of the Americans with Disabilities Act of 1990.     

Discrimination as a consequence of genetic testing.

Genetic discrimination refers to discrimination directed against an individual or family based solely on an apparent or perceived genetic variation from the "normal" human genotype. We describe here the results of a case history study designed to assess whether or not genetic discrimination exists. Using the above definition of genetic discrimination and applying stringent criteria for case selection, we find that genetic discrimination exists and is manifested in many social institutions, especially in the health and life insurance industries. Stigmatization, and denial of services or entitlements to individuals who have a genetic diagnosis but who are asymptomatic or who will never become significantly impaired, is noted. Follow-up comprehensive studies on the significance and varieties of genetic discrimination are needed. In order to avoid creating a new social underclass based on genetic discrimination (the "asymptomatic ill"), existing and future genetic testing or screening programs need review by medical, scientific, legal, and social policy experts, as well as the public, and may require modification.     

Genetic discrimination in insurance and employment

The results of the Human Genome project will eventually have a great impact on medicine. However, the expansion of genetic testing due to these results exacerbates ethical, legal, and economic problems related to the project even today. Virtually free access to the data of testing would present an encroachment on personal freedom, since it may lead to discrimination based on genetic characteristics, i.e., genetic discrimination. Examples of this discrimination are already known; they include unsubstantiated refusals to employ carriers of certain alleles and denials of life or health insurance coverage and the ability to adopt a child. The use of genetic information in the insurance and employment fields is of primary concern due to its economic importance. Consumers consider genetic discrimination in these areas to be intolerable moral and social injustice. Genetic discrimination may eventually lead to the formation of a class of people who cannot buy an insurance policy, and, in the employment field, rejection of persons with "undesirable" genes infringes on citizens' rights to equal opportunity. However, selection for genetic characteristics in employment is justified if these characteristics determine sensitivity to occupational hazards.     

Exploring the concept of genetic discrimination

The issue of genetic discrimination has attracted growing attention and has been the focus of a recent major Australian inquiry. It is, however, a complex and loaded notion, open to interpretation. This paper explores the concept of genetic discrimination in both its theoretical and practical dimensions. It examines its conceptual underpinnings, how it is understood, and how this understanding fits within the legal framework of disability discrimination. The paper also examines the phenomenon in practice, including the 'fear factor' and the resulting implications for the development of regulatory policy. The paper argues that we need to be discerning in our interpretation of this term, and differentiate between discriminatory conduct which is legal and illegal. Further, it is argued that a concerted societal response to the 'problem' of genetic discrimination is needed involving targeted legal reforms as well as broader non-legal strategies.     

Streitkultur and the governance of genetic testing and insurance in Germany

Rapid developments in genetic testing have given rise to fundamental ethical, legal, and social questions that need to be dealt with in society. Results of genetic tests may be of interest to third parties such as private insurance companies, leading to fears of genetic discrimination. In Germany, the Government adopted the Genetic Diagnosis Act (Gendiagnostikgesetz, GenDG) in 2009 to protect people from, inter alia, genetic discrimination in obtaining life or health insurance. Given the sensitivity of the topic, this legislation was continually revised between 2001 and 2009. In this article, we reconstruct the process of formulating the GenDG with regard to genetics and insurance. The article begins with the parliamentary Enquete Commission in 2000 to develop a strategy and recommendations for the governance of genetic diagnostics, and analyzes how these recommendations were applied during the legislative process. We demonstrate that the legislative process of GenDG was largely determined by conventional methods of governance, rather than Streitkultur called for by the Enquete Commission in 2002. We conclude that though Streitkultur was defined as a mechanism to develop a robust approach to the governance of genetic diagnostics, it failed to influence a crucial element in genetic testing and insurance; namely, to fully protect insurees from genetic discrimination.     

Genes,symptoms, and the “asymptomatic ill”: toward a broader understanding of genetic discrimination

Since the early 1990s, the term “genetic discrimination” has been used to designate adverse treatment on the grounds of genetic makeup. However, the full spectrum of possible disadvantage associated with genetic information has not been addressed by either the international scientific debate or statutory arrangements on genetic discrimination. Informed by legal contexts, they almost all focus on one specific group: the “asymptomatic ill.” On the basis of the sociological study, “Genetic Discrimination in Germany,” this article proposes to revise the terms of the debate and discusses some limitations of the concept. Drawing on the experiences reported by affected individuals, it advocates a more expansive social understanding which does not require that a person has to be healthy to be at risk of genetic discrimination.     

Genetic architecture underlying convergent evolution of egg-laying behavior in a seed-feeding beetle

Independent populations subjected to similar environments often exhibit convergent evolution. An unresolved question is the frequency with which such convergence reflects parallel genetic mechanisms. We examined the convergent evolution of egg-laying behavior in the seed-feeding beetle Callosobruchus maculatus. Females avoid ovipositing on seeds bearing conspecific eggs, but the degree of host discrimination varies among geographic populations. In a previous experiment, replicate lines switched from a small host to a large one evolved reduced discrimination after 40 generations. We used line crosses to determine the genetic architecture underlying this rapid response. The most parsimonious genetic models included dominance and/or epistasis for all crosses. The genetic architecture underlying reduced discrimination in two lines was not significantly different from the architecture underlying differences between geographic populations, but the architecture underlying the divergence of a third line differed from all others. We conclude that convergence of this complex trait may in some cases involve parallel genetic mechanisms.     

Absence of within-colony kin discrimination in behavioural interactions of swarm-founding wasps

Within-colony kin discrimination has not been demonstrated conclusively for any social insect, perhaps partly because highly polymorphic genetic markers necessary to assess within-colony relatednesses have only recently become available. We use microsatellite loci to investigate within-colony kin discrimination in behavioural interactions in the neotropical multiple-queen wasp, Parachartergus colobopterus. Within-colony kin discrimination would be particularly advantageous in this species since average genetic relatedness among colony members overall is low (0.32 =/- 0.06), compared to the relatedness value between full sisters of 0.75. Using seven colonies of individually marked females, we recorded behavioural interactions that were cooperative (222 grooming, 2438 feeding), aggressive (511 body or wing biting, 240 mandible biting) or neutral (1676 antennating). We expected cooperative behaviours to favour closer kin and aggressive behaviours to be directed towards more distant kin, but found that none of the behaviours we investigated showed discrimination on the basis of relatedness. We could have detected a difference in relatedness values of as little as between 0.03 and 0.12, depending on the behaviour being analysed. Thus, we found no evidence for kin discrimination in within-colony behaviour in this species.