长柄石杉居群遗传多样性和遗传结构的AFLP分析

石杉科植物因所含石杉碱甲(Huperzine A)对中老年痴呆等具有良好疗效,近年来倍受关注.利用AFLP分子标记对武夷山脉广布种长柄石杉[Huperzia serrata(Thunb.ex Murray)Trev.var.longipetiolata(Spring)H.M.Chang]7个居群112株个体进行遗传多样性和居群遗传结构分析.选用多态性高、分辨力强的8对选择性扩增引物组合共获得675个位点,其中多态位点比例为69.38%.居群内观测等位基因数(Na)为1.633,有效等位基因数(Ne)1.493;Nei’s基因多样性指数(He)与Shannon多态性信息指数(I)的平均值分别为0.272和0.392,多样性最高为地处武夷山脉中段的泰宁居群和建宁居群,最低为山脉北段的光泽居群.居群总基因多样性(Ht)为0.327 3,居群内基因多样性(Hs)为0.272 2,居群间的遗传分化系数(Gst)为0.168 1,表明居群内变异是长柄石杉遗传多样性的主要来源.由Gst估计,武夷山脉长柄石杉自然居群的基因流(Nm)为2.474 0.邻接树分析表明居群间遗传亲缘关系与地理位置相关.武夷山脉长柄石杉较高的遗传多样性和基因流水平表明其仍然具有相当的适应(生存)能力和进化潜力,这可能与其生物学(异交水平)、生态学特性及武夷山脉相对良好的生境条件有关.     

长江中下游及云贵高原芦苇居群的遗传变异特征

芦苇(Phragmites australis)为全球广泛分布的多型种,在水生生态系统,尤其是湿地生态系统中承担重要的生态功能,准确划分其生态型可为种质资源保护与利用提供理论依据,同时也为修复受损生态系统异地引种提供参考。利用叶绿体DNA rpl16基因序列,选取淡水湖泊较为集中且地理跨度较大的长江中下游和云贵高原,以地理位置为划分指标,探讨19个湖滨带芦苇居群的亲缘关系、遗传多样性和遗传结构,并分析了不同湖泊芦苇形态差异与遗传差异的异同,旨在分子水平为不同地理区域芦苇生态型的划分提供遗传分化方面的证据。结果表明:(1)rpl16基因序列分析显示:长江中下游芦苇居群基因水平上的变异程度较云贵高原的低;(2)基于rpl16基因序列重建的最简约系统发育树显示:以地理区域为划分指标,可将长江中下游和云贵高原的芦苇居群分为两个单系;(3)云贵高原芦苇居群的核苷酸多样性A=0.00047,单倍型多态性Hd=0.466,其遗传多样性显著高于长江中下游(Pi=0,Hd=0);(4)居群水平上的遗传分化系数F_(st)=0.535 37,其中53.54%的遗传变异来自于不同地理区域居群间,长江中下游芦苇居群的遗传差异指数F_(st)=0.537 24,基因流N_m=0.215 34,云贵高原芦苇居群的遗传差异指数F_(st)=0.532 78,基因流N_m=0.21923,居群间的遗传分化较强烈。分析结果证明了两种地理生态型芦苇具有遗传上的差异,且地理隔离可能是引起差异的主要原因。     

甘蔗光合系统Ⅰ亚基O基因的克隆与表达分析

光合系统Ⅰ亚基O(PhotosystemⅠsubunit O,Psa O)是光合系统Ⅰ中的蛋白亚基,在两个光合系统之间平衡激发能方面起着重要的作用.为研究Psa O基因的结构和功能,对甘蔗(Saccharum offi cinarum L.)叶片全长c DNA文库进行测序,获得光合系统Ⅰ亚基O基因的全长c DNA序列,命名为Sc Psa O(Gen Bank Accession Number:KF714498).生物信息学分析表明,该基因全长708 bp,开放阅读框435 bp,编码144个氨基酸;该基因所编码的蛋白定位于叶绿体基质,无信号肽,为疏水性非分泌碱性蛋白,二级结构多为无规则卷曲,含有PJN00046家族的保守结构域,参与能量新陈代谢及脂肪酸新陈代谢.同时,该基因在不同物种间具有较强的保守性,其中与同属C4植物的同源性较C3植物高.荧光定量PCR分析结果表明,甘蔗Sc Psa O基因在叶片中的相对表达量最高,具有一定的组织特异性;在氯化钠(Na Cl)、聚乙二醇(PEG)、氯化铜(Cu Cl2)、脱落酸(ABA)、水杨酸(SA)和茉莉酸甲酯(Me JA)等外源胁迫下,其表达量均呈下调趋势,且以PEG胁迫下的下调表现最为明显.这些结果表明这几种外源胁迫可能抑制甘蔗Sc Psa O基因的转录水平表达,为其进一步功能验证以及在甘蔗基因工程中的应用积累了基础资料.     

长柄石杉不同地理居群叶绿体DNA trnL-trnF序列变异与聚类分析

采用直接测序法,对石杉科广布种长柄石杉(Huperzia serrata var.longipetiolata)21个居群(福建省19个居群和江西省2个居群)共计126株个体的叶绿体DNA trnL-trnF序列进行测序,用Clustal X和MEGA5.0软件进行序列分析,UPGMA法构建聚类图,以期了解序列变异与地理分布的关系,为长柄石杉资源的保护及开发利用提供参考依据.扩增获得的序列全长在928-967 bp之间,经排序后两端切平,序列长925 bp,G+C平均含量为34.05%;21个长柄石杉居群的cpDNA trnL-trnF序列存在20个变异位点,5个信息位点.UPGMA聚类结果显示,福建省居群与江西居群存在较大遗传距离,分聚为不同分支;福建省内19个居群再分聚为3个分支,其中11个居群间的遗传距离为零.研究结果表明长柄石杉居群间存在较明显的遗传分化和一定强度的基因流,居群间的叶绿体DNA trnL-trnF序列变异与地理分布尤其是山脉形成的隔离和屏障有关,较高的遗传多样性和基因流水平有利于长柄石杉种群的生存与进化.     

氧化亚氮还原酶基因nosZⅡ及与环境的关系研究进展

温室效应在全球范围内日趋严重.其中氧化亚氮(N2O)作为最重要的温室气体之一,增温潜能是二氧化碳(CO2)的298倍,且其浓度仍呈逐年上升趋势.氧化亚氮还原酶(N2OR)能将N_2O还原成氮气(N_2),而nosZ基因是编码N_2OR的唯一基因.除了熟知的nosZⅠ基因外,新发现的分支nosZⅡ基因也能编码N_2OR.在国内外研究基础上本文总结nosZⅡ基因的基本概况,分析其与nosZⅠ的主要区别,阐述其相关反应机制,并归纳不同环境因素对nosZⅡ基因表达的影响.研究表明,nosZⅡ主要存在于ε-变形细菌、拟杆菌、产水菌中,含nosZⅡ的微生物中有部分种群缺乏nirS和nirK基因,且该类微生物在还原N_2O功能上具有较大潜能.影响含nosZⅡ微生物的丰度和种群结构的环境因子较多,主要包括土壤结构、土壤pH值、土壤C/N、温度、湖泊物理梯度等,其中pH值和C/N可能是主要的影响因素.随着分子生物技术手段的进步,对含nosZⅡ微生物的生态功能的探索已取得了重要进展,但还需进一步深入.今后还需对含nosZⅡ的微生物群落结构特征、微生物丰度和多样性影响因素以及对具有较强还原N_2O能力的含nosZⅡ菌株进行更深入的研究,为降低N_2O排放、有效减控温室气体提供理论依据.     

入侵植物加拿大一枝黄花和本地一枝黄花的遗传多样性比较

为了阐明加拿大一枝黄花成功入侵的机制,利用简单序列重复区间标记(ISSR)方法对加拿大一枝黄花和本地一枝黄花的遗传多样性进行比较研究。从100条引物中筛选出12条引物用于PCR扩增,利用POPGEN32软件对2种一枝黄花进行遗传多样性分析。结果表明,加拿大一枝黄花在物种水平上的多态位点百分率为95.19%,Nei’s基因多样性指数为0.308 5,Shannon’s信息指数为0.415 8;本地一枝黄花在物种水平上的多态位点百分率(89.80%)、Nei’s基因多样性指数(0.249 1)和Shannon’s信息指数(0.383 4)都比加拿大一枝黄花小。加拿大一枝黄花和本地一枝黄花居群间遗传分化系数分别为0.118 2和0.131 3,居群内变异分别为0.881 8和0.868 7,表明2个物种居群间的遗传分化不明显,遗传一致度高,且主要的遗传变异存在于居群内。入侵植物加拿大一枝黄花具有较高遗传多样性,且高于本地一枝黄花,这可能是加拿大一枝黄花成功入侵的原因之一。     

云南红豆杉的保护和开发利用

云南红豆杉(Taxus yunnanensis W. C. ChengL. K. Fu)是红豆杉属植物中紫杉醇含量最高的树种,同时也是我国红豆杉属中分布最广和资源蕴藏量最丰富的物种.然而,近年来由于人为大量砍伐,云南红豆杉野生资源遭到严重破坏,加上生长缓慢且天然更新困难,目前处于濒危状态.对云南红豆杉天然资源的保护和药用人工林的培育对于云南红豆杉资源的可持续利用具有重要的意义.对云南红豆杉资源分布现状及种群和繁殖生态学等方面的研究进行综述,发现云南红豆杉的濒危是由其自身繁殖机制引起的天然更新困难、对环境的适应能力较低以及人为因素共同导致的.同时从紫杉醇含量的影响因素及获取方式、人工药用林培育方面总结了云南红豆杉的开发利用现状,发现云南红豆杉中紫杉醇的含量受其自身特性及外在因素的综合影响;包含紫杉醇在内的紫杉烷类物质目前主要通过直接提取和半合成两种方式获取;此外在全合成、组织或细胞培养及真菌诱导等方式上也有了新的研究进展.最后,建议从就地保护、迁地保护及引种回归3个方面对云南红豆杉天然资源进行综合保护,同时大力推进紫杉烷工业化合成方式的研究和云南红豆杉人工药用原料林的培育,提高云南红豆杉的资源利用效率,在保护云南红豆杉天然林的同时实现资源的合理利用.(表1参129)     

猪场土壤中5种四环素抗性基因的检测和定量（Quantification of Five Tetracycline Resistance Genes in Soil from a Swine Feedlot）

近年来由于四环素在畜禽养殖业中的大量使用,诱导了环境中四环素抗性微生物的产生,然而有关四环素抗性基因在土壤中存在、迁移和扩散的研究目前还很少.论文提取了北京一规模化养猪场周边土壤的微生物DNA,利用普通PCR检测到5种四环素抗性基因(tet(B/P)、tet(M)、tet(O)、tet(T)、tet(W)),这5种基因都属于编码核糖体保护蛋白的一类.进一步建立了定量PCR程序对这5种基因进行了定量.结果显示,除tet(T)和tet(M)含量接近外(p=0.367),其余几种基因含量之间均差异显著(p<0.05),其中tet(W)的含量最高((2.16±0.20)×108copies·g-1(干土)),比含量最低的tet(B/P)((2.89±0.54)×106copies·g-(1干土))高出约两个数量级.抗性基因tet(W)、tet(T)、tet(M)、tet(O)含量均较高,为猪场土壤中优势抗性基因.这些基因曾被报道广泛存在于猪和牛的肠道中,显示抗性基因很可能是通过基因横向转移(HGT)等机制从养殖动物体内传播到周围土壤中土著微生物体内的.论文所建立的实验方法为进一步系统研究抗生素抗性基因在土壤中的环境行为及其生态风险提供了基础.     

麻疯树种质资源遗传多样性的ISSR分析

采用ISSR分子标记技术对大戟科属麻疯树9个自然居群的遗传多样性水平和遗传结构进行了研究.用10个引物对9个居群共135个样品进行了扩增,共获得169条清晰的扩增位点,其中多态性位点164个.POPGENE分析结果表明,麻疯树居群具有丰富的遗传多样水平(多态百分率PPB=97.04%,Neis遗传多样性H=0.2357,Shannon信息指数I=0.3760).AMOVA分析表明,9个自然居群间遗传多样性出现了较大的遗传分化(Gst=0.5398)可能与其有限的基因流(Nm=0.4667)和遗传漂变有关;而居群内有限的遗传分化可能是由于麻疯树自交、近交占主导方式的繁育方式有关.并进行了麻疯树居群的聚类分析.图3表4参19     

不同野生居群石蒜表型变异及物候期差异

石蒜是我国具有重要药用和园林应用价值的野生植物.以不同居群野生石蒜为材料,通过周年观察、切割繁殖、常规调查研究了其种质资源的基础特性.结果表明,不同野生居群石蒜单个鳞茎叶片数量多数为6片,叶长、叶宽、叶面积和叶绿素总量在18.36～21.56cm、0.50～0.70cm、9.22～12.95cm2和57.68～64.73SPAD之间比例最高.环境气候与表型性状相关性分析发现,热量及降雨量变化制约着叶宽、叶面积和叶片数量的变化,叶绿素总量与日照时数略微相关.方差分析发现,叶片数量呈显著水平差异(P0.05),而叶长、叶宽、叶面积和叶绿素总量均表现出极显著水平差异(P0.01).来自亚热带南部的居群物候期明显早于来自中北部的居群,南部居群的繁殖系数、子鳞茎质量也均优于中北部居群,所有居群繁殖系数均值为5.81,子鳞茎质量均值为1.64g.不同居群石蒜的子鳞茎5个生物学性状相比,根系数量差异最大(3.42～7.98根),其次是高度差异(1.27～2.01cm),再次是质量差异(1.33～1.94g),最后是直径差异(1.03～1.23cm).     

Protection versus culture-driven exploitation of wild plant resources: the case on Changbai Mountain

Under Chinese culture it is believed that herbal medicine is always safe and wild food is always healthy. Generally, the rarer a plant, the higher its value. The booming economy in China has promoted tourism development in wilderness areas and wild medicinal and food plants are part of the attraction to tourists. Conflicts between wild plant exploitation and protection have emerged in many parts of China, such as Changbai Mountain. Changbai Mountain supports numerous medicinal and food plants but many have become rare and endangered. This paper numerically evaluates 30 plant species that have relatively high conservation value for each type (medicinal, ornamental and food), and briefly describes the uses of four to five top ranked species per type that need more protection on Changbai Mountain. This paper also addresses some tree species with important timber values on Changbai Mountain. Over 90% of China's medicinal, ornamental and food plant species, as well as valuable timber trees are found in the conifer–broadleaf mixed forest zone across the boundary of Changbai Mountain Nature Reserve. It is a major challenge to protect the native biodiversity of mixed forest on Changbai Mountain and more efforts need to be made to protect rare and endangered plant species with high economic value.     

漂浮栽培美人蕉蕹菜等植物处理化粪池废水

采用漂浮栽培美人蕉、蕹菜、水稻、野生稻等４种植物进行了处理化粪池废水的静态试验。结果表明，废水中ＣＯＤ、ＢＯＤ５、总氮、总磷等污染物得到较好的净化，污染物去除率明显高于对照。４种植物的净化能力和净化效果综合评价结果为美人蕉＞蕹菜＞水稻＞野生稻，美人蕉、蕹菜对化粪池废水的净化能力强     

Critique of the Evolutionarily Significant Unit as a Definition for "Distinct Population Segments" under the U.S. Endangered Species Act

The U.S. Endangered Species Act grants protection to species, subspecies, and "distinct population segments" of vertebrate species. Historically, Congress included distinct population segments into endangered species legislation to enable the U.S. Fish and Wildlife Service to implement a flexible and pragmatic approach in listing populations of vertebrate species. Recently, the U.S. Fish and Wildlife Service and the National Marine Fisheries Service have proposed a policy that would narrowly define distinct population segments as evolutionarily significant units based on morphological and genetic distinctiveness between populations. Historically, the power to list species or populations as distinct population segments has been used to tailor management practices to unique circumstances; grant varied levels of protection in different parts of a species' range; protect species from extinction in significant portions of their ranges as well as to protect populations that are unique evolutionary entities. A strict redefinition of distinct population segments as evolutionarily significant units will compromise management efforts because the role of demographic and behavioral data will be reduced. Furthermore, strictly cultural, economic, or geographic justifications for listing populations as threatened or endangered will be greatly curtailed.     

Conservation Biology Framework for the Release of Wild-Born Orphaned Chimpanzees into the Conkouati Reserve, Congo

Abstract: Returning confiscated animals to their native habitats is desirable when it makes a positive contribution to the conservation of the species. Release of captive individuals is complex and controversial, however, particularly when risks are potentially high, as in the case of orphaned apes. We describe the decision-making process that led to the successive release of 20 wild-born orphan chimpanzees (    Pan troglodytes troglodytes ) into the Conkouati Reserve in the Republic of Congo. Recommendations of the Reintroduction Specialist Group of the World Conservation Union's Species Survival Commission were followed closely. The conservation status, ecology, and behavior of wild chimpanzees; the biological, social, economic and political context of the release site; and the health and genetic status of the candidates for release were all taken into account in the planning and execution of the project. Rigorous post-release monitoring of behavior and health allowed documentation of the outcome. The project was of benefit to the chimpanzees that were released but also brought broad benefits to the site through effective protection from poaching and deforestation, and direct and indirect benefits to local people. The genetic and behavioral diversity of chimpanzees require a variety of conservation strategies to reduce threats and maintain as many viable wild populations as possible.     

水稻根系吸收砷的动力学特征及硅的缓解机制（Uptake Kinetics of Arsenic Species in Two Genotypes of Rice Plants and Mitigatory Mechanism of Silicon）

采用室内模拟的方法,研究了两种基因型水稻(硅突变体和野生型)根系吸收As的动力学特征以及Si对水稻吸收As的影响.结果表明,两种基因型水稻根系对As(III)和As(V)的吸收均可以用Freundlich模型较好地模拟.外源Si的加入可显著降低水稻根系对As(III)和As(V)的吸收.在20μmol·L-1As处理下,与不加Si处理相比,加1.0mmol·L-1Si处理导致水稻根系吸收As(III)和As(V)分别降低94.1%和92.2%(野生型);74.4%和90.2%(突变体).在100μmol·L-1As处理下,水稻根系吸收As(III)和As(V)则分别降低64.5%和91.2%(野生型);76.1%和90.6%(突变体).     

Lyme Disease and Conservation

Lyme disease is a tick-borne illness that is widespread in North America, especially in the northeastern and northcentral United States. This disease could negatively influence efforts to conserve natural populations in two ways: (1) the disease could directly affect wild animal health; and (2) tick control efforts could adversely affect natural populations and communities. Lyme disease affects several domestic animals, but symptoms have been reported in only a few wild species. Direct effects of Lyme disease on wild animal populations have not been reported, but the disease should be considered as a possible cause in cases of unexplained population declines in endemic areas. Methods available to manage ticks and Lyme disease include human self-protection techniques, manipulation of habitats and host species populations, biological control, and pesticide applications. The diversity of available techniques allows selection of approaches to minimize environmental effects by (1) emphasizing personal protection techniques, (2) carefully targeting management efforts to maximize efficiency, and (3) integrating environmentally benign techniques to improve management while avoiding broad-scale environmentally destructive approaches. The environmental effects of Lyme disease depend, to a large extent, on the methods chosen to minimize human exposure to infected ticks. Conservation biologists can help design tick management programs that effectively lower the incidence of human Lyme disease while simultaneously minimizing negative effects on natural populations.     

Genetic Diversity and Tests of the Hybrid Origin of the Endangered Yellow Larkspur

Abstract: Delphinium luteum ( Ranunculaceae), an endangered larkspur, is restricted to two wild populations near Bodega Bay, California. The total number of individuals in these two populations is estimated to be <100. We used allozyme and random amplified polymorphic DNA ( RAPD) markers to (1) assess levels and patterns of genetic diversity in one wild population and two cultivated populations and (2) test the hypothesis that D. luteum is of hybrid origin between D. decorum and D. nudicaule . These data will be used to aid in developing a management plan to conserve the species. The wild population maintains high levels of genetic diversity. Genetic data indicate that both cultivated populations, especially the north Sonoma population, have several allozymes and RAPD markers not found in the wild population and could be used to establish new populations of D. luteum or to enhance the diversity and size of the wild population. The allozyme data did not reveal any fixed differences between D. decorum and D. nudicaule , although allele frequencies of the putative parental populations differed. At these loci, D. luteum resembled D. nudicaule more than D. decorum  . Many unique RAPD markers distinguish each of the three species. The diagnostic markers from populations of D. nudicaule and D. decorum were not additive in the putative hybrid, and these data indicate that D. luteum is not of recent hybrid origin. Conservation of the yellow larkspur should include strategies that use the cultivated populations of D. luteum , but hybridizing D. decorum and D. nudicaule to "recreate" D. luteum is not recommended.     

Analytical Solutions to Trade‐Offs between Size of Protected Areas and Land‐Use Intensity

Abstract: Land‐use change is affecting Earth's capacity to support both wild species and a growing human population. The question is how best to manage landscapes for both species conservation and economic output. If large areas are protected to conserve species richness, then the unprotected areas must be used more intensively. Likewise, low‐intensity use leaves less area protected but may allow wild species to persist in areas that are used for market purposes. This dilemma is present in policy debates on agriculture, housing, and forestry. Our goal was to develop a theoretical model to evaluate which land‐use strategy maximizes economic output while maintaining species richness. Our theoretical model extends previous analytical models by allowing land‐use intensity on unprotected land to influence species richness in protected areas. We devised general models in which species richness (with modified species‐area curves) and economic output (a Cobb–Douglas production function) are a function of land‐use intensity and the proportion of land protected. Economic output increased as land‐use intensity and extent increased, and species richness responded to increased intensity either negatively or following the intermediate disturbance hypothesis. We solved the model analytically to identify the combination of land‐use intensity and protected area that provided the maximum amount of economic output, given a target level of species richness. The land‐use strategy that maximized economic output while maintaining species richness depended jointly on the response of species richness to land‐use intensity and protection and the effect of land use outside protected areas on species richness within protected areas. Regardless of the land‐use strategy, species richness tended to respond to changing land‐use intensity and extent in a highly nonlinear fashion.     

Si对盐胁迫下水稻根系活力、丙二醛和营养元素含量的影响(Effects of Si on the Index of Root Activity, MDA Content and Nutritional Elements Uptake of Rice under Salt Stress)

为深入了解盐胁迫下外源硅(Si)对水稻生长的作用,采用溶液培养法研究了外源Si对盐(NaCl)胁迫下野生型水稻和硅突变体水稻(不能正常吸收Si)营养元素和生理指标的影响.结果表明:1)外源Si可显著提高盐胁迫下两种水稻的生物量.当NaCl的浓度为100mmol·L-1时,添加1.0mmol·L-1的Si可使野生型水稻茎叶和根系干重提高12.81%和15.25%,硅突变体水稻茎叶干重提高12.31%.2)外源Si可提高盐胁迫下两种水稻茎叶和根系中营养元素含量,降低Na的含量.3)外源Si可显著降低盐胁迫下野生型水稻叶片MDA含量,但对突变体水稻叶片MDA含量影响不大.4)外源Si可显著提高盐胁迫下两种水稻的根系活力.盐胁迫下,与不加Si的对照相比,添加1.0mmol·L-1的Si可分别使野生型和突变体水稻根系活力指数提高60.47%和42.42%.     

Phylogeographic study of the dwarf oyster, Ostreola stentina, from Morocco, Portugal and Tunisia: evidence of a geographic disjunction with the closely related taxa, Ostrea aupouria and Ostreola equestris

Despite the economic importance of oysters due to the high aquaculture production of several species, the current knowledge of oyster phylogeny and systematics is still fragmentary. In Europe, Ostrea edulis, the European flat oyster, and Ostreola stentina, the Provence oyster or dwarf oyster, are both present along the European and African, Atlantic and Mediterranean, coasts. In order to document the relationship not only between O. stentina and O. edulis, but also with the other Ostrea and Ostreola species, we performed a sequence analysis of the 16S mitochondrial fragment (16S rDNA: the large subunit rRNA-coding gene) and the COI fragment (COI: cytochrome oxidase subunit I). Oysters were sampled from populations in Portugal (two populations), Tunisia (two populations) and Morocco (one population), identified as O. stentina on the basis of shell morphological characters. Our data supported a high degree of differentiation between O. stentina and O. edulis and a close relationship between O. stentina and both Ostrea aupouria (from New Zealand) and Ostreola equestris (from Mexico Gulf/Atlantic). The status of this geographic disjunction between these closely related species is discussed. Furthermore, although identified in a separate genus Ostreola by Harry (Veliger 28:121–158, 1985), our molecular data on O. stentina, together with those available for the other two putative congeneric species, O. equestris and Ostreola conchaphila, would favour incorporation of Ostreola in Ostrea. Finally, a PCR-RFLP approach allowed the rapid identification of O. edulis and O. stentina.