丹顶鹤（Grusjaponensis）性别鉴定的分子标记方法

丹顶鹤(Grus japonensis)是我国一级保护动物,为单态性鸟,很难通过外观和形态来进行性别鉴定,给配对和人工繁殖造成了很大的困难.实验利用非伤害性取样方法,成功地从丹顶鹤羽毛中提取了基因组DNA,并筛选出3对引物组合对丹顶鹤EE0.6序列的相关片段进行了特异性扩增.结果显示,这3对引物组合从雄鹤中扩增出1条片段,从雌鹤中扩增出2条片段,均可以对丹顶鹤的性别作出了准确鉴定,解决了丹顶鹤人工繁育中的关键问题.     

粘类小麦雄性不育系线粒体DNA的AFLP标记及SCAR转化

为了能在分子水平上有效鉴定具有粘果山羊草(Aegilops kotschyi)、偏凸山羊草(A.ventricosa)、普通小麦变种斯卑尔脱(Triticum spelta)细胞质雄性不育系及其保持系90-110和8222,提高其在杂交小麦(Triticum aestivum L.)研究与应用中的定向遗传改良,本研究对其线粒体DNA进行了扩增片段长度多态性(amplified fragment length polymorphism,AFLP)标记和序列特征性片段扩增区域(sequence characterized amplified region,SCAR)标记。通过AFLP标记方法,应用64对引物组合EcoRⅠ-NNN/MseⅠ-NNN对小麦同核异质雄性不育系和保持系进行扩增,共扩增出682条带,其中113条为多态性条带。引物E-AGG/M-CTA组合在粘果山羊草细胞质雄性不育系中扩增出一条大小约300 bp的特异性条带,对该特异条带进行回收、测序,利用Primer Premier 5.0软件重新设计SCAR引物,并对这3种类型同核异质小麦细胞质雄性不育系和保持系进行扩增,其中引物YW1在3种细胞质雄性不育系和保持系中都扩增出条带,而引物YW2仅在粘果山羊草细胞质雄性不育系扩增出一条198 bp的特异性片段,结果表明,已成功地将AFLP标记转化为操作简便、表现稳定的SCAR标记。此片段与小麦线粒体基因组有很高的同源性(同源性为99%),为烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶基因(GenBank登录号:EU534409.1)上的序列,该酶是线粒体中氧化磷酸化的入口酶,与小麦细胞质雄性不育密切相关。本研究可以用于粘类小麦细胞质雄性不育系分子标记辅助育种,也为小麦细胞质种性鉴定提供了技术支撑和理论依据。     

应用PCR技术鉴定绵羊胎儿成纤维细胞系性别

根据牛、羊Y－染色体性别决定区域（SRY）的同源性设计了一对PCR引物，对8个绵羊胎儿成纤维细胞系SFF1－8进行了性别鉴定。阳性对照和细胞系SFF1、2、6、7扩增得到130bp片段，阴性对照、空白对照及细胞系SFF3、4、5、8没有扩增带。通过序列测定和同源性分析，证明PCR产物为SRY基因片段，说明有130bp扩增带的细胞系为雄性；无扩增带的为雄性。结果表明，该法具有简单、快速、准确的特点，可应用于转基因克隆动物研究中对体细胞系的早期性别鉴定。     

用过氧化物酶同工酶和SSR标记鉴定中油杂12种子纯度

杂交种的纯度鉴定是油菜种子生产的重要环节。对甘蓝型油菜（BrassicanapusL.）杂交品种中油杂12及其父母本的过氧化物酶同工酶分析结果显示,父本和杂种F1比母本多一条迁移率（Rf）为0.41的条带,该特征条带可用来进行粗放杂种纯度鉴定,但是难以鉴定出父本杂株。选用173对SSR引物对中油杂12的亲本进行扩增,筛选出了6对扩增效果好的引物,其扩增条带清晰且亲本间的差异能够重复,其中P052、P130、P131和P154这4对SSR引物在杂种F1中可分别扩增出父母本的特征条带。在同一反应体系中同时加入P131和P154两对引物,发现扩增产物带型是单独采用这两对引物分别扩增时产物带型的叠加。对于混杂程度较高的种子,采用P131＋P154引物组合可以在工作量不变的条件下提高检测外来花粉的灵敏度,从而更加准确地鉴定中油杂12的种子纯度。对100份大田制种的中油杂12单株DNA分别用引物P131、P154以及引物组合P131＋P154进行SSR分析,鉴定结果完全一致,并且与田间鉴定结果非常接近。     

不同种群草鱼遗传结构的TRAP分析

应用TRAP(targetregionamplifiedpolymorphism)技术,对草鱼(Ctenopharyngodonidella)野生种群和两个人工养殖种群的遗传结构进行了比较。从15个引物组合中,筛选出7个扩增效果比较好的引物组合,在3个种群中共检测到103个扩增位点,其中野生种群、淡水中心种群和前洲种群的多态性位点分别为67、55和46,表明人工养殖草鱼种群的遗传多样性有所降低。与野生种群相比较发现,人工养殖种群仅33.98%位点的基因频率保持不变,表明人工养殖种群的遗传结构发生了改变。淡水中心种群、前洲种群与野生种群间的遗传距离分别为0.0421和0.0809。引物组合Ga5-800-E5扩增的结果发现,人工养殖种群中存在扩增位点明显减少的区域,这为寻找鉴定草鱼野生种群和养殖种群的遗传标记提供了依据。     

Sry—PCR法快速鉴定体外培养兔胎儿成纤细胞系的性别

从怀孕17d母兔取得3个胎儿，分离培养得到3个胎儿成纤维细胞系RFF1、RFF2和RFF3。通过扩增兔sry基因核心序列片段对3个细胞系的性别进行鉴定（Sry-PCR），结果表明：RFF1和RFF3为雄笥，RFF2为雌性，进一步利用染色体核型分析法鉴定3个细胞系的性别，与Sry-PCR法的结果完全一致。因此表明本实验所用的Sry-PCR的引物、细胞基因组DNA的提取方法、PCR反应条件是合适的，可以快速准确地鉴定体外培养的兔胎儿成纤维细胞的性别。     

利用RAPD标记鉴定山葡萄种质

采用修改后的CTAB法获得了高质量的基因组DNA.利用随机扩增多态性即RAPD标记对山葡萄7份种质进行鉴定,用4个引物(从30个引物中筛选)对试材进行PCR扩增,共扩增出30条谱带,平均每条引物产生7.5条谱带,其中21条谱带为多态性谱带,占总谱带数的70%.不同引物扩增的谱带数不同,范围在6～9条之间.利用4个引物扩增出的多态性谱带可以将7份山葡萄种质区分.     

利用小麦微卫星引物建立偃麦草Ee染色体组特异SSR标记

选用40对小麦SSR引物对17份偃麦草(Thinopyrum sp．)、2份小麦(Triticum aestivum)材料进行了PCR扩增分析，从中筛选到引物Xgwm325能在不同偃麦草材料中扩增出4条长度分别为1400、440、120和100bp的特异DNA片段，可以作为偃麦草种质的特异SSR标记。利用小麦-二倍体长穗偃麦草(Th.elongatum)异代换系和异附加系对引物Xgwm325进行了扩增鉴定，结果只有100bp左右的片段出现在长穗偃麦草所有E^e组染色体上，该片段可以作为E^e染色体组的特异SSR标记。     

不同基因型刺梨及其近缘种亲缘关系的RAPD分析*

采用RAPD标记，对刺梨(Rosa roxburghii Tratt)7个基因型及8个近缘种(类型)进行鉴定和亲缘关系分析。结果表明，筛选出的16个随机引物可扩增出137条480bp到3.3kb大小的清晰。DNA片段，其中95条为多态性片段，为总数的69．3％；每个引物平均产生8．6条。由OPB-11、OPAF-16和OPW-02扩增的14条特异DNA片段，可有效地将普通刺梨7个基因型和无籽刺梨区分开来。基于遗传距离矩阵，采用IJPGMA法对15个供试样品的亲缘关系进行了聚类分析，文中还就无籽刺梨及重瓣刺梨的可能来源进行了探讨。     

海南薯蓣茎腐病病原菌的鉴定

为弄清海南薯蓣茎腐病病原,采用离体致病性测定、形态学和分子生物学鉴定等方法,对其进行了研究。从海南临高、海口等薯蓣生产基地采集病样89份,纯化出分离物28个。经致病性试验证实,分离物lp3—1、b16-5、bd6-4和lp2—5为薯蓣茎腐病病原菌。经形态学鉴定,4个分离物均为镰刀菌（Fusariumsp．）。利用真菌18SrRNA基因引物、镰刀菌特异性引物和轮枝镰刀菌特异引物进行PCR扩增,能扩增出510bp大小的镰刀菌DNA片段,不能扩增出轮枝镰刀菌DNA片段。序列分析表明：分子生物学和形态学鉴定结果一致,且镰刀菌Fu3／Fu4区序列分析能进一步确定薯蓣茎腐病是由尖孢镰刀菌薯蓣专化型（Fusariumoxysporumfsp．dioscorea）侵染引起的一类植物真菌病害。     

Introduced house mice Mus musculus: a significant predator of threatened and endemic birds on Gough Island, South Atlantic Ocean?

House mice Mus musculus have successfully colonized many temperate and sub-Antarctic islands that are the location for breeding colonies of millions of seabirds. Unlike other introduced mammals, the impact of house mice on seabirds and endemic birds is believed to have been negligible. The breeding ecology of seabirds breeding on Gough Island, central South Atlantic Ocean, was studied for the first time during September 2000 to September 2001. Breeding success of the endangered Tristan albatross Diomedea (exulans) dabbenena and endangered Atlantic Petrel Pterodroma incerta were 27.3 and 19.9% respectively. Mortality of large Tristan albatross and Atlantic petrel chicks was observed, and the pattern of wounds and observations of feeding indicate that introduced mice were responsible for this predation. Breeding numbers of the endemic Gough bunting Rowettia goughensis are mostly found in upland areas of Gough Island where mice are scarce and are restricted to inaccessible cliffs in the lowlands where mice are abundant. This pattern, together with the high predation rates of artificial-eggs in lowland habitats in comparison to the uplands, strongly suggests that mice constrain the distribution of Gough buntings. The results of this study provide the first evidence for the role of house mice as a significant predator of endangered and endemic birds. Further research is required to determine if the observed levels of mice predation are a regular occurrence.     

Migration routes and important resting areas of Siberian cranes (Grus leucogeranus) between northeastern Siberia and China as revealed by satellite tracking

In 1995 and 1996 thirteen Siberian cranes (Grus leucogeranus) were fitted with satellite transmitters on the breeding grounds in northeastern Siberia. Eleven of these 13 birds were successfully satellite tracked, and five of these 11 provided complete migratory information from their breeding grounds in Yakutia, Siberia, to their wintering area at Poyang Lake, in China. Several stopover sites were identified, the most important being in Qiqihar-Baicheng (China), Shuangtaizi River delta (China), and Yellow River delta (China). Birds rested more frequently in Russia than in China, suggesting availability of suitable wetland habitat in Russia and absence of adequate, suitable wetland habitat in China. Wintering habitat in China also faces numerous threats. Habitats utilized by Siberian cranes are also important to other threatened wetland birds that have been satellite-tracked recently. If Siberian crane habitat needs fail to be addressed, this critically endangered species will be further endangered.     

Large scale risk-assessment of wind-farms on population viability of a globally endangered long-lived raptor

Wind-farms receive public and governmental support as an alternative energy source mitigating air pollution. However, they can have adverse effects on wildlife, particularly through collision with turbines. Research on wind-farm effects has focused on estimating mortality rates, behavioural changes or interspecific differences in vulnerability. Studies dealing with their effects on endangered or rare species populations are notably scarce. We tested the hypothesis that wind-farms increase extinction probability of long-lived species through increments in mortality rates. For this purpose, we evaluate potential consequences of wind-farms on the population dynamics of a globally endangered long-lived raptor in an area where the species maintains its greatest stronghold and wind-farms are rapidly increasing. Nearly one-third of all breeding territories of our model species are in wind-farm risk zones. Our intensive survey shows that wind-farms decrease survival rates of this species differently depending on individual breeding status. Consistent with population monitoring, population projections showed that all subpopulations and the meta-population are decreasing. However, population sizes and, therefore, time to extinction significantly decreased when wind-farm mortality was included in models. Our results represent a qualitative warning exercise showing how very low reductions in survival of territorial and non-territorial birds associated with wind-farms can strongly impact population viability of long-lived species. This highlights the need for examining long-term impacts of wind-farms rather than focusing on short-term mortality, as is often promoted by power companies and some wildlife agencies. Unlike other non-natural causes of mortality difficult to eradicate or control, wind-farm fatalities can be lowered by powering down or removing risky turbines and/or farms, and by placing them outside areas critical for endangered birds.     

Decision analysis to guide recovery of the po‘ouli, a critically endangered Hawaiian honeycreeper

We used a probabilistic decision tree to help select among alternative recovery strategies for the Po‘ouli, a critically endangered forest bird endemic to the Hawaiian island of Maui. The Po‘ouli is one of the rarest birds in the world, with only three individuals known, and no breeding pairs. The most urgent conservation need for the species is to create a breeding pair and obtain eggs for captive propagation. Seven recovery strategies had been proposed, and there was disagreement among conservation workers about which strategy to pursue. In addition to lengthy discussions, a decision tree was introduced to provide an objective way of quantifying the chance of success under each alternative. All available information was used to attempt evaluation of each node in the decision tree. The overall chance of obtaining eggs was calculated by multiplying estimates at each node under each alternative. All options produced low estimates of potential success, but two options, removal of all birds to captivity and placement of the birds in a field aviary in an accessible location, produced similar estimates of success. The US Fish and Wildlife Service and the Hawaii Division of Forestry and Wildlife eventually agreed that removal to captivity was the preferred alternative because it could be implemented quickly, which was important considering the advanced age of the birds, and would make it easier to provide veterinary care and ensure their safety from predators, severe weather, and vandals. Decision trees and other structured models should not be relied on exclusively, but they can provide an objective method of helping to make difficult conservation decisions and provide a record of complex thought processes used in reaching a determination. The case of the Po‘ouli may serve as a template for navigation towards a decision to recover other species of extreme rarity.     

Extinction of an isolated population of the middle spotted woodpecker Dendrocopos medius (L.) in Sweden and its relation to general theories on extinction

A small population of middle spotted woodpeckers in southern Sweden became isolated at least 30 years ago. Since then the population has probably been rather stable at between 15 and 20 pairs. Censuses from 1967–1974 showed a stable population while a rapid decline, all the way to extinction, occurred in the period 1975–1983. In the period 1941–1978 no considerable changes in habitat area occurred. Losses of birds were correlated with number of cold days in winter. During the period 1975–1983 the reproduction of the birds was low, expecially because of many unsuccessful breeding attempts. Inbreeding depression is proposed to be the most probable factor causing low reproduction. If reduced fecundity is a general phenomenon in small and isolated populations, demographic models of extinction probabilities presented hitherto are too simple. Dispersal ability must be taken into account when planning reserve systems for endangered species.     

At-sea distribution of breeding Tristan albatrosses Diomedea dabbenena and potential interactions with pelagic longline fishing in the South Atlantic Ocean

The endangered Tristan albatross Diomedea dabbenena is restricted to Gough and Inaccessible Islands. The species is killed as bycatch by longline fisheries in the South Atlantic, but the impact of this mortality is unknown. We satellite tracked 38 breeding Tristan albatrosses and assessed the seasonal and annual at-sea distribution of these birds in relation to reported pelagic longline fishing effort. These birds ranged across the South Atlantic from 50°W to 15°E with most (97%) daytime satellite fixes between latitudes 30°S and 45°S. Considerable fishing effort occurred within the same latitudes. Although there was no correlation between their at-sea distributions, there was a broad overlap between birds and fishing effort. Estimated bycatch rates for Tristan albatross and other Diomedea species in the South Atlantic, and the spatio-temporal overlap between birds and hooks, yield a predicted annual mortality of 471-554 birds, sufficient to cause population decreases of 3.6-4.3% per year. An index of bird × hook interactions (proportional density of birds multiplied by number of hooks by decadal period for each 5° square of longitude and latitude) indicated that 47% of annual interactions occurred in areas around Gough Island, and 11% and 15% of interactions in areas of the west and east Atlantic, respectively. There were also within seasonal differences in the key areas of overlap. The fishing fleets of Taiwan and Japan are likely to be responsible for most interactions based upon the reported magnitude of effort expended in the South Atlantic by these fleets. Ensuring that licensed fishing vessels within the Tristan da Cunha Exclusive Economic Zone (EEZ) operate using best-practise mitigation measures and with fisheries observer programs, could reduce the potential bycatch mortality of breeding Tristan albatrosses in this region by nearly one third. Thorough implementation of international agreements is required in areas of the high seas where most remaining interactions are predicted to occur.     

The waldrapp, Geronticus eremita (Linnaeus, 1758): Historical review, taxonomic history, and present status

The Waldrapp ibis, Geronticus eremita Linnaeus, 1758, is known historically from Ancient Egypt, the Alps in southern Europe, the Near and Middle East, Morocco, and Algeria. This paper summarises what is known about the species from archaeological findings and historical expeditions to these regions, and outlines the taxonomic confusion which characterised this bird for four centuries—the 16th to 19th.

The ancient and recent decline of populations of the Waldrapp is discussed. The birds are at present restricted to about twelve breeding sites, totalling only about 400 individuals, in Turkey and Morocco, with an additional 400-plus specimens in captivity in zoos of the world. A plea is made for the conservation of this endangered species.     

The immediate effects of ditching a saltmarsh on nesting herring gulls Larus argentatus

We examined the effects of water management for mosquito control on the behaviour and breeding success of a resident colony of herring gulls. The colony resided on three close islands, one of which was ditched in March before the start of the second breeding season. The number of breeding pairs on the ditched island remained the same before and after ditching although the breeding number increased by 46% and 90% on the other two islands. Breeding chronology was similar on all three areas. However, birds nesting on spoil laid eggs a mean of 8 days later than non-spoil nesting birds. Behavioural observations on aggression and display rates indicated that birds on spoil behaved similarly to those in open grassy areas but differed from those nesting in the bushes. Nest site selection, breeding densities, and breeding success were similar on all three islands. Thus the differences noted were attributed to the appearance of the marsh. We postulated that pairs having nested on the experimental island in the previous year continued to do so after the island was ditched. However, pairs searching for new territory did not move onto the island that was ditched, but instead colonised the nearby islands. Similarly, those pairs breeding on spoil nested later because of the need to defend their nest sites which were situated in areas used for displaying by unmated birds.     

广西沿海红树林区的水鸟

广西沿海地区位于一条重要的候鸟迁徙通道上，那里的红树林区给为数众多的水鸟提供了繁殖、越冬和迁徙中途歇息的场所。在红树林区记录到115种水鸟。这些鸟类中，102种是候鸟，包括13种夏候鸟、64种冬候鸟、25种旅鸟，因此红树林区水鸟类的多样性表现出明显的季节性。迁徙季节水鸟的种类和数量都最多，繁殖季节则最少。黑脸瑟鹭（Platalea minor)是世界上最濒危的鸟类之一，一个越冬种群在该地区红树林中被发现。     

The proportion of immature breeders as a reliable early warning signal of population decline: evidence from the Spanish imperial eagle in Doñana

Methods to evaluate population trends have recently received particular attention because of perceived declines in several species during the 20th century. We investigated whether age at first breeding could be used as an “early warning signal” to detect possible changes in population trends in long-lived species with deferred maturity using data from the Spanish imperial eagle (Aquila adalberti) population in Doñana National Park (Spain). This bird of prey is an endangered species that has suffered a rapid decline in this population during the last 10 years. As a result of our 27-year monitoring (1976-2002) study, we detected that an increase in immature breeding birds occurred before population decline became evident. The proportion of immature-plumaged breeders in the population was significantly higher during the period of decline than during the period of stability. In our case, more than 10% of immature breeders can be considered as an “early warning signal” that anticipates population decline. Owing to the ignorance of this warning signal, urgent actions for the recovery of this eagle population started 10 years later than necessary, and when population size had been reduced.