七鳃鳗遗传多样性与演化研究进展

七鳃鳗(Petromyzonidae)是目前已知最古老的脊椎动物中惟一的幸存者.对其资源保护和演化发育生物学的研究正日益受到重视.本文从染色体、蛋白质和DNA水平总结近年来七鳃鳗遗传多样性与演化方面的研究进展.重点介绍了限制性酶切片段长度多态性、DNA随机扩增多态性、DNA扩增片段长度多态性、微卫星DNA标记等技术及线粒体DNA和功能基因研究应用于七鳃鳗种群遗传多样性、遗传分化、遗传结构、种质鉴定与渔业资源管理及系统进化等方面的新进展.     

鸡的遗传多样性的研究方法及研究进展

遗传多样性是生物学研究中的一个重要领域,研究鸡的遗传多样性,不仅能加强生物多样性的保护。同时对起源进化、分类鉴定及遗传育种等都有重要的意义。本文对目前DNA水平鸡的遗传多样性的研究方法和研究进展进行了详细的阐述。重点介绍了DNA分子标记的特征;概括了在鸡遗传多样性分子标记的方法,包括微卫星分子标记(SSR)、扩增片段长度多态性(AFLP)、随机扩增多态性标记(RAPD)、限制性片段长度多态性标记(RFLP)和单核苷酸多态性标记(SNP)。本文综述了最近有关鸡DNA水平的遗传多样性的研究方法在系统学、遗传结构、生物地理等研究中的应用情况;提出在研究鸡遗传多样性时,可根据研究的目的,选择合适的方法。     

朱鹮随机微卫星扩增多态DNA（RMAPD）研究

首次利用RMAPD标记对43只洋县人工饲养朱鹮群体的遗传多态性进行了分析。结果表明,RMAPD技术的稳定性与多态性比RAPD高。12对引物共扩增出2147条带纹,93条具有多态性。朱鹮的条带共享率为0.718,遗传多样性指数为3.664,反映群体内个体间相似程度较高,遗传多态性较低,需要加强对朱鹮的遗传多样性保护。     

DNA分子标记在实验动物遗传分析中的应用

DNA分子标记技术很多,基本都是建立在RFLP、PCR和重复顺序的基础上的。本文重点介绍了限制性片段长度多态性（RFLP）标记、随机扩增多态性DNA（RAPD）标记、微卫星DNA（STR）标记、DNA指纹（DFP）标记、扩增片段长度多态性（AFLP）标记等几种重要的DNA分子标记技术的定义、结构、分布、组成、保守性、优点及丰富的多态性等。并重点介绍了微卫星DNA（STR）标记在分子遗传监测、遗传多样性分析和遗传血缘关系及个体识别等领域的应用。     

DNA分子标记技术及其在植物遗传多样性研究中的应用

本文阐述了ＤＮＡ限制性片段长度多态性、ＤＮＡ指纹图谱、单位点小卫星、单位点微卫星及随机扩增多态ＤＮＡ等主要的ＤＮＡ遗传标记技术的基本原理和方法．综述了不同ＤＮＡ标记的优缺点及其在植物遗传多样性研究中的应用前景．     

基于粪便DNA的岩羊微卫星引物的筛选及个体识别

贺兰山保护区岩羊Pseudois nayaur种群数量近年来增长很快,为了解贺兰山保护区的岩羊种群的遗传健康状况,从而更好地保护和管理岩羊种群,需要对岩羊种群的遗传多样性进行研究。在其近缘物种中选出36对微卫星引物对收集到的岩羊粪便DNA样品进行PCR扩增。结果发现,有9对引物在岩羊粪便DNA中扩增出了多态性位点。各位点基因杂合度介于0.26~0.95之间,平均杂合度为0.48,各位点多态信息含量介于0.23~0.68之间,平均多态性为0.48。     

树鼩微卫星DNA的多态性研究

目的探索并建立一种检测树鼩群体遗传多样性的方法。方法利用聚合酶链反应（PCR）扩增技术对30只树鼩个体的11个微卫星位点进行了遗传检测。结果所选的11个微卫星DNA位点中,有9个具有高度多态性,2个微卫星DNA位点多态性较差。结论所研究的树鼩微卫星位点中,有9个符合遗传标记特点,可用于检测树鼩群体的遗传多样性。     

鲫鱼微卫星DNA的分离及多态性分析

目的：用近缘物种鲤微卫星引物来分离鲫鱼微卫星标记并对其多态性进行分析。方法：以鲫鱼基因组DNA为模板,采用6对鲤微卫星引物进行PCR扩增,PCR产物经8%的非变性聚丙烯酰胺凝胶电泳和银染色检测。结果：筛选出2个以AC和TA为重复单元的鲫鱼新的微卫星标记。多态性分析表明,这2个微卫星标记的遗传杂合度分别为0.611和0.644,多态信息含量为0.536和0.572,属于高度多态性标记。结论：该研究筛选的2个微卫星标记可应用于鲫鱼遗传多样性、遗传连锁图谱构建及分子标记辅助育种等方面的研究。     

大鸨东方亚种遗传多样性的微卫星分析

为了探索东方亚种数量正在减少的原因和制定科学、有效的保护措施,利用微卫星DNA对大鸨东方亚种(Otistardadybowskii)的遗传多样性进行了研究。应用3个大鸨指名亚种的微卫星位点和13个波斑鸨的微卫星位点扩增了47个个体的基因组DNA,筛选出8个具有多态性的微卫星。其中有3个微卫星的多态性较低,其余5个微卫星的多态性较高。各位点的观察杂合度为0.0435-1.0000,平均杂合度(h)为0.6595;各位点多态信息含量(PIC)为0.0416-0.8520,平均为0.5497;有效等位基因数(E)为1.04-7.46,平均为3.61。4个位点符合HardyWeinberg平衡,4个位点偏离了HardyWeinberg平衡。多方面比较发现,大鸨东方亚种遗传多样性很低,且低于指名亚种,这可能由于其种群较小、历史遗传瓶颈作用、生境破碎化、分布地域紧缩等原因造成的。     

一个人工朱曦种群的遗传多态性

利用蛋白质电泳、RAPD和微卫星DNA技术,对我国濒危保护动物———人工饲养朱进行遗传多态性研究。结果表明,40只朱在前白蛋白(Pre)、白蛋白(Alb)、后白蛋白(Pa)、运铁蛋白(Tf)共4个蛋白质位点上没有发现多态性。用40个随机引物对40只朱进行RAPD分析,其中35个引物扩增出完全相同的RAPD带纹,仅筛选到5个多态性引物,朱群体的平均带纹相似率为0.86。利用其它物种的22对微卫星引物对30只朱的DNA进行检测,共筛选出4对多态性微卫星引物,扩增出13个等位基因。朱在这4对微卫星位点的平均杂合度为0.3760,平均多态信息含量为0.3382,平均等位基因数为2.075。表明朱人工饲养群体的遗传多态性比发现之初得到了一定程度的恢复,但仍然比较贫乏     

中国珍稀濒危物种——朱鹮

朱曾广泛分布于中国、日本、朝鲜半岛和俄罗斯的远东地区。其它国家的朱先后灭绝,目前只栖息于我国,为我国所特有,是我国四大国宝之一,当今世界极濒危级鸟类。1981年朱在陕西洋县重新被发现时其种群数量只有7只,经20余年的保护,目前我国朱的总数已达到700余只,人工繁殖也取得巨大成功。笔者想通过本文唤起人们对生活在西部边远地区的我国珍稀鸟类———朱的关注与保护。     

Molecular Cloning and Biochemical Analysis of Tyrosinase from the Crested Ibis in China

The crested ibis, one of the most endangered birds in the world, could benefit from research into its genetic diversity as a tool for conservation in the future. Tyrosinase is thought to play a major role in the production of common yellow to black melanins in birds. We have cloned and sequenced four exons of the crested ibis tyrosinase gene and discovered that the amino acid sequence has high similarity to zebra finch tyrosinase (93?%), followed by chicken (91?%) and quail (91?%). Some functional and structural domains in the crested ibis tyrosinase coding area were found to be conserved during evolution. Nine sequence variants were found in the partial coding sequence, one in exon 1 and eight in exon 4. Sequence variant 1 (SV1) shows intermediate polymorphism (0.25?<?PIC?<?0.5), and further study is needed to determine whether it can be used as a potential molecular marker in crested ibis artificial breeding programs.     

Habitat evaluation for crested ibis: A GIS-based approach

We evaluated habitat quality for crested ibis (Nipponia nippon) using a geographic information system (GIS). First, we digitized the topography map, vegetation map, river map, road map and villages/towns map by ArcInfo, and gave each map layer a suitability index based on our perceptions of the needs of crested ibis. Second, we overlayed these maps to obtain an integrated map of habitat quality. Finally, we compared the calculated habitat quality with the actual distribution of crested ibis. We found that the birds were almost always located at the site of high quality (habitat suitability index [HSI]>0.6), which indicated that the factors we selected were important for crested ibis. We also found that crested ibis were never located at some sites of high quality, thus, we assume that other factors not considered in this study limit the distribution of crested ibis. Regression analysis indicated that crested ibis preferred lower elevation habitats and tolerated higher levels of human disturbance in recent years than previously reported. These results reflected a 20-year protection program for this species.     

基于MaxEnt模型的道路对朱鹮繁殖地的影响

道路对野生动物适宜繁殖地影响的定量研究是目前道路生态学领域有待深入的内容之一.本文以国家一级保护动物朱鹮为研究对象,利用物种分布模型(SDM)中MaxEnt模型,通过对影响朱鹮繁殖地的环境变量进行自相关分析,剔除自相关系数大于0.8的环境变量,最终选择了10个环境变量为朱鹮繁殖地的影响因子,分别为最冷季度平均气温、景观类型、归一化植被指数、坡度、坡向、距水体距离、距水田距离、距高等级公路(县级及以上道路)距离、距低等级公路(乡镇和林间道路)距离、距居住地距离.通过对环境变量的贡献率进行分析表明: 最冷季度平均气温、景观类型以及距水田距离、距高等级公路距离是影响朱鹮筑巢的主要因子.分别研究了含道路变量(情景Ⅰ)、不含高等级公路(情景Ⅱ)以及不含低等级公路(情景Ⅲ)3种情景下朱鹮繁殖地适宜区分布.结果表明:道路对朱鹮繁殖地干扰影响显著,高等级公路的影响比低等级公路大,高等级公路和低等级公路的存在分别使朱鹮繁殖地适宜区的面积减小了66.23和35.69 km².朱鹮筑巢对高等级公路有明显的回避效应,平均回避距离为1500 m.本研究对未来制定朱鹮的保护管理措施具有重要意义,同时可为工程建设项目对野生动物的环境影响定量评价提供应用示范.     

用组合模型综合比较的方法分析气候变化对朱鹮潜在生境的影响

气候变化的不确定性和物种与环境关系的不确定性使气候变化生物学的研究充满变数。为了降低不确定性,人们开始用组合模型综合比较的方法研究物种对气候变化的响应。以朱鹮(Nipponia nippon)为研究对象,介绍组合模型综合比较方法的特点。朱鹮曾经高度濒危,目前种群大小在迅速恢复中;然而其分布区依旧狭小,气候变化可能是朱鹮面临的新威胁。应用BIOMOD模型中的9种模型,选择了每年的最低温和最高温、温度的季节性变异、每年的总降水量和降水的季节性变异共5个气候因子,依据WorldClim气候数据的CGCM2气候模型的A2a排放情形,计算了朱鹮当前(1950—2000年)的适宜生境和2020年、2050年、2080年3个阶段的潜在生境范围。结果表明朱鹮潜在生境将逐渐北移,生境中心脱离现在的保护区。因此,制定朱鹮的长期保护策略是必要的。9个模型在预测结果上、变量权重上和拟合优度的指标上都有差异,反映了模型本身的不确定性。气候变化的生物学效应比较复杂,应用多个模型进行综合比较,可以尽可能地减少模型所导致的误差。     

Why the crested ibis declined in the middle twentieth century

The crested ibis (Nipponia nippon) had declined severely from a common species to only two pairs in last century. To analyze the declining process, we established a GIS database with historical occurrences of the crested ibis based on published literatures, and layers of environmental factors such as elevation, wetland, and human activities. We compared the environmental factors at the occurrence sites in different periods to quantify the changes of habitat use across time. To address the spatial deviation of the occurrences and check the effect of measuring scale on habitat use, we calculated the environmental factors in a serial measuring scales from 1 to 161 × 161 km². Our results indicated that the crested ibis traditionally lives in habitat with higher wetland density and higher human impact, then it gradually moved to areas with higher elevation and lower human impact in last century. In 1980–2000 the crested ibis stayed at a very high elevation, lower wetland density and lower human impact as possible consequences of human activities such as using fertilizer and pesticide, drying the over wintering rice paddies, and direct hunting. Our quantitative analysis of the habitat use matched well the previous published statements (which have no numerical evidences) on the declination of the crested ibis. We suggest to reestablish the habitat with traditional farming practice that the bird has adapted as a major solution for applications such as conservation planning and reintroduction.     

Current state and the future of the crested ibis (Nipponia nippon): A case study by population viability analysis

There is only one population of the crested ibis (Nipponia nippon) surviving in the wild, which has not exceeded 40 individuals since it was rediscovered in 1981. We used detailed data of the annual population size and natality collected by the Crested Ibis Conservation and Observation Station (CICOS) over the past 17years. We were able to obtain a more accurate census of age structure and mortality as all of the fledglings have been identified by leg bands in 1987. Consequently, we analyzed the current state of the crested ibis and predicted its future using Population Viability Analysis (PVA) with the help of the computer simulation model VORTEX (Version 7.2). The results indicated that the extinction probability of the crested ibis was 19.7% in 100years. Sensitivity analysis showed that the extinction probabilities were sensitive to catastrophes and environmental variation, and the long-term existence of the population was dependent on the carrying capacity of the habitat and inbreeding depression. Conservation efforts are critical for the survival of the crested ibis. The prior conservation project involves controlling hunting and predation, as well as improving the life level of these few remnant creatures.     

Genome-Wide SNP and STR Discovery in the Japanese Crested Ibis and Genetic Diversity among Founders of the Japanese Population

The Japanese crested ibis is an internationally conserved, critically threatened bird. Captive-breeding programs have been established to conserve this species in Japan. Since the current Japanese population of crested ibis originates only from 5 founders donated by the Chinese government, understanding the genetic diversity between them is critical for an effective population management. To discover genome-wide single nucleotide polymorphisms (SNPs) and short tandem repeats (STRs) while obtaining genotype data of these polymorphic markers in each founder, reduced representation libraries were independently prepared from each of the founder genomes and sequenced on an Illumina HiSeq2000. This yielded 316 million 101-bp reads. Consensus sequences were created by clustering sequence reads, and then sequence reads from each founder were mapped to the consensus sequences, resulting in the detection of 52,512 putative SNPs and 162 putative STRs. The numbers of haplotypes and STR alleles and the investigation of genetic similarities suggested that the total genetic diversity between the founders was lower, although we could not identify a pair with closely related genome sequences. This study provided important insight into protocols for genetic management of the captive breeding population of Japanese crested ibis in Japan and towards the national project for reintroduction of captive-bred individuals into the wild. We proposed a simple, efficient, and cost-effective approach for simultaneous detection of genome-wide polymorphic markers and their genotypes for species currently lacking a reference genome sequence.