以mtDNA细胞色素b基因探讨斑翅山鹑的分类地位

以线粒体DNA细胞色素b(mtDNA Cytb)基因作为分子标记,提取鸡形目雉科10个属18个样本的DNA,PCR扩增自动测序,获得序列长度828bp。将山鹑属中的斑翅山鹑(Perdix dauuricae)与其近缘属排序对比,分别以角叫鸭(Anhima cornuta)和海龟(Kachuga dhongoka)为外群,用邻接法、最大简约法和最小进化法构建分子系统发生树。邻接法构建的系统树显示斑翅山鹑与雉聚类,核苷酸序列差异值斑翅山鹑与环颈雉为15．2％,与石鸡为16．1％。分析认为在雉科中斑翅山鹑与雉类更接近,而与真正的鹑亲缘关系较远。同时以mtDNA分析鸟类系统发生时海龟更适合作为外群。以角叫鸭为外群加入西方松鸡(Tetrao urogallus)时,则斑翅山鹑的分类地位不能确定,这可能与鸡形目的适应辐射有关。     

高温环境样品总DNA直接和间接提取方法的比较

分别采用两种环境总DNA直接提取法和一种间接提取法从6种温泉菌席样品中提取总DNA,以DNA粗产物的纯度、能否用于后续PCR扩增及PCR-DGGE(变性梯度凝胶电泳)所反映的微生物多样性为评价指标对两类方法进行比较和评价。研究发现,虽然间接提取法效率低下,但对于高温极端环境中生物量较小的样品,间接法能得到有研究价值的、纯度较高的环境样品总DNA,而直接法得到的DNA量小且不适于PCR扩增操作。在使用这2类方法都能得到可用于研究操作的DNA的情况下,间接提取法能更好的体现环境样品中微生物的多样性。     

一种从大熊猫粪便中提取DNA的改进方法

本研究描述一个改进的方法，使从大熊猫粪便中提取DNA用于PCR扩增变得更加容易。在粪便DNA的提取过程中采用一个新的预处理方法，将粪便用预冷的丙酮洗2～3次，除去粪便中含有的大量PCR抑制物，然后用蛋白酶K裂解、酚氯仿抽提，能提取到纯度很高的DNA供PCR扩增。本实验PCR扩增了大熊猫脑源性神经营养因子(BDNF)基因和线粒体细胞色素6基因片段，并进行测序分析，证实了提取的可靠性。对比本方法和未经丙酮预处理的方法提取的DNA进行PCR扩增，前者的扩增结果明显优于后者。     

基于自动录音技术研究三种雉类 鸣叫特征和节律

自动录音机是一种低成本、高效、可以在较长时间和较大空间范围内对物种的活动进行有效监测的设备。2019年4至6月,在北京小龙门地区（40°00′ N,115°26′ E）褐马鸡（Crossoptilon mantchuricum）、勺鸡（Pucrasia macrolopha）和环颈雉（Phasianus colchicus）的栖息地内布设了songmeter自动录音机,对三种雉类的鸣声进行为期2个月的连续收集。利用Kaleidoscope软件（Wildlife Acoustics公司,美国）,对不同雉类鸣声进行了人工辅助的机器学习和自动识别,从录制的22 536 h录音数据中提取了褐马鸡、勺鸡和环颈雉的鸣声。优化鸣声提取的时频参数后,勺鸡鸣声提取的正确率为73.32%,探测率为52.91%;环颈雉鸣声提取的正确率为89.32%,探测率为67.36%;褐马鸡鸣声提取的正确率较低,仅为8.69%,探测率为58.54%。结合三种雉类的繁殖资料,揭示了三种雉类的鸣声节律：褐马鸡和环颈雉有早晚两个鸣叫高峰期,但高峰时段不尽相同,勺鸡只存在鸣叫早高峰;褐马鸡、勺鸡和环颈雉在交配期、孵卵前期具有一个鸣叫高峰。     

我国32种鸟类DNA条形码分析

DNA条形码是一种分子分类方法,近年来在物种鉴定方面得到迅速的发展和应用.本研究分析了我国27属32种鸟类(61只)的线粒体细胞色素c氧化酶亚基Ⅰ(COⅠ)基因的条形码片段,分别用阈值法、聚类法和诊断核苷酸进行了分析,探究DNA条形码鉴定我国鸟类的准确性.结果显示,种内CO Ⅰ序列变异很小,种间存在较多的变异位点,种间的遗传距离显著大于种内的遗传距离,DNA条形码序列能够鉴定所有鸟类.     

扬子鳄鞣制皮革和鳞片的DNA提取方法

运用一种改进的提取方法 ,作者从鞣制皮革中成功地提取了总DNA ,同时还对尾尖皮、鳞片、盐腌生皮等皮质进行了DNA提取 ;用 12SrRNA基因扩增的通用引物、扬子鳄鉴别引物、微卫星引物及RAPD引物进行PCR扩增 ,并对部分扩增结果进行测序 ,以检验提取效果。结果证明 ,几种皮质标本都可提取出DNA ,其中尾尖皮和鳞片的提取效果较好 ,用四种引物都可扩增出明显亮带 ;盐腌生皮和鞣制皮提取结果也很好 ,并且用12SrRNA通用引物、扬子鳄鉴别引物扩增的亮带较明显 ,可进行扬子鳄皮质用品等的分子鉴定及部分序列的扩增和测序研究     

保存方式和回软温度对蜜蜂标本DNA提取的影响北大核心CSCD

旨在探寻保存方式及制作干标本前回软温度对蜜蜂不同部位DNA的影响。采用酚-氯仿法对不同方式保存的蜜蜂以及不同温度回软后的蜜蜂干标本的总DNA进行提取,通过琼脂糖凝胶电泳和PCR扩增对DNA的提取结果进行鉴定。电泳结果显示,从新鲜标本、无水乙醇泡制或自然干燥保存半年的标本中均可提取到较高质量的总DNA,尤以头部与足部提取效果最佳。经不同水浴温度回软后保存半年的蜜蜂干标本总DNA提取结果显示,回软温度为65℃时对蜜蜂DNA的破坏性最小,DNA提取的最佳部位为蜜蜂足部。正交试验结果显示,55℃回软后的足部为蜜蜂干标本DNA提取的最优组合。PCR扩增结果显示,本实验提取的总DNA能成功地应用于蜜蜂线粒体基因16S rRNA和COI的扩增。从保存方式、提取部位和回软操作3个因素对蜜蜂标本DNA的提取进行了研究,提供了较好的选择方案。     

一种从毛发中提取DNA 的简易方法

用PCR 缓冲液及蛋白酶K 在PCR仪上对单支毛囊进行消化,　可获得用于PCR反应的足量的DNA。还对几种从毛发中提取DNA的不同方法进行了比较,并对从毛干中提取线粒体DNA进行了讨论。     

穿山甲标本和甲片的DNA提取及PCR扩增

为验证经处理后的穿山甲(Manis spp.)标本和甲片是否可以用于种间分子鉴定标记的开发及个体识别工作,本文在样品的预处理、消化、提取后纯化等方面对传统提取方法进行了改进,分别从穿山甲剥制标本、干皮标本及甲片中提取总DNA;然后用Cyt b基因扩增通用引物、12S rRNA基因全序列扩增引物、RAPD引物及微卫星引物进行了PCR扩增,并对部分扩增结果进行了序列测定.结果表明,除剥制标本的脚底皮张组织外,其他样品基本都可以提取出DNA.以此为模板的PCR扩增中,2种线粒体基因引物扩增出明显目的条带,RAPD引物扩增出种间特异条带,测序结果可用于种间特异性引物及SCAR引物的开发;微卫星引物在甲片样品中扩增稳定,可用于个体识别工作.     

中国珍稀濒危雉类研究现状和前景

据Delacour(1951)全世界共有雉类16属48种,我国就有12属(占全世界总属数75%)、26种(占55%)。我国雉资源居世界第一位。 (一)雉类现状令人忧虑据《野生动物保护法》附录中的国家重点保护野生动物名录,已列入保护名录的雉科鸟类有25种,占我国雉科鸟类总种数96.2%,其中属于国家一级保护动物有14种,占53.8%,二级保护动物有11种占,42.3%;世界鸟类“红皮书”(1982)上列举世界濒危雉类14种中,中国有九种,占全世界濒危总种数64.3%。上述数字表明我国雉类中的大多数已处于濒危状态,急待保护。雉类数量日趋下降的主要原因有:①森林是雉类赖以生存的主要条件,是栖息、繁衍生息的场所。如我国极为珍贵的一级保护动物绿孔雀、     

The distribution of endogenous chicken retrovirus sequences in the DNA of galliform birds does not coincide with avian phylogenetic relationships.

The chicken is a domesticated form of Red Jungle-fowl (Gallus gallus), which belongs to the Pheasant family (Phasianidae) within the order Galliformes. Domestic chickens carry the genome of the endogenous retrovirus RAV-O as DNA sequences integrated into host chromosomes transmitted through the germ line. We have examined the presence and distribution of RAV-O-related sequences in the DNA of Red Junglefowl and other closely related species of Junglefowl, as well as more distantly related Pheasants and Quail. DNA sequences homologous to RAV-O were analyzed by molecular hybridization in liquid and after electrophoresis of restriction endonuclease fragments. The presence of RAV-O-related sequences in avian DNA does not correlate with phylogenetic relationships. Under stringent conditions of hybridization in liquid, DNA sequences homologous to RAV-O cDNA were detected at high levels (greater than 80% homology( only in the genomes of the domestic chicken and its phylogenetic ancestor, the Red Junglefowl (Gallus gallus). The DNA of two other species of Gallus (G. sonnerati, Sonnerat's Junglefowl and G. varius, Green Junglefowl), of Ring-necked Pheasant and of Japanese Quail contained sequences with less than 10% homology to RAV-O cDNA. Under conditions permitting mismatching, however, Ring-necked Pheasant DNA hybridized up to 50% of the RAV-O cDNA, and Quail DNA 24%, whereas the extent of hybridization to Sonnerat's and Green Junglefowl DNA was not markedly increased. Analysis of restriction enzyme digests revealed several distinct fragments of DNA hybridizing to chick retrovirus cDNA in both Red Junglefowl and domestic chicken, and multiple fragments in DNA from two species of Phasianus. No fragments with sequences related to chicken retroviruses were found, however, in digests of DNA prepared from Sonnerat's, Ceylonese and Green Junglefowl, from two other Pheasant genera (Chrysolophus and Lophura), or from one Quail genus (Coturnix). Thus the DNA of three Junglefowl species closely related to Gallus gallus lacked RAV-O sequences while the DNA of more distantly related Phasianus species showed significant homology. These results show that RAV-O-related sequences have not diverged together with the normal host genes during the evolution of the Phasianidae. Although RAV-O sequences are endogenous in all domestic chickens and Red Junglefowl studied thus far, it appears that the RAV-O genome has been introduced relatively recently into the germ line of Gallus gallus, following speciation but before domestication, and independently of the related sequences found in members of the genus Phasianus.     

桂西南喀斯特山地雉类的生态分布和空间生态位分析

桂西南喀斯特地区位于中国广西的西南部,属于全球生物多样性热点地区。通过自2003年以来,对该地区雉类进行的调查,共记录到7种雉类,分别是中华鹧鸪(Francolinus pintadeanus)、褐胸山鹧鸪(Arborophila brunneopectus)、棕胸竹鸡(Bambusicola fytchii)、灰胸竹鸡(Bambusicola thoracica)、原鸡(Gallus gallus)、白鹇(Lophura nycthemera)和环颈雉(Phasianus colchicus)。对该地区雉类的生态分布状况及栖息地的植被类型和坡位等空间生态位进行分析和比较的结果表明,原鸡的综合生态位最宽,灰胸竹鸡第二,最窄为中华鹧鸪。综合生态位重叠值最大的是中华鹧鸪--环颈雉和灰胸竹鸡--原鸡。分布范围狭窄、种群数量相对较少及生态适应性较低的褐胸山鹧鸪应该是该地区最易受到威胁的种类。     

Public Access for Pheasant Hunters: Understanding an Emerging Need

Ring-necked pheasant (Phasianus colchicus; i.e., pheasant) hunting participation is declining across North America, reflecting a larger downward trend in American hunting participation and threatening benefits to grassland conservation and rural economies. To stabilize and expand the pheasant hunting population, we must first identify factors that influence pheasant hunter participation. We used an extensive in-person hunter survey to test the hypothesis that hunter demographics interact with social-ecological traits of hunting locations to affect hunter decisions, outcomes, and perceptions. We built a series of Bayesian mixed effects models to parse variation in demographics, perceptions, and hunt outcomes of pheasant hunters interviewed at public access hunting sites across 3 regions in Nebraska, USA, that varied in pheasant abundance and proximity to urban population centers. Among pheasant hunters in Nebraska, access to private lands was negatively related to the human population density of a pheasant hunter's home ZIP code and the distance a hunter had traveled to reach a hunting location. Pheasant hunters interviewed closer to metropolitan areas tended to be more urban and travel shorter distances, and their parties were more likely to include youth but less likely to include dogs. Hunter satisfaction was positively associated with seeing and harvesting pheasants and hunting with youth. Whereas youth participation and the number of pheasants seen varied by study region, hunter satisfaction did not differ across regions, suggesting that hunters may calibrate their expectations and build their parties based on where they plan to hunt. The variation in hunter demographics across hunting locations and disconnects between social and ecological correlates of hunter satisfaction suggests that diverse pheasant hunting constituencies will be best served by diverse pheasant hunting opportunities. © 2019 The Wildlife Society.     

Metagenomics analysis of the fecal microbiota in Ring-necked pheasants (Phasianus colchicus) and Green pheasants (Phasianus versicolor) using next generation sequencing

Pheasant reintroduction and conservation efforts have been in place in Pakistan since the 1980 s, yet there is still a scarcity of data on pheasant microbiome and zoonosis. Instead of growing vast numbers of bacteria in the laboratory, to investigate the fecal microbiome, pheasants (green and ring neck pheasant) were analyzed using 16S rRNA metagenomics and using IonS5TMXL sequencing from two flocks more than 10 birds. Operational taxonomic unit (OTU) cluster analysis and phylogenetic tree analysis was performed using Mothur software against the SSUrRNA database of SILVA and the MUSCLE (Version 3.8.31) software. Results of the analysis showed that firmicutes were the most abundant phylum among the top ten phyla, in both pheasant species, followed by other phyla such as actinobacteria and proteobacteria in ring necked pheasant and bacteroidetes in green necked pheasant. Bacillus was the most relatively abundant genus in both pheasants followed by Oceanobacillus and Teribacillus for ring necked pheasant and Lactobacillus for green necked pheasant. Because of their well-known beneficial characteristics, these genus warrants special attention. Bird droppings comprise germs from the urinary system, gut, and reproductive sites, making it difficult to research each anatomical site at the same time. We conclude that metagenomic analysis and classification provides baseline information of the pheasant fecal microbiome that plays a role in disease and health.     

The use of infrared‐triggered cameras for surveying phasianids in Sichuan Province,China

We report on the use of infrared‐triggered cameras as an effective tool to survey phasianid populations in Wanglang and Wolong Nature Reserves, China. Surveys at 183 camera‐trapping sites recorded 30 bird species, including nine phasianids (one grouse and eight pheasant species). Blood Pheasant Ithaginis cruentus and Temminck’s Tragopan Tragopan temminckii were the phasianids most often detected at both reserves and were found within the mid‐elevation range (2400–3600 m asl). The occupancy rate and detection probability of both species were examined using an occupancy model relative to eight sampling covariates and three detection covariates. The model estimates of occupancy for Blood Pheasant (0.30) and Temminck’s Tragopan (0.14) are close to the naïve estimates based on camera detections (0.27 and 0.13, respectively). The estimated detection probability during a 5‐day period was 0.36 for Blood Pheasant and 0.30 for Temminck’s Tragopan. The daily activity patterns for these two species were assessed from the time/date stamps on the photographs and sex ratios calculated for Blood Pheasant (152M : 72F) and Temminck’s Tragopan (48M : 21F). Infrared cameras are valuable for surveying these reclusive species and our protocol is applicable to research or monitoring of phasianids.     

Evolutionary shift in the site of cleavage of prelysozyme

Sequences are presented for the signal peptides of prelysozymes from 6 species of birds and compared to the known sequence for chicken prelysozyme c. The sequencing was done with synthetic oligonucleotides as primers and oviduct mRNA as the template, obviating the need to clone DNA from these species. Ring-necked pheasant prelysozyme c differs from all other prelysozymes c and pre-alpha-lactalbumins examined by being cleaved in vivo between amino acid residues 17 and 18 instead of between residues 18 and 19. The feature unique to the signal peptide of pheasant prelysozyme c is proline at position 17. Besides showing that proline is acceptable as the carboxyl-terminal amino acid of the signal peptide, our finding implies that it cannot occur as the penultimate amino acid in the signal peptide. This supports the view that unless a polypeptide has the proper secondary structure, signal peptidase will not cleave it, and that this secondary structure is a beta-turn. Another outcome of this comparative study is an estimate that the mean rate of sequence evolution in the prelysozyme signal peptide is 1%/two million years of divergence, similar to that calculated for the insulin signal peptide. Because this rate is a third of the silent substitution rate, it is likely that one out of every three amino acid substitutions is compatible with signal peptide function.     

Niche partitioning among three montane ground‐dwelling pheasant species along multiple ecological dimensions

Pheasants (order Galliformes) are typical ground‐dwelling birds, having a large body size and weak flight abilities. Sympatric pheasants are expected to share narrower niche space and face more extensive interspecific competition. However, little work has been undertaken simultaneously to investigate niche partitioning among sympatric pheasant species across multiple ecological dimensions. We compared microhabitat use, activity pattern and foraging strategy of sympatric Blood Pheasant Ithaginis cruentus, Buff‐throated Partridge Tetraophasis szechenyii and White Eared‐pheasant Crossoptilon crossoptilon on the Qinghai‐Tibet Plateau, China, to identify potential interspecific niche partitioning along different ecological dimensions in the breeding season. We found that the Buff‐throated Partridge differed significantly from the other two species in microhabitat use, and the three species showed different foraging strategies. It is likely that niche partitioning reduced potential interspecific competition, thus facilitating the species’ stable coexistence. Our study provides practical evidence of multidimensional niche theory within sympatric ground‐dwelling pheasant species, emphasizing that species interactions and coexistence within a guild are often not uni‐dimensional. Given global conservation concern for maintaining bird diversity, we recommend further restriction of yak grazing in these species’ habitat.     

Characterization of nine microsatellite loci for a globally vulnerable species, Reeves’s Pheasant (Syrmaticus reevesii)

Reeves’s Pheasant, Syrmaticus reevesii, is an endemic species of China. Due to habitat loss, poaching and human disturbance, its wild population has been decreased drastically and it is listed as a globally vulnerable species by IUCN/BirdLife/WPA (IUCN 2008). Here, we report nine new polymorphic microsatellite markers isolated from the Reeves’s Pheasant. The number of alleles per locus varies between four and fourteen, with expected heterozygosity ranging from 0.349–0.776 (n  = 90). These polymorphic loci provide a valuable tool for future population studies that relate to the conservation of this pheasant.     

白颈长尾雉栖息地的季节变化

１９９２年１０月至１９９３年５月,在浙江省乌岩岭自然保护区,利用无线电追踪技术和野外直接观察等采样方法对白颈长尾雉各季节栖息地及其食性进行了研究。