Introgression through rare hybridization: A genetic study of a hybrid zone between red and sika deer (genus Cervus) in Argyll, Scotland

In this article we describe the structure of a hybrid zone in Argyll, Scotland, between native red deer (Cervus elaphus) and introduced Japanese sika deer (Cervus nippon), on the basis of a genetic analysis using 11 microsatellite markers and mitochondrial DNA. In contrast to the findings of a previous study of the same population, we conclude that the deer fall into two distinct genetic classes, corresponding to either a sika-like or red-like phenotype. Introgression is rare at any one locus, but where the taxa overlap up to 40% of deer carry apparently introgressed alleles. While most putative hybrids are heterozygous at only one locus, there are rare multiple heterozygotes, reflecting significant linkage disequilibrium within both sika- and red-like populations. The rate of backcrossing into the sika population is estimated as H = 0.002 per generation and into red, H = 0.001 per generation. On the basis of historical evidence that red deer entered Kintyre only recently, a diffusion model evaluated by maximum likelihood shows that sika have increased at approximately 9.2% yr-1 from low frequency and disperse at a rate of approximately 3.7 km yr-1. Introgression into the red-like population is greater in the south, while introgression into sika varies little along the transect. For both sika- and red-like populations, the degree of introgression is 30-40% of that predicted from the rates of current hybridization inferred from linkage disequilibria; however, in neither case is this statistically significant evidence for selection against introgression.     

Introgression of exotic Cervus (nippon and canadensis) into red deer (Cervus elaphus) populations in Scotland and the English Lake District

Since the mid‐19th century, multiple introductions of Japanese sika deer (Cervus nippon nippon) and North American wapiti (C. canadensis) have taken place in the British Isles. While wapiti have generally been unsuccessful, sika have been very successful, especially in Scotland where they now overlap at least 40% of the range of native red deer (C. elaphus). Hybridization between these two species and red deer has been demonstrated in captivity and in the wild. Using a panel of 22 microsatellite loci that are highly diagnostic between red deer and sika, and moderately diagnostic between red deer and wapiti, we investigated the extent of introgression between these species in 2,943 deer sampled from around Scotland and from the English Lake District using the Bayesian clustering software STRUCTURE. We also used a diagnostic mitochondrial marker for red deer and sika. Our survey extends previous studies indicating little introgression of wapiti nuclear alleles into red deer, in particular in Northern Scotland, Kintyre, and the Lake District. We found a new area of extensive sika introgression in South Kintyre. In the North Highlands, we show for the first time geographically scattered evidence of past hybridization followed by extensive backcrossing, including one red‐like individual with sika introgression, two sika‐like individuals with red deer introgression, and six individuals that were apparently pure sika at the nuclear markers assessed but which carried red deer mitochondria. However, there has not been a collapse of assortative mating in this region. Similarly, in the English Lake District red deer, we found only traces of past sika introgression. No sika alleles were detected in the Central Highlands or the Hebridean red deer refugia. We make suggestions for management to prevent further spread of sika alleles into red deer and vice versa.     

Polymorphism and genetic control of erythrocyte 6-phosphogluconate dehydrogenase in the genus Cervus

A study of 11 enzyme systems in blood samples of Cervus dama, C. elaphus, C. nippon and hybrids C. elaphus X nippon has revealed an erythrocyte 6-phosphogluconate dehydrogenase polymorphism in the hybrid populations. Genetic analysis suggests that this enzyme is controlled by one gene locus with two codominant alleles, one specific for pure Japanese sika deer, the other for pure red deer as well as for fallow deer, while both alleles have been found in the red X sika hybrids.     

Evaluation of introgressive hybridization among Cervidae in Japan's Kinki District via two novel genetic markers developed from public NGS data

Hybridization and backcrossing of native populations with introduced species can lead to introgression and genetic alteration. In this study, we evaluated introgression in 43 deer from a potential hybrid zone around Okinoshima Island, Kinki District, Japan. This region witnessed the migration of a hybrid population (cross between the Formosan sika deer [Cervus nippon taiouanus] and other deer species) that could potentially breed with the native Japanese sika deer (C. n. centralis). We used an existing genetic marker for the mitochondrial cytochrome b gene and two novel markers for nuclear DNA, developed using publicly available next‐generation sequencing data. We identified one mainland deer with a mitochondrial haplotype identical to that of the Formosan sika deer as well as nuclear heterozygous sequences identical to those of Formosan and Japanese sika deer. This suggests that the mainland deer is a hybrid offspring of the Okinoshima population and native deer. However, only Japanese sika deer sequences were found in the other 42 samples, indicating limited introgression. Nevertheless, hybridization pre‐ and postintroduction in the Okinoshima population could cause multispecies introgression among Japanese sika deer, negatively affecting genetic integrity. We developed a simple test based on polymerase chain reaction–restriction fragment length polymorphism to detect introgression in natural populations. Our method can accelerate genetic monitoring of Japanese sika deer in Kinki District. In conclusion, to prevent further introgression and maintain genetic integrity of Japanese sika deer, we recommend establishing fences around Okinoshima Island to limit migration, besides a continued genetic monitoring of the native deer.     

A phylogenetic comparison of red deer and wapiti using mitochondrial DNA

A phylogeny was constructed for red deer/wapiti (Cervus elaphus) subspecies using sequence data from the control region of mitochondrial DNA (mtDNA). The tree was rooted using Cervus nippon (sika deer), Cervus albirostris (Thorold's white-lipped deer), and several Odocoileinae species. A division between the mtDNA haplotypes of red deer (European) and wapiti (Asian/North American) corresponds to subspecies found on opposite sides of the Himalayan Mountains and Gobi, which suggests wapiti should be reconsidered for the status of C. canadensis. Using parsimony and distance analysis, red deer and wapiti are derived from a single recent common ancestor, which is consistent with current taxonomy that recognizes the subspecies of Cervus elaphus as monophyletic group. However, maximum-likelihood analysis using weighted transitional substitutions caused red deer to form a sister group to sika deer (Cervus nippon) and wapiti. A phenetic comparison revealed wapiti also share more nucleotide similarities with sika deer, although approximately 5% sequence divergence separates wapiti, sika, and red deer. Phylogenetic evidence from the cytochrome b sequences corroborated observations from the control region. Observations from this study suggest that the species status of wapiti should be reinstated.     

白唇鹿的染色体组型、C带和G带带型

白唇鹿产于我国西藏东部和四川西部。它和大熊猫一样,是我国特有的珍贵动物。有关它的染色体组型至今尚未见报道。我们于去年八月,对两头雄性白唇鹿外周血及茸缸淋巴细胞进行了培养,并作了染色体组型观察和C带及G带带型的观察。 一、材料和方法 北京动物园的两头准性白唇廘,一头10岁,一头4岁。 每一头先后采样二次。用装有司可林(Succinylcholinum chloratum)麻醉弹的麻醉枪击中鹿后,在颈静脉处,用含有1毫升肝素(浓度为50微     

A genetic study of sika (Cervus nippon) in the New Forest and in the Purbeck region, southern England: is there evidence of recent or past hybridization with red deer (Cervus elaphus)?

Sika Cervus nippon are native to Japan and East Asia but are now naturalized in many parks of the world, including Britain. In contrast to the substantial body of research on the extent of hybridization between sika and red deer Cervus elaphus in Scottish populations, there has been little genetic analysis of the English populations of sika. Sika in England still have a patchy and discontinuous distribution; populations are thus still genetically isolated and may be expected to show higher variability in genetic type. The current paper uses DNA-based techniques to explore the genetic composition of sika in the New Forest (Hampshire) and that of sika from the largest population in England, in the Purbeck region (Dorset). The study aims were to determine whether New Forest sika show signs of recent interbreeding with New Forest red deer populations, whether New Forest sika are genetically distinct from Purbeck sika, and whether New Forest and Purbeck sika show evidence of past hybridization or whether they are pure (non-hybrid) strains. Microsatellite analysis was used to compare the genetic profiles of individual deer. Results showed that sika and red deer in the New Forest were genetically distinct, indicating that there is no large-scale in situ hybridization occurring between these feral populations. In terms of overall genetic composition, there was no significant difference between the sika in the New Forest and Purbeck. However, a more detailed analysis found that New Forest sika showed a lower level of introgression with red deer compared with the Purbeck sika. We conclude that, overall, the New Forest sika deer do appear to be more genetically pure bred than the Purbeck sika.     

Investigating temporal changes in hybridization and introgression in a predominantly bimodal hybridizing population of invasive sika (Cervus nippon) and native red deer (C. elaphus) on the Kintyre Peninsula,Scotland

We investigated temporal changes in hybridization and introgression between native red deer (Cervus elaphus) and invasive Japanese sika (Cervus nippon) on the Kintyre Peninsula, Scotland, over 15 years, through analysis of 1513 samples of deer at 20 microsatellite loci and a mtDNA marker. We found no evidence that either the proportion of recent hybrids, or the levels of introgression had changed over the study period. Nevertheless, in one population where the two species have been in contact since ～1970, 44% of individuals sampled during the study were hybrids. This suggests that hybridization between these species can proceed fairly rapidly. By analysing the number of alleles that have introgressed from polymorphic red deer into the genetically homogenous sika population, we reconstructed the haplotypes of red deer alleles introduced by backcrossing. Five separate hybridization events could account for all the recently hybridized sika‐like individuals found across a large section of the Peninsula. Although we demonstrate that low rates of F1 hybridization can lead to substantial introgression, the progress of hybridization and introgression appears to be unpredictable over the short timescales.     

东北马鹿和东北梅花鹿F_1杂种精母细胞联会复合体分析

作者以界面铺张-硝酸银染色技术,对东北马鹿和东北梅花鹿的F_1可育杂种的精母细胞联会复合体进行亚显微观察及分析。在减数分裂前期,杂种鹿精母细胞中形成31条完整的常染色体联会复合体、一个端着丝粒染色体/中着丝粒染色体的三价体和XY双价体。这进一步证明,两种亲本鹿的染色体具有高度的同源性,其差别仅在于一个罗伯逊易位。三价体的顺式构型可能和杂种鹿的可育性有关。     

Distribution and range expansion of deer in Ireland

• 1 Throughout Europe, the range of many deer species is expanding. We provide current distribution maps for red deer Cervus elaphus, sika Cervus nippon, fallow deer Dama dama and muntjac deer Muntiacus sp. in Ireland, and estimates of range expansion rates for red deer, sika and fallow deer.
• 2 There was a considerable expansion in the ranges of red deer, sika and fallow deer between 1978 and 2008. The compound annual rate of expansion was 7% for red deer, 5% for sika and 3% for fallow deer. The total range increase was 565% for red deer, 353% for sika and 174% for fallow deer. The potential implications of these expansions are discussed.
• 3 There are unknown numbers of red‐sika hybrid deer in some parts of Ireland. Range expansion is likely to lead to further hybridizations with implications for the genetic integrity of deer stocks.
• 4 Sightings of free‐roaming muntjac deer were first recorded in 2007. The distribution of confirmed sightings of single and multiple animals in the eastern region of Ireland suggests multiple releases.
• 5 Deer are already impacting on both the economic and biodiversity values of habitats in Ireland, where, at present, no sustainable deer management policy exists.

     

日本日光地区森林中梅花鹿的夏季巢区面积和内部利用

为了估算梅花鹿(Cervusnippon)巢区面积和内部利用以及雌雄个体间的比较,于1993-1997年在日本国日光国立公园使用无线电遥测定位法测定了8头成年雌鹿和4头雄鹿(包括1头亚成年)在森林生境中的活动定位点。按照当地梅花鹿离开和返回越夏地的日期,使用每年6月1日至10月25日的无线电遥测定位数据,采用定值核心法(Fixed kernel)计算了梅花鹿的夏季巢区面积和内部利用密度分布。梅花鹿雄鹿夏季巢区面积(192·5±50·5 hm2)显著大于雌鹿(115·7±56·7 hm2,P=0·017) ,其半家区面积(HRS0·50)和核心区面积(HRS0·25)也显著大于雌鹿(依次为P=0·027和P=0·042) ,但巢区内部利用集中度指标Ah(HRS0·50/HRS0·95)和Ac(HRS0·25/HRS0·50)在雌雄鹿之间没有显著性差异(依次为P=0·999和P=0·234)。在无线电遥测的12头梅花鹿中, 11头个体的巢区为单核心型,只有1头成年雄鹿的巢区为双核心型。11头梅花鹿个体的混合数据显示巢区面积和体重之间存在显著性相关(P=0·049) [动物学报52 (2) : 235 -241 , 2006]。     

Phylogenetic relationships among european red deer, wapiti, and sika deer inferred from mitochondrial DNA sequences

We determined the mitochondrial cytochrome b gene sequences (1140 bp) of one subspecies of the European red deer (Cervus elaphus in Europe), three subspecies of the wapiti (C. elaphus in Asia and North America), and six subspecies of the sika deer (C. nippon in Japan). Our phylogenetic analysis revealed the monophyly of the European red deer, that of the wapiti, and that of the sika deer. The wapiti, however, was shown to be more closely related to the sika deer than to the European red deer. This is in conflict with traditional morphological results, which suggest a close sister group relationship between the wapiti and the European red deer. The divergence time between the European red deer and the wapiti plus the sika deer was estimated to be approximately 0.80 Ma, and that between the wapiti and the sika deer was estimated to be 0.57 Ma. The sika deer was subdivided into two subspecies groups, and the wapiti was also found to consist of an Asian group and a North American group.     

Bovine microsatellite loci are highly conserved in red deer (Cervus elaphus), sika deer (Cervus nippon) and Soay sheep (Ovis aries)

We tested 174 bovine microsatellite primer pairs for use in a primitive breed of sheep and two species of deer. Of 173 markers, 127 (73·4%) gave a product in Soay sheep ( Ovis aries ) of which 54 (42·5%) were polymorphic. One hundred and twenty-nine of 174 (74·1%) markers gave a product in red deer ( Cervus elaphus ) of which 72 (55·8%) were polymorphic. In sika deer ( Cervus nippon ) 126 of 171 (73·7%) microsatellite primers gave a product with 47 (37·3%) polymorphic. The proportion of bovine microsatellite loci conserved across artiodactyl species was significantly greater in this study than previously reported. Reasons for this high degree of microsatellite conservation are discussed. We suggest that a high resolution comparative map of the artiodactyls can be constructed using microsatellites.     

Use of melengestrol acetate in feed for contraception in herds of captive ungulates

Herds of blackbuck antelope (Antilopa cervicapra) and barasingha (Cervus duvauceli), axis (Cervus axis), sambar (Cervus unicolor), and Formosan sika (Cervus nippon taiwanaus) deer at the Wildlife Conservation Society/Bronx Zoo (WCS/BZ) were fed melengestrol acetate (MGA) at a concentration of 0.000154% in pelleted feed for various periods of times during 1991–2001. The target dose per animal of MGA was 1–2 mg per day. Contraceptive rates during treatment were 100% for blackbuck antelope and barasingha, sambar, and sika deer, and approximately 93% for axis deer. There were no observed adverse effects from MGA treatment on gestation. Post‐treatment reproductive rates were lower than pretreatment rates. Zoo Biol 22:455–463, 2003. © 2003 Wiley‐Liss, Inc.     

圈养梅花鹿BDNF基因多态性与日常行为性状的关联分析

采用目标动物取样法(Focal Animal Sampling)和全事件记录方法(All-occurrence Recording),观察统计了江苏省扬州市平山堂养殖场及扬州市动物园共48只梅花鹿昼间日常行为节律。以脑源性神经生长因子(Brain-derived Neurotrophic Factor, BDNF)为目的基因,采用PCR-SSCP方法,研究梅花鹿行为性状与基因多态性之间的关联。结果表明,在P-4引物,AA与BB基因型在卧息行为上存在显著差异(P<0.05),在运动行为上存在极显著差异(P=0.001)。在P-5引物,CC/DD/CD基因型在观望行为上两两差异显著,并且CC和DD基因型在卧息与修饰行为均存在显著差异(P<0.05)。研究表明,BDNF基因多态性和动物日常行为性状存在一定相关。     

Phylogenomics of several deer species revealed by comparative chromosome painting with Chinese muntjac paints

A set of Chinese muntjac (Muntiacus reevesi) chromosome-specific paints has been hybridized onto the metaphases of sika deer (Cervus nippon, CNI, 2n = 66), red deer (Cervus elaphus, CEL, 2n = 62) and tufted deer (Elaphodus cephalophus, ECE, 2n = 47). Thirty-three homologous autosomal segments were detected in genomes of sika deer and red deer, while 31 autosomal homologous segments were delineated in genome of tufted deer. The Chinese muntjac chromosome X probe painted to the whole X chromosome, and the chromosome Y probe gave signals on the Y chromosome as well as distal region of the X chromosome of each species. Our results confirmed that exclusive Robertsonian translocations have contributed to the karyotypic evolution of sika deer and red deer. In addition to Robertsonian translocation, tandem fusions have played a more important role in the karyotypic evolution of tufted deer. Different types of chromosomal rearrangements have led to great differences in the genome organization between cervinae and muntiacinae species. Our analysis testified that six chromosomal fissions in the proposed 2n = 58 ancestral pecoran karyotype led to the formation of 2n = 70 ancestral cervid karyotype and the deer karyotypes is more derived compare with those of bovid species. Combining previous cytogenetic and molecular systematic studies, we analyzed the genome phylogeny for 11 cervid species.     

四川梅花鹿春季昼夜活动节律与时间分配

1997年4月,在四川铁布自然保护区用直接观察法对220头次四川梅花鹿群的昼夜活动节律和时间分配进行了观察。结果表明,鹿群春季昼夜活动的规律性较强。白昼,鹿群活动呈现明显的双峰型,2个高峰时段为08：30和19：30前后,活动频率分别为71．9％和94．13％,其他时间多处于休息状态;鹿群夜间活动强于昼间,活动频率均在6l％以上,仅在半夜01：30前后有一个相对不活跃期。同时发现,春季活动高峰期约90％的活动时间被四川梅花鹿用于采食和移动。这可能与光照、食物及人类活动的影响有关。     

Suppression of Regeneration in New Zealand Mountain Beech Forests is Dependent on Species of Introduced Deer

We compared the impacts on forest regeneration of introduced sika (Cervus nippon) and red (Cervus elaphus) deer in New Zealand. Plot data were used to compare mountain beech (Nothofagus solandri var. cliffortioides) regeneration between a region with sika deer, and four regions without sika deer. All regions surveyed had red deer present. In the region where sika deer had been present for more than a decade, there was evidence of poor mountain beech seedling regeneration. In the four regions without sika deer, there was evidence of a strong regenerative response at stands with low occupancy by trees. When compared to larger deer species, sika deer have a digestive morphology allowing greater dietary versatility, which may result in them impeding forest regeneration where red deer do not. In contrast to mountain beech, some small-leaved shrub species may have been competitively advantaged by intensive browsing from sika deer. This is contra to a current view that small-leaved shrub species with interlacing branches were able to tolerate browsing from extinct ratite birds, but not introduced deer. Sika deer have been introduced into countries where other deer species are indigenous, such as Canada, Denmark, Great Britain, Czech Republic, Ireland and the United States of America. Because of their dietary advantage, sika deer may have a greater potential to impede forest regeneration and competitively exclude larger deer species, particularly at low basal area sites where impacts on tree regeneration are likely to be greatest.     

Borrelia burgdorferi Sensu Lato in an Endemic Environment: Wild Sika Deer (Cervus nippon yesoensis) with Infected Ticks and Antibodies

Ticks and blood samples were collected from wild sika deer (Cervus nippon yesoensis) during a hunting season (August to October) of 1991 at a selected location in Hokkaido, Japan. Ixodes persulcatus (adult and nymph) and I. ovatus (adult) were the common ticks on sika deer. Spirochetes were detected in the midgut of the ticks by the indirect peroxidase-conjugated antibody staining method and by dark-field microscopy after cultivation. By the reactive pattern of monoclonal antibodies, isolates were considered to belong to Borrelia garinii or B. japonica. In an antibody test, the percentage of seropositive deer was 69.0%. Most of the adult sika deer were positive for antibodies to the spirochetes. There are significant age-dependency in antibody level and seropositive rate. The surveillance of deer should be valuable in monitoring the transmission risk of B. burgdorferi sensu lato in nature.     

Variability of Female Responses to Conspecific vs. Heterospecific Male Mating Calls in Polygynous Deer: An Open Door to Hybridization?

Males of all polygynous deer species (Cervinae) give conspicuous calls during the reproductive season. The extreme interspecific diversity that characterizes these vocalizations suggests that they play a strong role in species discrimination. However, interbreeding between several species of Cervinae indicates permeable interspecific reproductive barriers. This study examines the contribution of vocal behavior to female species discrimination and mating preferences in two closely related polygynous deer species known to hybridize in the wild after introductions. Specifically, we investigate the reaction of estrous female red deer (Cervus elaphus) to playbacks of red deer vs. sika deer (Cervus nippon) male mating calls, with the prediction that females will prefer conspecific calls. While on average female red deer preferred male red deer roars, two out of twenty females spent more time in close proximity to the speaker broadcasting male sika deer moans. We suggest that this absence of strict vocal preference for species-specific mating calls may contribute to the permeability of pre-zygotic reproductive barriers observed between these species. Our results also highlight the importance of examining inter-individual variation when studying the role of female preferences in species discrimination and intraspecific mate selection.