Phylogeny of Snub-Nosed Monkeys Inferred from Mitochondrial DNA,Cytochrome B,and 12S rRNA Sequences

Snub-nosed monkeys (Rhinopithecus spp.) are confined to isolated mountainous regions in China and North Vietnam. Their systematic classification and phylogenetic relationship has been controversial. The structures of mitochondrial DNA cytochrome b and 12S rRNA show that the 4 species of Rhinopithecus are quite different from other colobines. It is reasonable to regard them as an independent genus, as determined by external features, morphometric characters and behavior. However, whether or not there should be a subdivision between the Vietnamese and Chinese species at the subgeneric level remains to be clarified; more evidence from a large range of Asian colobine species is needed. The Guizhou species, Rhinopithecus brelichi, is a valid species, which is more closely related to Pygathrix than the other species ( R. roxellana, R. bieti and R. avunculus) are. Results also indicate that 3 species—Rhinopithecus roxellana, R. bieti and R. avunculus—might have diverged from R. brelichi, but the phylogenetic relationship of R. avunculus is not clear.     

Cranial morphology of the golden monkey (Rhinopithecus) and douc langur (Pygathrix nemaeus)

Because they have been less studied than most other non-human primates (partly due to the difficulties in accessing their habitat) the origins and phylogenetic relationships ofR. roxellana andR. bieti are controversial. These controversies may be clarified to some degree by adding information on the cranium. To this end, ten cranial dimensions analysed morphometrically here provide data about cranial differences among species of the genusRhinopithecus, and between species ofRhinopithecus andPygathrix nemaeus. Though more similar to each other than to any others in the same genus, the results show a significant separation betweenR. roxellana andR. bieti to the degree that they may be regarded as two different species. This confirms the conclusions of prior studies of external features, qualitative morphological characteristics and biochemical evidence (Yeet al, 1987; Zhang and Ryder, 1995; Jablonski, 1998; Penget al., 1988). The differences between these two species are mainly size-related, being highly correlated with cranial length. Other differences, probably non-size related shape differences, however, are highly correlated with cranial width. Sexual dimorphism plays a part in these findings. In relation to the other species, however, the results show that the Vietnam golden monkey (R. avunculus) has closer craniometric relationships with the douc langur (Pygathrix nemaeus) rather than with the three Chinese golden monkey species. Of these, the Gouzhou species (R. brelichi) shares more similarity withR. avunculus and is more separate fromRhinopithecus roxellana and R. bieti. The smaller differentiation between the two latter species could be due to their more recent separation following the dramatic elevation of Qinghai-Tibetan Plateau after the Middle Pleistocene.     

Sexual dimorphism in the snub-nosed langurs (Colobinae: Rhinopithecus)

Sexual dimorphism in the dentition and skeleton of the four extant species of snub-nosed langurs, Rhinopithecus (R.) bieti, R. (R.) brelichi, R. (R.) roxellana and R. (Presbytiscus) avunculus, was studied. The species shared a similar general pattern of sexual dimorphism, but were found to differ in respects that appear to reflect the influence of disparate socioecological and environmental factors. All the species showed marked canine dimorphism, but the very high degree of canine dimorphism in R. bieti appeared to be due to the intensity of intermale competition for mates during a temporally restricted breeding season, and possibly also to the intensity of competition between males for other resources during other times of the year. Sexual dimorphism in the postcranial skeleton of Rhinopithecus species was also most pronounced in R. bieti and may be related to the relatively higher frequency of terrestrial locomotion in males of the species. © 1995 Wiley-Liss, Inc.     

Resequencing and comparison of whole mitochondrial genome to gain insight into the evolutionary status of the Shennongjia golden snub‐nosed monkey (SNJ R. roxellana)

Shennongjia Rhinopithecus roxellana (SNJ R. roxellana) is the smallest geographical population of R. roxellana. The phylogenetic relationships among its genera and species and the biogeographic processes leading to their current distribution are largely unclear. To address these issues, we resequenced and obtained a new, complete mitochondrial genome of SNJ R. roxellana by next‐generation sequencing and standard Sanger sequencing. We analyzed the gene composition, constructed a phylogenetic tree, inferred the divergence ages based on complete mitochondrial genome sequences, and analyzed the genetic divergence of 13 functional mtDNA genes. The phylogenetic tree and divergence ages showed that R. avunculus (the Tonkin snub‐nosed monkey) was the first to diverge from the Rhinopithecus genus ca. 2.47 million years ago (Ma). Rhinopithecus bieti and Rhinopithecus strykeri formed sister groups, and the second divergence from the Rhinopithecus genus occurred ca. 1.90 Ma. R. roxellana and R. brelichi diverged from the Rhinopithecus genus third, ca. 1.57 Ma. SNJ R. roxellana was the last to diverge within R. roxellana species in 0.08 Ma, and the most recent common ancestor of R. roxellana is 0.10 Ma. The analyses on gene composition showed SNJ R. roxellana was the newest geographic population of R. roxellana. The work will help to develop a more accurate protection policy for SNJ R. roxellana and facilitate further research on selection and adaptation of R. roxellana.     

Reproductive parameters in Guizhou snub‐nosed monkeys (Rhinopithecus brelichi)

In this study, we present data on reproductive parameters and birth seasonality of Guizhou snub‐nosed monkeys (Rhinopithecus brelichi). Our analyses are based on data from a small captive population collected over 15 years and on 5 years of observations of free‐ranging snub‐nosed monkeys. Captive females (n=4) mature at an age of 70.8±6.7 months and reproduce for the first time at 103.4±7.5 months. The mean interbirth interval was 38.2±4.4 months if the infant survived more than 6 months, which is longer than that in R. roxellana and R. bieti. In the wild and in captivity, births are very seasonal and occur only in a period from the end of March to the end of April. Our data suggest that population growth in Guizhou snub‐nosed monkeys is slow compared with the other two Chinese snub‐nosed monkey species. The risk of extinction is therefore particularly high in this species, given the small overall population size and slow population recovery potential. Am. J. Primatol. 71:266–270, 2009. © 2008 Wiley‐Liss, Inc.     

A Phylogeny of Chinese Leaf Monkeys Using Mitochondrial ND3-ND4 Gene Sequences

The phylogeny of Chinese leaf monkeys, especially the snub-nosed monkeys (Rhinopithecus), has not been thoroughly investigated using molecular sequence data, perhaps due to their rarity in the wild and their poor representation in institutional collections. Despite several proposed classifications, systematic relationships of these species remain poorly defined and this has hindered their conservation. To clarify the phylogenetic relationships of the leaf monkey clade in China, we sequenced the mitochondrial ND3, ND4L, ND4, tRNA ^Arg , tRNA ^His , tRNA ^Ser , and tRNA ^Leu genes for Rhinopithecus bieti, R. roxellana, Trachypithecus francoisi, T. f. leucocephalus, and T. phayrei as well as Pygathrix nemaeus and Colobus guereza. We included a total of 2252 characters for each individual, excluding gaps in primary sequences. Our interpretation of the results from character-and distance-based phylogenetic analyses suggest that (1) Pygathrix nemaeus is sister to Rhinopithecus rather than to Trachypithecus though it is quite divergent from the former; (2) the Yunnan snub-nosed monkey, Rhinopithecus bieti, represents a valid species; (3) the white-headed leaf monkey is not a distinct species, but instead is a subspecies of Trachypithecus francoisi (T. f. leucocephalus), though it should still be considered a separate evolutionary significant unit (ESU); and (4) because two individuals of the Phayrei's leaf monkey, T. phayrei, are genetically distinct from one another, a more extensive revision of the taxonomy of this putative species in China is needed. These results, plus ongoing work on the molecular systematics of the entire Asian leaf monkey radiation, can provide a sound basis for identifying the appropriate units of conservation for this endangered group of primates.     

Identification of 13 Human Microsatellite Markers via Cross-species Amplification of Fecal Samples from Rhinopithecus bieti

Yunnan snub-nosed monkeys (Rhinopithecus bieti) are 1 of 3 snub-nosed monkey species endemic to China. Only ca. 1500 individuals remain in high-altitude forests 3000–4500 m above sea level on the Tibetan Plateau, making them the nonhuman primate living at the highest known elevation. It is one of the most endangered 25 primate species in the world. Proper knowledge of the population genetics and social system of Rhinopithecus bieti will contribute to more appropriate conservation management decisions. Cross-species amplification of human microsatellite loci has facilitated analysis of the population genetics and reproductive strategies of various primate species. We screened 72 human-derived markers to assess their utility in Yunnan snub-nosed monkeys. Thirteen of them produced reliable results and exhibited moderate levels of polymorphism.     

The men of Nelson's navy: a comparative stable isotope dietary study of late 18th century and early 19th century servicemen from Royal Naval Hospital burial grounds at Plymouth and Gosport, England

The Guizhou snub‐nosed monkey (Rhinopithecus brelichi) is a primate species endemic to the Wuling Mountains in southern China. With a maximum of 800 wild animals, the species is endangered and one of the rarest Chinese primates. To assess the genetic diversity within R. brelichi and to analyze its genetic population structure, we collected fecal samples from the wild R. brelichi population and sequenced the hypervariable region I of the mitochondrial control region from 141 individuals. We compared our data with those from the two other Chinese snub‐nosed species (R. roxellana, R. bieti) and reconstructed their phylogenetic relationships and divergence times. With only five haplotypes and a maximum of 25 polymorphic sites, R. brelichi shows the lowest genetic diversity in terms of haplotype diversity (h), nucleotide diversity (π), and average number of pairwise nucleotide differences (Π). The most recent common ancestor of R. brelichi lived ～0.36 million years ago (Ma), thus more recently than those of R. roxellana (～0.91 Ma) and R. bieti (～1.33 Ma). Phylogenetic analysis and analysis of molecular variance revealed a clear and significant differentiation among the three Chinese snub‐nosed monkey species. Population genetic analyses (Tajima's D, Fu's F_s, and mismatch distribution) suggest a stable population size for R. brelichi. For the other two species, results point in the same direction, but population substructure possibly introduces some ambiguity. Because of the lower genetic variation, the smaller population size and the more restricted distribution, R. brelichi might be more vulnerable to environmental changes or climate oscillations than the other two Chinese snub‐nosed monkey species. Am J Phys Anthropol, 2012. © 2011 Wiley Periodicals, Inc.     

A High Polymorphism Level in <Emphasis Type="Italic">Rhinopithecus roxellana</Emphasis>

The Sichuan golden monkey (Rhinopithecus roxellana) is a famous and beautiful endangered primate. Owing to the dearth of samples and technical limitations, previous studies could not adequately assess the levels of genetic polymorphisms in the species. To evaluate the polymorphisms further, we sequenced the entire mitochondrial control regions (D-loop) of 35 individuals from the 3 major habitats: Minshan, Qinglin, and Shennongjia. We observed many polymorphisms and estimated that the effective population size of the current total population is less than ca. 480–2300, and the time to the most recent common ancestor is ca. 4000–62,700 years. Through comparisons, we showed that the population of Rhinopithecus roxellana has a relatively higher polymorphism level than that of other endangered primates.     

Metrical dental analysis on golden monkey (Rhinopithecus roxellana)

Dental variation in the Chinese golden monkey (Rhinopithecus roxellana) is here evaluated by univariate, bivariate, and multivariate analyses. Allometric analyses indicate that canines and P3s are positively, but other dimensions negatively scaled to mandible and maxilla, and to body size. With the exception of the mesiodistal dimensions of I₁ and M₃, and the buccolingual dimension of P₄, mandibular dental variables show similar scaling relative to body size. Analysis of residuals shows that males have significantly larger canine, P³ and buccolingual dimensions of the postcanine teeth (M² and M³) than females. A significant difference in shape between the sexes is found in the buccolingual dimension of the upper teeth, but not in the mandible. Unlike the situation in some other species, female golden monkeys do not exhibit relatively larger postcanine teeth than males. In fact, the reverse is true, expecially for M²s and M³s. The fact that most of the dental variables show low negative allometry to body size might be related a cold environment that has led to the development of larger body size with reduced energy loss. When the raw data are examined by Discriminant Function Analysis the sexes are clearly distinguishable.     

金丝猴(RHINOPITHECUS)肌肉系统某些特征的比较研究

本文对金丝猴与其他灵长类之间和三种金丝猴之间特征不同的56条肌作了比较研究。结果表明:三种金丝猴的肌肉系统存在一些差异。纠正了Patterson(1942)对一例川金丝猴的一些错误观察记录。在金丝猴属的肌学方面除与猴超科共有的大部分特征外,还具有与类人猿和人相似的特征。在猴类中,金丝猴的肌肉系统更多地与叶猴相似,其中一些特征较叶猴更为进化。从进化观点看,金丝猴的肌学特征似介于叶猴与类人猿之间,故它们比其他猴超科动物更为进化。     

三种笼养灵长类活动时间分配的比较研究

采用焦点动物法和连续记录法对3种笼养灵长类活动时间分配进行比较研究。结果表明,川金丝猴Rhi-nopithecus roxellanae和熊猴Macaca assamensis用于休息的时间多于猕猴Macaca mulatta,移动时间则相反;川金丝猴和猕猴用于理毛的时间多于熊猴。不同性别年龄组之间的活动时间分配有差异,主要表现在川金丝猴母亲用于理毛的时间明显多于成年儿子,用于玩耍的时间则相反;猕猴成年雄性的移动时间和理毛时间多于成年雌性,玩耍时间则少于成年雌性;熊猴成年雌性的移动时间多于成年雄性,与成年个体相比,幼体花更多的时间用于玩耍,而相应的减少了其休息时间。     

九寨沟自然保护区川金丝猴的分布及种群数量

2014年2月至5月,对四川省九寨沟自然保护区野生川金丝猴的种群分布、数量及其社会结构进行调查,初步探讨该种群与其他种群在外部形态和社会结构方面的差异性。采用"V"型路线法调查,发现九寨沟保护区共有川金丝猴4群。然后结合长时间的持续跟踪观察,证实家庭单元19个,全雄群3个,能区分不同个体296只。OMU大小平均为12.7只,其中OMU内的成年个体占38%,青少年个体占53%,幼仔占9%。成年与青少年比例为1∶1.39,成年雌雄的比例为3.84∶1。与秦岭种群的外部形态相比,该种群的成年雄性更为壮大,毛发弯曲程度明显;成年雌性毛发略显灰黑色,金黄色暗淡,大部分个体嘴角长有很小的肉瘤;青少年个体毛色泛白。亦发现九寨沟种群倾向于选择海拔偏高(2400—3550 m)的针叶林带活动。     

川金丝猴秦岭与岷山社群社会结构与空间分布差异性比较

对不同地区川金丝猴社会结构和空间分布的比较有助于了解其种群与环境的相互应答关系。在秦岭山脉和岷山山脉的不同地区选取3个观察群,对比它们之间社群结构和空间分布的差异。研究结果表明,尽管同属于仰鼻猴属川金丝猴种,由于栖息地环境的不同,导致社群结构和空间分布既存在相同点又存在一定的差异。对于社会结构,扎如沟群,大坪峪群与玉皇庙群表现出同样的最高进化水平的重层社会体系;但是构成这一社会体系的基本单位一雄多雌（One-male unit,OMU）,其成员整体数量上没有差异,但是OMU内雌雄比例不仅在3个观察群间存在差异,而且与同属其他物种相比也存在差异;对于空间分布,由于食物竞争压力和天敌捕食压力在不同的栖息地内影响不同,导致3个观察群空间分布呈梯度变化关系。研究结果不仅为川金丝猴种群动态,种群与环境的相互应答关系以及种群进化提供基础资料,而且希望能藉此推进我国相关领域研究的开展,为灵长类社会与行为生态学研究者启发思路,并为保护这些濒危物种提供科学依据。     

Mitochondrial cytochrome b gene sequences of old world monkeys: With special reference on evolution of Asian colobines

The classification and phylogenetic relationships of the Old World monkeys are still controversial. For Asian colobines, from three to nine genera were recognized by different primatologists. In the present study, we have sequenced a 424 bp mitochondrial tRNA^Thr gene and cytochrome b gene fragment fromMacaca mulatta, Mandrillus sphinx, Mandrillus leucophaeus, Semnopithecus entellus, Trachypithecus vetulus, T. johnii, T. phayrei, T. francoisi, Pygathrix nemaeus, Rhinopithecus roxellanae, R. bieti, R. avunculus, Nasalis larvatus, andColobus polykomos in order to gain independent information on the classification and phylogenetic relationships of those species. Phylogenetic trees were constructed with parsimony analysis by weighting transversions 5 or 10 fold greater than transitions. Our results support the following conclusions: (1) the Old World monkeys are divided into two subfamilies; (2) that among the colobines,Colobus, the African group, diverged first, andNasalis andRhinopithecus form a sister clade toPygathrix; (3) that there are two clades within leaf monkeys, i.e. 1)S. entellus, T. johnii, andT. vetulus, and 2)T. phayrei andT. francoisi; (4) thatRhinopithecus avunculus, R. roxellanae, andR. bieti are closely related to each other, and they should be placed into the same subgenus; (5) thatRhinopithecus is a distinct genus; and (6) that the ancestors of Asian colobines migrated from Africa to Asia during the late Pliocene or early Pleistocene.     

云南拉沙山黑白仰鼻猴对种子传播潜力的评估

灵长类是森林生态系统中植物种子的主要传播者,有助于森林植被的更新,然而受研究方法的限制,灵长类种子传播潜力常被低估。为全面评估温带灵长类动物的种子传播潜力,采用直接观察法和粪便分析法评估珍稀濒危灵长类动物黑白仰鼻猴的种子传播潜力。于2018年11月—2019年10月采用直接观察法(瞬时扫描取样法)收集云岭省级自然保护区拉沙山黑白仰鼻猴的活动时间分配数据,获取每月取食果实的比例;同时每月收集黑白仰鼻猴的粪便,采用粪便分析法分拣猴粪中残留的植物种子,统计有完整种子残留的月份和粪便比例,应用这两种方法评估黑白仰鼻猴种子传播潜力及其差异。结果表明：直接观察法收集到黑白仰鼻猴取食果实的月份数为6个月(7—12月),月均取食果实的比例为(15.31±20.15)%,共取食13种果实;而粪便分析法发现黑白仰鼻猴粪粒内全年都有完整种子残留,粪便中月均完整种子残留比例(35.19±35.43)%,其中9月至第二年1月粪便中种子残留比例都大于50%,共取食18种果实;综合两种方法发现云南拉沙山黑白仰鼻猴共取食20种植物果实,具有较高的种子传播潜力。直接观察法可确定黑白仰鼻猴取食果实的物种数,而粪便分析法能...     

Diet and activity budget of <Emphasis Type="Italic">Rhinopithecus roxellana</Emphasis> in the Qinling Mountains,China

We collected data on diet and daytime activity budget, and investigated the phenology of food trees and food abundance for a group of Rhinopithecus roxellana on the East Ridge of Yuhuangmiao in the Qinling Mountains from November 2001 to December 2003. We calculated the seasonal activity budget using data collected by scan sampling from 84 full-day observations (winter 16, spring 18, summer 28, autumn 22 days). During scan sampling we recorded behavioral states, and the food items and species consumed. The subjects consumed 84 plant species, including trees and shrubs of 29 families, and lichens. Food species varied seasonally. The overall diet of R. roxellana consisted of 29.4% fruit/seeds, 29.0% lichens, 24.0% leaves, 11.1% bark, 4.2% buds, 1.3% twigs and 1.0% unidentified items. Because the abundance of different food items varied seasonally, the monkeys had to shift their major food items seasonally. The annual activity budget of R. roxellana was 36.2% time spent resting, 35.8% feeding, 22.9% moving, and 5.1% other behavior. Seasonal changes in activity budget were observed. R. roxellana spent more time moving in autumn, when the quality of the food might be highest, and least time moving in winter when the food quality might be lowest. Thus, this type of monkey has a passive foraging strategy.     

Comparative study of musculature in goldon monkey (Rhinopithecus)

Some differences between the muscular system of three species of golden monkeys were shown. The major features of the myology in these animals, besides that ofCercopithecoidea, are similar to the anthropoid apes and humans. Although most of the features between taxa of golden monkeys are largely homologous, there are also some differences. Our results differ slightly from thosePatterson (1942) obtained inR. roxellanae. Some conclusions reported by Patterson were obviously incorrect, since the specimen he used did not have a head. Many mylogical characters are similar to other species ofCercopithecoidea while some resemble those of apes. Many are similar to those of the leaf monkey, though some of them show much more development. The golden monkey is more advanced than any other species ofCercopithecoidea. We hypothesize that in terms of the myological characters the golden monkeys seem to occupy a position between the leaf monkeys and the apes.     

Assessment of Habitat Fragmentation and Corridors for an Isolated Subspecies of the Sichuan Golden Snub-Nosed Monkey, <Emphasis Type="Italic">Rhinopithecus roxellana hubeiensis</Emphasis>

Understanding habitat quality and landscape connectivity and exploring corridors connecting habitat patches are crucial for conservation, particularly for species distributed among isolated populations. The Sichuan golden snub-nosed monkey, Rhinopithecus roxellana, is an Endangered primate species endemic to mountainous forests in China. Its easternmost distribution lies in the Shennongjia area, which harbors an isolated subspecies, R. roxellana hubeiensis. Unfortunately, it has experienced significant habitat loss, fragmentation, and dramatic population decline in recent decades, primarily due to increased human disturbance. To quantify habitat quality, identify suitable habitat patches, and detect possible linkages among these patches for R. roxellana hubeiensis, we conducted habitat suitability assessments and landscape connectivity analyses in the Shennongjia area based on a set of environmental factors. We created a habitat quality model and a movement cost surface for the Shennongjia area based on a habitat suitability index, graph theory, expert knowledge, field experience, and information from the literature. Our results show that suitable habitat for R. roxellana hubeiensis in Shennongjia is fragmented and limited, and that this is particularly true for highly suitable habitats. We detected six core habitat patches and six least-cost paths and corridors. Our study does not provide accurate distributions of the monkeys and their habitat use. However, it identifies the most feasible and traversable habitats and corridors, which should be conservation priorities for this subspecies, and provides valuable guidance for reevaluating habitat conservation plans.     

Molecular phylogenetic relationships among Sichuan snub-nosed monkeys (Rhinopithecus roxellanae) inferred from mitochondrial cytochrome-b gene sequences

The Sichuan snub-nosed monkey (Rhinopithecus roxellanae) is a species endemic to China, and its distribution is the widest among all snub-nosed monkeys in China. To clarify whether there is subspecific differentiation within this species, we determined partial sequences of the cytochrome-b gene from four populations ofR. roxellanae. First, 402bps of the partial sequences fromR. roxellanae were compared with those fromR. bieti andR. avunculus, and the phylogenetic tree was constructed by the neighbor-joining method. The genetic distance was only 0–0.002 among the four populations, and their sequences constituted a monophyletic group. Further, comparison of longer sequences (735bp) among the four populations revealed that there were only four substitutions and the genetic distance was only 0.001–0.005 among them. Thus, we suggest that, at least on mtDNA phylogeny, the difference among the four populations does not reach the subspecies level, and that this species should be recognized as a monotypic species.