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Wang X.,Jilin University | Li L.,Jilin University | Zhao J.,Jilin University | Li F.,LIAONING Technical University | And 2 more authors.
Comptes Rendus - Biologies | Year: 2017

To evaluate the effects of different preservation methods (stored in a -20. °C ice chest, preserved in liquid nitrogen and dried in silica gel) on inter simple sequence repeat (ISSR) or random amplified polymorphic DNA (RAPD) analyses in various botanical specimens (including broad-leaved plants, needle-leaved plants and succulent plants) for different times (three weeks and three years), we used a statistical analysis based on the number of bands, genetic index and cluster analysis. The results demonstrate that methods used to preserve samples can provide sufficient amounts of genomic DNA for ISSR and RAPD analyses; however, the effect of different preservation methods on these analyses vary significantly, and the preservation time has little effect on these analyses. Our results provide a reference for researchers to select the most suitable preservation method depending on their study subject for the analysis of molecular markers based on genomic DNA. © 2017 Académie des sciences.


Liu Y.-F.,Jilin University | Liu Y.-F.,Beihua University | Xing M.,Jilin University | Zhao W.,Jilin University | And 6 more authors.
Plant Systematics and Evolution | Year: 2012

Rhododendron aureum Georgi (Ericaceae) is a perennial alpine shrub endemic to Changbai Mountain in China. We used ISSR and RAPD markers to describe the diversity and genetic structure within and among four natural populations located at different altitudes. DNA from 66 individuals was amplified with ten ISSR markers and seven RAPD markers. High genetic diversity was observed by these two techniques at the species level. The genetic diversity of populations increased with altitudinal gradients from low to high. The coefficient of gene differentiation (G ST 0.3652 in ISSR and 0.2511 in RAPD) and AMOVA analysis revealed that most genetic diversity was distributed within populations (61.96% in ISSR and 70.23% in RAPD). The estimate of gene flow based on G ST was 0.8690 in ISSR and 1.4910 in RAPD. The UPGMA clustering results using ISSR and RAPD showed that all individuals from the same altitude were gathered together, and the two populations (TYD2a and YHLa) from middle altitudes always clustered together. Compared with populations from different altitudes, similar genetic diversity and low genetic differentiation were obtained from populations at the same altitudes, as revealed by ISSR markers. In addition to the reproductive strategy of R. aureum, these data highlight that local environmental conditions may play an important role in shaping the diversity and genetic structure of this species. © 2012 Springer-Verlag.


Qi X.,Jilin University | Qi X.,Qiqihar University | Wang E.,Jilin University | Xing M.,Jilin University | And 3 more authors.
World Journal of Microbiology and Biotechnology | Year: 2012

To investigate bacterial communities between rhizosphere and non-rhizosphere soils of the wild medicinal plant Rumex patientia of Jilin, China, small subunit rRNAs (16S rDNA) from soil metagenome were amplified by polymerase chain reaction using primers specific to the domain bacteria and analysed by cloning and sequencing. The relative proportion of bacterial communities in rhizosphere soils was similar to non-rhizosphere soils in five phylogenetic groups (Proteobacteria, Actinobacteria, Acidobacteria, Chloroflexi and Planctomycetes). But there were differences in five other phylogenetic groups (Firmicutes, Bacteroidetes, Gemmatimonadetes, Verrucomicrobia and Unclassified bacteria). Over 97.24 % of the sequenced clones were found to be unique to rhizosphere and non-rhizosphere soils, while 2.76 % were shared by both of them. Our results indicate that there are differences in the composition and proportion of bacterial communities between rhizosphere and non-rhizosphere soils. Furthermore, the unique bacterial clones between rhizosphere and non-rhizosphere soils of the wild medicinal plant R. patientia have obvious differences. © 2012 Springer Science+Business Media B.V.


Xing M.,Jilin University | Du Y.,Jilin University | Wang X.,Jilin University | Niu L.,Changbai Mountain Academy of science | And 2 more authors.
Biotechnic and Histochemistry | Year: 2010

We present a simplified paraffin embedding method suitable for unsuberized or unlignified small botanical samples (diameter < 0.3 cm). Only 2 h are required to yield plant tissues embedded in paraffin for anatomical observation and molecular analysis. Our method achieved morphological preservation of cell structures and conservation of nucleic acids that were equivalent to the traditional protocol. Fourier transform infrared spectrometry showed that the degree of degradation of the cytoplasmic components (e.g., protein) resulting from our simplified protocol was similar to that of the traditional protocol. The DNA samples embedded using the simplified method was extractable and could be used for PCR analysis. The DNA quality was equivalent to that embedded using the traditional method. © 2010 Biological Stain Commission.


Zhao W.,Jilin University | Zhao W.,Changbai Mountain Academy of science | Shi X.,Jilin University | Li J.,Jilin University | And 4 more authors.
PLoS ONE | Year: 2014

Rhodiola sachalinensis is an endangered species with important medicinal value. We used inter-simple sequence repeat (ISSR) and methylation-sensitive amplified polymorphism (MSAP) markers to analyze genetic and epigenetic differentiation in different populations ofR. sachalinensis, including three natural populations and an ex situ population. Chromatographic fingerprint was used to reveal HPLC fingerprint differentiation. According to our results, the ex situ population of R. sachalinensis has higher level genetic diversity and greater HPLC fingerprint variation than natural populations, but shows lower epigenetic diversity. Most genetic variation (54.88%) was found to be distributed within populations, and epigenetic variation was primarily distributed among populations (63.87%). UPGMA cluster analysis of ISSR and MSAP data showed identical results, with individuals from each given population grouping together. The results of UPGMA cluster analysis of HPLC fingerprint patterns was significantly different from results obtained from ISSR and MSAP data. Correlation analysis revealed close relationships among altitude, genetic structure, epigenetic structure, and HPLC fingerprint patterns (R2 = 0.98 for genetic and epigenetic distance; R2 =0.90 for DNA methylation level and altitude; R2 =-0.95 for HPLC fingerprint and altitude). Taken together, our results indicate that ex situ population of R. sachalinensis show significantly different genetic and epigenetic population structures and HPLC fingerprint patterns. Along with other potential explanations, these findings suggest that the ex situ environmental factors caused by different altitude play an important role in keeping hereditary characteristic of R. sachalinensis. Copyright: © 2014 Zhao et al.


Li X.H.,Changbai Mountain Academy of science | Li X.H.,CAS Institute of Zoology | Yu J.J.,CAS Institute of Zoology | Zhang P.,Northeast Forestry University | And 2 more authors.
Shengtai Xuebao/ Acta Ecologica Sinica | Year: 2016

Using camera traps can provide valuable information for estimating wildlife density, and further contributes to conservation activities. For species that can be individually recognized, such as tigers and leopards, camera trapping combined with mark-recapture techniques can provide reliable estimates of population density. However, most species cannot be individually recognized, and no sophisticated models are available for accurate estimation of their densities. At present, the only available model is Rowcliffe et al.'s gas molecule movement model, which assumes animals behave like ideal gas particles, moving randomly and independently of one another. Such a model is not appropriate for either territorial or social species, or elusive species that usually move along trails. We developed a novel method to estimate the population density of animals that cannot be individually recognized. This method is based on the simulation of animal movement and pseudo camera trapping processes at a series of population densities. We matched results of real camera trapping with those of simulated camera trapping to estimate population density. The method was coded using the R language. We deployed 25 cameras (LTL5120) in the 25 hm2 forest dynamics plot in the Changbaishan National Nature Reserve, China for 41 days in the winter of 2011 and 40 days in the winter of 2012. The Siberian chipmunk (Eutamias sibiricus)and Korean field mouse (Apodemus peninsulae) are two dominant species in the plot. Animal movement was simulated by setting a starting location and a series of moving directions and step lengths. The starting location was a randomly selected point in the survey area. The direction of the first movement, θ, was also randomly selected from a range of 0-2π. The length and angle of deflection of subsequent movements followed normal distributions N (1 m, 0.1 m) and N (0, 30 degrees), respectively. We also defined a home range for each species by forcing the simulated animal to return to the starting location (assumed to be a mouse hole) at a rate of D/50, where D is the distance in meters between the current and original locations. The simulations of animal movement were run under a series of population densities. We matched the simulated results and the observed photo records using the random forest algorithm to estimate the population density and its confidence intervals. This analysis determined that the density of the Siberian chipmunk is 1.96hm2, and 2.71hm2 for the Korean field mouse. Our method has a number of limitations. First, the movement pattern of the target species must be known. In this study, we selected movement parameters (step length, angle of deflection, home range size, etc.) by visually checking the simulated footprint chains, which should be replaced by field tracking. Second, the cameras must be deployed in the field systematically, at regular intervals, so that virtual camera trapping can be simulated accordingly. In spite of the limitations, this method can provide reliable estimates for population density for animals that cannot be individually recognized. Our new method can be used for other camera trapping practices, as long as the movement pattern of the species is known. © 2016, Ecological Society of China. All rights reserved.


Zhou L.,CAS Shenyang Institute of Applied Ecology | Dai L.,CAS Shenyang Institute of Applied Ecology | Wang S.,Changbai Mountain Academy of science | Huang X.,Changbai Mountain Academy of science | And 6 more authors.
Annals of Forest Science | Year: 2011

Introduction The old-growth forests on Changbai Mountain have been well protected from human activities and provide a living laboratory for studying forest carbon sequestration under natural environmental conditions. • Objectives We used data from permanent plots measured periodically in 1981 and 2010 to quantify carbon densities for Korean pine-broadleaf mixed forest, coniferous forest and Erman's birch forest on Changbai Mountain. • Results Carbon pools were divided into tree stems, leaves, branches, coarse woody debris, tree roots, and soil. Although the mixed forest experienced minor wind damage, every forest component except for coarse woody debris experienced increases in carbon density, and the total forest carbon density increased from233 to 317 t C ha -1. The coniferous forest was severely damaged by wind, so carbon content in trees decreased but the total forest carbon density still increased from 298 to 327 t C ha -1. The birch forest gained much carbon in trees but the soil carbon pool remained relatively stable, and its total carbon density increased from 226 to 281 t C ha -1. The old-growth forest was more resilient to disturbance than previously thought. The positive increases in carbon for the three old-growth forests suggest that forest landscapes on Changbai Mountain are indeed carbon sinks. © INRA and Springer Science+Business Media B.V. 2011.


Wang Y.,China Academy of Transportation science | Guan L.,China Academy of Transportation science | Piao Z.-J.,Changbai Mountain Academy of science | Kong Y.-P.,China Academy of Transportation science
Chinese Journal of Ecology | Year: 2016

Recognition of barrier effect of road on wildlife is one of the main contents of road wildlife ecology research. In this paper, we selected the surrounding Ring Changbai Mountain Scenic Highway, which traverses the Changbai Mountain Nature Reserve, as a research area, and carried out sample line investigations from 2008 to 2012, to assess the barrier effect of highway on middle and large sized mammals. The results indicated that: (1) Twelve middle and large sized mammal species were found within the range of roadside 500 m, and five species were Chinese national protected species; (2) Species richness and the number of mammal signs in Korean pine broad-leaved forest were significantly higher than those in secondary white birch forest; (3) Species richness of inside reserve of the road was significantly higher than that of outside reserve of the road, and especially in snow season, species richness and the number of mammal signs of inside reserve of the road were significantly higher than those of outside reserve of the road; (4) The numbers of signs had no significant differences between two sides of the road and two vegetation types for wild boar (Sus scrofa), Siberian roe deer (Capreolus pygargus), Siberian weasel (Must el a sibirica), sable (Maries zibellina), yellow-throated marten (Maries flavigula), Asian badger (Meles leucurus), Eurasian red squirrel (Sciurus vulgaris) and Manchurian hare (Lepus mandshuricus); however, the number of signs of Siberian weasel in Korean pine broad-leaved forest was significantly higher than in secondary white birch forest in snow season, and the number of signs of wild boar and Siberian roe deer in Korean pine broad-leaved forest were significantly higher than in secondary white birch forest in non-snow season; (5) In snow season, in terms of the relationship of the number of signs and the crossing rate within the range of roadside 50 m, the crossing section of red deer (Cervus elaphus) through the highway was limited only in K25 to K27, meaning that the barrier effect of highway on red deer was serious. (6) The yearly rale of crossing highway of mammals was negatively correlated to daily traffic volume, although the relationship was not significant. This study indicated the barrier effect of the Ring Changbai Mountain Scenic Highway on middle and large sized mammals had appeared, and this situation showed a serious tendency over lime. Presently, human disturbance outside the nature reserve was serious, and we thus recommend building migration corridors for mammals between the nature reserves. © 2016, Editorial Board of Chinese Journal of Ecology. All rights reserved.


Wang Y.,China Academy of Transportation science | Piao Z.J.,Changbai Mountain Academy of science | Guan L.,China Academy of Transportation science | Wang X.Y.,China Academy of Transportation science | And 2 more authors.
North-Western Journal of Zoology | Year: 2013

The incidence of wildlife mortality on roads is a widespread phenomenon reported in many countries; despite this, it has received limited attention in China. To address this issue, a field investigation was conducted to assess the magnitude, composition, and temporal patterns of road mortality of native vertebrates on Ring Changbai Mountain Scenic Highway in north-east China from 2009 to 2012. A total of 3.475 vertebrates, belonging to 63 species were recorded as road mortalities. Amphibians were the most abundant taxa recorded (86.21%), followed by mammals (5.70%), birds (5.24%), and reptiles (2.85%). Regarding the number of species affected by the incidence of road kills, birds were the most affected taxa (31 species), followed by mammals (16 species) and reptiles (10 species), while amphibians were the least affected taxa (6 species). Three vertebrate species that were recorded as road mortalities are under second class protection in China. There were no differences between seasons or months for road mortality or richness for bird and reptile taxa, while significant differences between months were observed for mammals and amphibians. Mammal road mortality was highest in August, most road kills being encountered for the Siberian Chipmunk (Tamias sibiricus Laxmann, 1769) mortalities. Amphibian road mortality was highest in June, most road kills being encountered for the Chinese Brown Frog (Rana chensinensis David, 1875), Oriental Fire-bellied Toad (Bombina orientalis Boulenger, 1890), and Asiatic Toad (Bufo gargarizans Cantor, 1842). Each taxonomic group showed no distinct differences in species richness between months. This research recommends creating wildlife crossing structures for amphibians and small-sized mammals to reduce the incidence of road mortalities and adopting traffic management measures to mitigate amphibian mortality especially from April to September, and strictly controlling speed limitations to 60km/h on this highway. © NwjZ, Oradea, Romania, 2013.


Wang Y.,China Academy of Transportation science | Piao Z.-J.,Changbai Mountain Academy of science | Guan L.,China Academy of Transportation science | Kong Y.-P.,China Academy of Transportation science
Chinese Journal of Ecology | Year: 2013

With the large-scale expansion of highway construction in ecologically sensitive area, wildlife conservation has become an urgent and important topic during the highway infrastructure construction and operation. However, few studies have been made on the wildlife conservation along the highways in China. By using sample line method and infrared camera monitoring, a field survey from November 2008 to February 2012 was conducted on the road-kill, road effect zone for wildlife, wildlife crossing highway, and wildlife crossing structure along the Ring Changbai Mountain Scenic Highway, which is adjacent to the Changbai Mountain Nature Reserve. The results showed that within the 500 m roadside of the Highway, the species richness of mammals, birds, reptiles, and amphibians accounted for 42. 86%, 24. 78%, 66. 67%, and 66. 67% of the wildlife species richness in the Nature Reserve, respectively. Among the wildlife, 11 species were national protective species, 2 of which were of first-class, and 9 were of second-class. A total of 3475 wildlife belonging to 59 species were killed by the vehicles on this Highway, and the road-kill rate was averagely 61. 6 individuals · 100 km". In terms of magnitude, amphibians had the highest mortality (2996 individuals). The species richness of birds and mammals was not impacted by the Highway, but there was a concentrative trend about the frequency of these wildlife activities within the 200 m roadside. The 10 surveys in winter indicated that a total of 13 species were found crossing the Highway for 502 times, and Siberian Weasel had the highest crossing frequency (169 times). When crossing the Highway, the wildlife preferred to the highway section with the roadside vegetation being primitive Korean pine and broad-leaved forest. There were 6 medium and large sizes species using bridges and culverts to cross the Highway, and the utilization rate of the bridges and culverts was 88% and 44. 2%, respectively. Vegetation type, human disturbance, and size of wildlife crossing structure all had significant effects on the utilization rate. Some countermeasures for protecting the wildlife were provided correspondingly.

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