R-Band of Northeastern Sika Deer (Cervus nippon hortulorum) Chromosomes

Objeclive The aim of this study was to investigate R-band of Cervus nippon hortulorum chromosomes and to provide references for genetic variation and gene location of Cervus nippon hortulorum. [Metbod] Cell division was synchronized by the pepripheral blood lymphocyte culture and the excessive dosage of thymine deoxyribonucleoside, and R-band of Cervus nippon hortulorum chromosomes was also analyzed by RBG-banding technique. Result The number of haploid chromosome banding increased to 400. The R-band of No. 1, No. 2, No. 3, No. 4, chromosome X and Y were almost just opposite to the high-resolution G band of them. The terminal of chromosomes except No. 21, No. 24 and No. 28 were all pos- itive deeply stained. E Conclusion] R-band of Cervus nippon hortulorum chromosomes can be manifested by RBG-binding technique.

Exploring genetic diversity in Northeastern Sika deer population （Cervus nippon hortulorum） by AFLP molecular markers

The genetic diversity of 27 different Cervus nippon hortulorum was studied to provide theoretical evidence for their identification and utilization. The genomie DNA of 27 different Cereus nippon hortulorum were analyzed by amplified fragment length polymorphism （AFLP）. 11,443 bands associated with genetic polymorphism among total 15,169 bands were obtained with 9 kinds of primer pairs and restriction endonuclease EcoRI/Msel, percentage of polymorphie band was 78.43%, 1,271 polymorphic locus were shown per primer pair. The AFLP data showed that average genetic similarity was 0.7841 （0.6809-0.8648）. 27 samples were classified into Group I and Group II with cluster analysis, and Group II was divided into five subgroups. The result of AFLP and cluster analysis concluded that there was high genetic variation, which associated with orientated artificial breed selection and breeding in the population. Genetic similarity of Group II-4 was the highest, more than 0.82, while genetic distance in this group was the shortest, from 0.1354 to 0.1563, which was coordinated with breeding record. All these showed that there was great genetic polymorphism among the deer population. The results laid the foundation for main quantitative trait locus （QTLs） of Cervus nippon hortulorum.

Identification of sika deer and red deer using partial cytochrome b and 12s ribosomal RNA genes

A study was conducted on the identifications of the degraded samples of sika deer （Cervus nippon） and red deer （Cervus elaphus） by phylogenetic and nucleotide distance analysis of partial Cytb and 12s rRNA genes sequences. 402 bp Cytb genes were achieved by PCR-sequencing using DNA extracted from 8 case samples, and contrasted with 27 sequences of Cytb gene downloaded from GenBank database. The values of three nucleotide distance between three suspected samples and sika deer were identical （0.026±0.006）, which was smaller than the smallest nucleotide distance between eastern red deer and sika deer （0.036）. Furthermore, phylogenetic analysis of sika deer and red deer indicated that the evidences located within the same cluster as sika deer. The evidences were sika deer materials. As the same way, other three suspected samples were derived from red deer. The results were further confirmed by phylogenetic and nucleotide distance analysis of 387 bp 12s rRNA gene. The method was powerful and less time-consuming and helpful to reduce the related cases with wildlife.

Susceptible conditions for debarking by deer in subalpine coniferous forests in central Japan

Background： Recently, deer have expanded their distribution to higher altitude ranges including subalpine forests However, culling deer and construction of deer fence in subalpine forests are difficult because of steep slopes and complex topography. Thus it is necessary to clarify the factors which are associated with debarking by deer for the effective protection of subalpine forests. In this study, we examined which factors are associated with debarking b： sika deer （Cervus nippon） in subalpine coniferous forests. Methods： We conducted our survey in Minami-Alps National Park, central Japan. We established 24 10 m × 40 m plots and surveyed the occurrence of debarking on saplings 〉30 cm in height and 〈3 cm in diameter at breast heigh （DBH） and on trees 〉3 cm in DBH, as well as sapling density within each plot. Minimum distances to nearest grasslan（ of plots were calculated （tentatively assuming grassland would attract deer and would cause high debarking pressure in the surrounding subaipine forests）. Results： The mean percentage of debarked live saplings was higher than that of live trees. The mean percentage of debarked saplings which had already died was 81.6 %. Debarking of saplings increased with lower elevation taller sapling size, and marginally increased near grassland. Sapling density was lower in plots with low basa area of conspecific trees near grassland and differed among species. Sapling density marginally decreased with decreasing elevation and increasing stand tree density. Debarking of trees was positively related to small DBH and Io~ elevation, and marginally increased near grassland and differed among species. Conclusions： Our results suggest that tall saplings in subalpine forests of low elevation or near subalpine grassland were susceptible to debarking by deer and monitoring of these areas may permit the early detection of the impacts of deer in subalpine coniferous forests.

梅花鹿茸生长过程中顶端不同组织COL1A1基因启动子DNA甲基化模式及差异分析

为探究COL1A1基因表达的Ⅰ型胶原蛋白α1链在组成生物体结构和骨发育中的重要作用,以梅花鹿茸生长过程的小鞍子(前期)、二杠(中期)和三杈茸(后期)3个典型时期鹿茸顶端组织及其茸皮、间充质、前软骨和软骨4个组织层为试验材料,采用亚硫酸氢盐测序法(BSP技术),从时空角度研究鹿茸生长过程中顶端不同组织COL1A1基因启动子区DNA甲基化模式及其相互间DNA甲基化差异。结果显示:1)COL1A1基因在前、中、后期的茸皮组织中的甲基化率分别为(9.73±0.92)%、(7.60±0.69)%和(3.73±0.23)%;间充质组织中的甲基化率分别为(3.20±0.40)%、(1.33±0.23)%和(1.60±0.69)%;前软骨组织中的甲基化率分别为(4.67±0.83)%、(2.53±0.46)%和(2.67±0.23)%;软骨组织中的甲基化率分别为(5.60±0.40)%、(2.80±0.40)%和(2.27±0.61)%。2)COL1A1基因启动子区的甲基化区域共有25个CG位点,在17个CG位点上均发生了不同程度的甲基化。3)对COL1A1基因启动子区DNA甲基化差异分析发现,相同时期中,茸皮组织甲基化率最高,间充质组织甲基化率最低;相同组织中,中期比前期显著下调;CG位点的比较中,前期与中期的CG位点差异程度较大。综上,COL1A1基因在快速生长期,以及在间充质组织、前软骨组织、软骨组织中对鹿茸生长具有促进作用。在DNA甲基化水平上探究梅花鹿鹿茸不同组织的时空表观遗传差异,为鹿茸快速生长与骨化机制的研究,以及哺乳动物组织再生和器官修复等领域的研究提供了科学参考。