HbSNP12基因及HbCBF1基因在巴西橡胶树中的差异分析

以抗寒性较强和较弱的两组巴西橡胶树为材料,从NCBI中搜索HbSNP12、HbCBF1基因序列,用Primer Premier 5.0软件设计特异引物,PCR扩增后进行测序,比对两基因在两组材料中的碱基及氨基酸序列信息。结果表明,HbSNP12基因在105 bp、538 bp处,HbCBF1基因在92 bp、387 bp处,两组材料的碱基序列均存在差异,将其翻译为氨基酸序列后差异同样存在。本实验探讨了与抗性相关的HbSNP12、HbCBF1两基因在两组抗性相差较大材料中的单核苷酸多态性,为橡胶树抗寒性相关的SNP标记筛选提供依据。     

木聚糖酶基因xyn I的克隆和序列分析

以宇佐美曲霉E001总RNA为模板,采用RT-PCR等技术扩增了木聚糖酶基因(xynI)的cDNA全序列(GenBank登录号DQ302412)。该cDNA全长881 bp,其中5′和3′端非编码区分别为97和106 bp,信号肽编码区111 bp,成熟肽编码区567 bp编码188个氨基酸组成的木聚糖酶Xyn I。以E001基因组DNA为模板,采用单侧PCR等技术扩增了xynI的DNA全序列(GenBank登录号DQ302413)。该DNA全长1 098 bp,含启动子序列、内含子和外显子等核苷酸序列。将xynI cDNA所推导的Xyn I一级结构与GenBank中报道的其它相关序列进行了同源性比较。结果表明,E001 Xyn I是一种新的木聚糖酶,且具有第G/11家族木聚糖酶的共同特征。     

竹子捕光叶绿素a/b结合蛋白基因全长的克隆和序列分析

采用RT-PCR技术与RACE技术,从绿竹中克隆到捕光叶绿素a/b结合蛋白基因全长cDNA,命名为cab-BO1(GenBank登记号:EF061137).该基因长1 102 bp,从第64～861 bp含有1个开放阅读框,编码265个氨基酸.cab-BO1编码的氨基酸中第64～232位包括典型的捕光叶绿素a/b结合蛋白功能域,第9～11位为蛋白激酶C的磷酸化位点,第27～52位为泛素结构功能域信号,第55～58位为酪氨酸激酶(CK2)磷酸化位点.序列相似性分析结果表明:cab-BO1 DNA序列和编码的氨基酸序列与玉米、小果野蕉、小麦、水稻等的cab基因及其氨基酸序列的相似性分别在85%和89%以上,显示了cab家族基因在进化过程中的保守性.     

木聚糖酶基因xynⅠ的克隆和序列分析

以宇佐美曲霉E001总RNA为模板,采用RT- PCR等技术扩增了木聚糖酶基因(xyn I)的cDNA全序列(GenBank登录号DQ302412).该cDNA全长 881 bp,其中5′和3′端非编码区分别为97和 106 bp,信号肽编码区 111 bp,成熟肽编码区 567 bp 编码188个氨基酸组成的木聚糖酶Xyn I.以E001基因组DNA为模板,采用单侧PCR等技术扩增了xyn I的DNA全序列(GenBank登录号DQ302413).该DNA全长 1 098 bp,含启动子序列、内含子和外显子等核苷酸序列.将xyn I cDNA所推导的Xyn I一级结构与GenBank中报道的其它相关序列进行了同源性比较.结果表明,E001 Xyn I是一种新的木聚糖酶,且具有第G/11家族木聚糖酶的共同特征.     

利用核糖体小亚基RNA序列探讨四种腹毛目纤毛虫的系统进化

以四个在环境监测中比较有代表性的纤毛虫属－游仆虫属、尾柱虫属、棘尾虫属和伪角毛虫属纤毛虫(Euplotes, Urostyla, Stylonychia, Pseudokeronopsis)为材料,对其16s小亚基核糖体RNA(16s－like Small Subunit rRNA,SSrRNA)基因进行序列测定,并与其他种腹毛目纤毛虫相比较,以美国赭虫(Blepharisma americanum)为外群构建系统进化树.结果显示:游仆虫属最先从腹毛目纤毛虫中分化出来,尾柱虫属(Urostyla)与全列虫属(Holosticha)纤毛虫亲缘关系较近.贻贝棘尾虫(Stylonychia mytilus)和浮萍棘尾虫(Stylonychia lenmae)是姊妹种,在外观上及其相似,但可以用构建进化树的方法从分子角度加以区分.从构建的进化树拓扑结构上可知,红色伪角毛虫与全列虫科种类的亲缘关系较近,与尾柱虫科种类的亲缘关系较远.     

马尾松肉桂酰辅酶A还原酶基因(CCR)克隆及分析

对马尾松肉桂酰辅酶A还原酶基因(CCR)进行扩增、克隆和测序(GenBank登录号:EU753854).应用MEGA4.0.2软件对不同树种CCR基因的核苷酸和氨基酸序列进行比对,结果表明:马尾松CCR基因的外显子和内含子数目与火炬松、杨树、银合欢、光皮桦、冈尼桉、柳桉、蓝桉的外显子和内含子数目相同,都有5个外显子和4个内含子.马尾松CCR基因编码区975 bp,编码324个氨基酸.马尾松外显子Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ分别为133,155,186,353,148 bp,而内含子Ⅰ,Ⅱ,Ⅲ,Ⅳ分别为1 450,216,737,941 bp.马尾松CCR外显子和内含子连接处序列除内含子Ⅱ/外显子Ⅲ出现CTPuCG/外显子Ⅲ,其余遵循外显子/GTPuAG/外显子组成规律.马尾松外显子Ⅱ,Ⅲ,Ⅳ核苷酸数目与桉属、杨属、桦木属和松属树种相同,但外显子Ⅰ和外显子Ⅴ不尽相同.比对的树种间在该基因编码区核苷酸序列和氨基酸序列上相对保守,也存在一定差异.马尾松与火炬松、光皮桦、银合欢、杨树、冈尼桉、柳桉、蓝桉各物种间编码区核苷酸序列相似性大小分别为99.2%,67.8%,68.0%,68.9%,69.5%,69.3%,69.9%,而相应的氨基酸序列间相似性大小分别为99.1%,73.5%,72.5%,74.4%,75.0%,74.4%,75.0%.马尾松与其他13个树种CCR序列构建的系统进化树表明:3个针叶树种形成1个独立分支,而且最早进化.     

重金属胁迫下白骨壤数字基因表达谱分析

[目的]从转录组水平研究重金属污水胁迫处理下白骨壤的基因表达变化,以揭示白骨壤响应重金属污染胁迫的机制。[方法]采用基于高通量测序的数字基因表达谱(DGE)技术对重金属污水胁迫处理组和对照组白骨壤材料进行转录组测序分析。[结果]与对照组相比,重金属处理下白骨壤共有10 459个差异表达基因,其中,8 685个表达量上升,1 774个表达量下降。Me V聚类表明:大部分基因在重金属处理样本中的表达量受到极大的诱导。GO功能注释发现,差异表达基因主要定位于叶绿体以及细胞内各种膜结构,参与"细胞代谢"、"细胞组分的组合和生物合成"、"对刺激的响应"等过程。KEGG代谢通路分析显示,差异表达基因分布于126条Pathways,涉及"核糖体"、"光合作用"、"乙醛酸盐代谢"、"丙酮酸代谢"等途径。重金属胁迫还促进萜类、黄酮类化合物生物合成的关键酶基因(牻牛儿基焦磷酸合酶(0009238)、黄酮醇合成酶(0001833))的表达,进而促进白骨壤有效成分的积累;显著诱导细胞分裂素水解酶编码基因CKX7(0057124)、油菜素内脂信号转导组分基因BSK(0043741)等的表达,进而提高白骨壤对重金属胁迫的适应能力。此外,转录因子分析发现,b HLH、NAC、MYB-related和WRKY在重金属胁迫中发挥重要作用。实验选取8个与环境刺激响应密切相关的基因,通过qRT-PCR验证了它们在重金属污水胁迫处理下的表达差异,结果与数字基因表达谱分析的结果较一致,证实了差异表达基因数据的有效性。[结论]重金属处理影响了白骨壤大量的新陈代谢、光合作用、小分子酸合成、激素合成与信号转导及次生代谢产物合成相关基因的表达。     

油麦吊云杉叶绿体基因组特征及密码子偏性分析

【目的】分析油麦吊云杉cpDNA的特征和密码子偏好性,为云杉属植物叶绿体基因组学提供研究依据。【方法】通过Ilumina HiSeq测序平台对油麦吊云杉cpDNA进行测序,并利用Codon W 1.4.2和CUSP软件对其密码子偏性进行分析。【结果】油麦吊云杉cpDNA是一个全长为124 161 bp的环状DNA分子,与典型的具有保守的四分体cpDNA结构不同,油麦吊云杉cpDNA中存在反向重复区域的丢失,其cpDNA的GC含量为38.7%。油麦吊云杉cpDNA包含113个基因,其中蛋白质编码基因76个,核糖体RNA基因4个,转运RNA基因33个,系统发育分析显示,油麦吊云杉和西加云杉聚为一个独特的分支,油麦吊云杉cpDNA密码子不同位置上的GC含量从低到高依次是GC3(49.39%) 45的占86%,表明其密码子的偏性较弱;GC12和GC3的相关性不显著（r=0.166 4, b=-0.157 7）;ENC频数比值分布在-0.0...     

棕背(鼠平)巢区及对林木鼠害防治意义的研究

采用重捕标志法研究了棕背巢区,巢区面积7月份平均为250.89 m2,9月份平均为284.21 m2,雌雄间和季节间差异显著。棕背有集群分布现象。结合巢区面积和活动距离对鼠害防治布点规格与防治灭鼠率的关系进行了分析。     

基于形态特征和线粒体12S、Cyt b和ND4基因片段的横斑锦蛇和玉斑锦蛇的关系研究

本文通过对横斑锦蛇(Elaphe perlacea)肌肉线粒体DNA的提取,对其中的3个基因—12SRNA、ND4和细胞色素b进行扩增、测序,与玉斑锦蛇(Elaphe mandarinus)及锦蛇属(Elaphe)17个物种的相同基因片段重建系统发育树。系统发育结果表明横斑锦蛇与玉斑锦蛇系统发育关系最近,但是两者不是同一个物种。最后将横斑锦蛇和玉斑锦蛇的外部特点进行对比,发现横斑锦蛇与玉斑锦蛇在形态特征上存在稳定性差异。     

Over-winter weight dynamics in supplementally fed free-ranging sika deer (Cervus nippon)

Continuous body weight dynamics in supplementally fed free-ranging sika deer (Cervus nippon) during the three consecutive overwinters (from January to April), 1992–1994 were surveyed. Almost all individuals lost weight continuously throughout each winter (adult deer 5–13%; young deer 3–19%) and several deer lost weight even in April. Weight gain after loss began in the mid-April when ambient temperature increased and when plant growth began. Weight dynamics varied according to sex, age and reproductive status. Adult deer showed significant large weight loss than the young. Young deer lost weight (calves 18±11 g/day, yearling and two-year-olds 27±29 g/day) more slowly than the adults (65±23 g/day). Following this study of over-winter body weight dynamics, it seems that early springs are the most critical periods for the survival of free-ranging sika deer because of their poor nutritional condition. A portion of this report was presented at the 106th Annual Meeting of the Japanese Forestry Society (1994).     

Population density and range use of sika deer,Cervus nippon, on Mt. Ohdaigahara, central Japan

We investigated the population density and range use of sika deer,Cervus nippon, on Mt. Ohdaigahara, central Japan, from July 1996 to July 1997. The deer population density estimated by the block count was high at 17.5–30.9 deer/km². Although the deer remained this area from spring to fall, they moved out in winter, probably because of deep snow. A spotlight count showed that the deer prefer the eastern area as spring-summer range, where the forest consists of coniferous trees over a floor cover ofSasa nipponica, andS. nipponica grassland. The deer also use dense coniferous forest for cover, especially in summer and fall. Part of this study was presented at the 108th Annual Meeting of the Japanese Forestry Society (1997). This study was financially supported by the Environment Agency, Japan.     

Wintering habitat evaluation of Red deer (Cervus elaphus) in forest area of Northeastern China

Six ultimate factors (forage abundance, horizontal sheltering class, distance from human disturbance, snow depth, tree coverage, and edge effect) of Red deer (Cervus elaphus) were investigated in eastern Heilongjiang Province during winter from 1988 to 1991. On the basis of analysis of field data, we evaluated winter habitat conditions of Red deer. The results showed as follows: the wintering habitat quality of Red deer in poplar-birch stands was excellent in the Wanda Mountains, and was moderate in shrub-woods and Korean pine seed stands. According to habitat index (HI) values, the habitat quality in poplar-birch stands (HI=0.8185) was superior to that in shrub-woods (HI=0.4825) and in Korean pine seed stands (HI=0.4385). The wintering habitat quality of Red deer in the Wanda Mountains was superior to that of Dailing Forestry Region. According to the current situations of forestry areas in northeastern China, strengthening habitat management was important for the conservation and management of Red deer population in the next decades. This subject is supported by the Natural Science Foundation of Heilongjiang Province. (Responsible Editor: Zhu Hong)     

Food habits of sika deer as assessed by fecal analysis in Mt. Ohdaigahara,central Japan

Food habits of sika deer,Cervus nippon centralis temminck, in Mt. Ohdaigahara, central Japan, were investigated by the fecal analysis method from September, 1990, to June, 1991. The fecal analysis showed that the percentage ofSasa nipponica Makino et Shibata in the fecal composition was approximately 50% throughout the year, indicating that it was the main food plant. Bark and twigs were also found in the feces in all seasons, suggesting that the deer ate bark throughout the year. Even though the amount of the bark eaten per capita was small, the high density of deer may possibly cause serious bark damage. Barking can be one of the main causes of the mortality ofPicea jezoensis Sieb. et Zucc. var.hondoensis (Mayr) Rehder andAbies homolepis Sieb. et Zucc. in Mt. Ohdaigahara. A part of this study was presented at the 105th Annual Meeting of the Japanese Forestry Society (1994).     

Effects of 13-year fencing on browsing by sika deer on seedlings on Mt. Ohdaigahara, central Japan

We examined the effects of fencing on deer browsing on seedlings 13 years after the building the experimental fence on Mt. Ohdaigahara, central Japan, where the sika deer (Cervus nippon) population is high. There was no difference in the species number of seedlings between 1991 and 2004. The density of seedlings in the fenced plot in 2004 was three times higher than in 1991. Although the unfenced plots had no seedlings higher than 20 cm, there were seedlings up to 120 cm in the fenced plot. These results suggest that 13-year fencing promoted regeneration of seedlings in this area.     

Full length cDNA cloning and expression analysis of annexinA2 gene from deer antler tissue

ANXA2（AnnexinA2）, a calcium-dependent phospholipid bind- ing protein, is involved in various Ca2＋-related biological activities. In the present study, full-length cDNA of ANXA2 was isolated from the velvet antler tip tissue of sika deer （Cervus nippon hortulomm）; the amino acid sequence and gene expression was analyzed by using bioinformatics and real-time reverse transcdptase polymerase chain reaction （RT-PCR） techniques. Nucleotide sequence analysis reveals that the full-length cDNA of the ANXA2 gene was 1372 bp, of which 1020 bp was in the opan-reading frame （OR.F） encoding 339 amino acids; its relative mo- lecular weight was 38.3 kDa; and isoelectrie point was 6.72. Sequence analysis indicates that the protein includes four conserved tan- dem-duplication ANX domains. The gene-aceession nucleotide sequence number in GenBank is JX315571. Expression analysis by RT-PCR re- veals that ANXA2 gene expression has a significant positive correlation with the antler-tissue mineralization process, indicating that this gene may play an important role in the regulation of antler-tissue mineraliza- tion.     

Development of a method for forest cafeteria tests for evaluating chemical deterrents against bark stripping by sika deer (Cervus nippon yesoensis)

Damage caused by sika deer (Cervus nippon yesoensis) is a serious problem in commercial and environmental (non-harvested) forests in Hokkaido, northern Japan. Cafeteria tests in forests may be useful for evaluating the efficacy of chemical deterrents against bark stripping by deer. To develop a method for forest cafeteria tests in the continuous snow cover period, two experiments were carried out. In the experiments, logs were produced from tree trunks, and used as carriers of chemical deterrents. Carriers were installed in forests and fed to deer. The first experiment was to find suitable sites and installation methods for carriers. Criteria for the local suitability and the installation methods were as follows: a) Sites where deer are active should be selected; b) Carriers should be installed along actively used deer trails; c) Installation sites of carriers should be changed in response to deer movement; d) Carriers should be produced from tree species that deer naturally prefer; and e) Each carrier should be partially buried in the snow. The second experiment evaluated the feasibility of a cafeteria test method based on the results of the first experiment. The method was used for 13 sets of the cafeteria test, in which the deterrent effectiveness of 5 chemicals (wood tar, rosin, wood vinegar, and 2 pyroligneous liquors) was examined. We obtained results from all the sets. The chemicals tested did not deter bark stripping by deer. Nevertheless, the method used in the present study was practical for the cafeteria tests.     

Expansion of dwarf bamboo, Sasa nipponica, grassland under feeding pressure of sika deer, Cervus nippon, on subalpine coniferous forest in central Japan

The long-term changes of the area of Sasa nipponica grassland, bark stripping damage to trees by sika deer, Cervus nippon, and sika deer densities on Mt. Ohdaigahara were assessed with aerial photographs, information from previous studies, and field experiments. The grasslands expanded year after year. The expansion rate was highest from 1982 to 1992, and lowest from 1992 to 1997. Bark-stripping damage on coniferous trees was heavier than that on broadleaf trees in both the early 1980s and 1998. The deer density in the eastern part of the area had been high (14.4–64.3 km⁻²) from 1984 until 1998. We presume that rich growth of S. nipponica and heavy feeding pressure by sika deer over the long term caused expansion of S. nipponica grasslands by killing canopy trees by bark stripping and preventing forest regeneration.     

Wintering Habitat Evaluation of Red Deer (Cervus elaphus) in Forest Area of Northeastern China

IntroductionTheprofessionalsworkofwildlifemanagementisatargetforpopulationmanagement,butmoreandmoreprofessionalsworkersareinterestedinthemanagementofthewildlifehabitat(Belley,1984).ItisoneofessentiafworksforustoappraisehabitatwiththescientificmanagementontheReddeerpopulation.Soitisusefultoconservation,developmentandutilizationoftheReddeerinananimal'sperspective.Sincethe1970'smanyscientistshavebeenstudyingthehabitatevaluationfordeer.Severalmodelsandmethodssuchasthelineardiagram,thematrixmodel…     

Bark stripping by sika deer on veitch fir related to stand age, bark nutrition, and season in northern Mount Fuji district, central Japan

We carried out this study to clarify the relationships between the seasonal changes of bark stripping and food quality, and between bark-stripping intensity and bark nutrition with age of veitch fir (Abies veitchii) by sika deer (Cervus nippon) in the northern Mount Fuji district, from June 2000 to July 2003. We found that sika deer gnawed and ate all of the bark stripped, and a seasonal change in bark stripping occurred from December to May or June with a peak in March–April. The stripping period largely overlapped with the periods of low food availability and poor food nutrition (indicated by fecal chemical component index) of sika deer, from January to April. Both bark nutritional quality and bark-stripping intensity related to stand age and tree size of veitch fir negatively. Consequently, bark-stripping intensity related to bark nutritional quality positively. February is the worst forage period in terms of both quantity (due to deep snow) and quality (indicated by fecal chemical components). There was a time lag of 1 to 2 months in the peak of bark stripping in March–April when compared with the poor forage period in February. This time lag may suggest that sika deer need more nutritious and easily digested food from March due to increased nutrient demands that result from depleted body condition in both sexes, gestation of pregnant females, and the recovery of active metabolism.