采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性

Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) methods were applied to detect genetic variation of 20 bulking samples and two individually sampled populations of Oryza granulata (Nees et Arn. ex Watt.) from China (M5 from Hainan Province and M27 from Yunnan Province). For the bulking sampled populations, 20 RAPD and 12 ISSR primers generated 209 and 122 bands, of which 134 (64.11%) and 89 (72.95%) were polymorphic respectively. For population M5, 146 and 95 bands were generated with 21.48% and 34.78% being polymorphic (PPB). For population M27, 151 (PPB = 17.22%) and 94 (PPB = 19.15%) bands of RAPD and ISSR were obtained. The results indicated that the level of genetic variation of O． granulata was lower than other species detected in the genus Oryza. UPGMA cluster based on genetic similarity and principle component analyses (PCA) based on band patterns divided the populations into two groups corresponding to their sources, which revealed that the majority of genetic variation of O. granulata occurred between Hainan and Yunnan. Consequently, more populations of the species should be considered for a reasonable conservation management. Comparison between the two marker systems showed that ISSR was better than RAPD in terms of reproducibility and ability of detecting genetic polymorphism. Mantel test revealed that the goodness of fit between them was significant (r=0.917, t=12.718) when detecting genetic diversity at species level, but was poor at population level (r<0.200). The discrepancy was considered as the facts that different fragments were targeted by RAPD and ISSR, and that evolutionary process of O. granulata varied at different hierarchical levels.     

Male spike strobiles with Gnetum affinity from the Early Cretaceous in western Liaoning,Northeast China

A fossil with Gnetum affinity was found in the Jianshangou Member （Barremian Age） of the Yixian Formation （Lower Cretaceous Epoch） of the Jehol Group in western Liaoning, northeastern China. The single fossil specimen is represented by both elongate-cylindrical male spike strobiles which borne within a nodal bract of cauliflorous branch. The spike strobiles have apparent nodes, invisible internodes, and numerous verticillate involucral collars. The microsporangiate units within involucral collars are not seen. The male spike strobiles with verticillate involucral collars occur exclusively in Gnetum; hence, the fossil strobiles are attributed to a new taxon, Khitania columnispicata gen. ＆ sp. nov., being closely related to Gnetum. The general isotopic dating suggests an age of Barremian, ca. 125-122 million years （Myr） ago for the Jianshangou Member. The palaeoecological and palaeoclimatic inference based on the compositions of flora and fauna, and lithological characters of the fossil locality suggests that the fossil plants grew in a subtropical mesophytic forest and under a warmer climate. The remains of male spike strobiles are the first record of gnetalean macrofossil. It documents the evolution of the distinct gnetoid morphology and indicates a wider range of distribution of Gnetaceae in the Early Cretaceous than present day.     

Tet调控CYP2E1基因表达细胞系的代谢能力分析

应用Tet-On基因表达系统,调控CYP2E1基因在NIH 3T3细胞中的表达水平,探讨CYP2E1在化学致癌物二甲基亚硝胺(N-nitrosodimethylamine, NDMA)代谢中的作用．先后将 Tet-on基因表达系统的调控质粒pRevTet-on和反应质粒pRevTRE-2E1转染NIH 3T3细胞,分别用G418和潮霉素筛选,并通过RT-PCR和Western印迹检测,成功获得了3个具有良好诱导性Tet调控的CYP2E1基因表达细胞系（Tet/3T3-2E1）．应用不同浓度强力霉素(doxycycline, Dox)处理Tet/3T3-2E1细胞诱导CYP2E1表达,HPLC分析细胞对CYP2E1特异性药物探针氯唑沙腙的原位代谢能力及MTT法分析CYP2E1介导的NDMA细胞毒性作用．结果显示,Tet/3T3-2E1细胞CYP2E1的表达及其代谢能力有明显的Dox浓度依赖性,而且NDMA对Dox诱导组细胞有明显浓度依赖性细胞毒性作用,其IC₅₀值为12.06 μmol/L,无Dox诱导组未见明显的NDMA细胞毒性作用．该细胞模型的成功建立对于开展与CYP2E1相关的毒物、致癌物代谢研究,筛选抗毒物、抗癌物等具有重要价值．     

Floral-dip转化法将PEPC基因ihpRNA干扰表达载体导入甘蓝型油菜

根据磷酸烯醇丙酮酸羧化酶(PEPC)基因控制油菜籽中蛋白质与脂肪酸合成的原理,构建了PEPC基因片段ihpRNA干扰表达载体,该载体包含了一个PEPC基因片段正反向序列的回文结构．表达RNA干扰的载体结构（ihpRNA）,在大肠杆菌体内进行克隆的过程中易被大肠杆菌自身携带的某种酶剪切,剪切位点不确定,可能是一种随机性的剪切．通过反复实验,最终获得了一个与我们设计一致的PEPC基因片段正反向序列结构, RNA干扰表达载体构建获得成功．利用花序浸渍法(floral-dip)转基因将构建的PEPC基因ihpRNA干扰表达载体转移到甘蓝型油菜中,并通过抗性筛选、PCR特异扩增、PCR-Southern杂交和克隆测序,对获得的转基因植株进行了鉴定,转化率达到了069%,说明floral-dip方法有效地实现了ihpRNA干扰表达载体的遗传转化     

Utility of iPBS retrotransposons markers for molecular characterization of African Gnetum species

Abstract

Species of African Gnetum are lianas used as vegetables, medicines and for generating income. Despite the taxonomic confusion, identification of new species and diverse morphological characters in African Gnetum, molecular markers on these plants are lacking. However, the inter-primer binding site (iPBS) retrotransposons markers could be simple and excellent molecular markers for African Gnetum. The objective of this study was to determine the efficiency of iPBS markers in detecting genetic differentiation in African Gnetum species. A set of 21 iPBS markers were analysed on 14 accessions including G. africanum Welw., G. buchholzianum Engl. and the recently identified species G. latispicum. Six best selected primers generated 103 bands in G. africanum, 95 in G. buchholzianum and 24 in G. latispicum. Cluster analysis divided the accessions into two major groups. The first group contained all the accessions of G. africanum, whereas the second group was further divided in two subgroups representing accessions of G. buchholzianum and G. latispicum. Additionally, the Jaccard similarity coefficient indicated a close relationship between accessions of G. buchholzianum and G. latispicum. The iPBS marker system revealed genetic differentiation within African Gnetum and could be useful for evaluating genetic diversity, conservation, taxonomy and evolution studies.     

不同坡位下巨桉人工林土壤动物群落特征

为了解坡位对巨桉人工林土壤动物群落结构的影响,2009年1-10月,调查了四川盆地低山丘陵区不同坡位巨桉人工林的土壤动物群落.从3个巨桉人工林共获取土壤动物34741只,鉴定出146个类群,隶属于7门16纲31目;巨桉人工林下土壤动物的群落组成、营养类群、多样性及其季节动态基本都存在坡位变化,大型和中小型干生土壤动物的个体数、大型土壤动物的腐食性类群和中小型干生土壤动物的杂食性类群数量表现出随坡位的下降而逐渐增多的趋势,说明土壤动物响应了坡位引起的土壤生态因子的变化.其中大型土壤动物的腐食性类群和中小型湿生土壤动物的多样性显示出显著的坡位差异,表明这两个指标能指示巨桉人工林不同坡位生境的异质性.总的看来,巨桉人工林坡位因子对土壤动物群落的分布及结构特征具有一定影响,但作用尚不显著.     

基于广义可加模型的时空和环境因子对东南太平洋智利竹筴鱼渔场的影响

根据2001-2007年我国渔船在东南太平洋海域的智利竹筴鱼生产统计数据、时空因子以及海表温度、叶绿素浓度、海表温度梯度等环境数据,利用广义可加模型（GAM）分析了智利竹筴鱼资源变动情况及其与时空和环境因子的关系.结果表明：GAM模型对单位捕捞努力量渔获量(catch per unit effort,CPUE)总偏差解释率为31.47%;竹筴鱼生产具有明显的季节变化,主要集中在第17～41周,其中第25～31周的CPUE较高;东南太平洋智利竹筴鱼作业渔场集中在78°-98° W,30°-435° S之间,适宜海表温度为12 ℃～18 ℃,14 ℃～16 ℃最适宜,适宜叶绿素浓度在0.09～0.18 mg·m^-3,海表温度梯度在0.3 ℃～1.9 ℃.逐步建模法结果表明,影响竹筴鱼资源丰度的因子按重要性从大到小依次为周次、年份、海表温度、经度、纬度、叶绿素浓度和海表温度梯度.     

增强UV-B辐射对暖温带落叶阔叶林土庄绣线菊水分利用效率、气孔导度、叶氮素含量及形态特性的影响

在北京东灵山暖温带森林生态系统中,选择常见灌丛土庄绣线菊(Spiraea pubescens),建立UV-B控制实验。连续3个生长季每天增补9.4 kJ·m^-2的辐射剂量,模拟臭氧衰减17%时近地表面UV-B辐射的增强。该实验的目的是在野外环境下观测,长时间人工增强UV-B辐射对土庄绣线菊叶片的气孔导度、碳同位素比率(δ¹³C)、叶含水量、叶面积、特别是水分利用效率(WUE)和叶片全氮含量等指标的影响。实验结果表明,增强UV-B辐射显著减少了土庄绣线菊的叶面积(50.1%),提高了叶片全氮含量(102%)。同时,UV-B辐射还在一定程度上(尽管统计显示不显著)降低了气孔导度(16.1%)、胞间CO₂浓度与大气CO₂浓度之比(C_i/C_a) (4.0%)、提高了碳同位素比率(δ¹³C)(20.5‰)、叶含水量(3.1%)及比叶重(SLW)(5.2%),从而导致WUE的增加(4.1%)和植物的抗旱能力增强。值得注意的是,深层土壤(30~40 cm)含水量变化会影响气孔导度、δ¹³C和WUE对紫外辐射的响应程度:在土壤干旱的季节(6月和9月),气孔导度、δ¹³C、WUE这些指标处理和对照的差异很小,但是当土壤水分充足时(7月和8月),处理和对照的差异就较为显著。另外,随着实验处理时间的延长,UV-B的效应变得不显著。相关分析表明,UV-B辐射降低了土壤含水量(30~40 cm)与土庄绣线菊叶含水量、δ¹³C、C_i/C_a和气孔导度的相关系数,增强了WUE与土壤含水量的相关性,这也许是由于UV-B辐射增强了WUE对土壤水分变化的敏感性。该研究的结果表明UV-B辐射对土庄绣线菊的形态和生长有显著的影响,但对主要水分生理指标影响不显著。     

游仆虫重组核糖体蛋白L11与肽链释放因子的体外相互作用分析

核糖体蛋白L11(ribosome protein L11)是一种高度保守的蛋白质．为研究真核生物的核糖体蛋白L11的功能,从八肋游仆虫（Euplotes octocarinatus）大核基因组中克隆到核糖体蛋白L11基因,构建了重组表达质粒pGEX-6p1-L11,通过谷胱甘肽-Sepharose 4B亲和层析,纯化了重组融合蛋白GST-L11．Pull down 分析显示,八肋游仆虫的核糖体蛋白L11与第一类肽链释放因子eRF1a可以在体外相互作用．这一结果提示,与原核生物一样,低等真核生物的核糖体蛋白L11在肽链终止过程中可能起一定的作用．