茶树CsMYB123转录因子的克隆及表达特性研究

该实验以茶树品种‘紫娟’为试验材料,利用RT-PCR方法,从茶树cDNA中克隆得到一个R2R3-MYB型基因(CsMYB123)。生物信息学分析显示,CsMYB123基因的开放阅读框为915bp,编码304个氨基酸,蛋白分子量约34.07kD,理论等电点为8.69,含有2个保守的MYB结构域,编码1个R2R3-MYB蛋白;CsMYB123蛋白与拟南芥(Arabidopsis thaliana)MYB转录因子家族第五亚组的AtMYB123亲缘关系最近;CsMYB123属于亲水性蛋白,无N端信号肽,可能定位于细胞核上。荧光定量PCR分析表明,CsMYB123基因在茶树各组织的表达量大小依次为:一芽一叶第二叶第三叶第四叶老茎嫩茎,且在一芽一叶中的表达量是嫩茎的15.68倍;但CsMYB123的表达受IAA、ABA、ETH和GA3的抑制。花青素含量检测显示,茶树‘紫娟’各组织中花青素的含量高低依次为:第二叶一芽一叶第三叶第四叶嫩茎老茎,且第二叶和一芽一叶的含量分别为老茎的15倍和11倍。研究发现,CsMYB123基因在茶树‘紫娟’的新稍中高水平表达,且其表达模式与不同组织中的花青素含量呈较好的正相关关系,推测CsMYB123基因与茶树花青素合成的调控相关。     

基于pufM基因的乌梁素海富营养化湖区好氧不产氧光合细菌系统发育多样性分析

好氧不产氧光合细菌(AAPB)的多样性在海洋中已经广泛研究,但在富营养化湖泊中却研究甚少。通过构建和分析AAPB光合中心合成中的关键基因pufM克隆文库,以期揭示乌梁素海富营养化湖区AAPB分布及其系统发育多样性,探讨其在富营养化湖泊中的功能和作用。对乌梁素海红圪卜湖区水体文库中的52个克隆子进行分析,产生了28个OTU,文库覆盖度达到71.4%,反映出文库有较好的代表性。同源性和系统发育分析结果表明,乌梁素海红圪卜湖区AAPB有较高的多样性,与我们之前所发现的同一湖区总细菌多样性较低形成鲜明对比。所获得的序列分属7个亚群,即γ-Proteobacteria(44.2%,含Group-1,-2和-3共3个亚群)、β-Proteobacteria(21.2%)、Rhodobacter-like(7.69%)及2个未知亚群unknown Group-1(21.2%)和Group-2(5.77%)。其中γ-Proteobacteria占到总克隆的44.2%,在低盐的乌梁素海环境中出现高比例的γ-Proteobacteria之前并未见类似报道,并且乌梁素海中存在一些可能是富营养化湖泊特有的AAPB。这表明AAPB可能在富营养化湖泊生态系统的维持和稳定中有重要作用。     

PCP degradation is mediated by closely related strains of the genus Sphingomonas

There have been numerous reports in the literature of diverse bacteria capable of degrading pentachlorophenol (PCP). In order to gain further insight into the phylogenetic relationships of PCP-degrading bacteria, we examined four strains: Arthrobacter sp. strain ATCC 33790, Flavobacterium sp. strain ATCC 39723, Pseudomonas sp. strain SR3, and Sphingomonas sp. strain RA2. These organisms were isolated from different geographical locations and all of them degrade high concentrations (100–200 mg/L) of PCP. Southern blot analyses determined that these bacteria all harbour DNA that encodes similar, if not identical, genes involved in PCP degradation. Comparison of the 16S rRNA nucleotide sequences revealed that these organisms were very closely related and, in fact, represent a monophyletic group. The 16S rRNA analyses together with fatty acid and sphingolipid analyses strongly suggest that the four strains are members of the genus Sphingomonas . The close relationship of the four organisms is supported by nucleotide sequence analysis data of the pcpB locus encoding PCP-4-monooxygenase, the first enzyme in the PCP degradative pathway.     

Diagnostic microbial microarrays in soil ecology

Soil microbial communities are responsible for important physiological and metabolic processes. In the last decade soil microorganisms have been frequently analysed by cultivation-independent techniques because only a minority of the natural microbial communities are accessible by cultivation. Cultivation-independent community analyses have revolutionized our understanding of soil microbial diversity and population dynamics. Nevertheless, many methods are still laborious and time-consuming, and high-throughput methods have to be applied in order to understand population shifts at a finer level and to be better able to link microbial diversity with ecosystems functioning. Microbial diagnostic microarrays (MDMs) represent a powerful tool for the parallel, high-throughput identification of many microorganisms. Three categories of MDMs have been defined based on the nature of the probe and target molecules used: phylogenetic oligonucleotide microarrays with short oligonucleotides against a phylogenetic marker gene; functional gene arrays containing probes targeting genes encoding specific functions; and community genome arrays employing whole genomes as probes. In this review, important methodological developments relevant to the application of the different types of diagnostic microarrays in soil ecology will be addressed and new approaches, needs and future directions will be identified, which might lead to a better insight into the functional activities of soil microbial communities.     

昆虫发育进度的非正态描述──分段指数曲线

提出了分段连续指数曲线转化为多元线性回归方程的方法。利用取剩余平方和最小的方法确定未知折点。给出了在描述昆虫发育进度的模拟模型。     

不同降水年型下长期施肥旱地小麦产量效应

以黄土高原30年长期肥料定位试验为基础,依据降水将生育年划分为干旱年、平水年和丰水年,研究不同施肥方式对黄土高原冬小麦产量、肥料贡献率和降水利用率的影响.结果表明: 小麦连续种植30年中,氮磷配施和氮磷钾配施下小麦产量、肥料贡献率和降水利用率显著高于不施肥和单施肥处理,氮磷钾配施下小麦产量、肥料贡献率和降水利用率分别达到3480 kg·hm^-2、61.45 kg·kg^-1、6.13 kg·mm^-1·hm^-2;不同降水年型下,丰水年小麦产量、肥料贡献率和降水利用率相对较高;使用逐步回归分析可知,不同降水年型下小麦产量主要受氮磷肥施用量、休闲期降水和越冬期降水影响.黄土旱塬可以通过提高氮磷用量同时适当施用钾肥,以及在休闲期做好蓄水保墒工作来提高小麦产量.     

暖温带乔木和灌木物候变化及对气候变化的响应

根据2003-2014年气象数据和暖温带3种乔木(辽东栎、五角枫和核桃楸)和3种灌木(土庄绣线菊、毛叶丁香和六道木)的物候观测数据资料, 采用气候倾向率和回归分析等方法, 观察乔木和灌木物候变化特征的差异, 分析温度、降水以及乔木、灌木的物候变化趋势, 同时对气象因子与乔木和灌木物候期的相关关系进行研究。结果表明: ①研究期间, 北京东灵山平均气温呈不显著的上升趋势, 气候倾向率为0.200℃·10a–1, 春季(3–5月)和夏季(6-8月)温度显著上升; 降水量呈下降趋势, 平均减少71.630 mm·10a–1, 总体呈暖、干的趋势。②3种乔木的生长季长度都缩短, 辽东栎、五角枫和核桃楸平均生长季长度分别缩短50.70 d·10 a–1、29.83 d·10a–1和22.36 d·10a–1。3种灌木的生长季长度也都缩短, 土庄绣线菊、毛叶丁香和六道木的平均生长季长度分别缩短42.55 d·10a–1、42.76 d·10a–1和38.15 d·10a–1。乔木和灌木的物候变化趋势相同, 整体表现为春季物候推迟, 秋季物候提前, 生长季长度都缩短且生长季长度相差不大。乔木和灌木都表现出芽期推迟最明显, 每10年推迟达19天以上。③乔木和灌木各物候期与气温总体表现为负相关, 即气温升高, 物候期提前, 其相关性显示出夏季(6-8月)温度对植被物候期影响较大, 夏季温度与各物候期表现为正相关, 即夏季温度升高, 物候期推迟。同时乔木和灌木与总体降水没有明显的相关关系, 但秋季物候与不同时段降水表现不同的相关性, 由此可知夏季温度变化对木本植物春季物候(出芽期、展叶期和首花期)的影响更大, 而秋季物候(叶变色期和落叶期)受温度和降水共同影响。     

Environmental factors function as constraints on soil nitrous oxide fluxes in bioenergy feedstock cropping systems

Nitrous oxide (N₂O) is a potent greenhouse gas and major component of the net global warming potential of bioenergy feedstock cropping systems. Numerous environmental factors influence soil N₂O production, making direct correlation difficult to any one factor of N₂O fluxes under field conditions. We instead employed quantile regression to evaluate whether soil temperature, water‐filled pore space (WFPS), and concentrations of soil nitrate () and ammonium () determined upper bounds for soil N₂O flux magnitudes. We collected data over 6 years from a range of bioenergy feedstock cropping systems including no‐till grain crops, perennial warm‐season grasses, hybrid poplar, and polycultures of tallgrass prairie species each with and without nitrogen (N) addition grown at two sites. The upper bounds for soil N₂O fluxes had a significant and positive correlation with all four environmental factors, although relatively large fluxes were still possible at minimal values for nearly all factors. The correlation with was generally weaker, suggesting it is less important than in driving large fluxes. Quantile regression slopes were generally lower for unfertilized perennials than for other systems, but this may have resulted from a perpetual state of nitrogen limitation, which prevented other factors from being clear constraints. This framework suggests efforts to reduce concentrations of in the soil may be effective at reducing high‐intensity periods—”hot moments”—of N₂O production.     

Fluorescence polarization assay and inhibitor design for MDM2/p53 interaction

MDM2 is an important negative regulator of the tumor suppressor protein p53 which regulates the expression of many genes including MDM2. The delicate balance of this autoregulatory loop is crucial for the maintenance of the genome and control of the cell cycle and apoptosis. MDM2 hyperactivity, due to amplification/overexpression or mutational inactivation of the ARF locus, inhibits the function of wild-type p53 and can lead to the development of a wide variety of cancers. Thus, the development of anti-MDM2 therapies may restore normal p53 function in tumor cells and induce growth suppression and apoptosis. We report here a novel high-throughput fluorescence polarization binding assay and its application in rank ordering small-molecule inhibitors that block the binding of MDM2 to a p53-derived fluorescent peptide.     

The evolution of Queensland spiny mountain crayfish of the genus Euastacus. I. Testing vicariance and dispersal with interspecific mitochondrial DNA

The upland mesic rainforests of eastern Australia have been described as a "mesothermal archipelago" where a chain of cool mountain "islands" arise from a warm "sea" of tropical and subtropical lowlands. An endemic freshwater crayfish belonging to the genus Euastacus is found on each of these mountain "islands." The Euastacus are particularly suitable for the study of evolution because each mountain harbors a unique species, there are many taxa present providing replication within the group and, most importantly, their distribution is linear, extending along a south-north axis. This group could have evolved by "simultaneous vicariance" where there was one vicariant separation event of a widespread ancestor, or by "south to north stepping stone dispersal" where there were long distance dispersal events from neighboring mountain islands, starting in the south and proceeding north in a dispersal-colonization wave. We used pairwise genetic distances between nearest geographic neighbors as a novel way to test the two hypotheses. If diversification was due to "south to north stepping stone dispersal," then pairwise genetic distances between nearest geographic neighbors should decrease progressively the farther north the taxon pairs are found, reflecting the decreasing periods of isolation. In this case there should be a negative correlation between the south to north rank order of nearest neighbors and pairwise genetic distances. A Spearman's correlation on 16S mtDNA pairwise genetic distances and geographic rank order was not significant, indicating there was no support for the south to north stepping stone dispersal hypothesis. If simultaneous vicariance was responsible for diversification then all nearest geographic neighbor taxon pairs should have similar genetic distances and, therefore, the variance in nearest neighbor distances should be zero, or close to it. To test if the observed variance was tending towards zero we developed a randomization test where nearest neighbor taxon pairs were assigned random genetic distances and the variances calculated. The observed variance lay in the < 0.05 range of the simulated variances, providing support for the simultaneous vicariance hypothesis. The data also suggest there was simultaneous vicariance of at least two ancestral Queensland lineages. The timing of this vicariant event was probably in the Pliocene, which is consistent with the divergence times reported for other Australian mesic rainforest restricted taxa.