RNA editing of 10 Didymium iridis mitochondrial genes and comparison with the homologous genes in Physarum polycephalum

Regions of the Didymium iridis mitochondrial genome were identified with similarity to typical mitochondrial genes; however, these regions contained numerous stop codons. We used RT-PCR and DNA sequencing to determine whether, through RNA editing, these regions were transcribed into mRNAs that could encode functional proteins. Ten putative gene regions were examined: atp1, atp6, atp8, atp9, cox1, cox2, cytb, nad4L, nad6, and nad7. The cDNA sequences of each gene could encode a functional mitochondrial protein that was highly conserved compared with homologous genes. The type of editing events and editing sequence features were very similar to those observed in the homologous genes of Physarum polycephalum, though the actual editing locations showed a variable degree of conservation. Edited sites were compared with encoded sites in D. iridis and P. polycephalum for all 10 genes. Edited sequence for a portion of the cox1 gene was available for six myxomycetes, which, when compared, showed a high degree of conservation at the protein level. Different types of editing events showed varying degrees of site conservation with C-to-U base changes being the least conserved. Several aspects of single C insertion editing events led to the preferential creation of hydrophobic amino acid codons that may help to minimize adverse effects on the resulting protein structure.     

Cis-regulatory characterization of sequence conservation surrounding the Hox4 genes

Hox genes are key regulators of anterior-posterior axis patterning and have a major role in hindbrain development. The zebrafish Hox4 paralogs have strong overlapping activities in hindbrain rhombomeres 7 and 8, in the spinal cord and in the pharyngeal arches. With the aim to predict enhancers that act on the hoxa4a, hoxb4a, hoxc4a and hoxd4a genes, we used sequence conservation around the Hox4 genes to analyze all fish:human conserved non-coding sequences by reporter assays in stable zebrafish transgenesis. Thirty-four elements were functionally tested in GFP reporter gene constructs and more than 100 F1 lines were analyzed to establish a correlation between sequence conservation and cis-regulatory function, constituting a catalog of Hox4 CNEs. Sixteen tissue-specific enhancers could be identified. Multiple alignments of the CNEs revealed paralogous cis-regulatory sequences, however, the CNE sequence similarities were found not to correlate with tissue specificity. To identify ancestral enhancers that direct Hox4 gene activity, genome sequence alignments of mammals, teleosts, horn shark and the cephalochordate amphioxus, which is the most basal extant chordate possessing a single prototypical Hox cluster, were performed. Three elements were identified and two of them exhibited regulatory activity in transgenic zebrafish, however revealing no specificity. Our data show that the approach to identify cis-regulatory sequences by genome sequence alignments and subsequent testing in zebrafish transgenesis can be used to define enhancers within the Hox clusters and that these have significantly diverged in their function during evolution.     

Latitudinal variation in blue tit and great tit nest characteristics indicates environmental adjustment

Aim The laying of eggs and the building of a nest structure to accommodate them are two of the defining characteristics of members of the class Aves. Nest structures vary considerably across avian taxa and for many species the structure of the completed nest can have important consequences both for parents and their offspring. While nest characteristics are expected to vary adaptively in response to environmental conditions, large‐scale spatial variation in nest characteristics has been largely overlooked. Here, we examine the effects of latitudinal variation in spring temperatures on nest characteristics, including insulatory properties, and reproductive success of blue tits, Cyanistes caeruleus, and great tits, Parus major. Location Great Britain. Methods Nests and reproductive data were collected from seven study sites, spread over 5° of latitude. The nest insulatory properties were then determined before the nests were separated into nest base material and cup lining material. Results As spring temperatures increased with decreasing latitude, the mass of the nest base material did not vary in either species, while the mass of the cup lining material and nest insulatory properties decreased in both species. This suggests that in response to increasing temperatures the breeding female reduces the mass of the cup lining material, thereby maintaining an appropriate microclimate for incubating and brooding. The mean first egg date of both species advanced with decreasing latitude and increasing spring temperatures, although clutch size and brood size at hatching and fledging did not vary. Main conclusions This is the first study to demonstrate that the nest‐construction behaviour of birds varies in response to large‐scale spatial variation in ambient temperatures. Therefore, nest composition reliably indicates environmental conditions and we suggest that studies of nest structure may be sentinels for the early signs of rapid climate change.     

Climatic stability in the Brazilian Cerrado: implications for biogeographical connections of South American savannas,species richness and conservation in a biodiversity hotspot

Aim To investigate the historical distribution of the Cerrado across Quaternary climatic fluctuations and to generate historical stability maps to test: (1) whether the ‘historical climate’ stability hypothesis explains squamate reptile richness in the Cerrado; and (2) the hypothesis of Pleistocene connections between savannas located north and south of Amazonia. Location The Cerrado, a savanna biome and a global biodiversity hotspot distributed mainly in central Brazil. Methods We generated occurrence datasets from 1000 presence points randomly selected from the entire distribution of the Cerrado, as determined by two spatial definitions. We modelled the potential Cerrado distribution by implementing a maximum‐entropy machine‐learning algorithm across four time projections: current, mid‐Holocene (6 ka), Last Glacial Maximum (LGM, 21 ka) and Last Interglacial (LIG, 120 ka). We generated historical stability maps (refugial areas) by overlapping presence/absence projections of all scenarios, and checked consistencies with qualitative comparisons with available fossil pollen records. We built a spatially explicit simultaneous autoregressive model to explore the relationship between current climate, climatic stability, and squamate species richness. Results Models predicted the LGM and LIG as the periods of narrowest and widest Cerrado distributions, respectively, and were largely corroborated by palynological evidence. We found evidence for two savanna corridors (eastern coastal during the LIG, and Andean during the LGM) and predicted a large refugial area in the north‐eastern Cerrado (Serra Geral de Goiás refugium). Variables related to climatic stability predicted squamate richness better than present climatic variables did. Main conclusions Our results indicate that Bolivian savannas should be included within the Cerrado range and that the Cerrado’s biogeographical counterparts are not Chaco and Caatinga but rather the disjunct savannas of the Guyana shield plateaus. Climatic stability is a good predictor of Cerrado squamate richness, and our stability maps could be used in future studies to test diversity patterns and genetic signatures of different taxonomic groups and as a higher‐order landscape biodiversity surrogate for conservation planning.     

Phylogeography of the chestnut‐tailed antbird (Myrmeciza hemimelaena) clarifies the role of rivers in Amazonian biogeography

Aim We examined patterns of spatial and temporal diversification of the Amazonian endemic chestnut‐tailed antbird, Mymeciza hemimelaena (Thamnophilidae), to evaluate the diversification of a widespread avian taxon across rivers that potentially represent major natural barriers. Location Lowland Amazonia. Methods Sequences of the mitochondrial ND2 and cytochrome b genes were investigated from 65 individuals distributed throughout the entire range of M. hemimelaena, and including the two currently valid subspecies M. h. hemimelaena and M. h. pallens. Based on a combination of phylogeographic tools, molecular dating, and population genetic methods, we reconstructed a spatio‐temporal scenario of diversification of M. hemimelaena in the Amazon. Results The data revealed three genetically divergent and monophyletic groups in M. hemimelaena, which can also be distinguished by a combination of morphological and vocal characters. Two of these clades correspond to the previously described taxa M. h. hemimelaena and M. h. pallens, which are separated by the upper Madeira River, a main Amazonian tributary. The third clade is distributed between the middle reaches of the Madeira River and the much smaller tributaries Jiparaná and Aripuanã, and, although currently treated as M. h. pallens, clearly constitutes an independent evolutionary lineage probably deserving separate species status. Molecular clock and population genetic analyses indicate that diversification in this group occurred throughout the Pleistocene, with demographic fluctuations assumed for M. h. hemimelaena and M. h. pallens. Main conclusions The findings implicate rivers as barriers driving diversification in the M. hemimelaena complex. Levels of mitochondrial DNA divergence and associated morphological and vocal traits support its division into at least three separate species with comparatively small ranges. The existence of a previously unrecognized lineage in the M. hemimelaena complex, and the high degree of population structuring found in M. h. hemimelaena underscore the pervasiveness of cryptic endemism throughout Amazonia and the importance of DNA‐based taxonomic and phylogeographic studies in providing the accurate estimates of diversity that are essential for conservation planning.     

肝素钠的生产及其存在的问题及解决的方法（Ⅱ）

本文就粗品肝素钠生产的原料控制硬件设施管理和环保等方面进行了论述，介绍了一些改进的方法和措施，并就该方面的的一些问题进行了探讨，提出了解决的方法。     

A new species of Galearis (Orchidinae: Orchidaceae) from Sichuan,China

A new species, Galearis huanglongensis Q.W.Meng & Y.B.Luo, is described and illustrated. It is similar to Galearis cyclochila (Franch. & Sav.) Soó and Galearis diantha (Schltr.) P.F.Hunt, but differs in having a short spur, two elliptical lateral stigma lobes and distinctly separated bursicles. This new species is known only from the type locality, the Huanglong Valley, Songpan County, western Sichuan, China, growing amongst mosses under alpine shrubs at an elevation of about 3000 m. Based on two years of observations of its population size, the species was categorized as critically endangered CR (B1a, B2a) according to the World Conservation Union (IUCN) Red List Categories and Criteria, Version 3.1. The micromorphology of pollinia and seeds was observed by scanning electron microscopy and compared with that of G. cyclochila and G. diantha. The results supported G. huanglongensis Q.W.Meng & Y.B.Luo as a new species. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 689–695.     

珠江口伶仃洋中华白海豚栖息地利用对海岸线等变迁的响应

沿岸鲸豚类栖息地易受人类活动的干扰，导致其分布和核心栖息地发生变化。珠江口-漠阳江口中华白海豚种群是目前所知全球范围内最大的种群，其中伶仃洋水域是其重要的栖息地。近年来，珠江口伶仃洋周边城市发展带来的人类活动增加，白海豚的生存压力日益增大，分析伶仃洋中华白海豚对栖息地环境变化的响应，研究对应的保护策略显得非常迫切。以多源陆地资源卫星Landsat为数据源，通过影像分析近43年珠江口伶仃洋围填海造成的海域流失，结合近20年来采用截线抽样法收集的海豚观测数据，运用含障碍核插值（Kernel interpolation with barriers）方法，分析白海豚的分布及核心栖息地的变化。结果显示：1986-2015年期间，研究区域内流失的海域面积为344.08km²；目击分布离人工海岸线的平均距离大于自然海岸线的平均距离，目击分布到自然海岸线和人工海岸线的平均距离均在减小，表明过去20年白海豚的栖息地使用选择发生了一些变化，被迫适应人类活动的干扰；1997-2016年白海豚的分布范围呈现先增加后减小，白海豚栖息地使用的重心偏向伶仃洋东部水域，核心栖息地趋向主航道和无人海岛附近水域萎缩，可能是海豚因海域食物资源减少而迫不得已的选择。不同时期，珠江口中华白海豚国家级自然保护区所覆盖的核心栖息地比例呈递减趋势，占比由79.9%下降到49.4%，当前有必要对保护区范围和功能区作出一些优化调整，以适应栖息地使用的变化格局。