Persistence of Common Alleles in Two Related Populations or Species

Mathematical studies are conducted on three problems that arise in molecular population genetics. (1) The time required for a particular allele to become extinct in a population under the effects of mutation, selection, and random genetic drift is studied. In the absence of selection, the mean extinction time of an allele with an initial frequency close to 1 is of the order of the reciprocal of the mutation rate when 4Nv << 1, where N is the effective population size and v is the mutation rate per generation. Advantageous mutations reduce the extinction time considerably, whereas deleterious mutations increase it tremendously even if the effect on fitness is very slight. (2) Mathematical formulae are derived for the distribution and the moments of extinction time of a particular allele from one or both of two related populations or species under the assumption of no selection. When 4Nv << 1, the mean extinction time is about half that for a single population, if the two populations are descended from a common original stock. (3) The expected number as well as the proportion of common neutral alleles shared by two related species at the tth generation after their separation are studied. It is shown that if 4Nv is small, the two species are expected to share a high proportion of common alleles even 4N generations after separation. In addition to the above mathematical studies, the implications of our results for the common alleles at protein loci in related Drosophila species and for the degeneration of unused characters in cave animals are discussed.     

Total Polysaccharide Content of Developing Pollen in Two Angiosperm Species

The PAS reaction was used to stain total insoluble polysaccharides in whole pollen grains of anthers of Borago officinalis and Lycopersicum peruvianum from early microspore to ripe pollen stage. Total polysaccharide content was quantified with a microdensitometer: in Borago officinalis values showed two peaks, one during the microspore and the other during the binucleate stage; in Lycopersicum, there was a single peak during the latter stage. In both species total polysaccharide content decreased markedly in the last stage of pollen development before anthesis, when starch was hydrolysed and the polysaccharide reserves of the pollen transferred to the cytoplasm. Pollen grain volume was also determined at the various stages. It was found to increase, though with a different pattern in the two species, and to decrease before dehiscence. The results are discussed in terms of cytoembryological data of pollen grain development.     

Low Inbreeding and High Pollen Dispersal Distances in Populations of Two Amazonian Forest Tree Species

Recent studies suggest that tropical tree species exhibit low inbreeding and high gene dispersal levels despite the typically low density of conspecifics in tropical forests. To examine this, we undertook a study of pollen gene dispersal and mating system of two Amazonian tree species. We analyzed 341 seeds from 33 trees at four microsatellite loci in a Carapa guianensis population from Brazil, and 212 seeds from 22 trees at four microsatellite loci in a Sextonia rubra population from French Guiana. Differentiation of allele frequencies among the pollen pool of individual trees was Φ_FT= 0.053 (95% CI: 0.027–0.074) for C. guianensis and Φ_FT= 0.064 (95% CI: 0.017–0.088) for S. rubra. The mean pollen dispersal distances were estimated at 69–355 m for C. guianensis , and 86–303 m for S. rubra , depending on the pollen dispersal model and the estimate of reproductive tree density used. The multi-locus outcrossing rate was estimated at 0.918 and 0.945, and the correlation of paternity at 0.089 and 0.096, for C. guianensis and S. rubra , respectively, while no significant levels of biparental inbreeding were detected. Comparing trees with high and low local density of conspecifics, we found no evidence for differences in inbreeding levels. The results are discussed within the framework of the emerging picture of the reproductive biology of tropical forest trees.     

两种山鹪莺的春季换羽及尾羽变化

换羽是鸟类为保证持续生存的重要过程。换羽策略与鸟类进化及对环境的适应紧密相关,研究鸟类换羽特征,对于了解鸟类的分类、系统发育、进化历史及其对环境的适应性等方面都有重要意义。2007年3月至9月,在广东肇庆市江溪村对黄腹山鹪莺(Prinia flaviventris)和纯色山鹪莺(P.inornata)的春季换羽进行了研究。通过设置雾网捕捉2种山鹪莺,对捕捉到的成体进行体重及身体量度的测量;对飞羽及尾羽进行标记:初级飞羽以翅尖的第一枚羽毛标记为"P1",次级飞羽以翅中部最外一枚标记为"S1",向内依次递增标记;尾羽以中央两根最长尾羽为"T1",分别向两侧递增标记为"T2~T5"。采用单因素方差分析(One way ANOVA)对不同月份山鹪莺的体重值进行差异性检验,对体重与月份进行Pearson相关分析,对尾羽的长度和宽度进行Pearson偏相关分析(控制变量:体长)。研究结果表明:1)两种山鹪莺换羽期为3至5月,持续时间约为60 d;2)两种山鹪莺春季换羽仅更换尾羽,换羽模式均为离心型,即中央一对尾羽最先开始替换,然后向两侧由内到外逐次更替;3)两种山鹪莺的尾羽长度和宽度同步变化,但绝大部分山鹪莺非繁殖期尾羽长度与繁殖期尾羽长度之比大于非繁殖期尾羽宽度与繁殖期尾羽宽度之比,即繁殖期尾羽相对较宽;4)两种山鹪莺换羽期间体重大致呈现下降趋势,但变化不显著(P0.05)。推测两种山鹪莺通过增加食物的摄入来抵抗换羽期和繁殖期重叠而导致的能量消耗,这可能与该地区丰富的食物资源有关,并在一定程度上体现了两种山鹪莺换羽策略对环境的适应性。     

Gene Flow among Populations of Two Rare Co-Occurring Fern Species Differing in Ploidy Level

Differences in ploidy levels among different fern species have a vast influence on their mating system, their colonization ability and on the gene flow among populations. Differences in the colonization abilities of species with different ploidy levels are well known: tetraploids, in contrast to diploids, are able to undergo intra-gametophytic selfing. Because fertilization is a post-dispersal process in ferns, selfing results in better colonization abilities in tetraploids because of single spore colonization. Considerably less is known about the gene flow among populations of different ploidy levels. The present study examines two rare fern species that differ in ploidy. While it has already been confirmed that tetraploid species are better at colonizing, the present study focuses on the gene flow among existing populations. We analyzed the genetic structure of a set of populations in a 10×10 km study region using isoenzymes. Genetic variation in tetraploid species is distributed mainly among populations; the genetic distance between populations is correlated with the geographical distance, and larger populations host more genetic diversity than smaller populations. In the diploid species, most variability is partitioned within populations; the genetic distance is not related to geographic distance, and the genetic diversity of populations is not related to the population size. This suggests that in tetraploid species, which undergo selfing, gene flow is limited. In contrast, in the diploid species, which experience outcrossing, gene flow is extensive and the whole system behaves as one large population. Our results suggest that in ferns, the ability to colonize new habitats and the gene flow among existing populations are affected by the mating system.     

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

The investigation of the intracellular protein levels of bacterial species is of importance to understanding the pathogenic mechanisms of diseases caused by these organisms. Here we describe a procedure for protein extraction from Burkholderia species based on mechanical lysis using glass beads in the presence of ethylenediamine tetraacetic acid and phenylmethylsulfonyl fluoride in phosphate buffered saline. This method can be used for different Burkholderia species, for different growth conditions, and it is likely suitable for the use in proteomic studies of other bacteria. Following protein extraction, a two-dimensional (2-D) gel electrophoresis proteomic technique is described to study global changes in the proteomes of these organisms. This method consists of the separation of proteins according to their isoelectric point by isoelectric focusing in the first dimension, followed by separation on the basis of molecular weight by acrylamide gel electrophoresis in the second dimension. Visualization of separated proteins is carried out by silver staining.     

Identification of Two Distinct Populations of Protein Kinase C in Rat Brain Membranes

The regulatory enzyme protein kinase C (PKC) is proposed to be activated on its translocation from the cytosol to the membrane. However, a portion of the native activity is always associated with the membrane fraction. Using a noninvasive procedure to extract this endogenous activity from rat brain membranes, it has been possible to characterize the activity in a partially purified reconstituted system bearing resemblance to the in vivo system. Two subpopulations of membrane-associated PKC were identified and characterized at the level of activation, inhibition, and isozyme immunologic characteristics and chromatographic properties. One peak had properties similar to those of cytosolic PKC, whereas the second population, extracted as protein-lipid complexes, had considerable constitutive activity that could be stimulated further on addition of PKC activators. This latter activity was relatively resistant to staurosporine inhibition and phorbol ester treatment, but it phosphorylated the exogenous PKC substrates, histone 1 and the epidermal growth factor receptor peptide KTRLRR. The constitutive activity was totally dependent on its endogenous associated lipids coextracted by the solubilization procedure. The ratio between these two populations was ontogenetically regulated and modulated by phorbol ester treatment, suggesting that different PKC populations may serve unique functions in the rat brain regulated by the lipid environment. Analyses of the phospholipids extracted in these protein-lipid complexes showed differences in the major classes correlating to age. However, apart from a markedly lower cholesterol content in these complexes, no direct relationship between a specific lipid composition and the amount of constitutive PKC activity was evident.     

Protein Profiling in Hepatocellular Carcinoma by Label-Free Quantitative Proteomics in Two West African Populations

Background

Hepatocellular Carcinoma is the third most common cause of cancer related death worldwide, often diagnosed by measuring serum AFP; a poor performance stand-alone biomarker. With the aim of improving on this, our study focuses on plasma proteins identified by Mass Spectrometry in order to investigate and validate differences seen in the respective proteomes of controls and subjects with LC and HCC.

Methods

Mass Spectrometry analysis using liquid chromatography electro spray ionization quadrupole time-of-flight was conducted on 339 subjects using a pooled expression profiling approach. ELISA assays were performed on four significantly differentially expressed proteins to validate their expression profiles in subjects from the Gambia and a pilot group from Nigeria. Results from this were collated for statistical multiplexing using logistic regression analysis.

Results

Twenty-six proteins were identified as differentially expressed between the three subject groups. Direct measurements of four; hemopexin, alpha-1-antitrypsin, apolipoprotein A1 and complement component 3 confirmed their change in abundance in LC and HCC versus control patients. These trends were independently replicated in the pilot validation subjects from Nigeria. The statistical multiplexing of these proteins demonstrated performance comparable to or greater than ALT in identifying liver cirrhosis or carcinogenesis. This exercise also proposed preliminary cut offs with achievable sensitivity, specificity and AUC statistics greater than reported AFP averages.

Conclusions

The validated changes of expression in these proteins have the potential for development into high-performance tests usable in the diagnosis and or monitoring of HCC and LC patients. The identification of sustained expression trends strengthens the suggestion of these four proteins as worthy candidates for further investigation in the context of liver disease. The statistical combinations also provide a novel inroad of analyses able to propose definitive cut-offs and combinations for evaluation of performance.     

Changes in Protein Synthesis in Rapeseed (Brassica napus) Seedlings during a Low Temperature Treatment

Changes induced by cold treatment in young rapeseed (Brassica napus) seedlings were investigated at the molecular level. Following germination at 18°C for 48 hours, one half of the seedlings was transferred to 0°C for another 48 hour period, the other half being kept at 18°C as a control. Newly synthesized proteins were labeled for the last 6 hours of incubation with [³⁵S]methionine. The different polypeptides were separated by two-dimensional electrophoresis in polyacrylamide gels. Newly synthesized proteins were revealed by fluorography. Protein synthesis clearly continues at 0°C and some polypeptides preferentially accumulate at this temperature. On the other hand, synthesis of several others is repressed while many are insensitive to cold treatment. Similar changes are also observed when mRNA is prepared from cold treated seedlings, translated in vitro in a reticulocyte cell free system and compared with the products of mRNA extracted from control samples. Among the genes which are repressed we identified the small subunit of ribulose 1,6-bisphosphate carboxylase. These changes are also detectable after shorter treatments.     

Changes in Whistle Structure of Two Dolphin Species During Interspecific Associations

Dolphin communicative signals show great plasticity. Dolphins modify signal structure to cope with their environment, in response to stress, and in some species to mimic group members. Hence, whistle structure variations may offer insights to interspecific associations among dolphin species, which although temporal and opportunistic are common. In this study, I test the hypothesis that interspecific interactions influence dolphin whistle structure, particularly during social events. The study took place in the Southern Caribbean coast of Costa Rica, where interspecific associations of the distantly related Guyana and Bottlenose dolphins occur on daily basis. The results indicate that interspecific groups emit whistles that show intermediate whistle structure compared to whistles emitted in intraspecific groups. This pattern is seen during social interactions between species, but not when interspecific groups are traveling. Social events in interspecific groups were of antagonistic nature, where Bottlenose dolphins isolated and harassed one or two Guyana dolphins. Contour data suggest that the most vocal species during these encounters was the Guyana dolphin. Therefore, the observed modifications in whistles structure likely reflect a stress response by the Guyana dolphins. Another alternative explanation includes signal convergence between interacting species. However, to understand the nature of these potential modifications, future studies should combine acoustic tags and directional recording systems to follow the vocalizing animals. Despite the shortcomings of this study, it provides some of the first insights into dolphin interspecific communication, providing evidence of overall signal change during interspecific interactions.     

Admixture in Humans of Two Divergent Plasmodium knowlesi Populations Associated with Different Macaque Host Species

Human malaria parasite species were originally acquired from other primate hosts and subsequently became endemic, then spread throughout large parts of the world. A major zoonosis is now occurring with Plasmodium knowlesi from macaques in Southeast Asia, with a recent acceleration in numbers of reported cases particularly in Malaysia. To investigate the parasite population genetics, we developed sensitive and species-specific microsatellite genotyping protocols and applied these to analysis of samples from 10 sites covering a range of >1,600 km within which most cases have occurred. Genotypic analyses of 599 P. knowlesi infections (552 in humans and 47 in wild macaques) at 10 highly polymorphic loci provide radical new insights on the emergence. Parasites from sympatric long-tailed macaques (Macaca fascicularis) and pig-tailed macaques (M. nemestrina) were very highly differentiated (FST = 0.22, and K-means clustering confirmed two host-associated subpopulations). Approximately two thirds of human P. knowlesi infections were of the long-tailed macaque type (Cluster 1), and one third were of the pig-tailed-macaque type (Cluster 2), with relative proportions varying across the different sites. Among the samples from humans, there was significant indication of genetic isolation by geographical distance overall and within Cluster 1 alone. Across the different sites, the level of multi-locus linkage disequilibrium correlated with the degree of local admixture of the two different clusters. The widespread occurrence of both types of P. knowlesi in humans enhances the potential for parasite adaptation in this zoonotic system.     

Parallel Patterns of Morphological and Behavioral Variation among Host-Associated Populations of Two Gall Wasp Species

A powerful approach to address the general factors contributing to ecological speciation is to compare distantly related taxa that inhabit the same selective environments. In this design, similarities among taxa can elucidate general mechanisms of the process whereas differences may uncover specific factors important to the process for individual taxa. Herein, we present evidence of parallel patterns of morphological and behavioral variation among host-associated populations of two species of cynipid gall wasps, Belonocnema treatae and Disholcaspis quercusvirens, that each exhibit a life cycle intimately tied to the same two host plant environments, Quercus geminata and Q. virginiana. Across both gall-former species we find consistent differences in body size and gall morphology associated with host plant use, as well as strong differences in host plant preference, a measure of habitat isolation among populations. These consistent differences among taxa highlight the important role of host plant use in promoting reproductive isolation and morphological variation among herbivorous insect populations–a prerequisite for ecological speciation.     

两种亚热带榕属乔木植冠损伤后的位置依赖性枝发生

三峡库区需要大量植株用于三峡工程所涉及的诸多建设(如铁路、公路、建筑)之后的植被恢复和绿化.由于具有优美树冠和耐瘠薄土壤的能力,榕(Ficus microcarpa L.)和黄桷树(Fvirens Ait.var.sublanceolata(Miq.)Cornor) 在三峡库区作为绿化和行道树木大量栽植.在三峡库区,这两种树种苗木的培育主要通过切枝扦插的方式进行.大量切枝损伤植株植冠并且使叶组织数量减少,对植株生长有很大影响.植株生长与植株的枝发生格局有很大关系,为明确植冠损伤对植株生长的可能影响,对榕和黄桷树植冠损伤后的枝发生进行了研究.实验发现,切枝造成的植冠损伤对榕和黄桷树植株侧枝上的枝发生没有影响,不同损伤强度之间和不同损伤发生时间之间都不存在明显的差异.但是,植冠损伤后,榕和黄桷树植株主茎上的枝发生是位置依赖性的.植冠损伤不影响榕和黄桷树植株损伤处理后新生主茎段和具侧枝主茎段上的枝发生,却促进了损伤处理后裸露主茎段上的枝发生,并且裸露主茎段上枝发生的数量和密度随植冠损伤强度的升高而增大.此外,实验结果表明,秋季植冠损伤植株裸露主茎段上的枝发生要高于春季植冠损伤植株裸露主茎段上的枝发生;在榕和黄桷树的枝发生中,仅有裸露主茎段上增强的枝发生会促进植冠损伤植株对叶组织的生物量投资并有利于受损植株的恢复和生长.     

两种亚热带榕属乔木植冠损伤后的位置依赖性枝发生

三峡库区需要大量植株用于三峡工程所涉及的诸多建设(如铁路、公路、建筑)之后的植被恢复和绿化。由于具有优美树冠和耐瘠薄土壤的能力,榕(Ficus microcarpa L.)和黄桷树(F. virens Ait. var. sublanceolata (Miq.) Cornor)在三峡库区作为绿化和行道树木大量栽植。在三峡库区,这两种树种苗木的培育主要通过切枝扦插的方式进行。大量切枝损伤植株植冠并且使叶组织数量减少,对植株生长有很大影响。植株生长与植株的枝发生格局有很大关系,为明确植冠损伤对植株生长的可能影响,对榕和黄桷树植冠损伤后的枝发生进行了研究。实验发现,切枝造成的植冠损伤对榕和黄桷树植株侧枝上的枝发生没有影响,不同损伤强度之间和不同损伤发生时间之间都不存在明显的差异。但是,植冠损伤后,榕和黄桷树植株主茎上的枝发生是位置依赖性的。植冠损伤不影响榕和黄桷树植株损伤处理后新生主茎段和具侧枝主茎段上的枝发生,却促进了损伤处理后裸露主茎段上的枝发生,并且裸露主茎段上枝发生的数量和密度随植冠损伤强度的升高而增大。此外,实验结果表明, 秋季植冠损伤植株裸露主茎段上的枝发生要高于春季植冠损伤植株裸露主茎段上的枝发生; 在榕和黄桷树的枝发生中,仅有裸露主茎段上增强的枝发生会促进植冠损伤植株对叶组织的生物量投     

两种热带雨林树苗对环境光强变化的生理响应和适应机制

干季末雨季初以西双版纳热带雨林中木奶果和玉蕊2种树苗为材料,研究了将生长于12.5%自然光（相当于小林窗的光强）和36%自然光（相当于大林窗的光强）下的这2种树苗分别移至36%自然光和12.5%自然光下之后各自叶片最大净光合速率（Pmax）、叶绿素荧光参数、光合色素含量、比叶重（LMA）以及叶片悬挂角（MA）的变化过程,探讨了2种植物幼苗在生长环境光强改变后其形态和生理生态特性做出的相应调整以适应新的光环境的过程与机制。结果表明,这2种树苗均不适宜生长在大林窗的强光环境下,但木奶果对光环境的增强表现出一定的耐受力,光合适应潜力强于玉蕊,2种树苗对低光环境都能较好地适应;新叶的生成在整株植物对生长光环境变化的适应过程中也起到至关重要的作用。     

SDS-PAGE Pattern Polymorphism of X-Chromosomal Glue Proteins in Natural Populations of Two Drosophila nasuta Subgroup Species

For the first time, the glue protein patternpolymorphism in natural populations of D. n.nasuta and D. s. neonasuta has beenanalyzed by SDS-PAGE. The study involving 200 and 185isofemale lines comprising 2028 and 1900 individuals of D. n.nasuta and D. s. neonasuta, respectively,revealed the occurrence of eight variant glue proteinphenotypes in D. n. nasuta and seven in D.s. neonasuta. The number and frequency of variant phenotypes in differentpopulations of both species were found to vary. Analysisof glue protein patterns in the F₁ progeny ofcrosses involving parents with variant glue proteinphenotypes revealed that the polymorphic fractions areproduced by co-dominant genes located on the Xchromosome. More than 87% of the naturally inseminatedadult females were found to produce polymorphic progeny. The heterozygous female larvae were found toexceed the homozygotes in the isofemale line progeny ofmost of the populations.