Genetic variation in Lumnitzera racemosa, a mangrove species from the Indo-West Pacific

The genetic structure of 18 populations of Lumnitzera racemosa from the Indo-West Pacific, including South China, Malay Peninsula, Sri Lanka, and North Australia, was assessed by inter simple sequence repeat (ISSR) markers. Our results showed a relatively high level of genetic variation at the species level (P = 87.04%, H_e = 0.260). The value of G_st was 0.642, suggesting significant genetic differentiation among populations. At the population level, however, genetic diversity was low (P = 32.17%, H_e = 0.097). When populations were grouped according to geographic regions, i.e., South China Sea, the East Indian Ocean, and North Australia, it was inferred from AMOVA that more than half the total variation (55.37%) was accounted for by differentiation between regions. A UPGMA dendrogram based on genetic distance also revealed a deep split between populations from these regions, indicating that Malay Peninsula and the Indonesia archipelago may play an important part on the genetic differentiation in L. racemosa. The high degree of population differentiation between regions and low genetic variation within populations recorded here highlights the need for appropriate conservation measures for this species, both in terms of incorporating further populations into protected areas, and the restoration strategies for separate regions.     

High genetic diversity vs. low genetic differentiation in Nouelia insignis (Asteraceae), a narrowly distributed and endemic species in China, revealed by ISSR fingerprinting

BACKGROUND AND AIMS: Nouelia insignis Franch., a monotypic genus of the Asteraceae, is an endangered species endemic in Yunnan and Sichuan Provinces of China. Most of the populations are seriously threatened. Some of them are even at the brink of extinction. In this study, the genetic diversity and differentiation between populations of this species were examined in two drainage areas. METHODS: DNA fingerprinting based on inter-simple sequence repeat polymorphisms was employed to detect the genetic variation and population structure in the species. KEY RESULTS: Genetic diversity at species level was high with P=65.05% (percentage of polymorphic loci) and Ht=0.2248 (total genetic diversity). The coefficient of genetic differentiation among populations, Gst, which was estimated by partitioning the total gene diversity, was 0.2529; whereas, the genetic differentiation between populations in the Jinsha and Nanpan drainage areas was unexpectedly low (Gst=0.0702). CONCLUSIONS: Based on the genetic analyses of the DNA fingerprinting, recent habitat fragmentation may not have led to genetic differentiation or the loss of genetic diversity in the rare species. Spatial apportionment of fingerprinting polymorphisms provides a footprint of historical migration across geographical barriers. The high diversity detected in this study holds promise for conservation and restoration efforts to save the endangered species from extinction.     

长筒石蒜种质资源的RAPD及ISSR研究

借助RAPD及ISSR分子标记对长筒石蒜的种质资源进行了初步的研究,结果表明：RAPD扩增得到的77个位点中,53个位点具有多态性,约占总数的68．8％;ISSR扩增得到的67个位点中,其中62个位点具有多态性,约占总数的92．5％。因此,长筒石蒜遗传多样性是十分丰富的,可以作为育种材料储备种质资源。从UPGMA聚类图来看,依据花色区分的3种类型,被聚在一起,表明亲缘关系较近。因此,在未来的长筒石蒜种质资源利用中,不同花色的长筒石蒜可以作为品系进行开发。     

Marker-assisted selection based on a multi-trait economic index in chicken: Experimental results and simulation

A method proposed herein allows simultaneous selection for several production traits, taking into consideration their marginal economic values (i.e. the economic value of a trait's additional unit). This economic index-marker assisted selection (EI-MAS) method is based on the calculation of the predicted economic breeding value (BV), using information on DNA markers that have previously been found to be associated with relevant quantitative trait loci. Based on the proposed method, results with real birds showed that sire progeny performance was significantly correlated with expected performance (r = 0.61-0.76; P = 0.03-0.01). Simulation analysis using a computer program written specifically for this purpose suggested that the relative advantage of EI-MAS would be large for traits with low heritability values. As expected, the response to EI-MAS was higher when the map distance between the marker and the quantitative trait gene was small, and vice versa. A large number of distantly located markers, spread 10 cM apart, yielded higher response to selection than a small number of closely located markers spread 3 cM apart. Additionally, the response to EI-MAS was higher when a large number (ca.150) of progeny was used for the prediction equation.     

A genome scan for loci affecting pork quality in a Duroc-Landrace F population

A genome scan was conducted on 370 F2 Duroc-Landrace pigs. Microsatellite markers (n = 182) were genotyped across the entire F2 population, all F1 parents and the paternal grandparents. Breed of origin of all chromosomal segments inherited in F2 progeny were predicted using GenoProb, where genotypic data, genetic maps and extended pedigrees were used as inputs. Statistical tests for quantitative trait loci (QTL) associations were conducted on 41 phenotypes with SAS using output from GenoProb for genotypic data. Fixed effects included sex and age at slaughter. For certain analyses carcass weight, RYR1 genotype and/or PRKAG3 genotype were also included as covariates. Subjective and objective measures of pork colour, marbling and tenderness were recorded, as well as measures of carcass fatness and muscularity. Test results were adjusted to a genome-wide level of significance. Five genomic regions presented significant evidence for QTL at chromosome 1 positions 6 cM (intramuscular fat) and 67 cM (Hunter L*), chromosome 2 position 62 cM (taste panel tenderness), chromosome 17 position 50 (loineye area and image analysis estimated loineye area) and X position 87 cM (carcass weight). Sixty-six suggestive associations were detected. Fourteen of these associations were within the regions with significant QTL on chromosomes 2, 17 and X, and the remaining 52 associations resided in 29 other regions on 13 different chromosomes of the porcine genome. The chromosome 2 region of 60-66 cM was associated with all measures of pork tenderness and the region on chromosome 17 (32-39 cM) was associated with both measures of intramuscular fat and loineye area. After verification, the QTL for marbling and tenderness should be useful in commercial production to improve pork quality as the population was developed from two of the three most utilized breeds of swine in the USA.     

Genetic diversity in European pigs utilizing amplified fragment length polymorphism markers

The use of DNA markers to evaluate genetic diversity is an important component of the management of animal genetic resources. The Food and Agriculture Organisation of the United Nations (FAO) has published a list of recommended microsatellite markers for such studies; however, other markers are potential alternatives. This paper describes results obtained with a set of amplified fragment length polymorphism (AFLP) markers as part of a genetic diversity study of European pig breeds that also utilized microsatellite markers. Data from 148 AFLP markers genotyped across samples from 58 European and one Chinese breed were analysed. The results were compared with previous analyses of data from 50 microsatellite markers genotyped on the same animals. The AFLP markers had an average within-breed heterozygosity of 0.124 but there was wide variation, with individual markers being monomorphic in 3-98% of the populations. The biallelic and dominant nature of AFLP markers creates a challenge for their use in genetic diversity studies as each individual marker contains limited information and AFLPs only provide indirect estimates of the allelic frequencies that are needed to estimate genetic distances. Nonetheless, AFLP marker-based characterization of genetic distances was consistent with expectations based on breed and regional distributions and produced a similar pattern to that obtained with microsatellites. Thus, data from AFLP markers can be combined with microsatellite data for measuring genetic diversity.     

HPLC analysis of discrete haptoglobin isoform N-linked oligosaccharides following 2D-PAGE isolation

Glycosylation is a common but variable modification that regulates glycoprotein structure and function. We combined small format 2D-PAGE with HPLC to analyse discrete human haptoglobin isoform N-glycans. Seven major and several minor haptoglobin isoforms were detected by 2D-PAGE. N-Glycans released from Coomassie-stained gel spots using PNGase were labeled at their reducing termini with 2-aminobenzamide. HPLC analysis of selected major isoform N-glycans indicated that sialic acid composition determined their separation by isoelectric focussing. N-Glycans from two doublets of quantitatively minor isoforms were also analysed. Although separation of each pair of doublets was influenced by sialylation, individual spots within each doublet contained identical N-glycans. Thus, heterogeneity in minor haptoglobin isoforms was due to modifications distinct from N-glycan structure. These studies describe a simple method for analysing low abundance protein N-glycans and provide details of discrete haptoglobin isoform N-glycan structures which will be useful in proteomic analysis of human plasma samples.     

Analysis of neuron-like differentiation of human bone marrow mesenchymal stem cells

The objective of the study was to evaluate differentiation of human bone marrow mesenchymal stem cells into true or pseudo neurons after treating with chemical induction medium in vitro. The morphological changes were assessed using interference contrast microscopy. Immunocytochemistry and Western blotting were performed using neuronal markers. Further evaluation was conducted with proteomic profiling, DNA microarray analysis and the whole-cell patch clamp test. After three hours of treatment with chemical induction medium, nearly three-fourths of the hMSCs changed to cells with a neuronal phenotype. The results of immunocytochemistry and Western blotting showed a high expression of neuronal markers in these cells at 3 h which decreased at 24 h. The proteomics analysis showed no change of proteins related to neuronal differentiation. DNA microarray showed downregulation of neuron related genes. The patch clamp test was unable to demonstrate any similarity to true neurons. Our findings suggest that neuron-like cells derived from chemical induction of hMSCs are not the genuine neurons as they resemble true neurons phenotypically but are different in genotypic and electrophysiological characteristics.     

A Study of Conservation Genetics in Cupressus chengiana,an Endangered Endemic of China,Using ISSR Markers

ISSR markers were used to analyze the genetic diversity and genetic structure of eight natural populations of Cupressus chengiana in China. ISSR analysis using 10 primers was carried out on 92 different samples. At the species level, 136 polymorphic loci were detected. The percentage of polymorphic bands (PPB) was 99%. Genetic diversity (H _e) was 0.3120, effective number of alleles (A _e) was 1.5236, and Shannon’s information index (I) was 0.4740. At the population level, PPB = 48%, A _e=1.2774, H _e=0.1631, and I=0.2452. Genetic differentiation (G _st) detected by Nei’s genetic diversity analysis suggested 48% occurred among populations. The partitioning of molecular variance by AMOVA analysis indicated significant genetic differentiation within populations (54%) and among populations (46%; P < 0.0003). The average number of individuals exchanged between populations per generation (N _m ) was 0.5436. Samples from the same population clustered in the same population-specific cluster, and two groups of Sichuan and Gansu populations were distinguishable. A significantly positive correlation between genetic and geographic distance was detected (r=0.6701). Human impacts were considered one of the main factors to cause the rarity of C. chengiana, and conservation strategies are suggested based on the genetic characters and field investigation, e.g., protection of wild populations, reestablishment of germplasm bank, and reintroduction of more genetic diversity.     

Genetic diversity among geographically separated populations of Nepenthes mirabilis

The distribution of Nepenthes mirabilis ranges from Northeast (NE) to South (S) Thailand. Eleven individuals from NE, S and Suen Jatujak market in Bangkok, Central (C) Thailand, were collected and divided into four populations according to their geographical areas. These four populations were analyzed to determine a genetic diversity profile using thirteen inter-simple sequence repeat markers. The individuals produced 75.18% polymorphic banding profiles. The Shannon’s index was used to estimate genetic diversity. Total genetic diversity (H _T) and inter-population genetic diversity (H _S) were 0.854 and 0.678, respectively. The degree of genetic differentiation (G _ST) of the species populations is 0.206, whereas the gene flow (Nm) among all the various geographical area populations is 1.016. Both the dendrogram and the results of the Shannon’s diversity index suggest great genetic diversity. These results support the broad range of distribution sites of Nepenthes mirabilis, which would require high genetic diversity to adapt to the environmental variations that can be found between NE, C, and S Thailand. ANOVA shows that the genetic diversity in each population is not significantly different (P > 0.05). Mantel tests reveal that geographical distance is an important factor for affecting the genetic distances among populations.