Assessment of genetic diversity in Calamus thwaitesii BECC. (Arecaceae) using RAPD markers

Calamus thwaitesii Becc. is a potentially useful rattan found in the Western Ghats of India and Sri Lanka. The wild stock of this rattan species is greatly diminished due to overexploitation for the furniture industry and increasingly rare. Genetic diversity was estimated in 80 samples representing eight populations from the Western Ghats and Sri Lanka using Random Amplified Polymorphic DNA (RAPD) markers. RAPDs generated a total of 120 markers with 10 decamer primers, of which 85% were found to be polymorphic. The percentage of polymorphic loci varied from 40.00 to 60.83 and genetic distance between populations ranged from 0.0332 to 0.2777. Among the analysed populations, Goa was found to be genetically superior followed by Achenkovil, Sinharaja and Talakkaveri. Majority of the genetic diversity was distributed within populations (70.79%) and only (29.21%) among populations. Genetic relationships estimated by the unweighted pair-group method with arithmetic averaging (UPGMA) cluster analysis and principal co-ordinate analysis failed to separate Indian and Sri Lankan populations geographically into two distinct groups.     

Comparative Assessment of Genetic Variability in the Populations of Endemic and Endangered Yellow Catfish, Horabagrus brachysoma (Teleostei: Horabagridae), Based on Allozyme, RAPD, and Microsatellite Markers

The comparative assessment of genetic diversity using allozymes, random amplified polymorphic DNA (RAPD), and microsatellite markers was conducted in endemic and endangered yellow catfish (Horabagrus brachysoma) sampled from three locations in Western Ghats river systems of India. Among the three markers, microsatellites show more polymorphism, having 100% polymorphic loci, whereas allozymes show the least (56%). In RAPD, 60.5% of fragments were polymorphic. Observed heterozygosity and F(ST) values were very high in microsatellites, compared with the other markers. Microsatellite and RAPD markers reported a higher degree of genetic differentiation than allozymes among the populations depicted by pairwise F(ST)/G(ST), AMOVA, Nei's genetic distance, and UPGMA dendrogram. The three classes of markers demonstrated striking genetic differentiation between pairs of H. brachysoma populations. The data emphasize the need for fishery management, conservation, and rehabilitation of this species.     

Genetic variation and population structure of endemic yellow catfish, <Emphasis Type="Italic">Horabagrus</Emphasis><Emphasis Type="Italic">brachysoma</Emphasis> (Bagridae) among three populations of Western Ghat region using RAPD and microsatellite markers

Random amplified polymorphic DNA (RAPD) and microsatellite markers were applied to evaluate the genetic variation in endemic and endangered yellow catfish, Horabagrus brachysoma sampled from three geographic locations of Western Ghat, South India river systems. In RAPD, of 32 10-mer RAPD primers screened initially, 10 were chosen and used in a comparative analysis of H. brachysoma collected from Meenachil, Chalakkudy and Nethravathi River systems. Of the 124 total RAPD fragments amplified, 49 (39.51%) were found to be shared by individuals of all 3 populations. The remaining 75 fragments were found to be polymorphic (60.48%). In microsatellites, six polymorphic microsatellite loci were identified by using primers developed for Pangasius hypophthalmus, Clarias macrocephalus and Clarias gariepinus. The identified loci were confirmed as microsatellite by sequencing after making a clone. The nucleotide sequences of 6 loci were published in NCBI genbank. The number of alleles across the six loci ranged from 4 to 7 and heterozygosities ranged from 0.07 to 0.93. The mean number of alleles and effective number of alleles per locus were 5.00 and 3.314, respectively. The average heterozygosity across all investigated samples was 0.72, indicating a significant deficiency of heterozygotes in this species. RAPD and microsatellite methods reported a high degree of gene diversity and genetic distances depicted by UPGMA dendrograms among the populations of H. brachysoma.     

Genetic diversity of common carp from two largest Chinese lakes and the Yangtze River revealed by microsatellite markers

Six polymorphic microsatellites (eight loci) were used to study the genetic diversity and population structure of common carp from Dongting Lake (DTC), Poyang Lake (PYC), and the Yangtze River (YZC) in China. The gene diversity was high among populations with values close to 1. The number of alleles per locus ranged from 2 to 11, and the average number of alleles among 3 populations ranged from 6.5 to 7.9. The mean observed (H _O) and expected (H _E) heterozygosity ranged from 0.4888 to 0.5162 and from 0.7679 to 0.7708, respectively. Significant deviations from Hardy–Weinberg Equilibrium expectation were found at majority of the loci and in all three populations in which heterozygote deficits were apparent. The analysis of molecular variance (AMOVA) indicated that the percent of variance among populations and within populations were 3.03 and 96.97, respectively. The Fst values between populations indicated that there were significant genetic differentiations for the common carp populations from the Yangtze River and two largest Chinese freshwater lakes. The factors that may result in genetic divergence and significant reduction of the observed heterozygosity were discussed.     

Assessment of Genetic Relatedness of Crossbred Chicken Populations Using Microsatellite Markers

To measure genetic relatedness between populations, for breeding purposes, we analyzed 170 birds from six crossbred populations of three pure lines of White Leghorn chickens, using 14 microsatellite markers. All the microsatellites were polymorphic, with 2–6 alleles. The mean number of alleles per locus was 3.21. The effective number of alleles varied from 1.14 to 3.94. The observed heterozygosity varied from 0.133 to 1.00, with a mean of 0.748. The F _IS values were mostly negative, with an average of ?0.345. The mean F _ST value was 0.056. The Nm values ranged from 1.91 to 42.17. The highest genetic identity was observed between IWI × IWK and IWK × IWI. The relation between any two groups of crosses was more than 85%. The results suggest that the crossbred populations were very closely related.     

Genetic variation and phylogenetic relationship between two species of yellow catfish, <Emphasis Type="Italic">Horabagrus brachysoma</Emphasis> and <Emphasis Type="Italic">H. nigricollaris</Emphasis> (Teleostei: Horabagridae) based on RAPD and microsatellite markers

The two species of yellow catfish, Horabagrus brachysoma and H. nigricollaris are categorized as ‘endangered’ and ‘critically endangered’ respectively in their wild habitat. Proper knowledge of genetic structure and variability of these endangered species are highly essential for the management, conservation and improvement of fish stocks. Therefore, genetic variation and phylogenetic relationships between these species of yellow catfish sampled from Chalakkudy River in the hot spot of biodiversity-Western Ghats region, Kerala, India were analyzed by using Random amplified polymorphic DNA (RAPD) and microsatellite markers. 85 RAPD and five microsatellites loci were detected to analyze the genetic variation and phylogenetic relationships among these species. Out of 85 RAPD loci produced only 52.94% were polymorphic whereas in microsatellite, all 5 loci were polymorphic (100%). Species-specific RAPD bands were found in both species studied. In microsatellite, the number of alleles across the five loci ranged from 1 to 8. The observed heterozygosities in H. brachysoma and H. nigricollaris were 0.463 and 0.443, respectively. Here, both RAPD and microsatellite methods reported a low degree of gene diversity and lack of genetic heterogeneity in both species of Horabagrus which strongly emphasize the need of fishery management, conservation and rehabilitation of these species.     

Genetic structure and diversity of Coscinium fenestratum: a critically endangered liana of Western Ghats,India

Coscinium fenestratum is a critically endangered medicinal plant, well-known for its bioactive isoquinoline alkaloid berberine. The species has been over harvested from its natural habitats to meet the huge requirement of raw drug market and industrial consumption. This has lead to a rapid decline in the population size and has also led to local population extinction at few locations in the Western Ghats, India. In this study, inter-simple sequence repeat markers were used to investigate the genetic variation and population structure of seven extant populations of C. fenestratum from the central Western Ghats, India. Eight primer combination produced a total of 57 unambiguous bands, of which (47.1 %) were polymorphic. The species exhibited a moderate to low level of intra population genetic diversity (H _s = 0.347 ± 0.008; H _t = 0.378 ± 0.006 (POPGENE) and H _s = 0.262 ± 0.0028; H _t = 0.204 ± 0.020 (HICKORY)). The populations were low to moderately differentiated from one another (G _ST = 0.221) and geographical distance was not significantly correlated with genetic distance, suggesting that these long-lived, geographically distant remnant populations were once connected through gene flow. There was a significant amount of genetic variation among populations (19.85 %). The Bayesian software STRUCTURE and HICKORY were used to further reveal the genetic structure of C. fenestratum. The results revealed weak population structure (K = 2) with one single widespread gene pool, and indicated that gene flow and inbreeding are likely to be the major driving force in shaping current population genetic structure of C. fenestratum. Thus, an understanding of the genetic diversity and population structure of C. fenestratum can provide insight into the conservation and management of this species.     

Isolation and characterization of ten polymorphic microsatellite loci in Ixodes arboricola, and cross-amplification in three other Ixodes species

We characterized ten polymorphic microsatellite loci from the tree-hole tick, Ixodes arboricola. Loci were screened in 11–18 individuals from three Belgian populations and five to ten alleles were found at each locus. Seven loci did not show deviations from Hardy–Weinberg equilibrium conditions and there were no indications for null alleles at these loci. The three other loci showed significant heterozygote deficiencies in at least one population, and a high potential for the occurrence of null alleles. We observed no effect of potential host DNA on the scoring of the microsatellites. Cross-amplification of the microsatellites was tested in eight specimens of three congeneric species: I. ricinus, I. hexagonus and I. frontalis. Depending on the species, six or seven of the loci were amplified in ≥4 of the 8 specimens and were polymorphic in each of these species (except for Ixaf 11 in I. frontalis and I. ricinus). These loci thus provide a tool for population genetic analysis of I. arboricola. The suitability of these markers needs to be further investigated in its congeners.     

Development of compound microsatellite markers in the harmful dinoflagellate Cochlodinium polykrikoides (Dinophyceae)

We isolated 15 polymorphic microsatellites from Cochlodinium polykrikoides. These loci provide a class of highly variable genetic markers, as the number of alleles ranged from two to 15, and the estimate of gene diversity was from 0.083 to 0.880 across the 15 microsatellites. We consider that these loci have a potential to reveal the genetic structure and gene flow among C. polykrikoides populations.     

Development of polymorphic microsatellite loci in Nothapodytes nimmoniana, a medicinally important tree from the Western Ghats, India

Nothapodytes nimmoniana is a medicinally important tree species that occur in the Western Ghats, a megadiversity hotspot in southern India. Inner stem bark of the tree contains an important anti-cancer alkaloid, camptothecin for which the natural population of the tree is heavily harvested. In this paper, we report the isolation and characterization of eight polymorphic microsatellite loci using enrichment hybridization protocol. Analysis of 36 individuals representing two populations revealed three to 12 alleles per locus. Observed heterozygosity ranged from 0.21 to 0.94 for the two populations. None of the loci tested showed linkage disequilibrium. These markers are invaluable for evaluating the genetic structure and assessing the genetic impacts of harvesting of N. nimmoniana in the Western Ghats to formulate strategies for conservation of the species.     

THIS ARTICLE HAS BEEN RETRACTEDDevelopment of microsatellite markers in the noxious red tide‐causing dinoflagellate Heterocapsa circularisquama (Dinophyceae)

We isolated 15 polymorphic microsatellites from one of the most noxious red tide‐causing dinoflagellate species, Heterocapsa circularisquama. These loci provide one class of highly variable genetic markers, as the number of alleles ranged from two to six, and the estimate of gene diversity was from 0.205 to 0.684 across the 15 microsatellites. These loci have the potential to reveal genetic structure and gene flow among H. circularisquama populations.     

Isolation and characterization of microsatellite loci in the striped mullet,Mugil cephalus

Forty‐three microsatellites were isolated from grey mullet (Mugil cephalus) of Mediterranean origin using the standard enrichment methods followed by enhanced chemiluminescence for the rapid identification of clones containing microsatellites. Eleven microsatellites were characterized to assess genetic diversity of different populations surrounding the island of Sardinia in Italy. The versatility of these markers was also assessed using an Australian population. All 11 microsatellites were highly polymorphic with a mean heterozygosity of 0.834. The number of alleles ranged from 14 to 38 in the Sardinian populations and from 11 to 25 in the Australian samples.     

Development of microsatellite markers for the Manila clam Ruditapes philippinarum

We isolated nine polymorphic microsatellites from the Manila clam Ruditapes philippinarum. These loci provide one class of highly variable genetic marker as the number of alleles ranged from six to 22, and the observed and expected heterozygosity ranged from 0.136 to 0.909 and from 0.553 to 0.954, respectively. We consider that these loci are potentially useful for detailing the genetic structure and gene flow among R. philippinarum populations.     

Development and characterization of microsatellite markers in Indian sesame (Sesamum indicum L.)

The genetic characterization of Indian sesame cultivars and related wild species was analysed using 102 simple sequence repeat (SSR; microsatellite) markers. Of these, 62 were novel sesame-specific microsatellites isolated in the course of the present investigation by constructing genomic libraries. Characterization of the 68 sesame accessions and three related wild species using 72 polymorphic SSR primers resulted in the detection of 170 alleles. The number of alleles ranged from two to four with an average of 2.5 alleles per locus. Polymorphic information content of the markers ranged from 0.43 to 0.88 with an average of 0.66. UPGMA cluster analysis grouped all the accessions into two major clusters with a genetic similarity ranging from 0.40 to 0.91. A moderate to high level of genetic variability was observed. The three wild accessions used in the study formed separate clades and distant genetic relationships were observed between the cultivar lines and wild species. Differentiation of genotypes according to geographical region was not observed. Analysis of molecular variance (AMOVA) analysis revealed that a high percentage of variation was within populations (87.1 %). An overall F _st of 0.11 among the populations indicated low population differentiation. The SSR markers developed will be useful for further genetic analysis, linkage mapping and selection of parents in future breeding programmes.     

Development of microsatellite markers for the crown‐of‐thorns starfish Acanthaster planci

We isolated nine polymorphic microsatellites from the crown‐of‐thorns starfish, Acanthaster planci. These loci provide one class of highly variable genetic marker as the number of alleles ranged from three to 12 and the observed and expected heterozygosities ranged from 0.130 to 0.783 and from 0.163 to 0.862, respectively. We consider that these loci are potentially useful for detailing the genetic structure and gene flow among A. planci populations.     

Microsatellite diversity and population genetic structure of yellowcheek, Elopichthys bambusa (Cyprinidae) in the Yangtze River

Yellowcheek carp (Elopichthys bambusa) is the only species of genus Elopichthys. It is widely distributed in Chinese freshwaters but currently its populations have declined to threatening level. We examined the genetic diversity and population structure of E. bambusa in the Yangtze River basin. A total of nine polymorphic microsatellite markers were employed to study five populations occurring in middle and lower reaches of the river. The results revealed low-to-moderate genetic diversity. The number of alleles per locus varied between 3 and 8 with an average of 4.8. Observed heterozygosity ranged from 0.15 to a maximum of 1.00. Significant deviations (P < 0.01) from Hardy–Weinberg equilibrium were observed for all the tested locus-population combinations with clear heterozygosity deficits. AMOVA indicated that majority of the variance lies within populations (93.81%) than among the populations (7.05%). Pairwise F_ST and unbiased genetic distance pointed out significant differentiation among the samples from populations with different connections to the Yangtze River. In the UPGMA dendrogram, clustering pattern of populations indicated that most of the populations are reproductively isolated due to anthropogenic interventions. Clustering of PYL and DTL populations shows ongoing gene flow through the mainstream. The recent hydrological alterations and overfishing are major factors shaping the current genetic structure. These results can be helpful for effective management and sustainable conservation of E. bambusa populations.     

Twenty-one new microsatellite markers for the ecologically important midge Heterotrissocladius marcidus,and their use in studies of alpine lakes

Heterotrissocladius marcidus (Walker, 1856) (Diptera: Chironomidae) inhabits aquatic habitats across the Holarctic and Oriental regions, including remote alpine glacial lakes. It is thus a promising species for studies of population structure and genetic diversity in these highly vulnerable habitats. Here, 21 polymorphic microsatellites for H. marcidus are described, obtained using 454 GS-FLX pyrosequencing. The number of alleles per locus varied from 2 to 9 and no significant linkage disequilibrium was observed. To test applicability, 40 individuals were used from two lakes in the Tatra Mts (Western Carpathians). Observed and expected heterozygosity varied from 0 to 0.9 and 0.102 to 0.888, respectively. In both populations, three loci deviated significantly from Hardy–Weinberg equilibrium after Bonferroni correction, probably due to presence of null alleles or undetected biological processes. Application of microsatellites was tested on six alpine lake populations. Bayesian cluster analysis assigned individuals to lakes of their origin and revealed limited gene-flow between them. Five loci were successfully cross-amplified in the related midge Macropelopia sp. (Tanypodinae) and two in Pseudodiamesa branickii (Nowicki, 1873) (Diamesinae). The microsatellites described herein proved to be useful for genetic studies of alpine populations, and can provide important data for management and conservation of these threatened habitats.     

Reproductive biology of the endemic cyprinid fish Hypselobarbus thomassi (Day, 1874) from Kallada River in the Western Ghats,India

The study provides new information on the reproductive biology of Hypselobarbus thomassi (Day, 1874) an endemic cyprinid fish in the Western Ghats–a biodiversity hotspot of India. A total of 384 specimens (males = 244 and females = 140) were collected monthly from April 2009 to March 2011 from Kallada River of Southern part of Western Ghats, India. Monthly sex ratio indicated that males predominated in the fishery, and an overall sex ratio was significantly different from the expected value of 1:1 (male:female = 1:0.57, x² = 28.17, p < .01). Based on the monthly analysis of gonadosomatic index (GSI) and the proportion of mature individuals, the spawning season of H. thomassi occurred between May to October with a peak in June to August. The length at first maturity (L₅₀) recorded was 290 mm TL in males and 330 mm TL in females. The absolute fecundity ranged between 305 (234 mm TL) to 1,089 (414 mm TL) and relative fecundity oscillated between 77.3–220.69 oocytes/g of fish. The fecundity established a linear relationship with total length, body weight and ovary weight. The findings of this study would be very effective to impose sustainable conservation plan for this threatened species in Kallada River and other Western Ghats river systems.     

Isolation and cross-amplification of microsatellites in pink abalone (Haliotis corrugata)

Ten novel microsatellite loci were isolated in pink abalone, Haliotis corrugata, using (GT)₁₅ and (CT)₁₅ enriched genomic libraries. Two previously reported Haliotis kamtschatkana microsatellites cross‐amplified in H. corrugata. A set of 12 polymorphic microsatellites were evaluated in a wild population sample (N = 49). The number of alleles ranged from two to 55, and the observed and expected heterozygosities ranged from 0.104 to 0.939 and from 0.213 to 0.982, respectively. Significant deviations from Hardy–Weinberg equilibrium at three loci and no linkage disequilibrium were observed. Haliotis corrugata microsatellites cross‐amplified in other abalone species, two in H. fulgens, and seven in H. rufescens.     

Development and characterization of 13 microsatellite markers of Chanodichthys mongolicus (Cypriniformes: Cyprinidae) by RAD-seq

This study developed and characterized 13 novel polymorphic microsatellite markers from Mongolian redfin Chanodichthys mongolicus by restriction-site associated DNA sequencing (RAD-seq). Variability was tested on 36 individuals randomly collected from Qiantang River, Zhejiang Province, China. The number of alleles per locus ranged from 2 (MGB_1990, MGB_3456, MGB_6007, MGB_6539, MGB_7273, MGB_8632) to 9 (MGB_8632). Observed heterozygosity ranged from 0.000 (MGB_7273) to 0.972 (MGB_5057), whereas the expected heterozygosity ranged from 0.028 (MGB_6007) to 0.795 (MGB_4875). Polymorphism Information Content ranged from 0.027 (MGB_6007) to 0.755 (MGB_4875). Moreover, seven microsatellites were in agreement with Hardy-Weinberg equilibrium. Four pairwise tests in seven microsatellite loci (MGB_1498, MGB_1990, MGB_4875, MGB_5057, MGB_6539, MGB_7273, MGB_8632) indicated linkage disequilibrium among loci. These microsatellites can evaluate Mongolian redfin populations’ genetic parameters to provide precious information for a conservation management system.