Isolation and characterisation of eighteen microsatellite markers from the sea cucumber Holothuria scabra (Echinodermata : Holothuriidae)

2012 ◽  
Vol 60 (6) ◽  
pp. 368 ◽  
Author(s):  
Alison J. Fitch ◽  
Grant Leeworthy ◽  
Xiaoxu Li ◽  
Will Bowman ◽  
Luke Turner ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Sea Cucumber ◽  
Asia Pacific ◽  
Asia Pacific Region ◽  
Single Population ◽  
Important Species ◽  
Holothuria Scabra ◽  
Commercially Important ◽  
Polymorphic Microsatellite ◽  
Generation Sequencing

We isolated 18 new, polymorphic, microsatellite markers from Holothuria scabra, a commercially important species of sea cucumber found throughout the Asia-Pacific region. Next-generation sequencing was used to identify 206 unique loci for which primers were designed. Of these unique loci, we trialled 65 primer pairs in the target species, and 48 (74%) amplified a product of the expected size. Eighteen loci were found to be polymorphic and reliable, and were screened for variation in 50 individuals, from a single population from Croker Island, Northern Territory, Australia. Observed heterozygosity ranged from 0.00 to 0.96 (mean = 0.46) and the number of alleles per locus from 2 to 28 (mean = 9.61). These loci will be useful for the investigation of population structure and mating systems in H. scabra and may also be of use in other holothurian species.

scholarly journals Microsatellite development via next-generation sequencing in Acacia stenophylla (Fabaceae) and Duma florulenta (Polygonaceae): two ecologically important plant species of Australian dryland floodplains

2018 ◽  
Author(s):  
Bruce F Murray ◽  
Michael A Reid ◽  
Shu-Biao Wu
Keyword(s):  
Gene Flow ◽  
Next Generation Sequencing ◽  
Microsatellite Markers ◽  
Information Content ◽  
Polymorphic Information Content ◽  
Extreme Variability ◽  
Polymorphic Microsatellite ◽  
Generation Sequencing ◽  
Semi Arid

Duma florulenta and Acacia stenophylla are two ecologically important but understudied species that naturally occur on the floodplains and riverbanks of Australia’s arid and semi-arid river systems. This paper describes the discovery and characterization of 12 and 13 polymorphic microsatellite markers for D. florulenta and A. stenophylla respectively. The number of alleles per locus for D. florulenta ranged from 2-12 with an average of 6.1. Across all samples, observed and expected heterozygosities ranged from 0.026 to 0.784 and 0.026 to 0.824 respectively and mean polymorphic information content was equal to 0.453. For A. stenophylla, the number of alleles per locus ranged between 2 and 8 with an overall mean of 4.8. Across all samples, observed and expected heterozygosities ranged from 0.029 to 0.650 and 0.029 to 0.761 respectively and mean polymorphic information content was 0.388. The developed suites of 12 and 13 microsatellite markers for D. florulenta and A. stenophylla respectively provide opportunity for novel research into mechanisms of gene flow, dispersal and breeding system and how they operate under the extreme variability these species are exposed to in the environments in which they live.

scholarly journals Highly Polymorphic Microsatellite Markers in Pulsatilla vulgaris (Ranunculaceae) Using Next-Generation Sequencing

2015 ◽  
Vol 3 (7) ◽  
pp. 1500031 ◽  
Author(s):  
Michelle F. DiLeo ◽  
René Graf ◽  
Rolf Holderegger ◽  
Yessica Rico ◽  
Helene H. Wagner
Keyword(s):  
Next Generation Sequencing ◽  
Microsatellite Markers ◽  
Next Generation ◽  
Polymorphic Microsatellite ◽  
Generation Sequencing

Isolation and characterisation via 454 sequencing of microsatellites from the tawny frogmouth, Podargus strigoides (Class Aves, Family Podargidae)

2012 ◽  
Vol 60 (2) ◽  
pp. 133
Author(s):  
Fiona E. Hogan ◽  
Marian Weaving ◽  
Gregory R. Johnston
Keyword(s):  
Population Genetics ◽  
Gene Flow ◽  
Microsatellite Markers ◽  
Reproductive Ecology ◽  
454 Sequencing ◽  
Urban Environments ◽  
Shotgun Sequencing ◽  
Target Species ◽  
Single Population ◽  
Polymorphic Microsatellite

We isolated 24 novel polymorphic microsatellite markers from the tawny frogmouth, a nocturnal bird endemic to Australia, which has successfully adapted to urban environments. Initially, 454 shotgun sequencing was used to identify 733 loci with primers designed. Of these, we trialled 30 in the target species of which all amplified a product of expected size. Subsequently, all 30 of these loci were screened for variation in 25 individuals, from a single population in Melbourne, Victoria, Australia. Twenty-eight loci were polymorphic with observed heterozygosity ranging from 0.03 to 0.96 (mean 0.58) and the number of alleles per locus ranged from 2 to 18 (average of 6.5); we confirmed that 24 loci conformed to Hardy–Weinberg expectations. The 24 loci identified here will be sufficient to unequivocally identify individuals and will be useful in understanding the reproductive ecology, population genetics and the gene flow amongst localities in urban environments where this bird thrives.

16 microsatellite loci for the Australian Great Artesian Basin spring amphipod, Wangiannachiltonia guzikae

2013 ◽  
Vol 61 (2) ◽  
pp. 109 ◽  
Author(s):  
Hannah L. Robertson ◽  
Nicholas P. Murphy
Keyword(s):  
Next Generation Sequencing ◽  
Microsatellite Markers ◽  
Microsatellite Loci ◽  
Single Population ◽  
Artesian Basin ◽  
Great Artesian Basin ◽  
Hardy Weinberg Equilibrium ◽  
Stochastic Events ◽  
Generation Sequencing ◽  
454 Next Generation Sequencing

454 Next Generation sequencing was used to develop a set of microsatellite markers for the Great Artesian Basin (GAB) amphipod, Wangiannachiltonia guzikae. Primers were designed for 42 microsatellite loci. A total of 22 loci were successfully amplified and 16 characterised using 30 individuals from a single GAB spring population. Across these 16 loci, observed heterozygosity ranged from 0.000 to 0.818 (mean = 0.445) and the number of alleles per locus ranged from 2 to 12 (mean = 6.688). Of these 16 loci, however, only 10 were in Hardy–Weinberg equilibrium, though all 16 loci should be retained for further studies in the event that stochastic events affected equilibrium of this single population.

scholarly journals Development and characterization of 29 polymorphic microsatellite loci of Megalobrama pellegrini by next-generation sequencing technology and cross-species amplification in related species

2016 ◽  
Author(s):  
Wen Song ◽  
Dongmei Zhu ◽  
Yefeng Lv ◽  
Weimin Wang
Keyword(s):  
Microsatellite Markers ◽  
Microsatellite Loci ◽  
Related Species ◽  
Polymorphic Information Content ◽  
Sequencing Technology ◽  
Next Generation Sequencing Technology ◽  
Expected Heterozygosity ◽  
Polymorphic Microsatellite ◽  
Generation Sequencing

Megalobrama pellegrini is one of the economically important freshwater fish in China. Here, we developed 29 polymorphic microsatellite loci of M. pellegrini. The number of alleles (Na), effective number of alleles (Ne), observed heterozygosity (HO), expected heterozygosity (HE) and polymorphic information content (PIC) ranged from 3 to 11 (mean±SD 5.4828±1.9571), 2.8708 to 9.6257 (mean±SD 5.0865±1.6681), 0.4333 to 0.9333 (mean±SD 0.7874±0.1213), 0.6627 to 0.9113 (mean±SD 0.7946±0.0751) and 0.5785 to 0.8868 (mean±SD 0.7439±0.0950), respectively. Cross-species amplification was successful at most loci for related species such as M. amblycephala, M. hoffmanni, M. skolkovii and Parabramis pekinensis. The transferability rate of the 29 polymorphic microsatellite markers in M. amblycephala, M. hoffmanni, M. skolkovii and P. pekinensis were 96.55%, 86.21%, 86.21% and 75.86%, respectively. These polymorphic microsatellites are not only useful in genetic study and conservation of M. pellegrini, but also an effective tool for identifying the related species. We could use 5 microsatellite markers (HHF-63, HHF-104, HHF-113, HHF-148, HHF-163) to distinguish the 5 species.

scholarly journals Comparative genome-wide polymorphic microsatellite markers in Antarctic penguins through next generation sequencing

2017 ◽  
Vol 40 (3) ◽  
pp. 676-687 ◽  
Author(s):  
Juliana A. Vianna ◽  
Daly Noll ◽  
Isidora Mura-Jornet ◽  
Paulina Valenzuela-Guerra ◽  
Daniel González-Acuña ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Microsatellite Markers ◽  
Next Generation ◽  
Comparative Genome ◽  
Genome Wide ◽  
Polymorphic Microsatellite ◽  
Generation Sequencing

scholarly journals Sex steroids and steroidogenesis-related genes in the sea cucumber, Holothuria scabra and their potential role in gonad maturation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tipsuda Thongbuakaew ◽  
Saowaros Suwansa-ard ◽  
Arada Chaiyamoon ◽  
Scott F. Cummins ◽  
Prasert Sobhon
Keyword(s):  
Sex Steroids ◽  
Sea Cucumber ◽  
Ovarian Development ◽  
Regulatory Protein ◽  
Asia Pacific ◽  
Pcr Analysis ◽  
Gonad Maturation ◽  
Gene Expressions ◽  
Sea Cucumbers ◽  
Holothuria Scabra

AbstractThe sea cucumber Holothuria scabra is an economically valuable marine species which is distributed throughout the Asia–Pacific region. With the natural population declining due to over fishing, aquaculture of this species is deemed necessary. Hence, it is essential to understand the mechanisms regulating the reproduction in order to increase their populations. Sex steroids, including estrogens, androgens and progestogens, play an important role in reproduction in most vertebrates and several invertebrates. It has been proposed that sea cucumbers have the same sex steroids as vertebrates but the steroidogenic pathway in the sea cucumbers is still unclear. In this study, we demonstrated by using liquid chromatography-tandem mass spectrometry (LC–MS/MS) that sex steroids (estradiol, progesterone, and testosterone) were present in H. scabra neural and gonadal tissues. In silico searches of available sea cucumber transcriptome data identified 26 steroidogenesis-related genes. Comparative analysis of encoded proteins for the steroidogenic acute regulatory protein (HscStAR), CYP P450 10, 17 and 3A (HscCYP10, HscCYP17, HscCYP3A) and hydroxysteroid dehydrogenases (Hsc3β-HSD, Hsc17β-HSD) with other species was performed to confirm their evolutionary conservation. Gene expression analyses revealed widespread tissue expression. Real-time PCR analysis revealed that HscStAR, HscCYP10, Hsc3β-HSD, and Hsc17β-HSD gene expressions were similar to those in ovaries and testes, which increased during the gonad maturation. HscCYP17 mRNA was increased during ovarian development and its expression declined at late stages in females but continued high level in males. The expression of the HscCYP3A was high at the early stages of ovarian development, but not at other later stages in ovaries, however it remained low in testes. Moreover, a role for steroids in reproduction was confirmed following the effect of sex steroids on vitellogenin (Vtg) expression in ovary explant culture, showing upregulation of Vtg level. Collectively, this study has confirmed the existence of steroids in an echinoderm, as well as characterizing key genes associated with the steroidogenic pathway. We propose that sex steroids might also be associated with the reproduction of H. scabra, and the identification of biosynthetic genes enables future functional studies to be performed.

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