Isolation and characterization of 28 polymorphic microsatellite loci in black carp (Mylopharyngodon piceus)

This study reports the isolation and characterization of 28 polymorphic microsatellite loci developed from black carp (Mylopharyngodon piceus), which is an important freshwater fish in China. The polymorphism was assessed with 32 individuals. Results showed that the numbers of loci alleles ranged from 2 to 19, and the values of observed and expected heterozygosities were from 0.2609 to 1.0000 and from 0.2417 to 0.9385, respectively. These markers are potentially useful for black carp population genetics analysis.This study reports the isolation and characterization of 28 polymorphic microsatellite loci developed from black carp (Mylopharyngodon piceus), which is an important freshwater fish in China. The polymorphism was assessed with 32 individuals. Results showed that the numbers of loci alleles ranged from 2 to 19, and the values of observed and expected heterozygosities were from 0.2609 to 1.0000 and from 0.2417 to 0.9385, respectively. These markers are potentially useful for black carp population genetics analysis

Differential gene expression in response to brown planthopper feeding in rice.

Summary Plant responses to herbivores are complex. 108 cDNA clones representing genes relating to plant responses to chewing insect-feeding, pathogen infection, wounding and other stresses were collected. Northern blot and cDNA array analysis were employed to investigate gene expression regulated by piercing-sucking insect, brown planthopper (BPH), Nilaparvata lugens (Homoptera: Dephacidae) on both the resistant and susceptible rice genotypes. After BPH feeding in rice for 72 h, the expression of most tested genes was affected. 14 genes in resistant rice variety B5 and 44 genes in susceptible MH63 were significantly up-or down-regulated. Most of the well-regulated genes were grouped in the categories of signaling pathways, oxidative stress/apoptosis, wound-response, drought-inducible and pathogen-related proteins. Those related to the flavonoid pathway, aromatic metabolidsm and the octadecanoid pathway were mostly kept unchanged or down-regulated. Our results indicate that BPH feeding induces plant responses which would take part in a jasmonic acidindependent pathway and crosstalk with those related to abiotic stress, pathogen invasion and phytohormone signaling pathways

The identification of metallothionein in rare minnow (Gobiocypris rarus) and its expression following heavy metal exposure

Heavy metal, such as cadmium (Cd), lead (Pb) and copper (Cu) poses serious toxin to aquatic organisms. These exogenous materials affect biological processes including physiology, biochemistry and development. Metallothionein (MT), one of the metal-regulated genes, participates in regulating essential and detoxifying non-essential metals in living animals. In this study, MT EST in rare minnow (Gobiocypris rarus) (GrMT) was obtained from the cDNA subtraction library and the GrMT cDNA was firstly cloned by RACE with a sequence of 379 bp, which can code 60 amino acids. After Cd exposure, the GrMT expression levels dramatically changed in liver, spleen, gill, kidney, intestine, but moderately in muscle. Significantly positive relationships were found between Cd dosages and MT expression levels in liver. And there distinction existed at the GrMT transcript level in fish gender and developmental stages during Cd exposure. While Pb exposure, the expression alteration happened in spleen, gill, intestine, muscle, and moderately in liver. In the Cu treatment test, there were slight changes in intestine and kidney, but more significant changes occurred in spleen, gill, muscle, and liver. This study had investigated the effects of diverse heavy metals on GrMT expression patterns. The results suggest that GrMT would be a potential biomarker to metal contamination monitoring in aquatic environments and rare minnow could be one of the perfect experimental fishes for surveying the freshwater pollution in China. (C) 2014 Elsevier B.V. All rights reserved

MicroRNA-944 Affects Cell Growth by Targeting EPHA7 in Non-Small Cell Lung Cancer

MicroRNAs (miRNAs) have critical roles in lung tumorigenesis and development. To determine aberrantly expressed miRNAs involved in non-small cell lung cancer (NSCLC) and investigate pathophysiological functions and mechanisms, we firstly carried out small RNA deep sequencing in NSCLC cell lines (EPLC-32M1, A549 and 801D) and a human immortalized cell line 16HBE, we then studied miRNA function by cell proliferation and apoptosis. cDNA microarray, luciferase reporter assay and miRNA transfection were used to investigate interaction between the miRNA and target gene. miR-944 was significantly down-regulated in NSCLC and had many putative targets. Moreover, the forced expression of miR-944 significantly inhibited the proliferation of NSCLC cells in vitro. By integrating mRNA expression data and miR-944-target prediction, we disclosed that EPHA7 was a potential target of miR-944, which was further verified by luciferase reporter assay and microRNA transfection. Our data indicated that miR-944 targets EPHA7 in NSCLC and regulates NSCLC cell proliferation, which may offer a new mechanism underlying the development and progression of NSCLC

The identification of apolipoprotein C-I in rare minnow (Gobiocypris rarus) and its expression following cadmium exposure

Cadmium (Cd) is a ubiquitous environment contaminant that has been detected in a variety of terrestrial and aquatic organisms. In our study, rare minnow were exposed to 0.4 mg/L Cd2+ solution, fish tissues including gill, intestine, kidney, liver, muscle, and spleen were collected at 1, 6, 12,24 and 48 h after exposure. Differential expression sequence tags (ESTs) were screened and an EST similar to Hemibarbus mylodon apolipoprotein C-I (ApoC-I) was identified. The ApoC-I cDNA had been obtained with the sequence of 583 bp, which can code 84 amino acids. The transcription expression patterns of ApoC-I were compared after Cd exposure with the real-time PCR in the above tissues. ApoC-I regulation was prominent in spleen, muscle, gill and liver and moderate in kidney and intestine. This suggests a complex transcriptional regulation of ApoC-I expression following Cd exposure. (c) 2013 Elsevier B.V. All rights reserved.Cadmium (Cd) is a ubiquitous environment contaminant that has been detected in a variety of terrestrial and aquatic organisms. In our study, rare minnow were exposed to 0.4 mg/L Cd2+ solution, fish tissues including gill, intestine, kidney, liver, muscle, and spleen were collected at 1, 6, 12,24 and 48 h after exposure. Differential expression sequence tags (ESTs) were screened and an EST similar to Hemibarbus mylodon apolipoprotein C-I (ApoC-I) was identified. The ApoC-I cDNA had been obtained with the sequence of 583 bp, which can code 84 amino acids. The transcription expression patterns of ApoC-I were compared after Cd exposure with the real-time PCR in the above tissues. ApoC-I regulation was prominent in spleen, muscle, gill and liver and moderate in kidney and intestine. This suggests a complex transcriptional regulation of ApoC-I expression following Cd exposure. (c) 2013 Elsevier B.V. All rights reserved

Novel insights into the reproductive strategies of wild Chinese sturgeon (Acipenser sinensis) populations based on the kinship analysis

Understanding the reproductive strategy of an organism is important in conservation ecology as it directly affects the population performance under changing environmental conditions. Chinese sturgeon (Acipenser sinensis) are the largest anadromous fish in the Yangtze River, China. Currently, the species has only one spawning ground and has failed to spawn in recent years, leading it to the brink of extinction. To develop effective conservation measures, a further understanding of its reproductive strategy is needed. In our study, we conducted kinship analyses by using mitochondrial and microsatellite DNA data from 216 wild juveniles collected over nine years (2006−2013, 2015) to understand the mating system, breeding interval, effective number of breeding adults, and reproductive success. The results from these analyses suggested polygynandry, with some parents contributing up to eight half-sibling juvenile genotypes. Although the spawning ground was restricted to a limited area, genetic diversity was maintained at a relatively high level (observed heterozygosity from 0.698 to 0.787 and expected heterozygosity from 0.763 to 0.787) and inbreeding coefficients in each year-class ranged from −1% to 9% (low to modest detrimental effects on offspring). A parental inference analysis revealed that Chinese sturgeon have a breeding interval of 2–6 years, indicating that it has the potential to feed, accumulate nutrition in the ocean, and then migrate back to the Yangtze River for iteroparous reproduction. The annual effective number of breeders in the Yangtze River ranged from 14 to 161 during the study period, and it decreased by 62.1% from the 2011−2014 year-classes. This sharp population decline likely contributes to the reproduction failure. However, the ratios of effective to census population size (Ne/Nc) were all larger than 0.20 after the 2010 year-class, indicating relatively even reproductive success. Based on these results, a suggested approach to protect this species is to restock parent fish to increase the reproductive stock size and optimize the discharge of the Three Gorges Dam to reduce the unsuitable hydrological conditions and rehabilitate spawning ground habitats

Genetic diversity in population of largemouth bronze gudgeon (Coreius guichenoti Sauvage et Dabry) from Yangtze River determined by microsatellite DNA analysis

Largemouth bronze gudgeon (Coreius guichenoti Sauvage et Dabry 1874), one of the endemic fish species in the upper reaches of the Yangtze River in China, is a benthic and potamodromous fish that is typically found in rivers with torrential flow. Three dams in the Yangtze River, Ertan Dam, Three Gorges Dam and Gezhouba Dam, may have had vital impacts on the habitat and spawning behaviors of largemouth bronze gudgeon, and could ultimately threaten the survival of this fish. We studied the population genetic diversity of C. guichenoti samples collected at seven sites (JH, GLP, BX, HJ, MD, SDP and XB) within the Yangtze River and one of its tributaries, the Yalong River. Genetic diversity patterns were determined by analyzing genetic data from 11 polymorphic microsatellite loci. A high genetic diversity among these largemouth bronze gudgeon populations was indicated by the number of microsatellite alleles (A) and the expected heterozygosity (HE). No significant population variation occurred among GLP, BX, HJ and MD populations, but dramatic population differentiation was observed among JH and XB, two dam-blocked populations, versus other populations. Tests for bottlenecks did not indicate recent dramatic population declines and concurrent losses of genetic diversity in any largemouth bronze gudgeon populations. To the contrary, we found that dams accelerated the population differentiation of this fish

High genetic diversity in population of Lepturichthys fimbriata from the Yangtze River revealed by microsatellite DNA analysis

Lepturichthys fimbriata (Gunther) is one of the benthic and rock-attached fish species that is typically found in torrential flows of the upper reaches of the Yangtze River in China. Several dams in the Yangtze River (the Ertan Dam, the Three Gorges Dam, the Gezhouba Dam, the Xiluodu Dam and the Xiangjiaba Dam (the latter two dams are under construction)) may have significant effects on the habitat and spawning behaviors of L. fimbriata, and could ultimately threaten the survival of this fish. We studied the population genetic diversity of L. fimbriata samples collected at three sites within the Yangtze River and one of its tributaries, the Yalong River. Genetic diversity patterns were determined by analyzing genetic data from 14 polymorphic microsatellite loci isolated in Jinshaia sinensis (Sauvage et Dabry). A high genetic diversity among these L. fimbriata local populations was indicated by the number of microsatellite alleles (A) and the expected heterozygosity. No reductions of genetic diversity in any L. fimbriata population were observed. However, significant population differentiations were observed among three local populations by pairwise comparisons (P<0.001). We deduced that L. fimbriata local populations were not small ones. In addition, the habitat behaviors of rock-attachment and possible residence of L. fimbriata could account for the genetic differences found in local populations.Lepturichthys fimbriata (Gunther) is one of the benthic and rock-attached fish species that is typically found in torrential flows of the upper reaches of the Yangtze River in China. Several dams in the Yangtze River (the Ertan Dam, the Three Gorges Dam, the Gezhouba Dam, the Xiluodu Dam and the Xiangjiaba Dam (the latter two dams are under construction)) may have significant effects on the habitat and spawning behaviors of L. fimbriata, and could ultimately threaten the survival of this fish. We studied the population genetic diversity of L. fimbriata samples collected at three sites within the Yangtze River and one of its tributaries, the Yalong River. Genetic diversity patterns were determined by analyzing genetic data from 14 polymorphic microsatellite loci isolated in Jinshaia sinensis (Sauvage et Dabry). A high genetic diversity among these L. fimbriata local populations was indicated by the number of microsatellite alleles (A) and the expected heterozygosity. No reductions of genetic diversity in any L. fimbriata population were observed. However, significant population differentiations were observed among three local populations by pairwise comparisons (P<0.001). We deduced that L. fimbriata local populations were not small ones. In addition, the habitat behaviors of rock-attachment and possible residence of L. fimbriata could account for the genetic differences found in local populations