Genetic diversity within two Tunisian wild jirds: Meriones shawi and Meriones libycus (Rodentia,Gerbillinae)

Three Meriones species inhabit Tunisia, namely M. shawi, M. libycus and M. crassus, but little genetic data exist on these gerbils. We collected Meriones from eight localities in Tunisia, and obtained mitochondrial (cytochrome b) and nuclear (IRBP) gene sequence data for 37 and 13 specimens, respectively, belonging to two species: M. shawi and M. libycus. We also optimised three microsatellite markers previously described in M. unguiculatus to obtain a finer analysis of their genetic diversity and geographic structure, given their wide distribution. Phylogenetic inferences of cyt b and IRBP data for these species, in the context of other gerbillin data, corroborate their taxonomic affinities reported by previous studies. High cyt b haplotype diversity was observed in both species (25 haplotypes in 29 and 27 sequences for M. shawi and M. libycus, respectively) with little geographical structure for M. shawi but three divergent groups in M. libycus. The average microsatellite diversity within each population was high (H_O ≥ 0.6, H_E ≥ 0.8) with M. libycus populations attaining the highest values. Population differentiation was moderate for several population pairs (F_st ≥ 0.1), the highest being between M. shawi populations. However, genetic distance among populations was not significantly correlated with geographic distance in either M. shawi or M. libycus. Our results contribute to a better characterisation of Tunisian Meriones species, suggesting high geographic structure in mtDNA of M. libycus populations within North Africa.     

Analysis of DNA Methylation and mRNA Expression Level of MSTN Gene in Xinjiang Bashiby Sheep

In order to investigate DNA methylation and expression levels of myostatin (MSTN) gene in mscule and fat, 5 months of age of Bashiby sheep were selected, the promoter region and exon 1 methylation levels of MSTN gene was analyzed using bisulfite sequencing PCR (BSP). Real-time PCR was used to detect the expression level of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat of Bashiby sheep. The results showed that the methylation probability of muscle was higher than fat, the methylation probability of biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat were 74.2%, 74.2%, 83.2%, 83.7%, 82.1% and 25.3%, respectively. The expression levels of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle were significantly lower than tail fat in Bashiby sheep (P < 0.05), but there were no significant difference among biceps femoris, femoral triceps, semitendinosus, semimembranosus and longissimus dorsi muscle (P > 0.05).The DNA methylation was negatively correlated with the expression levels in muscle and fat of Bashiby sheep (r=-0.886, P < 0.05).     

哺乳动物生殖细胞冷冻保存对其DNA甲基化影响的研究进展

哺乳动物生殖细胞和胚胎冷冻保存在辅助生殖技术中得到了广泛的应用,同时在保存物种的多样性方面发挥着巨大的作用。但是,冷冻引起的DNA甲基化对其印记基因缺陷性疾病的发生和后代生理功能的差异会有很大的影响。作者就冷冻保存技术对配子（精子和卵子）和胚胎的DNA甲基化的影响以及冷冻引起的DNA甲基化对后代的影响进行了综述,以期对配子的冷冻保存和辅助生殖技术的发展提供参考。     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Chlorogenic acid supplementation during in vitro maturation improves maturation,fertilization and developmental competence of porcine oocytes

Chlorogenic acid (CGA) is a quinic acid conjugate of caffeic acid, and a phytochemical found in many fruits and beverages that acts as an antioxidant. The present study investigated the effects of CGA supplementation during in vitro maturation (IVM), on in vitro development of porcine oocytes, to improve the porcine in vitro production (IVP) system. Oocytes were matured either without (control) or with CGA (10, 50, 100 and 200 μM). Subsequently, the matured oocytes were fertilized and cultured in vitro for 7 day. The rates of maturation, fertilization and blastocyst formation of oocytes matured with 50 μM CGA were significantly (p < .05) higher than those of the control oocytes. Hydrogen peroxide (H₂O₂) is one of the reactive oxygen species and induces DNA damage in porcine oocytes. When oocytes were matured with 1 mM H₂O₂ to assess the protective effect of CGA, 50 μM CGA supplementation improved the maturation rate and the proportion of DNA‐fragmented nuclei in oocytes compared with control oocytes matured without CGA. Moreover, when oocytes were matured with either 50 μM CGA (control) or caffeic acid (10, 50 and 100 μM), the rates of maturation, fertilization and the blastocyst formation of oocytes matured with 50 μM CGA were similar to those of oocytes matured with 10 and 50 μM caffeic acid. Our results suggest that CGA has comparable effects to caffeic acid, and IVM with 50 μM CGA is particularly beneficial to IVP of porcine embryos and protects oocytes from DNA damage induced by oxidative stress. Supplementation of CGA to the maturation medium has a potential to improve porcine IVP system.     

基于BP神经网络的三峡库区重庆段水资源安全评价

三峡库区水资源安全关系到库区的生态安全,对库区水资源安全利用进行评价,找出问题较为严重的区域,明确其限制因子,从而为库区水资源可持续发展和水资源安全提供科学的决策依据。结合2000—2014年三峡库区水资源安全数据,选取BP神经网络构建水资源安全利用评价模型,结合ARIMA模型进行指标预测,分析了三峡库区重庆段水资源安全利用的时空差异。研究表明:(1)三峡库区重庆段2014年水资源安全利用总体分布在较不安全到较安全之间,渝中区和大渡口区为较不安全等级,沙坪坝区等7个区县为基本安全,其他区县为较安全等级;在子系统的评价中,主城区的社会安全与供需安全等级均为最低,为极不安全和较不安全等级。(2)2000—2014年水资源安全利用等级主要受生态安全因子制约,供需安全因子次之;2015—2020年水资源安全利用等级主要受供需安全因子限制,生态安全因子次之。2000—2020年水资源安全利用等级总体呈上升趋势。     

与GyrA相互作用的GyrB结构域突变对结核分枝杆菌螺旋酶功能的影响

DNA螺旋酶中的GyrA与GyrB亚基只有互作重组后才有酶活性。为寻找GyrB亚基和GyrA亚基相互作用的关键区域,构建了不同的GyrB亚基突变体,分析GyrB各突变体与GyrA亚基的相互作用对全酶活性的影响。结果显示:GyrB亚基C端是其与GyrA相互作用的主要结构域,结合GyrB的二维结构,提出GyrB中第531~550位氨基酸是影响螺旋酶功能的关键区域,并可能是理想的新药设计靶标。     

宜良县云南松林生物量遥感估测方法研究

利用landsat5影像和通过蓄积量-生物量转换方法获得的样地生物量数据,从宜良县云南松林遥感特征、地形特征、郁闭度等方面选取自变量,采用随机森林回归和偏最小二乘法进行生物量遥感估测建模。结果表明:随机森林回归估测模型调整决定系数R2为0.881,模型估测精度为82.3%;偏最小二乘模型的调整决定系数R2为0.753,模型估测精度为78.43%。随机森林回归在模型拟合效果和检验精度上均优于偏最小二乘模型。     

文冠果转录组SSR特征分析及EST-SSR标记开发

【目的】利用高通量测序技术分析文冠果转录组中SSR位点分布类型与特征,并设计开发EST-SSR标记,为文冠果的遗传多样性分析提供标记资源。【方法】利用MISA软件筛选文冠果51 867条unigenes中的SSR(2~6bp)位点,统计分析SSR位点的分布特征、发生频率和平均分布距离。利用Prime 3在线软件随机设计合成60对EST-SSR标记,用我国7个省份的16份文冠果材料对所设计的标记进行多态性筛选;用Popgene 32软件分析标记的等位基因数(NA)、观测杂合度(HO)、期望杂合度(HE)和多态性信息含量(PIC)等。【结果】从文冠果unigenes中筛选出6 707个SSR位点,其平均发生频率为12.93%,平均分布距离5.38kb,平均长度17.30bp,其中2、3核苷酸重复单元的发生频率分别为6.24%和4.93%。设计的60对标记中有47对能扩增出与目的片段长短相符的产物,其中14对多态性较高,共扩增到52个等位基因,HO和HE分别为0~0.813和0.353~0.762,PIC为0.283~0.701。【结论】文冠果转录组SSR重复单元以2、3核苷酸重复为主;新开发的14个EST-SSR标记可作为文冠果种质资源多样性分析的标记资源。     

DNA在黑土胶体微界面的吸附与解吸特性

为了揭示DNA在黑土微生物的遗传进化和基因转移机制方面的重要意义,采用平衡法研究了含有机质黑土胶体(Coarse organic colloid,COC)和去有机质黑土胶体(Coarse inorganic colloid,CIC)对DNA的吸附与解吸特性。结果表明:CIC在快速吸附期吸附DNA的速率和数量都小于COC,且更快达到吸附平衡;随着pH值升高,黑土胶体表面吸附DNA数量逐渐减少;Langmuir吸附方程可较好地描述COC和CIC对DNA的等温吸附,COC和CIC的决定系数(R~2)分别为0.972和0.991,且COC的吸附平衡常数(K)大于CIC,说明COC对DNA的吸附亲和能力大于CIC;CIC和COC表面DNA的解吸率分别为63.5%和47.4%。研究结果证明,有机质在黑土胶体吸附DNA的数量和亲和力方面贡献较大,pH是影响黑土胶体吸附DNA的重要因子。