Unfaithful maintenance of methylation imprints due to loss of maternal nuclear Dnmt1 during somatic cell nuclear transfer

The low success rate of somatic cell nuclear transfer (SCNT) in mammalian cloning is largely due to imprinting problems. However, little is known about the mechanisms of reprogramming imprinted genes during SCNT. Parental origin-specific DNA methylation regulates the monoallelic expression of imprinted genes. In natural fertilization, methylation imprints are established in the parental germline and maintained throughout embryonic development. However, it is unclear whether methylation imprints are protected from global changes of DNA methylation in cloned preimplantation embryos. Here, we demonstrate that cloned porcine preimplantation embryos exhibit demethylation at differentially methylated regions (DMRs) of imprinted genes; in particular, demethylation occurs during the first two cell cycles. By RNAi-mediated knockdown, we found that Dnmt1 is required for the maintenance of methylation imprints in porcine preimplantation embryos. However, no clear signals were detected in the nuclei of oocytes and preimplantation embryos by immunofluorescence. Thus, Dnmt1 is present at very low levels in the nuclei of porcine oocytes and preimplantation embryos and maintains methylation imprints. We further showed that methylation imprints were rescued in nonenucleated metaphase II (MII) oocytes. Our results indicate that loss of Dnmt1 in the maternal nucleus during SCNT significantly contributes to the unfaithful maintenance of methylation imprints in cloned embryos.     

Root size and soil environments determine root lifespan: evidence from an alpine meadow on the Tibetan Plateau

We used a minirhizotron system to investigate the influence of three major factors—root morphology, root depth, and season of root emergence—on root survivorship and longevity in a Kobresia humilis meadow on the Tibetan Plateau during the growing season of 2009. Root longevity was assessed by survival analysis, Kaplan–Meier analysis, and Cox proportional hazards regression. Root longevity was correlated positively with root diameter. A 17.5 % decrease in the risk of mortality was associated with a 0.1-mm increase in diameter. Roots distributed in the top 10 cm of the soil had significantly shorter longevities than roots at greater depths, with a 48 % decrease of mortality risk for each 10-cm increase in soil depth from the surface to 40 cm. Of all the factors examined, the season of root emergence had the strongest effect on root lifespan. Roots that emerged in May and June had shorter longevity than roots that emerged later in the year, and roots that emerged in September and October were more likely to survive over winter. Our findings indicated that life-history traits of roots in K. humilis meadows are highly heterogeneous, and this heterogeneity should be considered when modeling the contribution of roots to carbon and nitrogen fluxes in this type of meadow ecosystem. Moreover, temporal, spatial, and compositional variations in root longevity must be considered.     

三亚岩相潮间带底栖海藻群落结构及其季节变化

为了解三亚潮间带大型海藻生态现状,布设6条岩相断面,于2008年1—11月进行了四季野外的调查,并使用物种多样性指数与聚类分析研究了潮间带大型海藻四季的群落结构和多样性变化。调查共鉴定出大型底栖藻类130种,隶属于4门75属,其中红藻门41属67种、褐藻门16属31种、绿藻门15属29种、蓝藻门3属3种。潮间带海藻种类数季节变化不大,并呈低潮带>中潮带>高潮带的垂直分布特征,且中低潮带有共同分布种,夏季尤为普遍。潮间带海藻夏冬两季平均生物量高于春秋两季,但并无显著差异。优势种季节变化明显:春季有波利团扇藻Padina boryana、日本仙菜Ceramium japonicum、半叶马尾藻Sargassum hemiphyllum、海柏Polyopes polyideoides,夏季有波状软凹藻Chondrophycus undulates、冠叶马尾藻Sargassum cristaefolium、宽扁叉节藻Amphiroa dilatata、石花菜Gelidium amansii,秋季有冠叶马尾藻、波状软凹藻、叶状铁钉菜Ishige foliacea,冬季有瓦氏马尾藻Sargassum vachellianum、苔状鸭毛藻Symphyocladia marchantioides、珊瑚藻Corallina officinalis、波利团扇藻、日本仙菜。群落多样性冬高夏低:Shannon多样性和Margalef丰富度指数最大值出现在冬季,Shannon多样性和Pielou均匀度指数最低值出现在夏季。区域底栖藻类以暖水性热带-亚热带种为主,其次为温水性的暖温带种类,基本符合南海南区暖水区系特点。研究表明,尽管三亚潮间带底栖海藻种类数与生物量上并无显著的季节差异,但群落结构与多样性均显示出一定的季节变化。     

UHRF1通过miR-206调控ERα表达促进甲状腺乳头状癌细胞增殖、侵袭和迁移

该研究探讨了泛素样含PDH和环指域1(UHRF1)对甲状腺乳头状癌(papillary thyroid carcinoma,PTC)细胞增殖、侵袭和迁移的影响。应用Real-time PCR检测正常甲状腺细胞Nthyori3-1、甲状腺乳头状癌细胞BCPAP和K1中UHRF1 mRNA、miR-206和ERαmRNA的表达水平;Real-time PCR检测UHRF1过表达或干扰对miR-206和ERαmRNA的表达影响;MTT、Transwell检测UHRF1过表达或干扰对Nthy-ori3-1、BCPAP、K1细胞增殖、侵袭和迁移影响;Western blot和双荧光素酶报告实验分析miR-206与ERα的靶向关系。结果表明,与Nthy-ori3-1细胞相比,BCPAP和K1细胞中UHRF1 mRNA和ERαmRNA表达水平显著增高(P<0.05),而miR-206表达水平显著降低(P<0.05);UHRF1过表达或干扰处理细胞后,miR-206表达水平与之变化趋势相反,而ERα表达水平与之变化趋势相同;UHRF1促进Nthy-ori3-1、BCPAP和K1细胞增殖、侵袭和迁移;Western blot和双荧光素酶报告实验证实,miR-206靶向ERα基因并抑制其表达。以上结果说明,UHRF1可通过miR-206调控ERα表达,促进甲状腺乳头状癌细胞增殖、侵袭和迁移。     

Comprehensive assessments of root biomass and production in a Kobresia humilis meadow on the Qinghai-Tibetan Plateau

Alpine meadow covers ca. 700,000 km² with an extreme altitude range from 3200 m to 5200 m. It is the most widely distributed vegetation on the vast Qinghai-Tibetan Plateau. Previous studies suggest that meadow ecosystems play the most important role in both uptake and storage of carbon in the plateau. The ecosystem has been considered currently as an active “CO₂ sink”, in which roots may contribute a very important part, because of the large root biomass, for storage and translocation of carbon to soil. To bridge the gap between the potential importance and few experimental data, root systems, root biomass, turnover rate, and net primary production were investigated in a Kobresia humilis meadow on the plateau during the growing season from May to September in 2008 and 2009. We hypothesized that BNPP/NPP of the alpine meadow would be more than 50%, and that small diameter roots sampled in ingrowth cores have a shorter lifespan than the lager diameter roots, moreover we expected that roots in surface soils would turn over more quickly than those in deeper soil layers. The mean root mass in the 0–20 cm soil layer, investigated by the sequential coring method, was 1995?±?479 g?m^?2 and 1595?±?254 g?m^?2 in growing season of 2008 and 2009, respectively. And the mean fine root biomass in ingrowth cores of the same soil layer was 119?±?37 g?m^?2 and 196?±?45 g?m^?2 in the 2 years. Annual total NPP was 12387 kg?ha^?1?year^?1, in which 53% was allocated to roots. In addition, fine roots accounted for 33% of belowground NPP and 18% of the total NPP, respectively. Root turnover rate was 0.52 year^?1 for bulk roots and 0.74 year^?1 for fine roots. Furthermore, roots turnover was faster in surface than in deeper soil layers. The results confirmed the important role of roots in carbon storage and turnover in the alpine meadow ecosystem. It also suggested the necessity of separating fine roots from the whole root system for a better understanding of root turnover rate and its response to environmental factors.     

黄龙钙化滩流地物种-面积关系

黄龙沟钙化滩流地由于溪流的作用,在滩流地之间形成面积大小不一的植物群落斑块。这些小尺度斑块面积的大小对植物物种数量的影响尚不清楚。应用回归分析法和幂函数方程对黄龙沟钙化滩流地内的物种-面积关系进行了研究。结果表明黄龙沟钙化滩流地中斑块的大小对物种数(含兰科植物)具有强烈的影响,而调查的其他因子对总的植物物种数量的影响不显著。在所调查的环境因子中,斑块面积对物种数量的影响达到79.5%,即斑块越大,所包含的物种数量越多。兰科植物种类数量除了受斑块面积的影响外,还与距离林缘的距离有关(负相关)。物种-面积关系符合幂函数方程S=cA^Z的规律。不同的尺度下,z值略有差异,在中等尺度下 (1-10 m²)最大,为0.2616,较大尺度下(10-100 m²)的最小,z值为0.2050,小尺度下(<1 m²),z值为0.2382。表明中等尺度的斑块(1-10 m²)包含的物种数(含兰科植物)的增长速度最快,而在斑块面积大于10 m²时,物种数增长速度最小。     

人乳铁蛋白cDNA 基因乳腺表达载体的构建与鉴定

为了构建人乳铁蛋白基因 (hLF) 的乳腺表达载体并验证其在乳腺细胞中的表达情况,本载体以山羊β-casein基因上游包括启动子、外显子1、内含子1、部分外显子2作为5′端调控序列,下游包括部分外显子7、内含子7、外显子8、内含子8、外显子9及3′部分基因组片段作为3′端调控序列,长度分别为6.2 kb和7.1 kb,将hLF基因 (目的基因) 和Neo基因 (筛选标记) 分别插入到5′端调控序列和3′端调控序列的下游,构建成pBC1-hLF-Neo载体,其全长为25.348 kb。为了检测该载体的生物学     

Wind-dragged corolla enhances self-pollination: a new mechanism of delayed self-pollination

BACKGROUND AND AIMS: Delayed self-pollination is a mechanism that allows animal-pollinated plants to outcross while ensuring seed production in the absence of pollinators. This study aims to explore a new mechanism of delayed self-pollination facilitated by wind-driven corolla abscission in Incarvillea sinensis var. sinensis. METHODS: Floral morphology and development, and the process of delayed self-pollination were surveyed. Experiments dealing with pollinator and wind exclusion, pollination manipulations, and pollinator observations were conducted in the field. KEY RESULTS: Delayed self-pollination occurs when the abscising corolla driven by wind drags the adherent epipetalous stamens, thus leading to contact of anthers with stigma in late anthesis. There is no dichogamy and self-incompatibility in this species. The significantly higher proportion of abscised corolla under natural conditions as compared with that in wind-excluding tents indicates the importance of wind in corolla abscission. When pollinators were excluded, corolla abscission significantly increased the number of pollen grains deposited on the stigma and, as a result, the fruit and seed set. Only half of the flowers in plots were visited by pollinators, and the fruit set of emasculated flowers was significantly lower than that of untreated flowers in open pollination. This species has a sensitive stigma, and its two open stigmatic lobes closed soon after being touched by a pollinator, but always reopened if no or only little pollen was deposited. CONCLUSIONS: This delayed self-pollination, which involved the movement of floral parts, the active participation of the wind and sensitive stigma, is quite different from that reported previously. This mechanism provides reproductive assurance for this species. The sensitive stigma contributes to ensuring seed production and reducing the interference of selfing with outcrossing. The pollination pattern, which combines actions by bees with indirect participation by wind, is also a new addition to ambophily.     

区域尺度无人机涡动相关通量观测系统的应用研究

陆地生态系统的水热循环与碳循环是陆地表层系统中物质能量循环的核心,其中区域尺度地表水、热、碳通量的直接观测是当下陆地生态系统通量观测与模拟研究中的热点与难点。机载涡动相关方法能够直接观测区域尺度生态系统通量,基于无人机平台的涡动相关通量观测技术同时兼具了区域覆盖性与经济灵活性等优点,是机载通量观测技术的最新发展方向。在介绍机载涡动相关通量观测方法的主要技术原理、观测特点以及无人机通量观测系统组成的基础上,通过在相对均匀的区域开展无人机与地面通量观测对比试验,采用谱分析、观测结果对比以及源区分析等方式对无人机通量观测系统的性能进行了初步评价。结果表明：无人机通量观测系统能够实现对大气高频湍流信号的有效采样;无人机与地面观测的湍流通量具有较好的一致性,但是感热和CO₂通量出现了低估、潜热和摩擦风速出现了高估;观测平台与仪器的差异、垂直通量辐散、大气边界层条件、不同的地面源区及地表异质性的影响是造成二者差异的潜在主要因素。最后对未来研究目标进行了展望,以进一步推动该技术在相关领域中的应用。     

Long noncoding RNA OIP5-AS1 causes cisplatin resistance in osteosarcoma through inducing the LPAATβ/PI3K/AKT/mTOR signaling pathway by sponging the miR-340-5p

The abnormal expression of long noncoding RNAs (lncRNAs) plays an important role in the regulation of human cancer progression and drug resistance. The lncRNA OPI5-AS1 is a crucial regulator in some cancers; however, its role in cisplatin resistance of osteosarcoma remains unclear. We found that OIP5-AS1 was significantly upregulated in cisplatin-resistant (CR) osteosarcoma cells MG63-CR and SaOS2-CR compared with the corresponding parental cells. OIP5-AS1 silencing suppressed cell growth in vitro and in vivo, and promoted apoptosis of MG63-CR and SaOS2-CR cells, indicating that knockdown of OIP5-AS1 significantly decreased cisplatin resistance in MG63-CR and SaOS2-CR cells. This conclusion was supported by the decreased expression of the drug resistance-related factors multidrug resistance-associated protein 1 (MRP1) and P-glycoprotein (P-gp) upon OIP5-AS1 silencing. In addition, OIP5-AS1 downregulation suppressed the PI3K/AKT/mTOR signaling pathway. Importantly, we demonstrated that OIP5-AS1 functions as a competing endogenous RNA of miR-340-5p and regulates the expression of lysophosphatidic acid acyltransferase (LPAATβ), which is a target of miR-340-5p. Moreover, downregulation of miR-340-5p partly reversed the inhibitory effect of OIP5-AS1 knockdown on the PI3K/AKT/mTOR pathway and therefore counteracted cisplatin resistance in MG63-CR and SaOS2-CR cells. In conclusion, OIP5-AS1 causes cisplatin resistance in osteosarcoma through inducing the LPAATβ/PI3K/AKT/mTOR signaling pathway by sponging the miR-340-5p. Our results contribute to a better understanding of the function and mechanism of OIP5-AS1 in osteosarcoma cisplatin resistance.