Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta

AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.     

苦瓜转录因子McEIL2的克隆及其表达分析

本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。     

鸡Perilipin1基因的克隆及亚细胞定位

本研究旨在克隆鸡(Gallus gallus)脂滴包被蛋白基因(Perilipin1)的cDNA序列,并检测其在鸡前脂肪细胞分化过程中的亚细胞定位.本研究以7周龄肉鸡腹部脂肪组织为材料,采用RT-PCR和RACE的方法扩增并克隆了鸡Perilipin1基因的5'UTR、CDS和3'UTR片段,分析了该基因的结构;以鸡原代前脂肪细胞为材料,利用免疫荧光及激光共聚焦技术,在诱导分化的不同时间点(12～120 h),检测了鸡Perilipin1在前脂肪细胞中的表达位置.结果表明,本研究获得的鸡Perilipin11基因5'UTR、CDS和YUTR序列总长度为2 379 bp,该基因由9个外最子、8个内含子组成(ATG位于第二外显子上);在鸡原代前脂肪细胞诱导分化过程中,Perilipin1始终包被在脂滴周围.综上,本研究成功克隆了鸡Perilipinl基因的完整cDNA序列并确定了Perilipin1在鸡前脂肪细胞分化过程中与脂滴的空间位置关系,该结果为深入开展鸡Perilipinl基因的功能研究,揭示鸡脂肪代谢的分子遗传机理提供了重要的理论依据.     

Aggregate-related changes in network patterns of nematodes and ammonia oxidizers in an acidic soil

Nitrification plays a central role in global nitrogen cycle, which is affected by biological interaction between soil microfauna and microorganisms. However, the complexity of soil biotic communities made it difficult to reveal organizational principles of the community and the interactions among species. Here, we used the network analysis to decipher the interactions between nematodes and ammonia oxidizers within aggregate fractions under 10-year manure application, and examine their associations with soil variables and potential nitrification activity (PNA). Three aggregate fractions included large macroaggregates (>2000 μm, LA), small macroaggregates (250–2000 μm, SA), and inter-aggregate soil and space (<250 μm, IA). Aggregate factions showed a remarkable effect on association networks of nematodes and ammonia oxidizers. The average connectivity (avgK) and the number of edges in overall networks increased with increasing aggregate sizes, while the average geodesic distance (GD) followed the opposite trend. The LA network could be viewed as a better organized or a better operational soil food web with more functional interrelated members than the SA and IA networks. The modules related to PNA were significantly correlated and clustered together as meta-modules in networks of aggregate fractions. The role-shifts prevailed among the network members such as significant module memberships (MMs) and generalist/specialist operational taxonomic units (OTUs). A half of shared nodes were further identified as shared MMs, dominated by ammonia-oxidizing bacteria (AOB) especially for Nitrosospira cluster 3a and 10. Soil pH could explain partly the shift of module hubs in different networks, while grazing by bacterivores might account for three exclusively connecters related to Nitrososphaera clusters 1.1. The strongly coupled modules correlated positively to pH and total carbon (TC), regardless of aggregate fractions. The network analysis approach provided new insights into potential importance of network interactions between nematodes and ammonia oxidizers in soil nitrogen cycling.     

水稻OsZ314基因克隆及转录激活验证

穗是水稻(Oryza sativa L.)重要的生殖器官,其生长发育直接影响水稻的产量和品质。前期研究中,我们对水稻生殖生长时期穗发育各阶段的转录组、蛋白组数据进行联合分析,筛选到一个MYB基因家族的转录因子OsZ314基因。生物学信息分析表明,该基因位于1号染色体上,拥有典型的R2R3-MYB型转录因子结构域,且在水稻幼穗时期表达量最高;亚细胞定位结果显示,OsZ314基因定位于细胞核中;转录激活试验表明,OsZ314基因具有转录激活功能。本研究为深入探索OsZ314基因在水稻穗发育时期的分子生物学功能奠定了良好的基础。     

拟南芥DUF647家族成员RUS4植物表达载体的构建及亚细胞定位

ROOT UV-B SENSITIVE4 (RUS4)是拟南芥DUF647蛋白家族的一个功能未知的成员。RUS1和RUS2曾经报道和根UV-B-感应途径相关联,并在拟南芥早期幼苗形态发生和发育中发挥重要作用。为了探究RUS4的分子功能,本研究对RUS4蛋白进行了亚细胞定位分析。首先,我们通过Gateway TOPO载体系统,构建了融合蛋白表达载体pMDC83-RUS4;然后,通过农杆菌介导的花苞转化法,获得了转化pMDC83-RUS4的转基因拟南芥株系;对转基因植物叶肉原生质体的荧光显微镜观察显示,RUS4-GFP信号与叶绿体的自发荧光共定位,说明RUS4蛋白定位叶绿体中。该结果为进一步研究RUS4的分子功能奠定了良好的基础。     

Soil organic and inorganic carbon sequestration by consecutive biochar application: Results from a decade field experiment

Biochar addition can expand soil organic carbon (SOC) stock and has potential ability in mitigating climate change. Also, some incubation experiments have shown that biochar can increase soil inorganic carbon (SIC) contents. However, there is no direct evidence for this from the field experiment. In order to make up the sparseness of available data resulting from the long‐term effect of biochar amendment on soil carbon fractions, here we detected the contents and stocks of the bulk SIC and SOC fractions based on a 10‐year field experiment of consecutive biochar application in Shandong Province, China. There are three biochar treatments as no‐biochar (control), and biochar application at 4.5 Mg ha^?1 year^?1 (B4.5) and 9.0 Mg ha^?1 year^?1 (B9.0), respectively. The results showed that biochar application significantly enhanced SIC content (3.2%–24.3%), >53 μm particulate organic carbon content (POC, 38.2%–166.2%) and total soil organic carbon content (15.8%–82.2%), compared with the no‐biochar control. However, <53 μm silt–clay‐associated organic carbon (SCOC) content was significantly decreased (14%–27%) under the B9.0 treatment. Our study provides the direct field evidence that SIC contributed to carbon sequestration after the biochar application, and indicates that the applied biochar was allocated mainly in POC fraction. Further, the decreased SCOC and increased microbial biomass carbon contents observed in field suggest that the biochar application might exert a positive priming effect on native soil organic carbon.     

高羊茅光周期调控基因FaCONSTANS亚细胞定位与差异表达分析

构建高羊茅光周期调控基因FaCONSTANS与绿色荧光蛋白基因GFP融合的植物表达载体P-FaCONSTA NS-hGFP,基因枪转化法转入洋葱表皮细胞,荧光显微镜检测融合基因的瞬时表达.并运用实时荧光定量PCR研究光周期调控基因FaCONSTANS在长日照与短日照条件下昼夜24 h的表达差异.结果表明:FaCONST...     

黄芩纤维构成及黄酮含量的差异分析

为分析不同粒度黄芩颗粒纤维构成及黄酮含量的差异性,以5种黄芩为研究对象,分别对不同品种、不同粒度的黄芩样品中木质素、半纤维素和纤维素等纤维构成及黄芩甙、汉黄芩甙、黄芩素和汉黄芩素等主要黄酮含量进行了测定。通过方差分析发现,不同粒度黄芩中纤维构成、黄芩甙、黄芩素和总黄酮含量差异显著(P<0.05),汉黄芩甙和汉黄芩素含量无显著性差异(P>0.05)。