BENT UPPERMOST INTERNODE1 encodes the class II formin FH5 crucial for actin organization and rice development

The actin cytoskeleton is an important regulator of cell expansion and morphogenesis in plants. However, the molecular mechanisms linking the actin cytoskeleton to these processes remain largely unknown. Here, we report the functional analysis of rice (Oryza sativa) FH5/BENT UPPERMOST INTERNODE1 (BUI1), which encodes a formin-type actin nucleation factor and affects cell expansion and plant morphogenesis in rice. The bui1 mutant displayed pleiotropic phenotypes, including bent uppermost internode, dwarfism, wavy panicle rachis, and enhanced gravitropic response. Cytological observation indicated that the growth defects of bui1 were caused mainly by inhibition of cell expansion. Map-based cloning revealed that BUI1 encodes the class II formin FH5. FH5 contains a phosphatase tensin-like domain at its amino terminus and two highly conserved formin-homology domains, FH1 and FH2. In vitro biochemical analyses indicated that FH5 is capable of nucleating actin assembly from free or profilin-bound monomeric actin. FH5 also interacts with the barbed end of actin filaments and prevents the addition and loss of actin subunits from the same end. Interestingly, the FH2 domain of FH5 could bundle actin filaments directly and stabilize actin filaments in vitro. Consistent with these in vitro biochemical activities of FH5/BUI1, the amount of filamentous actin decreased, and the longitudinal actin cables almost disappeared in bui1 cells. The FH2 or FH1FH2 domains of FH5 could also bind to and bundle microtubules in vitro. Thus, our study identified a rice formin protein that regulates de novo actin nucleation and spatial organization of the actin filaments, which are important for proper cell expansion and rice morphogenesis.     

Sonic Hedgehog dependent phosphorylation by CK1α and GRK2 is required for ciliary accumulation and activation of smoothened

Hedgehog (Hh) signaling regulates embryonic development and adult tissue homeostasis through the GPCR-like protein Smoothened (Smo), but how vertebrate Smo is activated remains poorly understood. In Drosophila, Hh dependent phosphorylation activates Smo. Whether this is also the case in vertebrates is unclear, owing to the marked sequence divergence between vertebrate and Drosophila Smo (dSmo) and the involvement of primary cilia in vertebrate Hh signaling. Here we demonstrate that mammalian Smo (mSmo) is activated through multi-site phosphorylation of its carboxyl-terminal tail by CK1α and GRK2. Phosphorylation of mSmo induces its active conformation and simultaneously promotes its ciliary accumulation. We demonstrate that graded Hh signals induce increasing levels of mSmo phosphorylation that fine-tune its ciliary localization, conformation, and activity. We show that mSmo phosphorylation is induced by its agonists and oncogenic mutations but is blocked by its antagonist cyclopamine, and efficient mSmo phosphorylation depends on the kinesin-II ciliary motor. Furthermore, we provide evidence that Hh signaling recruits CK1α to initiate mSmo phosphorylation, and phosphorylation further increases the binding of CK1α and GRK2 to mSmo, forming a positive feedback loop that amplifies and/or sustains mSmo phosphorylation. Hence, despite divergence in their primary sequences and their subcellular trafficking, mSmo and dSmo employ analogous mechanisms for their activation.     

Three main genetic regions for grain development revealed through QTL detection and meta-analysis in maize

Grain weight is one of the three direct yield components, being developed through a dynamic process of grain filling in maize. In this study, 258 recombinant inbred lines derived from a cross between a dent corn and a popcorn inbred were evaluated for grain fresh and dry weight at 10, 20, 30, and 40?days after pollination (DAP) and the activities of ADP-Glc pyrophosphorylase (AGPP), granule-bound starch synthase (GBSS), and soluble starch synthase (SSS) at 30 DAP. Grain-filling rate (GFR) and increasing rate of fresh weight (FWIR) were calculated during all periods. Quantitative trait locus (QTL) mapping was conducted for all traits. Meta-QTL (mQTL) was revealed by meta-analysis using BioMercator. Totally, 161 QTL were detected for six traits. QTL on chromosomes 1, 7, and 10 were detected in most cases, with 43, 54, and 28 QTL, respectively. For each trait, 1?C4 QTL were detected but no QTL for GBSS. Three mQTL at bins 7.02?C7.03, 1.03?C1.04, and 10.05?C10.06 included 47, 24, and 23 QTL detected in this study. Together with 28 QTL for grain weight detected in our previous research, they included 53, 28, and 25 QTL, respectively. Five identified expressed sequence tags (EST), five candidate genes with related functions, and QTL for grain weight in other research were co-located in these regions. It is worth concentrating further research on these regions to develop near-isogenic lines (NILs) of common QTL and their chromosome segment substitution lines (CSSL). Also, cloning and function validation for co-located EST and candidate genes could facilitate identification of genes for grain development and final weight.     

Genotoxicity of silver nanoparticles evaluated using the Ames test and in vitro micronucleus assay

Silver nanoparticles (AgNPs) have antimicrobial properties, which have contributed to their widespread use in consumer products. A current issue regarding nanomaterials is the extent to which existing genotoxicity assays are useful for evaluating the risks associated with their use. In this study, the genotoxicity of 5 nm AgNPs was assessed using two standard genotoxicity assays, the Salmonella reverse mutation assay (Ames test) and the in vitro micronucleus assay. Using the preincubation version of the Ames assay, Salmonella strains TA102, TA100, TA1537, TA98, and TA1535 were treated with 0.15-76.8 μg/plate of the AgNPs. Toxicity limited the doses that could be assayed to 2.4-38.4 μg/plate; no increases in mutant frequency over the vehicle control were found for the concentrations that could be assayed. Human lymphoblastoid TK6 cells were treated with 10-30 μg/ml AgNPs, and additional cells were treated with water and 0.73 gy X-rays as vehicle and positive controls. Micronucleus frequency was increased by the AgNP treatment in a dose-dependent manner. At a concentration of 30 μg/ml (with 45.4% relative population doubling), AgNPs induced a significant, 3.17-fold increase with a net increase of 1.60% in micronucleus frequency over the vehicle control, a weak positive response by our criteria. These results demonstrate that the 5 nm AgNP are genotoxic in TK6 cells. Also, the data suggest that the in vitro micronucleus assay may be more appropriate than the Ames test for evaluating the genotoxicity of the AgNPs.     

Dopamine Affects the Change of Pain-Related Electrical Activity Induced by Morphine Dependence

Morphine is among the most effective analgesics. However, many evidences suggest that, besides the well-know analgesic activity, repeated opioids treatment can induce some side effects such as dependence, hyperalgesia and tolerance. The mechanism of noxious information transmission in the central nervous system after dependence is not clear. An important neurotransmitter, dopamine (DA) participates not only in the process of opioid dependence but also in pain modulation in the central nervous system. In the present study we observed changes of electrical activities of pain-excitation neurons (PENs) and pain-inhibition neurons (PINs) in the caudate nucleus (Cd) following the development of morphine dependence. We also observed the role of DA on these changes. Our results revealed that both the latency of PEN discharges and the inhibitory duration of PIN discharges decreased, and the net increased values of PEN and PIN discharges increased in the Cd of morphine dependent rats. Those demonstrated that electrical activities of both PENs and PINs increased in morphine dependent rats. DA inhibited the electrical activities of PENs and enhanced those of PINs in morphine dependent rats.     

生态基因组学研究进展

生态基因组学是一个整合生态学、分子遗传学和进化基因组学的新兴交叉学科。生态基因组学将基因组学的研究手段和方法引入生态学领域,通过将群体基因组学、转录组学、蛋白质组学等手段与方法将个体、种群及群落、生态系统不同层次的生态学相互作用整合起来,确定在生态学响应及相互作用中具有重要意义的关键的基因和遗传途径,阐明这些基因及遗传途径变异的程度及其生态和进化后果的特征,从基因水平探索有机体响应天然环境(包括生物与非生物的环境因子)的遗传学机制。生态基因组学的研究对象可以分为模式生物与非模式生物两大类。拟南芥、酿酒酵母等模式生物在生态基因组学领域发挥了重要作用。随着越来越多基因组学技术的开发与完善,越来越多的非模式生物生态基因组学的研究将为生态学的发展提供重要的理论与实践依据。生态基因组学最核心的方法包括寻找序列变异、研究基因差异表达和分析基因功能等方法。生态基因组学已广泛渗透到生态学的相关领域中,将会在生物对环境的响应、物种间的相互作用、进化生态学、全球变化生态学、入侵生态学、群落生态学等研究领域发挥更大的作用。     

苏云金芽孢杆菌Cry1A(b)抗虫基因LAMP检测方法的建立与应用

以转基因玉米MON810为模板,针对Cry1A(b)抗虫基因核酸保守序列设计特异性引物,建立LAMP检测体系。对该体系的可行性、灵敏性、特异性进行分析,并应用于转基因产品的检测。研究结果显示该方法快速简单、灵敏度特异性高、结果可视化,可应用于转基因产品中Cry1A(b)基因的初步筛选。     

Deubiquitinase USP39 and E3 ligase TRIM26 balance the level of ZEB1 ubiquitination and thereby determine the progression of hepatocellular carcinoma

Emerging evidence suggests that USP39 plays an important role in the development of hepatocellular carcinoma (HCC). However, the molecular mechanism by which USP39 promotes HCC progression has not been well defined, especially regarding its putative ubiquitination function. Zinc-finger E-box-binding homeobox 1 (ZEB1) is a crucial inducer of epithelial-to-mesenchymal transition (EMT) to promote tumor proliferation and metastasis, but the regulatory mechanism of ZEB1 stability in HCC remains enigmatic. Here, we reveal that USP39 is highly expressed in human HCC tissues and correlated with poor prognosis. Moreover, USP39 depletion inhibits HCC cell proliferation and metastasis by promoting ZEB1 degradation. Intriguingly, deubiquitinase USP39 has a direct interaction with the E3 ligase TRIM26 identified by co-immunoprecipitation assays and immunofluorescence staining assays. We further demonstrate that TRIM26 is lowly expressed in human HCC tissues and inhibits HCC cell proliferation and migration. TRIM26 promotes the degradation of ZEB1 protein by ubiquitination in HCC. Deubiquitinase USP39 and E3 ligase TRIM26 function in an antagonistic pattern, but not a competitive pattern, and play key roles in controlling ZEB1 stability to determine the HCC progression. In summary, our data reveal a previously unknown mechanism that USP39 and TRIM26 balance the level of ZEB1 ubiquitination and thereby determine HCC cell proliferation and migration. This novel mechanism may provide new approaches to target treatment for inhibiting HCC development by restoring TRIM26 or suppressing USP39 expression in HCC cases with high ZEB1 protein levels.Subject terms: Gene regulation, Prognostic markers     

俄罗斯杨水培苗对硼胁迫的生长和生理响应

该研究采用水培试验,分析了俄罗斯杨(Populus russkii)对短期硼胁迫(1、5、10mmol/L硼酸)的生长和生理响应特征,探明其对硼胁迫的耐受范围,初步揭示其耐硼的生理基础。结果显示:(1)与对照相比,1mmol/L硼酸处理俄罗斯杨水培苗的生长情况更优,叶片光合色素含量和净光合速率显著上升,SOD和APX活性及抗坏血酸含量显著上调。(2)5mmol/L硼酸处理幼苗有轻微胁迫受害症状,叶片褪绿变黄,叶绿素b含量比对照显著上升,而Chla/Chlb比值显著下降,其SOD和APX活性显著上调。(3)10mmol/L硼酸处理幼苗表现出严重毒害症状,叶片发黄、茎尖和根尖生长受抑制;叶片PSⅡ原初光能转换效率(F_v/F_m)和Chla/Chlb比值比对照显著下降;其叶片MDA含量、POD活性、抗坏血酸含量以及游离脯氨酸和可溶性糖含量均比对照显著上升。研究表明,在不同浓度硼酸短期胁迫下,俄罗斯杨水培苗能通过上调抗氧化酶活性和抗氧化物质、渗透调节物质含量来有效清除体内胁迫产生的活性氧,减轻膜脂过氧化伤害,使其在低于5mmol/L硼酸胁迫环境下正常生长,表现出较强的耐硼性。