西瓜含有JmjC结构域的组蛋白去甲基化酶家族的生物信息学分析

利用生物信息学方法,对西瓜（Citrullus lanatus（Thunb.）Matsum.&Nakai）JmjC基因家族的成员进行鉴定,对该基因家族的染色体定位、基因结构、蛋白结构域、选择压力和酶活位点进行分析,并对该基因家族与其它物种的系统进化及共线性关系进行研究。结果显示：西瓜全基因组含有17个JmjC候选基因,核苷酸序列长度为1209~5541 bp;这些基因均含有JmjC结构域,分别位于9条染色体上,归属8个亚族。系统进化、选择压力以及共线性分析结果表明,西瓜与黄瓜（Cucumis sativus L.）亲缘关系较近,JmjC家族基因数量相同,其中14个成员呈现一对一的共线性关系;而西瓜与拟南芥（Arabidopsis thaliana（L.）Heynh）亲缘关系较远,但西瓜和拟南芥同一亚族中JmjC基因间K_a/K_s的比值均小于1,推测西瓜各个亚族成员的编码蛋白功能与同一亚族的拟南芥成员功能极为相似。酶活位点分析结果表明西瓜JmjC基因家族中有10个成员具有潜在的组蛋白去甲基化酶活性。     

西伯利亚蝗抗菌肽基因的克隆及表达特性研究

[目的]扩增西伯利亚蝗Gomphocerus sibiricus抗菌肽(attacin)基因,并对其进行生物信息学分析,同时分析西伯利亚蝗不同组织中抗菌肽基因的差异表达.[方法]基于RACE技术从西伯利亚蝗中克隆得到抗菌肽,利用生物信息学软件对西伯利亚蝗抗菌肽的一级结构、二级结构、三级结构、系统进化和亚细胞定位进行分析.采用实时荧光定量PCR技术检测了西伯利亚蝗抗菌肽基因在马氏管、中肠、后肠、脂肪体和唾液腺中的相对表达量.[结果]克隆获得的抗菌肽基因的开放阅读框为282 bp,编码93个氨基酸,其编码蛋白预测理论分子量为9.98 ku,等电点为9.26.抗菌肽氨基酸序列中存在信号肽序列.基因定量分析显示,抗菌肽基因在西伯利亚蝗的中肠中表达量最高.[结论]attacin在西伯利亚蝗成虫不同组织中差异表达,提示其消化道在防御病原微生物的入侵中起着重要的作用.     

A Highlights from MBoC Selection: Protein kinase C activation decreases peripheral actin network density and increases central nonmuscle myosin II contractility in neuronal growth cones

Protein kinase C (PKC) can dramatically alter cell structure and motility via effects on actin filament networks. In neurons, PKC activation has been implicated in repulsive guidance responses and inhibition of axon regeneration; however, the cytoskeletal mechanisms underlying these effects are not well understood. Here we investigate the acute effects of PKC activation on actin network structure and dynamics in large Aplysia neuronal growth cones. We provide evidence of a novel two-tiered mechanism of PKC action: 1) PKC activity enhances myosin II regulatory light chain phosphorylation and C-kinase–potentiated protein phosphatase inhibitor phosphorylation. These effects are correlated with increased contractility in the central cytoplasmic domain. 2) PKC activation results in significant reduction of P-domain actin network density accompanied by Arp2/3 complex delocalization from the leading edge and increased rates of retrograde actin network flow. Our results show that PKC activation strongly affects both actin polymerization and myosin II contractility. This synergistic mode of action is relevant to understanding the pleiotropic reported effects of PKC on neuronal growth and regeneration.     

Molecular properties and immune defense of two ferritin subunits from freshwater pearl mussel,Hyriopsis schlegelii

Ferritin is a conserved iron-binding protein involved in cellular iron metabolism and host defense. In the present study, two distinct cDNAs for ferritins in the freshwater pearl mussel Hyriopsis schlegelii were identified (designated as HsFer-1 and HsFer-2) by SMART RACE approach and expressed sequence tag (EST) analysis. The full-length cDNAs of HsFer-1 and HsFer-2 were of 760 and 877 bp, respectively. Both of the two cDNAs contained an open reading frame (ORF) of 522 bp encoding for 174 amino acid residues. Sequence characterization and homology alignment indicated that HsFer-1 and HsFer-2 had higher similarity to H-type subunit of vertebrate ferritins than L-type subunit. Analysis of the HsFer-1 and HsFer-2 untranslated regions (UTR) showed that both of them had an iron response element (IRE) in the 5′-UTR, which was considered to be the binding site for iron regulatory protein (IRP). Quantitative real-time PCR (qPCR) assays were employed to examine the mRNA expression profiles. Under normal physiological conditions, the expression level of both HsFer-1 and HsFer-2 mRNA were the highest in hepatopancreas, moderate in gonad, axe foot, intestine, kidney, heart, gill, adductor muscle and mantle, the lowest in hemocytes. After stimulation with bacteria Aeromonas hydrophila, HsFer-1 mRNA experienced a different degree of increase in the tissues of hepatopancreas, gonad and hemocytes, the peak level was 2.47-fold, 9.59-fold and 1.37-fold, respectively. Comparatively, HsFer-2 showed up-regulation in gonad but down-regulation in hepatopancreas and hemocytes. Varying expression patterns indicate that two types of ferritins in H. schlegelii might play different roles in response to bacterial challenge. Further bacteriostatic analysis showed that both the purified recombinant ferritins inhibited the growth of A. hydrophila to a certain degree. Collectively, our results suggest that HsFer-1 and HsFer-2 are likely to be functional proteins involved in immune defense against bacterial infection.     

Molecular cloning,characterization of one key molecule of teleost innate immunity from orange-spotted grouper (Epinephelus coioides): Serum amyloid A

The orange-spotted grouper (Epinephelus coioides), a favorite marine food fish, is widely cultured in China and Southeast Asian countries. However, little is known about its acute phase response (APR) caused by viral diseases. Serum amyloid A (SAA) is a major acute phase protein (APP). In this study, a new SAA homologous (EcSAA) gene was cloned from grouper, E. coioides, by rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA sequence of SAA was 508 bp and contained a 363 bp open reading frame (ORF) coding for a protein of 121 aa. Similar to other fish known SAA genes, the EcSAA gene contained four exons and three introns. Quantitative real-time PCR analysis revealed that EcSAA mRNA is predominately expressed in liver and gill of grouper. Furthermore, the expression of EcSAA was differentially up-regulated in liver after infection with Staphyloccocus aureus, Vibrio vulnificus, Vibrio parahaemolyticus, Saccharomyces cerevisiae and Singapore grouper iridovirus (SGIV). Recombinant EcSAA (rEcSAA) was expressed in Escherichia BL21 (DE3) and purified for mouse anti-EcSAA serum preparation. The rEcSAA fusion protein was demonstrated to bind to all tested bacteria and yeast, and inhibit the replication of SGIV. Overexpression of EcSAA in grouper spleen (GS) cells could also inhibit the replication of SGIV. These results suggest that EcSAA may be an important molecule in the innate immunity of grouper.     

The effects of post-stroke upper-limb training with an electromyography (EMG)-driven hand robot

Loss of hand function and finger dexterity are main disabilities in the upper limb after stroke. An electromyography (EMG)-driven hand robot had been developed for post-stroke rehabilitation training. The effectiveness of the hand robot assisted whole upper limb training was investigated on persons with chronic stroke (n = 10) in this work. All subjects attended a 20-session training (3–5 times/week) by using the hand robot to practice object grasp/release and arm transportation tasks. Significant motor improvements were observed in the Fugl-Meyer hand/wrist and shoulder/elbow scores (p < 0.05), and also in the Action Research Arm Test and Wolf Motor Function Test (p < 0.05). Significant reduction in spasticity of the fingers as was measured by the Modified Ashworth Score (p < 0.05). The training improved the muscle co-ordination between the antagonist muscle pair (flexor digitorum (FD) and extensor digitorum (ED)), associated with a significant reduction in the ED EMG level (p < 0.05) and a significant decrease of ED and FD co-contraction during the training (p < 0.05); the excessive muscle activities in the biceps brachii were also reduced significantly after the training (p < 0.05).     

A Comparison of Vegetation and Ground‐Dwelling Ants in Abandoned and Restored Gullies and Landslide Surfaces in the Western Colombian Andes

Landslides and gullies are two common manifestations of land degradation in the densely populated Colombian Andes. In these unstable areas, further mass movements pose a serious threat to local populations and cause off‐site environmental damage through sedimentation, pollution, and increased flooding. A novel approach for restoring severely eroded slopes combines the use of stabilization structures made with stalks of Guadua angustifolia Kunth, Poaceae (bamboo), with high‐density planting of species that exhibit quick growth and sprouting. This study compared the vegetation and ground‐dwelling ant assemblages of 10 pairs of gullies, each pair formed by one enhanced and one untreated or control gully, 6–8 years after restoration or abandonment. The restoration treatment had significant effects on the complexity of vegetation. Average values for plant species richness, basal area, stem density, foliage density index, and total vegetation volume were 11.6, 140, 30, 11.5, and 15.6 times larger, respectively, in enhanced than in control gullies. Mirroring the differences in vegetation, average ant species richness was significantly larger (13 vs. 7.6 species per gully), and a higher proportion of ant species nested within enhanced than control gullies (52.5 vs. 30%). While control gullies were dominated by generalist ants such as Ectatomma ruidum and Linepithema angulatum, enhanced gullies had more specialized ground‐dwelling species, normally associated with high plant cover and abundant leaf litter such as Octostruma balzani and Heteroponera inca. We conclude that this restoration strategy promotes a fast recovery of vegetation and the ground‐dwelling ant fauna in these tropical mountains.     

Establishing Restoration Strategy of Eastern Oyster via a Coupled Biophysical Transport Model

For marine fish and invertebrates, larval dispersal plays a critical role in determining connections among source and sink habitats, and the lack of a predictive understanding of larval dispersal is a fundamental obstacle to the development of spatially explicit restoration plans for marine populations. We investigated larval dispersal patterns of eastern oyster in an estuary along the Northern Gulf of Mexico under different simulation scenarios of tidal amplitude and phase, river discharge, wind direction, and larval vertical migration, using a coupled biophysical transport model. We focused on the dispersal of larvae released from the commercially exploited (Cedar Point, CP) and non‐exploited (Bon Secour Bay, BSB) oyster populations. We found that high flushing rates through the dominant inlet prevented larval exchange between the commercially exploited and non‐exploited populations, resulting in negligible connectivity between them. Variations in tidal amplitude, river discharge and wind direction played a more important role in the amount of larvae retained in Mobile Bay when they are released from CP than from BSB. Under most of the scenarios, larvae from BSB were retained around the spawning area, while larvae from CP showed a predominant westward flow. Net sinking behavior of late‐stage larvae increased larval retention in the bay, but physical transport showed a higher impact in the amount of larvae retained. These findings have enhanced our understanding of larval dispersal of eastern oyster in a wide, shallow estuarine system, and been used to establish spatially explicit strategies for oyster restoration in the Mobile Bay system, Alabama.     

Integration of Biological Control and Native Seeding to Restore Invaded Plant Communities

Disturbed natural areas frequently experience invasion by introduced plant species that can reduce native biodiversity. Biological control can suppress these introduced species, but without restoration another introduced species can invade. Integration of biological control with concurrent revegetation can both aid in weed reduction via interspecific plant competition and establish a restored native plant community. This 3‐year study investigated an integrated approach to controlling the introduced annual Mile‐a‐minute weed (Persicaria perfoliata [L.] H. Gross [Polygonaceae]) using the biocontrol weevil Rhinoncomimus latipes Korotyaev (Coleoptera: Curculionidae) and restoration planting using a native seed mix. A fully factorial design tested weevils and seeding, separately and together, using insecticide to eliminate weevils. The weevils together with the native seed mix reduced P. perfoliata percent cover in 2009 and 2010, and peak seed cluster production in 2010, compared to the insecticide ? no seed control treatment. Persicaria perfoliata final dry biomass was reduced by 75% in 2010 and by 57% in 2011 in the weevils plus seed treatment compared to the control, with weevils having the greatest effect in 2010 and the seed treatment having the greatest impact in 2011. Results suggest an additive effect of biocontrol and seeding in suppressing P. perfoliata. Seeded treatments also developed the highest native plant species richness and diversity, comprised of spontaneous recolonization in addition to species from the seed mix. Results support the use of integrated management of this invasive weed, with suppression through biological control and native revegetation together helping prevent reinvasion while restoring native plant biodiversity.