水稻齿叶矮缩病毒Pns10蛋白在水稻原生质体内的表达

【目的】水稻齿叶矮缩病毒(Rice ragged stunt virus,RRSV)Pns10蛋白在介体昆虫细胞内可形成类似病毒原质(viroplasm)的内含体,是RRSV侵染介体所必需。然而Pns10蛋白在水稻寄主中是否具有类似功能及其表达情况如何未见报道。【方法】利用大肠杆菌系统表达Pns10蛋白,免疫家兔制备多克隆抗体;通过水稻原生质体病毒侵染体系,利用免疫荧光技术分析Pns10蛋白在水稻原生质体内的分布情况,利用实时定量PCR技术和Western blot技术分别检测Pns10 RNA和Pns10蛋白在水稻原生质体内的积累情况。【结果】将Pns10基因克隆到Gateway系统原核表达载体p DEST17中,IPTG诱导表达成功后,制备融合蛋白抗血清。Western blot检测显示,该抗血清可检测感病水稻叶片中的Pns10蛋白。病毒侵染水稻原生质体后,Pns10蛋白可形成类似病毒原质的内含体;Pns10 RNA在病毒接种8 h后开始积累,24 h后达到最大值,随后开始下降;Pns10蛋白在24 h后开始表达,之后维持较高水平,60 h后略有下降。【结论】成功获得了Pns10抗血清;Pns10在水稻原生质体内成功表达,可形成类似病毒原质的内含体,并且Pns10 RNA的表达先于其蛋白的表达。     

RNA干扰对大鲵蛙病毒主要功能基因表达及其增殖的影响

采用q PCR与病毒滴度测定观察RNA干扰(RNA interference,RNAi)对大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)主要衣壳蛋白(major capsid protein,MCP)、甲基转移酶(methyltransferases,MTases)、DNA多聚酶(DNA polymerase)基因表达与病毒增殖的影响。结果表明,小干扰RNA(small interfering RNA,siRNA)能推迟鲤鱼上皮瘤细胞(epithelioma papillosum cyprini,EPC)出现病变,且病变程度也较对照组轻。在各功能基因表达量上,NC-FAM对照组与阴性对照组差异不显著(P0.05);而干扰组的干扰率极显著高于阴性对照组,其中siR-DP-1组siRNA对MCP基因的干扰率为79%,极显著高于其余组(P0.01);siR-MT-1、siR-MT-2组siRNA对MTases基因的干扰率为77%,极显著高于其余组(P0.01);siR-DP-1组siRNA对DNA polymerase基因的干扰率为79%,极显著高于其余组(P0.01)。干扰实验组病毒滴度与NC-FAM对照组和阴性对照组相比也有不同程度的降低。阴性对照组lg TCID50为8.362,NC-FAM对照组lg TCID50为7.848,siR-MCP、siR-MT、siR-DP实验组最低lg TCID50分别为5.764、5.317、5.362。证实RNAi能够抑制CGSRV主要功能基因的表达并影响病毒的复制。     

水稻苗期根系铵钾离子吸收的交互作用研究

【目的】为探讨水稻幼苗根系NH_4~+、K~+吸收的交互作用,深化水稻养分吸收理论,【方法】采用溶液培养的方法,对低钾及高钾浓度下水稻在有铵和无铵时的K~+吸收动力学特征进行了研究,对不同钾浓度下水稻根系NH_4~+的吸收速率进行了比较。【结果】1)当K~+0.2 mmol/L时,水稻根系通过高亲和转运系统吸收K~+服从Michaelich-Menten动力学方程;NH_4~+的存在显著降低K~+的最大吸收速率(Vmax),且降幅随着NH_4~+浓度的增加而增大;NH_4~+对水稻根表载体与K~+的亲和力(Km)影响较小,在1.62 mmol/L NH_4~+浓度下,水稻品种齐粒丝苗和沪科3号的Km分别下降了12.33%和16.46%,远低于Vmax 47.30%和39.21%的降幅。2)当K~+0.5 mmol/L时,水稻根系K~+低亲和转运系统发挥作用,K~+吸收速率随浓度的增加而不断增加,呈不饱和特征;但在相同K~+浓度下,水稻根系的K~+吸收速率随NH_4~+浓度的增加而下降。3)水稻根系对NH_4~+的吸收速率随着NH_4~+浓度的增加而增加;在相同NH_4~+浓度下,水稻根系对NH_4~+的吸收速率受K~+浓度的影响很小。【结论】NH_4~+抑制水稻苗期根系K~+的高亲和转运和低亲和转运,NH_4~+对K~+高亲和吸收的影响主要是由于铵竞争细胞膜上的钾载体所致;外界K~+浓度的变化对水稻幼苗的NH_4~+吸收速率影响很小。水稻铵钾的交互作用主要表现在NH_4~+对K~+吸收的抑制作用。     

Transgenic sorghum with suppressed synthesis of kafirin subclasses: Effects on flour and dough rheological characteristics

Arising from work showing that conventionally bred high protein digestibility sorghum types have improved flour and dough functionality, the flour and dough properties of transgenic biofortified sorghum lines with increased protein digestibility and high lysine content (TG-HD) resulting from suppressed synthesis of several kafirin subclasses, especially the cysteine-rich γ-kafirin, were studied. TG-HD sorghums had higher flour water solubility at 30 °C (p < 0.05) and much higher paste viscosity (41% higher) than their null controls (NC). TG-HD doughs were twice as strong as their NC and dynamic rheological analysis indicated that the TG doughs were somewhat more elastic up to 90 °C. CLSM of doughs and pastes indicated that TG-HD had a less compact endosperm protein matrix surround the starch compared to their NC. The improved flour and dough functional properties of the TG-HD sorghums seem to be caused by reduced endosperm compactness resulting from suppression of synthesis of several kafirin subclasses which modifies protein body and protein matrix structure, and to improved protein-starch interaction through hydrogen bonding specifically caused by reduction in the level of the hydrophobic γ-kafirin. The improved flour functionality of these transgenic biofortified sorghums can increase their commercial utility by complementing their improved nutritional quality.     

Experimental investigation on the transport of different fish species in a jet fish pump

A jet fish pump with a throat of ø60 mm was designed to study its performance in the transport of different fish species and the physiological changes in fish thereafter. Experiments were conducted to investigate the fish transport rate and energy required to transport each ton of fish when transporting Carassius auratus, commonly known as the Chinese goldfish, Megalobrama amblycephala, or Wuchang bream, and Ctenopharyngodon idella, the grass carp. Fish were examined for external injuries as well as for several important enzymes and hormones which are indicators of tissue injury and stress. The results showed that the transport rate for all three species of fish rose dramatically with an increase in the primary stream rate. In this experiment, the transport rates of C. auratus, M. amblycephala and C. idella reached 2357 ± 37.2 kg ∙ h⁻¹, 2888 ± 41.6 kg ∙ h⁻¹, and 2060 ± 40.2 kg ∙ h⁻¹, respectively. However, both injury rate and energy required to transport each ton of fish increased no matter whether the primary stream rate was too low or too high. Considering both transport rate and injury rate, the mean primary stream rate of 80 m³ ∙ h⁻¹ was determined to be the optimal operating condition in this experiment. Fish were stressed and most likely some of their organs were damaged. However, most physiological indexes almost fully recovered after several hours.     

乌鄯线冬季冷热油交替输送运行方案

为保障西部原油管道乌鄯线安全平稳运行,实现节能降耗输送,分析了乌鄯线和阿独乌线外输哈国油物性、沿线油温、地温数据,进而探讨了乌鄯线冬季运行冷热油交替输送的可行性。研究结果表明：长输原油管道冬季低输量运行,其进站油温差距不大并与地温的变化趋势一致,沿线地温为冬季冷热油交替输送的重要参数;采用冷热油交替输送工艺,哈国油常温输送对北疆油物性影响不大;乌鄯线冬季运行采用冷热油交替输送,节约燃料天然气约51.6%,节能降耗效果明显,说明乌鄯线冬季运行采用冷热油交替输送具有可行性。     

ApoB RNA编辑高效蛋白检测体系的构建

[目的]建立一个载脂蛋白B(apoB)RNA编辑蛋白检测体系。[方法]在慢病毒表达质粒载体pCSII-CMV-IRES-Neor中插入apoB RNA编辑保守序列,并在其2端嵌入红色荧光蛋白(DsRed)、绿色荧光蛋白(GFP)表达序列,以此慢病载体转染大鼠肝癌细胞系CBRH-7919获得稳定表达。采用共聚焦显微镜测序方法检测apoB RNA的编辑。[结果]目的指示基因能够在CBRH-7919细胞中稳定表达,表现出指征RNA编辑的多色特征。[结论]试验成功构建了在细胞水平上以颜色变化指示apoB RNA编辑的检测体系,该体系为快速检测以apoB RNA编辑为靶的降胆固醇药物筛选提供了基础。     

Exploring connectivity between spawning and nursery areas of Mullus barbatus (L., 1758) in the Mediterranean through a dispersal model

Connectivity between spawning and nursery areas plays a major role in determining the spatial structure of fish populations and the boundaries of stock units. Here, the potential effects of surface current on a red mullet population in the Central Mediterranean were simulated using a physical oceanographic model. Red mullet larvae were represented as Lagrangian drifters released in known spawning areas of the Strait of Sicily (SoS), which represents one of the most productive demersal fishing‐grounds of the Mediterranean. To consider the effect of inter‐annual variability of oceanographic patterns, numerical simulations were performed for the spawning seasons from 1999 to 2012. The main goal was to explore connectivity between population subunits, in terms of spawning and nursery areas, inhabiting the northern (Sicilian‐Maltese) and southern (African) continental shelves of the SoS. The numerical simulations revealed a certain degree of connectivity between the Sicilian–Maltese and the African sides of the SoS. Connectivity is present in both directions, but it is stronger from the Sicilian–Maltese spawning areas to the African nurseries owing to the marine circulation features of the region. However, because the majority of the larvae are transported to areas unsuitable for settlement or outside the SoS, the dispersal process is characterized by a strong loss of potential settlers born in the spawning areas. These results are in agreement with the low genetic heterogeneity reported for this species in the Mediterranean Sea and support the existence of a metapopulation structure of red mullet in the SoS and the adjacent areas.     

Breeding for increased grain protein and micronutrient content in wheat: Ten years of the GPC-B1 gene

To provide food and nutrition security for a growing world population, continued improvements in the yield and nutritional quality of agricultural crops will be required. Wheat is an important source of calories, protein and micronutrients and is thus a priority to breed for improvements in these traits. The GRAIN PROTEIN CONTENT-B1 (GPC-B1) gene is a positive regulator of nutrient translocation which increases protein, iron and zinc concentration in the wheat grain. In the ten years since it was cloned, the impacts of GPC-B1 allelic variation on quality and yield traits have been extensively analyzed in diverse genetic backgrounds in field studies spanning forty environments and seven countries. In this review, we compile data from twenty-five studies to summarize the impact of GPC-B1 allelic variation on fifty different traits. Taken together, the results demonstrate that the functional copy of the GPC-B1 gene is associated with consistent positive effects on grain protein, Fe and Zn content with only marginally negative impacts on yield. We conclude that the GPC-B1 gene has the potential to increase nutritional and end use quality in a wide range of modern cultivars and environments and discuss the possibilities for its application in wheat breeding.     

Conservation needs for the endangered New Zealand sea lion,Phocarctos hookeri

1. Understanding population size and trend is critical information in species management and conservation. To enable accurate population trend estimates, consistent robust monitoring of a species is essential, particularly for a species such as the New Zealand (NZ) sea lion, Phocarctos hookeri , which has experienced an almost continuous decline in pup production since the late 1990s.
2. This research examines the pup production estimates for all known breeding sites for this species, and using a stage‐structured matrix population model, estimates population size and trend between 1995 and 2015.
3. Overall, it is estimated that 2,316 pups were born in 2015, a decrease of 13% since 1995 and a 27% decline since the highest pup production estimate in 1998. This decline has been driven by the significant decline of 48% at the main breeding area, the Auckland Islands since 1998.
4. Using the stage‐structured matrix population model a total species population size of 11,767 sea lions (95% CrI: 10,790–12,923) was estimated. This is the lowest population size of any sea lion species. Trend data for the Auckland Islands indicated that pup and population numbers have decreased at 1.9% yr^?1 in the last 20 yr, while total species population decline is 0.6% yr^?1.
5. Estimates of population trends for this species have been hindered by inconsistent monitoring at most breeding sites. This study strengthens the growing field of research highlighting the need for consistent long‐term monitoring for the conservation management of endangered species.