siRNA-mediated depletion of endogenous protein phosphatase 2Acalpha markedly attenuates ceramide-activated protein phosphatase activity in insulin-secreting INS-832/13 cells

The sphingolipid ceramide (CER) and its metabolites have been recognized as important mediators of signal transduction processes leading to a variety of cellular responses, including survival and demise via apoptosis. Accumulating evidence implicates key regulatory roles for intracellularly generated CER in metabolic dysfunction of the islet beta cell. We have previously reported localization of an okadaic (OKA)-sensitive CER-activated protein phosphatase (CAPP) in the islet beta cell. We have also reported immunological identification of the structural A subunit, the regulatory B56alpha subunit, and the catalytic C subunit for CAPP holoenzyme complex in insulin-secreting INS-1 cells. Herein, we provide the first evidence to suggest that siRNA-mediated knockdown of the alpha isoform of the catalytic subunit of PP2Ac (PP2Acalpha) markedly reduces the CAPP activity in INS 832/13 cells. Potential significance of the functional activation of CAPP holoenzyme in the context of lipid-and glucose-induced metabolic dysfunction of the islet beta cell is discussed.     

Interaction of nucleoredoxin with protein phosphatase 2A

A trimeric protein phosphatase 2A (PP2A(T55)) composed of the catalytic (PP2Ac), structural (PR65/A), and regulatory (PR55/B) subunits was isolated from rabbit skeletal muscle by thiophosphorylase affinity chromatography, and contained two additional proteins of 54 and 55 kDa, respectively. The 54 kDa protein was identified as eukaryotic translation termination factor 1 (eRF1) and as a PP2A interacting protein. The 55 kDa protein is now identified as nucleoredoxin (NRX). The formation of a complex between GST-NRX, PP2A(C) and PP2A(D) was demonstrated by pull-down experiments with purified forms of PP2A, and by immunoprecipitation of HA-tagged NRX expressed in HEK293 cells complexed endogenous PP2A subunits. Analysis of PP2A activity in the presence of GST-NRX showed that NRX competed with polycations for both stimulatory and inhibitory effects on different forms of PP2A.     

Spatial and temporal dynamics of oviposition behavior of bollworm and three of its predators in Bt and non-Bt cotton fields

Host plants exhibiting insect resistance traits have long been known to influence within‐plant distributions of pests and their natural enemies. Sites and timing of egg deposition are particularly important for synchrony of predators and their prey in the field. Temporal and spatial distribution of eggs of the cotton bollworms [Heliothis virescens (F.) and Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] and that of the predators Geocoris punctipes (Say) (Heteroptera: Geocoridae), Chrysoperla rufilabris (Burmeister) (Neuroptera: Chrysopidae), and Micromus spec. (Neuroptera: Hemerobiidae) were determined during three cotton seasons, from 2002 to 2004, by collecting and examining plants throughout each season. Comparisons also were made between Bt and non‐Bt cotton to investigate possible changes in oviposition behavior on Bt cotton. The study was conducted in commercial fields with insecticide use to manage pests when economic thresholds were exceeded in both cotton types. Egg densities for predators and bollworms varied among years, but were similar on Bt and non‐Bt cottons. Oviposition of bollworms and G. punctipes correlated spatially within plants, with most eggs laid on structures in the top five nodes of cotton plants and on the three outermost leaves on lateral branches regardless of cotton type. Bollworm oviposition dynamics exhibited two peaks within the season (early July and early August). Eggs of all predators and bollworms collected from the field and incubated in the laboratory had high hatching rates throughout each season (74–100%). Temporal association of predator with bollworm oviposition showed a significant correlation with green lacewings, a delay of 10 days for big‐eyed bugs, and no correlation with brown lacewings. Furthermore, Bt cotton plants exerted no significant effect on temporal or spatial patterns of oviposition of bollworms or the predators, indicating no change in oviposition behavior of bollworm females within plant structures after almost one decade of widespread planting of Bt cotton.     

Cry2Ab及Cry1Ac杀虫蛋白对棉铃虫中肠蛋白酶活性的影响

为明确Cry2Ab和Cry1Ac2种Bt杀虫蛋白单用与混用对棉铃虫Helicoverpa armigera（Htibner）中肠主要蛋白酶活性的影响,本文测定了取食含不同Bt蛋白人工饲料后棉铃虫中肠总蛋白酶、类胰蛋白酶和类胰凝乳蛋白酶活性的差异。结果发现：Cry2Ab处理12h后对棉铃虫中肠总蛋白酶影响不大;对类胰蛋白酶的影响最大,除最高浓度处理外,其他浓度处理后棉铃虫类胰蛋白酶的活性明显高于对照;但对类胰凝乳蛋白酶活性的影响呈倒“V”字型,只有6．67ug／gCry2Ab处理后的棉铃虫酶活力显著高于对照,其他浓度处理与对照差异不显著或略低于对照;随着取食含Cry2Ab饲料时间的增加,棉铃虫中肠类胰蛋白酶和类胰凝乳蛋白酶的活性比对照显著增加;与对照相比,处理36h后类胰蛋白酶活性最高可增加到6．43倍。Cry1Ac处理棉铃虫12h后总蛋白酶、类胰蛋白酶和类胰凝乳蛋白酶活性都明显增加,而且与处理浓度呈正相关;但是24h后,处理后棉铃虫的总蛋白酶和类胰凝乳蛋白酶活性明显降低,只有类胰蛋白酶活性仍高于对照,但活性增长倍数低于12h时的处理。Cru2Ab和Cry1Ac2种蛋白混用处理棉铃虫后,2种酶的酶活力基本低于Cry1Ac和Cry2Ab单用的酶活力之和;只有2种蛋白浓度均为2．22ug／g混用时,处理12h后类胰蛋白酶和类胰凝乳蛋白酶的活性高于2种蛋白单用时酶活力之和,且都显著的高于对照。     

转cry1Ac＋cry2Ab基因棉与转cry1Ac＋剧咯邢基因棉荒地的生存竞争能力

【背景】此研究为“十二五”转基因生物新品种培育国家项目中创建新的转基因棉花品种环境安全评价技术而设。【方法】以转双价双成抗虫基因（cry1Ac＋cry2Ab）棉和转双价抗虫、抗除草剂基因（cry1Ac＋肼强阳）棉为观察品种,非转基因棉赣棉11号为对照品种,在荒地用撒播和3cm深度播种2种方式,于2011年5月一2012年3月对棉花出苗率、株高、生育进程、棉吐絮瓣数、絮瓣脱落率、自生苗等生存竞争能力进行比较,检测、评价其杂草化的风险,并探讨、验证检测技术的可行性。【结果】在荒地条件下,以2种方式播种的转cry1Ac＋cry2Ab基因棉和转cry1Ac＋EP5P5基因棉与非转基因棉相比,上述各项指标的竞争能力总体上未表现显著优势。【结论与意义】转cry1Ac＋cry2Ab基因棉和转cry1Ac＋EPSPS基因棉在荒地条件下生长无杂草化风险。同时,研究证明,在荒地自然生态条件下,可以采用撒播和3cm深度播种方法检测新的转基因棉花品种在生存竞争能力上的杂草化风险,在测评上有互为参照效应,为定性评价新的转基因棉花品种的杂草化风险提供了保障。     

Ac/Ds(GUS)结构介导的水稻启动子捕获系统的建立

构建了基于Activator/Dissociation(β-glucuronidase)[简称Ac/Ds(GUS)]结构的捕获质粒p13B,用于分离水稻基因启动子.以此质粒用衣杆菌介导的方法转化粳稻品种中花11的胚性愈伤组织,对获得的18个独立转化株的T2代植株进行了抗除草剂筛选,从141个抗除草剂转基因植株中用PCR方法检测到其中37株是Ds因子发生了转座的植株,而且这种转座到新位置上的Ds因子是遗传的.初步观察到其中5株的GUS染色呈阳性.     

Studies on the mobilization of a maize transposable family,Ac/Ds, in pepper using In Vivo transient assay system

A maize transposable family, Ac/Ds, has been successfully utilized as a mutagenizing agent not only in monocot but also in dicot. In order to develop insertional mutagenesis system in pepper, the mobility of Ac/Ds has been examined. In this study, the excision of the elements was monitored via transient assay system with protoplasts. Two different systems were developed and compared; one- and two-elements systems. In a one-element system, Ac alone was introduced into cells. As a two-element system, Ac and Ds were cloned into a single vector and were expressed in protoplasts. Our data showed that both Ac and Ds elements were highly mobile in pepper cells. This is the first report suggesting that Ac/Ds mediated gene tagging system could be successfully operated in pepper.     

棉铃虫单核衣壳核多角体病毒囊膜蛋白odv—e66基因及其邻近区域的序列分析

杆状病毒ODV－E66蛋白是包涵体来源病毒（occlusion-derived virus,ODV）囊膜的结构蛋白,ODV囊膜对ODV的稳定性和感染性具有重要作用。本文报道了HaSNPV odv-e66基因及其邻近区域共4237bp的核苷酸序列及其分析结果。HaSNPV的odv-e66基因编码区全长2019bp,推测编码一个由672个氨基酸残基组成的,分子量为74.5kD的蛋白质。在起始密码子ATG上游具有杆状病毒晚期转录起始信号ATAAG。与其他杆状病毒ODV－E66的氨基酸序列比较分显示HaSNPV ODV-E66蛋白具有多个保守区域,包括N末端的强疏水功能区、序列中部的一个可能的核定位信号RKIW,两个Leu-xipper以及5个跨膜区。odv-e66基因上游的两个ORF分别与AcMNPV的orf108和orf109具有同源性,下游的ORF与LsNPV的p13基因具有同源性。     

Species-specific differences in oak foliage affect preference and performance of gypsy moth caterpillars

Laboratory feeding experiments using two transgenic Bacillus thuringiensis (Bt) rape cultivars (Bt‐Westar and Bt‐Oscar) both expressing the Cry1Ac protein, and the corresponding untransformed lines, were carried out to study the effects of transgenic Bt rape on the non‐target herbivore Athalia rosae (L.) (Hymenoptera: Tenthredinidae). Furthermore, Cry1Ac protein concentration in Bt rape leaves, A. rosae larvae fed Bt rape, their faeces, eonymph instars, pupae, and adults were quantified using an enzyme‐linked immunosorbent assay (ELISA). There were no significant differences in mortality, larval development, and weight between transgenic Bt rape and non‐transgenic rape fed A. rosae. Additionally, we did not detect any significant differences in the fecundity and fertility of adult females either fed as larvae with transgenic Bt or with non‐transgenic rape. However, results of the ELISA indicated that Cry1Ac protein was detectable in larvae and faeces (Bt‐Westar 1.1 ± 0.2 and Bt‐Oscar 0.3 ± 0.2 µg Cry1Ac protein/g fresh weight) although this was less than in the leaf material, where concentrations were 2.2 ± 0.8 µg Cry1Ac protein/g fresh weight for Bt‐Westar and 7.5 ± 2.9 µg Cry1Ac protein/g fresh weight in Bt‐Oscar. In contrast, Cry1Ac protein could not be detected in eonymphs, pupae, or adults of A. rosae. Our results suggest that Cry1Ac protein in Bt rape does not have a significant effect on the herbivore A. rosae but the protein is still detectable after ingestion and excretion by these herbivores, thus providing the possibility of exposure to organisms other than herbivores.     

转cry1Ab/cry1Ac基因籼稻对稻田节肢动物群落影响

将稻田节肢动物群落按营养关系划分为5个功能团,即植食类、寄生类、捕食类、腐食类和其它类,从功能团优势度、功能团内科组成及其优势度、群落主要参数及群落相异性等方面,经两年四点的调查就2个转cry1Ab/cry1Ac基因籼稻（Bt水稻）品系TT9.3和TT9.4对稻田节肢动物群落的影响作了较系统评价。植食类、寄生类和腐食类功能团内某些优势科的优势度在Bt水稻田与对照（IR72）田之间有时呈显著或极显著差异,如Bt水稻田中茧蜂或姬蜂科的优势度有时明显低于对照。但是,在大多情况下Bt水稻田与对照田之间功能团优势度、功能团内科组成及其优势度、群落主要参数（物种丰富度、Shannon-Wiener多样性指数、均匀性指数、优势集中性指数）及其时间动态基本无明显差异;Bt水稻田与对照田间植食类、寄生类、捕食类亚群落和整个节肢动物群落的相异性大多较低。可见,Bt水稻对稻田节肢动物群落基本无明显的负面影响。