胡椒4-香豆酸：辅酶A连接酶Pn4CL基因的克隆及表达分析     

范睿  胡丽松  伍宝朵  郝朝运 《热带作物学报》2020,41(4):737-744

根据胡椒4-香豆酸：辅酶A连接酶（4-coumarate:coenzyme A ligase, 4CL）基因的部分序列设计引物,运用RACE方法获得其家族成员的1个全长cDNA,命名为Pn4cl,长度2130 bp,开放阅读框1638 bp,编码545个氨基酸。预测Pn4CL分子量为59.57 kDa,理论等电点为5.70。该基因含有AMP-binding（AMP-binding enzyme）、CaiC[Acyl-CoA synthetase (AMP-forming) /AMP-acid ligaseⅡ]、PLN02246、AFD-class I等结合域,具有植物4CL所共有的保守结构域。系统进化分析表明,Pn4CL与北细辛的同源性最高,同时与木兰分支类植物的4CL聚类在一起,与菊分支的进化距离较近,与蔷薇分支的进化距离较远。亚细胞定位表明,该蛋白定位在细胞膜上。Real-time RT-PCR结果表明,该基因受外援激素SA和MeJA诱导表达,同时接种辣椒疫霉菌后,Pn4CL基因的表达量在抗/感2种胡椒中均出现先增加后减少的现象,并且在抗病种质中表达量较高。研究结果为Pn4CL的功能研究提供了理论依据。  相似文献   

Verrucosamide,a Cytotoxic 1,4-Thiazepane-Containing Thiodepsipeptide from a Marine-Derived Actinomycete     

Vimal Nair  Min Cheol Kim  James A. Golen  Arnold L. Rheingold  Gabriel A. Castro  Paul R. Jensen  William Fenical 《Marine drugs》2020,18(11)

A new cytotoxic thiodepsipeptide, verrucosamide (1), was isolated along with the known, related cyclic peptide thiocoraline, from the extract of a marine-derived actinomycete, a Verrucosispora sp., our strain CNX-026. The new peptide, which is composed of two rare seven-membered 1,4-thiazepane rings, was elucidated by a combination of spectral methods and the absolute configuration was determined by a single X-ray diffraction study. Verrucosamide (1) showed moderate cytotoxicity and selectivity in the NCI 60 cell line bioassay. The most susceptible cell lines were MDA-MB-468 breast carcinoma with an LD₅₀ of 1.26 µM, and COLO 205 colon adenocarcinoma with an LD₅₀ of 1.4 µM. Also isolated along with verrucosamide were three small 3-hydroxy(alkoxy)-quinaldic acid derivatives that appear to be products of the same biosynthetic pathway.  相似文献   

Roles of protein phosphatase 4 in down-regulation of eNOS Ser633 phosphorylation induced by palmitic acid in human umbilical vein endotheli?al cells     

QIN Si  ZHANG Qian  WANG Jing-jie  YU Min  LUO Yan-bei  DING Jing  LU De-qin 《园艺学报》2020,36(5):803-809

AIMTo investigate the roles of protein phosphatase 4 (PP4) in down-regulation of endothelial nitric oxide synthase (eNOS) Ser633 phosphorylation induced by palmitic acid (PA). METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with PA at 25 μmol/L, 50 μmol/L, 100 μmol/L and 200μmol/L for 36 h, or treated with PA at 100 μmol/L for 12 h, 24 h, 36 h and 48 h. Protein phosphatase 2A (PP2A) family inhibitor fostriecin (FST, 20 nmol/L) or okadaic acid (OA, 5 nmol/L) was selected to pretreat the HUVECs for 30 min. Protein phosphatase 4 catalytic subunit (PP4c) siRNA or protein phosphatase 2A catalytic subunit (PP2Ac) siRNA was transfected into the HUVECs. The protein expression levels of of eNOS, PP4c and PP2Ac, as well as the level of eNOS Ser633 phosphorylation, were detected by Western blot. The intracellular nitric oxide (NO) content was measured by DAF-FM DA. RESULTS(1) Compared with control group, the levels of eNOS Ser633 phosphorylation were decreased in PA groups in which the HUVECs were treated with 25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L PA for 36 h (P<0.05) and 100 μmol/L PA for 24 h, 36 h and 48 h (P<0.05). No significant difference in the level of total eNOS protein expression among all the groups was observed. (2) Compared with control group, both FST and OA pretreatment reversed the reduction of eNOS Ser633 phosphorylation (P<0.05) and the decrease in intracellular NO content (P<0.05) induced by PA. No significant difference in the level of total eNOS protein expression among all the groups was observed. (3) Compared with si-Control group, the PP4c protein expression was significantly reduced (P<0.05), while the level of eNOS Ser633 phosphorylation was significantly increased in si-PP4c group (P<0.05). Although the levels of PP2Ac protein expression declined significantly (P<0.05), the level of eNOS Ser633 phosphorylation remained unchanged in si-PP2Ac group. No significant differencein the level of total eNOS protein expression among all the groups was found. CONCLUSION PA significantly reduces the level of eNOS Ser633 phosphorylation and the content of NO in the HUVECs, which may be due to PA inducing the activation of the PP2A family member PP4 rather than PP2A.  相似文献   

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway     

LI Ting  HE Xue-min  PENG Rong-dong  LI Jing-ren  FU Xia-nan  ZHOU Li  SHI Guo-jun  CHEN Yan-ming 《园艺学报》2020,36(4):577-587

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.  相似文献   

Role of IP₃R in LH-EGFR-induced mouse oocyte meiotic resumption     

HAO Xiao-qiong  LI Yan-kang  FU Xu-yang  JIA Fang-yi 《园艺学报》2020,36(3):532-538

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.  相似文献   

陕西关中地区猕猴桃溃疡病菌对CuSO₄的抗性评价     

韩宁  高小宁  陈子璨  王娜娜  秦虎强  黄丽丽 《西北林学院学报》2020,35(2):125-130

为明确陕西省猕猴桃细菌性溃疡病菌（Pseudomonas syringae pv.actinidiae,Psa）对CuSO₄的抗性水平及与copA和copB基因表达量的相关性,使用最小抑菌浓度法（MIC）对分离自陕西省关中地区的84株Psa菌株对于CuSO₄的抗性进行检测。在此基础上,使用实时荧光定量PCR方法,对3株不同抗性水平Psa菌株的基因copA和copB与抗铜性的关系进行了分析。结果表明,供试的84株Psa菌株MIC分布在3.75~6.25 mmol·L^-1范围内;不同地区来源的菌株对CuSO₄的抗性表现出明显的差异,其中分离自周至的Psa菌株的抗铜水平明显高于眉县。实时荧光定量PCR分析结果显示,不同浓度CuSO₄条件诱导下,3株不同抗性水平的菌株抗铜基因copA和copB表达水平与CuSO₄浓度成正相关,且菌株的抗性水平越高,基因copA和copB相对表达水平亦越高。陕西省关中地区的Psa菌株对CuSO₄表现出较高的抗性水平,且不同地区的Psa菌株的抗性表现出明显差异;copA和copB基因参与了Psa菌株对铜离子的适应性反应。  相似文献   

稻田转为菜地初始阶段温室气体排放特征     

邬磊  何志龙  汤水荣  吴限  张文菊  胡荣桂 《中国农业科学》2020,53(24):5050-5062

【目的】近年来,随着我国社会经济的快速发展和人们生活水平的提高及膳食结构的改善,越来越多的稻田被转为蔬菜种植,影响了土壤碳氮转化过程及其引起的温室气体排放。因此有必要探究稻田转为蔬菜种植,特别是该土地利用方式转变初始阶段的温室气体（CH₄和N₂O）排放特征及其关键影响因素。【方法】试验选取了长期种植水稻的双季稻田,将其中一部分转为蔬菜种植,另一部分继续种植水稻,每个处理设置了3个重复,按照当地常规模式进行管理。采用静态暗箱—气相色谱法连续3年进行田间原位观测,比较分析稻田和由稻田转变的菜地CH₄和N₂O排放特征及其年际变化差异,明确稻田转为菜地初始阶段CH₄和N₂O排放的关键影响因素。【结果】稻田是重要的CH₄排放源,其第一年的排放强度（183.91 kg CH₄-C·hm^-2?a^-1）明显低于后续两年（241.56—371.50 kg CH₄-C·hm^-2?a^-1）,这主要归功于后两年降雨量的增加引起了土壤水分含量的升高。稻田转为菜地显著减少了CH₄排放,减少量相当于稻田CH₄年累积排放量的83%—100%。菜地第一年的CH₄累积排放量（31.22 kg CH₄-C·hm^-2）显著高于第二年（0.45 kg CH₄-C·hm^-2）和第三年（0.89 kg CH₄-C·hm^-2）,表明稻田转菜地对CH₄排放的影响具有时间滞后效应。稻田是弱的N₂O排放源（1.35—3.49 kg N₂O-N·hm^-2?a^-1）,其转为菜地显著增强了N₂O排放。菜地第一年的N₂O累积排放量（95.12 kg N₂O-N·hm^-2）显著高于第二年（38.28 kg N₂O-N?hm^-2）和第三年（40.07 kg N₂O-N·hm^-2）。菜地土壤异养呼吸对N₂O排放的影响在第一年明显高于第二、三年,表明稻田转为蔬菜种植的第一年,有机质矿化对N₂O排放有重要贡献。在100年尺度CO₂当量下,稻田转为蔬菜种植第一和第二年的综合增温潜势（GWP）相对于稻田分别显著增加了390%和98%,主要是由于增加的N₂O增温潜势超过了减少的CH₄增温潜势。但是,稻田转为菜地的第三年,菜地的GWP（(16.72±3.25) Mg CO₂-eq·hm^-2）与稻田（(14.84±1.39) Mg CO₂-eq·hm^-2）相比无显著差异,主要是由于减少的CH₄ 增温潜势完全抵消了增加的N₂O增温潜势。这些研究结果表明稻田转菜地对GWP的影响主要集中在该土地利用方式转变的第一年。【结论】稻田转为菜地显著减少了CH₄排放,增加了N₂O排放,增强了菜地第一和第二年的综合增温潜势。有机质矿化过程对新转菜地第一年较高的N₂O排放有重要贡献。这些研究结果表明了评价土地利用方式转变初始阶段温室气体排放特征的重要性,便于及时采取有效管理措施缓解温室气体排放,实现环境友好型农业可持续生产。  相似文献   

辛基酚对家蚕（Bombyx mori L.）生长发育的影响     

袁红霞  秦粉菊  顾华杰 《农业环境科学学报》2020,39(8):1661-1666

为探讨辛基酚对鳞翅目模式昆虫家蚕生长发育的影响,使用添加0.01～0.08 g·kg~(-1)辛基酚(4-tert-octylphenol,4-t-OP)的人工饲料饲养家蚕(Bombyx mori L.),研究了4-t-OP对家蚕幼虫和蛹的生长发育速度、生长量、发育整齐度和生命力等影响。结果表明:连续取食0.01 g·kg~(-1)以上浓度4-t-OP的饲料,家蚕幼虫和蛹的生长发育速度(幼虫期和蛹期发育经过时间)、生长量(幼虫期体质量和蛹期茧层量)、生命力(3龄起蚕绝食生命时数、4龄起蚕结茧率、蚕茧的死笼率和羽化率)等都明显下降,抑制眠蚕体质量。4-t-OP对家蚕幼虫生长发育后期、化蛹及羽化等发育变态期的影响更加明显,其在家蚕体内可能有累积效应。添食0.08 g·kg~(-1)4-t-OP的家蚕不能完成生活史。研究表明,4-t-OP对家蚕生长发育毒性影响有显著的浓度效应和时间效应。  相似文献   

聚对氯苯胺修饰电极的制作及应用研究     

滕占才  王波  宋晓丹  毕洪梅  徐凤英  毕红梅  张玉瑛 《黑龙江八一农垦大学学报》2006,18(4):66-68

通过对聚对氯苯胺膜修饰电极的制作和实验特性的研究,发现聚对氯苯胺膜表面均匀、光滑、牢固,在pH3～11范围内均有响应,测定的重现性好。电极使用长久且稳定性高。该电极和参比电极可直接插入家兔等活体内测定血乳酸的变化情况,对生命科学的活体检测和诊断有其实用意义。  相似文献   

小麦条锈菌鉴别寄主中4抗条锈性的转录组学分析     

汤亚琪  苏文文  李莹  王保通  李强 《麦类作物学报》2021,41(7):806-815

为解析中4的抗条锈性分子机制,通过转录组测序,以侵染0 h为对照,分析条锈菌侵染12、24和48 h后差异基因的表达情况。结果共获得5 151个持续上调表达差异基因,其中,4 482个基因获得定位信息（A、B、D染色体组各占30.75%、35.27%、33.98%）,669个为新基因（无法确定其染色体位置）。GO功能注释分类将这些基因分成3大类46个亚类,主要涉及催化活性、转运活性、细胞部件、细胞、细胞器、代谢过程、细胞过程、单有机过程等;KEGG通路分析结果显示,获得定位信息的上调表达差异基因主要涉及剪接体、苯丙氨酸代谢、过氧化物酶体、半乳糖和氨基酸代谢等通路;新基因主要涉及糖的合成与代谢、氨基酸的代谢等通路。植物与病原菌互作通路共发现33个持续上调表达差异基因,编码抗病蛋白、MLO类蛋白、NAC和WRKY转录因子等。本研究结果为进一步解析中4抗条锈病的作用机制奠定了基础。  相似文献