不同辣椒种质对象耳豆根结线虫的抗性评价

近年来象耳豆根结线虫的危害日益严重.为筛选抗象耳豆根结线虫的辣椒种质,本试验利用种属特异性引物对根结线虫病原物开展分子鉴定,确定为象耳豆根结线虫.接着采用室内人工接种法,对10份一年生辣椒种质和10份中国辣椒种质进行象耳豆根结线虫抗性鉴定,计算根结指数和卵粒指数,并通过比较隶属函数,发现其中3份辣椒种质对象耳豆根结线虫表现为高抗,抗病性最强的是L518M和L525-1M,L42M次之;10份表现为中抗;7份表现为感病,抗病能力最弱的是L69-1M;未发现免疫品种.本研究发现中国辣椒种质的抗病性整体高于一年生辣椒,是重要的抗病种质来源,可用于象耳豆根结线虫抗病育种和后续抗病机理的研究.     

小球藻遗传转化技术研究进展

小球藻( Chlorella )是一种单细胞真核藻类,属绿藻门、绿藻纲、绿球藻目、小球藻科、小球藻属 [1] 。作为最早开发的真核微藻之一,具有高营养价值、生长快速、结构简单、易工业化集成等显著优点。其细胞形态为球形或椭圆形,直径3~12 μm,呈单生或聚集成群状生长 [2] ,分布广泛,多见于淡水、咸水和土壤中。作为地球上最早的生命之一,小球藻基因比较稳定,至今未见有关其基因自发突变的报道。因其富含蛋白质、脂质、维生素、活性代谢产物等多种营养物质而被公认为具有高附加值和医疗保健作用,已经被广泛应用于保健食品 [3] 、水产养殖 [4] 、生物能源 [5] 等方面,关于小球藻生物技术的研究主要集中在基因组学 [6] 、分子遗传学 [7] 、代谢机理 [8] 、大规模培养 [9] 等方向。     

基于格网的GCM数据修订分析未来海南岛农业水热资源的变化特征

以海南岛为研究区域，选用5个大气环流模式（GCMs）1970−1999年的逐日输出数据和同期地面气象观测数据，使用空间插值降尺度到0.5°×0.5°格网。以格网单元为基础，应用系统误差修订（修正值法或比值法）和多模式集合平均方法（贝叶斯模型平均法BMA或等权重平均法EW），训练与验证GCMs输出值并进行综合修订。在此基础上，分析RCP2.6、RCP4.5和RCP8.5情景下，未来海南岛近期（2020−2059年）和远期（2060−2099年）农业水热资源，包括年平均气温、1月平均气温、≥10℃积温、≥20℃积温、年降水量、1月降水量和≥20℃界限温度生长期间降水量的变化特征。结果表明：GCMs输出值的系统误差和BMA权重系数在格网间存在较大的空间差异，且GCMs输出值低估逐日最高气温约3.55℃，高估逐日最低气温约1.19℃，逐日降水量仅为观测值的54.35%。基于格网的综合修订，可有效降低GCMs输出值在空间上的不确定性，BMA与EW的修订结果相似，均优于单一GCM模式。通过格网BMA综合修订后，最高气温、最低气温和降水量在验证期的相关系数r分别约提升0.10、0.07和0.06；均方根误差RMSE分别约降低2.38℃、1.01℃和1.01mm；较单一GCM相对观测值的偏差平均约减少3.25℃、1.13℃和25.67mm。未来海南岛农业热量资源在空间上主要表现为从中部向外围逐渐升高，高温主要分布在南部至西部沿海地区，年平均气温的增幅全岛较为接近，1月平均气温、≥10℃积温和≥20℃积温的增幅分别表现为由东向西、由北向南和由中部向外围递减。在时间上，RCP8.5情景下所有农业热量资源均为极显著增加且增温最快，RCP4.5情景为先增加后平缓，RCP2.6情景较为平缓，远期无显著增温。未来海南岛降水资源在空间上转为由东向西逐步递减的格局，南部和北部沿海地区降水变率增加，西部和中部降水变率减少，在时间上无显著变化趋势。随着未来海南岛气候变暖和降水格局的改变，农作物适宜种植面积扩大，会对农业生产带来巨大挑战，应提前布局，做好趋利避害。     

称重式降水传感器与翻斗雨量传感器测雨性能对比分析

为了观测降水数据的准确性,选取了南通市4个台站2018年4月—10月份的DCS1型称重式降水传感器和SL3-1型翻斗雨量传感器观测的降水数据,分析两者观测的日降水量和降水总量等方面的差异。结果显示：在选取的观测样本中,以人工观测的降雨日数为参考,称重传感器的平均一致率为91.2%;以翻斗传感器观测的降雨日为参考,称重传感器的一致率为94.5%。称重传感器观测的累积降水量与翻斗传感器观测的累积降水量相对平均误差为-1.55%,符合中国气象局规定的相对误差不超过±4%的规范要求。在分析日降水量方面,发现称重传感器比翻斗传感器观测量平均偏小0.29 mm;有明显性降水时,两者在日降水量等级判别上基本一致。称重传感器观测的数据小于翻斗传感器观测的数据概率要高,两种传感器观测的差值分布较为一致。称重传感器在捕捉微量降水的能力不比翻斗传感器差,甚至更好。称重传感器观测的日降水量与翻斗传感器观测的日降水量相关系数为0.99826,达到0.01显著相关的水平。     

玉米秸秆消化率全基因组关联分析

48h体外干物质消化率（48h in vitro dry matter digestibility, 48h IVDMD）是衡量青贮玉米品质的重要指标。为了初步探究玉米秸秆消化率的分子遗传机理,以341份玉米自交系为材料,于2018年在沈阳和通辽种植,收获后测定秸秆48h IVDMD。利用全基因组重测序获得的6 276 612个高质量SNPs进行全基因组关联分析,共检测到153个与玉米秸秆消化率显著相关的SNPs位点（P<1.0×10^-6）,4个SNPs显著水平在P<1.0×10^-8以上;共找到38个秸秆消化率的候选基因,主要涉及细胞生长发育、防御反应和信号转导等生物学功能。     

Effect of Jieyuwan on depression-like behaviors and expression of BDNF in hippocampus and prefrontal cortex of WKY rats

AIM:To investigate the mechanism of depression and its development, and to study the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus and prefrontal cortex of Wistar-Kyoto (WKY) rats treated with Jieyuwan. METHODS:Adult male WKY rats were used as an animal model of endogenous depression. Wistar rats of the same strain were selected as control group. WKY rats were randomly divided into model group, citalopram group and Jieyuwan group. After intragastric administration for 21 d, the changes of depression-like behaviors were observed by sucrose preference test and forced swimming test. Immunofluorescence and Western blot were used to detect the expression of BDNF in the hippocampus and prefrontal cortex. RESULTS:WKY rats showed significant depression-like behaviors, and the expression of BDNF was significantly decreased in the hippocampus and prefrontal cortex (P<0.01). The reduction of neuronal axons in hippocampus was also observed. After drug treatment, the depression-like behaviors of WKY rats were significantly attenuated, and the expression of BDNF and the number of axons were increased (P<0.01). CONCLUSION:Jieyuwan effectively attenuates the depression-like behaviors of WKY rats, and BDNF is a key factor in its antidepressant effect. Our findings further confirm the involvement of BDNF in the development of depression.     

Salvianolate attenuates oxidative stress injury of human endothelial EA.hy926 cells induced by hydrogen peroxide

AIM: To investigate the effect of salvianolate on oxidative damage induced by hydrogen peroxide in human endothelial EA.hy926 cells.METHODS: EA.hy926 cells were cultured in vitro and divided into the following groups:control group, damage group, and anti-damage groups (salvianolate+damage groups). The cell viability was measured by CCK-8 assay. The migration ability of the EA.hy926 cells was detected by Transwell assay. The content of nitric oxide (NO) in the culture supernatant of the EA.hy926 cells was examined. The levels of vascular endothelial growth factor (VEGF) were detected by ELISA. The apoptosis,mitochondrial membrane potential and intracellular superoxide anion content of the EA.hy926 cells were analyzed by flow cytometry. The protein levels of caspase-3, cleaved caspase-3, Bcl-2, Bax, NF-κB and p53 were determined by Western blot. RESULTS: Compared with damage group, the viability of EA.hy926 cells pretreated with salvianolate at different concentrations was significantly increased (P<0.05). The apoptotic rate was significantly decreased (P<0.05). Savianolate enhanced the migration ability of the cells. The levels of VEGF, NO and mitochondrial transmembrane potential were increased (P<0.05), and the intracellular ROS level was significantly decreased (P<0.05). The protein levels of NF-κB, p53, Bax and cleaved caspase-3 were significantly decreased, and the protein level of Bcl-2 was markedly increased(P<0.05). CONCLUSION: Savianolate reduces the damage of EA.hy926 cells by hydrogen peroxide exposure, and its mechanism may be related to the blocking of NF-κB signaling pathway.     

山羊卵泡发育相关基因的筛选及分析

【背景】山羊第一卵泡波中的优势卵泡（dominant follicles, DF）和从属卵泡（subordinate follicles, SF）是整个卵泡发育过程中最为关键的两个阶段。随着卵泡的进一步发育,最终DF可能发育成为成熟卵泡,直到排卵;SF将走向闭锁,其中颗粒细胞的凋亡是导致卵泡发生闭锁的关键因素。然而目前对促进卵泡的优势化或导致其闭锁的分子机理尚不清楚。【目的】通过对山羊第一卵泡波中DF和SF颗粒细胞进行高通量测序,旨在筛选影响卵泡发育的关键基因,为深入探究卵泡发育的调控机制提供理论依据。【方法】选取10只1岁龄健康的贵州白山羊分别注射前列腺素F_2α,使其同期发情,此后每天用B超检测并记录卵泡的生长情况,发情3 d后,统一屠宰并采集第一卵泡波中DF (直径4.5—6 mm)与SF (直径3 —4.5 mm),分别分离其中的颗粒细胞,提取总RNA、构建文库后通过Illumina Hiseq 2500平台进行测序。利用FastQC对测序产出raw reads进行质量评估并经过过滤后,获得品质较高的clean reads;使用Trinity对得到的clean reads进行重新组装,从而获得unigenes;使用CLC Genomics Workbench将unigenes与山羊RefSeq数据库进行比对获得mRNA;使用DESeq2 软件对获得的mRNA进行差异表达分析;分别采用goseq和kobas软件对得到的差异表达基因进行GO分析及KEGG信号通路分析;最终通过qRT-PCR对筛选出的可能影响卵泡发育的关键基因进行验证。【结果】分别对测序得到的raw reads进行过滤后,在DF颗粒细胞中获得43 217 934条clean reads,占raw reads的比例为95.19%;SF颗粒细胞中获得40 766 348条clean reads,占raw reads的比例为95.35%。将得到的unigenes与山羊的RefSeq 数据库进行比对后,共得到33 896条带有注释的转录本,再通过设定FPKM>1, q value<0.05,共在两种卵泡颗粒细胞中获得13 644个基因。设定参数：FPKM≥1,SF-FPKM/DF-FPKM>1,P<0.05,获得695个差异表达mRNA,其中233个在SF颗粒细胞中表达显著上调,462个表达显著下调;对所获得695个差异表达mRNA进行GO功能富集分析,共分为三大类42组：其中生物学过程占47.6%,细胞组分占47.6%,分子功能占4.8%;KEGG信号通路分析,发现20条通路,其中与核糖体通路相关的基因富集最为显著。通过在Genecard中进行功能分析后,筛选6个可能与山羊卵泡发育密切相关的基因,其中PRLR、PTX3、RGN在SF颗粒细胞中表现为上调;DKK3、ALDH1A2、RARRES1则表现为下调。qRT-PCR显示PRLR、RGN、DKK3、ALDH1A2、RARRES1的表达趋势与高通量测序结果一致,且RGN在从属卵泡颗粒细胞中的表达量极显著地高于优势卵泡（P<0.01）;DKK3、ALDH1A2、RARRES1在优势卵泡颗粒细胞中的表达量极显著地高于从属卵泡（P<0.01）。【结论】DKK3、ALDH1A2、RARRES1和RGN在优势卵泡和从属卵泡中表达量存在极显著差异,推测在山羊卵泡发育过程中可能促进卵泡的优势化或导致闭锁,对深入探究卵泡发育的调控机制具有重要意义。     

沙柳多元醇液化产物流变性能的研究

沙柳木粉在液化剂和催化剂的作用下制成的液化产物可生产制作聚氨酯、环氧树脂、胶黏剂等。研究沙柳液化产物的流变性能,可探索宏观流变性质与液体微观内部反应机理之间的关系,优化设备结构和加工工艺条件,对其高效利用有着重大意义。本试验将沙柳木粉在浓硫酸催化条件下进行多元醇液化,通过改变液化处理条件(反应时间、反应温度和催化剂用量)制备具有不同流变性能的沙柳液化产物。利用旋转型流变仪对所制备的沙柳液化产物进行流变性能测试和分析。沙柳木粉液化条件的单因素试验和正交试验分析结果表明:影响沙柳液化产物黏度的主要因素是反应时间,其次是反应温度和催化剂用量,最佳工艺条件为反应时间70 min、反应温度170℃、催化剂用量5%。在最佳工艺条件下,剪切速率为78.87 s-1时,黏度为0.26 Pa·s。红外光谱(FT-IR)分析得出,液化物中纤维素被大量降解,半纤维素和木质素部分降解,羟基增加,生成更多的反应活性官能团,此条件下液化反应更加充分,流体黏度较大。流变性能测试结果显示:稳态扫描测试时,黏度随剪切速率的增加逐渐减小,表现出剪切变稀的现象;剪切应力随着剪切速率的增加逐渐升高,表现出假塑性流体的性质。通过动态频率扫描曲线变化规律分析,储能模量和损耗模量随着角频率的升高而逐渐增加,复数黏度却随之减小。     

Role of PERK pathway in morphine-induced myocardial protection of H9c2 cells

AIM: To explore whether morphine protects oxidative stress-damaged myocardial cells by inhibiting the PERK pathway to reduce endoplasmic reticulum stress and prevent mitochondrial permeability transition pore (mPTP) opening. METHODS: Rat myocardial H9c2 cells were cultured to establish an oxidative stress model, and then randomly divided into control group, H₂O₂ group, H₂O₂+morphine group, H₂O₂+morphine+PERK pathway inhibitor GSK2656157 group, morphine group and GSK2656157 group. Immunohistochemical method was used to detect the effects of morphine on expression of glucose-regulated protein (GRP) 78 and GRP94 induced by oxidative stress. The protein levels of PERK signaling pathway-related molecules were determined by Western blot. Confocal microscopy was used to observe the effects of morphine on mPTP opening and endoplasmic reticulum induced by oxidative stress. Cellular toxicity was detected by lactate dehydrogenase (LDH) kit and cell viability was measured by MTT assay. RESULTS: Compared with control group, GRP78 and GRP94 proteins in H₂O₂ group were strongly expressed, and the brown-yellow particles were significantly increased, but morphine significantly inhibited this process. Compared with control group, the phosphorylation of PERK was significantly reduced with GSK2656157 treatment at different concentrations, among which 2 μmol/L had the most significant effect (P < 0.05). Oxidative stress significantly increased the protein levels of GRP78, GRP94, p-PERK and CHOP, but significantly decreased p-GSK-3β level. These changes were inhibited by morphine, and the effects of morphine were further enhanced by GSK2656157 (P < 0.05). Compared with control group, oxidative stress significantly reduced the fluorescence intensity of mitochondrial TMRE and ER-Tracker Red. Morphine significantly inhibited this effect even when mitochondrial membrane potential was reduced, mPTP was open, and endoplasmic reticulum was damaged, while GSK2656157 further enhanced the effect of morphine (P < 0.05). Compared with control group, H₂O₂ significantly increased cellular toxicity and decreased the cell viability. Morphine inhibited this effect and GSK2656157 significantly enhanced the effect of morphine (P < 0.05). CONCLUSION: Morphine protects cardiac H9c2 cells under oxidative condition by inhibiting endoplasmic reticulum stress through PERK pathway and preventing the mPTP opening via GSK-3β inactivation.