水牛卵母细胞孤雌激活及孤雌胚与体外受精胚发育比较

比较了水牛卵母细胞体外成熟时间对孤雌激活胚发育能力的影响,以及不同化学激活方法对水牛卵母细胞孤雌激活效果的影响,并在相同条件下,对孤雌激活胚与体外受精胚的发育能力进行了比较.结果表明,水牛卵母细胞体外成熟27 h或30 h的囊胚发育率(19.0%、17.7%)明显高于体外成熟21 h或24h的囊胚发育率(12.3%、13.8%);Ion联合6-DMAP激活水牛卵母细胞的效果优于其他几组激活方法;在相同条件下,孤雌激活胚与体外受精胚的发育能力存在着差异,其中卵裂率差异不显著,但孤雌激活胚的囊胚发育率显著高于体外受精胚.     

Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.     

Effects of selective cutting and timing of herbicide application on growth and development of Cirsium arvense in spring barley

Recently, a new implement for controlling weeds in cereals (CombCut) has been developed. It cuts weeds in growing cereals without damaging them by using the physical differences (in height, stem thickness, straw stiffness and branching pattern) between crops and weeds. To evaluate and compare the effects of selective cutting with different timings of herbicide application on Cirsium arvense in spring barley, a randomised block experiment was conducted in Sweden in 2015–2017, in a field with a naturally occurring C. arvense population. Treatments consisted of control (C), herbicide application at 4–5‐leaf stage of C. arvense (H1), herbicide application at 8–10‐leaf stage (H2) and selective cutting at 10‐leaf stage (S). The treatments were performed in 2015 and repeated in 2016 in the same plots, and a final evaluation was performed in 2017. Compared to the control, S, H1 and H2 were equally efficient in reducing above‐ground biomass production of C. arvense and increasing spring barley grain yield per unit area. The number of C. arvense shoots per area was, however, higher in S compared to H1 and H2. No differences in control effects on shoot number were observed between H1 and H2. Our study indicates that (i) selective cutting (S) reduces C. arvense equally efficient as herbicide application and (ii) early herbicide spraying is as efficient as spraying later in the season.     

马铃薯卷叶病毒PLRV RT-LAMP检测方法优化

马铃薯卷叶病毒Potato leafroll virus(PLRV)是目前严重影响马铃薯产量与品质的主要病毒之一,给马铃薯产业造成巨大损失。本研究采用环介导等温核酸扩增(loop-mediated isothermal amplification, LAMP)技术建立PLRV的RT-LAMP检测方法。采取单因素变化试验,对RT-LAMP反应体系中多个因素包括引物组合、温度条件及Mg~(2+)、betaine、Bst 3.0 DNA聚合酶、dNTPs、UNG、SYBR GreenⅠ和引物组合的浓度进行一系列试验和优化。采用RT-PCR检测方法进行平行比对试验,对优化后的RT-LAMP反应体系进行了验证。结果表明,最佳引物组合为P3,最适反应温度62℃,25μL反应体系中,Mg~(2+)、betaine、Bst 3.0 DNA聚合酶和UNG的最佳终浓度分别为4 mmol/L、0 mmol/L、0.64 U/μL和0.08 U/μL,dNTPs的最佳用量为1μL(dATP、dGTP、dCTP各0.4 mmol/L,dUTP 1.2 mmol/L),SYBR GreenⅠ(20×)的最佳用量1μL,primer mix的最佳用量2.5μL(PLRV-FIP/BIP、PLRV-F3/B3和PLRV-LF/LB的浓度分别为0.8、0.2μmol/L和0.6μmol/L),RNA模板1μL(2 ng/μL),加DEPC-H_2O至25μL,反应时间50 min。优化后的RT-LAMP检测结果与RT-PCR一致,且可视化判读结果。因此,建立的PLRV RT-LAMP检测方法为进一步开发RT-LAMP检测试剂盒及其实际应用奠定了基础。     

氯虫苯甲酰胺和氟虫腈对黏虫细胞色素P450基因表达的影响

为筛选出黏虫Mythimna separata参与杀虫剂解毒代谢的主效细胞色素P450基因,采用叶片浸渍法测定了用于处理黏虫3龄幼虫的亚致死浓度,通过构建转录组测序文库并结合数字基因表达(digital gene expression,DGE)对不同处理的黏虫进行测序,并运用实时荧光定量PCR(realtime quantitative polymerase chain reaction,RT-qPCR)技术验证12个P450基因的表达情况。结果表明,用于处理黏虫的氯虫苯甲酰胺和氟虫腈亚致死浓度LC_(10)分别为0.15、13.66 mg/L;对照样品、氟虫腈处理样品和氯虫苯甲酰胺处理样品分别获得59 521 504、64 838 148和41 722 990个原始序列数据,分别获得57 441 216、62 368 912和40 285 164个过滤后的序列数据;过滤后的序列长度分别为8.62、9.36和6.04 G;碱基错误率均为0.02%;Phred数值大于20、30的碱基占总碱基的百分比均高于90.59%;鸟嘌呤+胞嘧啶(guanine cytosine,GC)含量分别为47.16%、48.94%和47.55%,表明转录组测序质量较高;黏虫受氯虫苯甲酰胺胁迫后,29个P450基因表达量上调,27个P450基因表达量下调;黏虫受氟虫腈胁迫后,23个P450基因表达量上调,26个P450基因表达量下调;12个P450基因表达量的RT-qPCR技术检测结果与DGE测序文库显示的结果基本一致。     

不同生境黑果枸杞实生苗生长及土壤养分空间差异的研究

研究不同生境黑果枸杞实生苗数量、生长状况、生物量分配及土壤养分空间差异,探讨环境因子对黑果枸杞实生苗生长的影响,为黑果枸杞种群恢复及持续经营提供理论依据。采用固定样地调查法,于2016年对张掖市甘州区、临泽县、肃南县及周边地区的农田地埂、水渠边、盐化沙地、盐碱荒地等各生境黑果枸杞进行调查。研究结果表明：不同生境均有黑果枸杞实生苗分布,农田地埂生境实生苗数量相对水渠边、盐化沙地、盐碱荒地分别增加66.67%、122.22%、215.79%。因环境条件差异,不同生境的黑果枸杞实生苗采取了不同的生存策略适应环境：农田地埂生境的黑果枸杞主要为增高生长,盐化沙地的黑果枸杞主要为冠幅的增长,而盐碱荒地的黑果枸杞主要为增粗生长。农田地埂生境0~20 cm土层有机质含量相对水渠边、盐化沙地、盐碱荒地分别增加28.50%、2.62%、10.13%;全氮含量分别增加44.08%、33.16%、72.41%;有效氮含量分别增加46.81%、0%、50%。空气湿度、土壤含水量对黑果枸杞幼苗数量及幼苗生长有显著影响。在今后的黑果枸杞栽培管理中,幼苗期应加强水分管理。     

咸蛋黄快速腌制过程中理化性质变化规律

该研究以新鲜蛋黄为原料,利用快速腌制模具,探究在咸蛋黄的上表面添加食盐单侧腌制过程中,食盐添加量和腌制时间对咸鸡蛋黄快速腌制过程中形貌特征和理化性质变化规律的影响。借助多种仪器分析手段对蛋黄腌制过程中形貌与物性的变化、水分及盐分的迁移规律进行了表征。低场核磁及成像结果表明：在腌制过程中,蛋黄中的水分不断向外迁移,含水率显著降低,当增加食盐的添加量和延长腌制时间,会加快水分的迁移速率;原子吸收结果表明：增加食盐添加盐量越多和腌制时间越长盐分迁移速率越快,质构、色差结果共同表明咸蛋黄腌制过程中,由于水分的向外迁移和盐分的向内渗入,使得蛋黄的蛋白质发生聚集使颜色加深;同时与市售整个腌制后分离的鸡蛋黄产品相比,当腌制时间为7d,添加盐量为3%;腌制时间为3d,添加盐量为5%时,所得的样品与市面的成品咸鸡蛋黄的感官品质及量化指标差异不显著（P<0.05）,为咸蛋黄单侧腌制技术提供理论的可行性。     

烤烟主要物理性状与化学成分的典型相关分析

以烤烟品种K 326为供试材料,采用典型相关分析方法研究了烤烟不同部位烟叶的主要物理性状和化学成分的关系.结果表明,烤烟物理性状和化学成分所具有的显著性指标因部位而异;上、中、下3个部位物理性状共有的显著性指标是叶长、叶厚和叶面密度,化学成分共有的显著性指标是氯含量、钾含量和钾氯比,且后者对典型变量的相对作用大小一致,表现为钾>钾氯比>氯;烤烟不同部位烟叶叶长、叶厚和叶面密度与钾、钾氯比和氯的关系最为密切.典型负载系数可以作为确定各部位物理特性和化学成分显著性指标的重要依据.     

甘露糖—小清蛋白美拉德反应产物的免疫活性

为探讨以甘露糖为还原糖进行美拉德反应对小清蛋白免疫活性的影响,通过将鲢重组小清蛋白(r PV)和甘露糖混合物在干热条件下(100°C)反应100 min制得糖化的r PV(Mr PV),采用Tricine-SDS-PAGE和斑点杂交(Dot-blotting)分析糖化前后r PV的聚合特性、免疫活性及消化稳定性的变化;采用圆二色谱仪和扫描电子显微镜分析糖化对r PV的二级结构及微观结构的影响。结果显示,r PV糖化后形成大分子量的片状聚集体;甘露糖糖化修饰的r PV免疫活性及消化稳定性均显著下降;美拉德反应导致r PV二级结构中的α-螺旋和无规则卷曲的含量有一定程度的减少,而β-折叠的含量显著增加。研究表明,以甘露糖为还原糖进行美拉德反应可有效降低r PV的免疫活性及消化稳定性,这可能与糖化后r PV聚集体的形成及二级结构的改变有关。     

水产动物6种主要病原菌与抗血清的免疫交叉反应

采用ELISA、试管凝集、Western-blot等方法,分析了鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(V. harveyi)、溶藻胶弧菌(V. alginolyticus)、副溶血弧菌(V. paraheamolyticus)、迟钝爱德华氏菌(Edwardsiella tarda)和荧光假单胞菌(Psedomonas fluorescens)等水产养殖中主要病原细菌与抗血清之间的免疫交叉反应.结果表明,弧菌属细菌之间的交叉反应程度比较大,而与其他两属的细菌之间存在的交叉反应程度小,或不存在交叉反应;Western-blot分析结果显示,哈维氏弧菌、溶藻胶弧菌和副溶血弧菌抗血清分别与其他3种弧菌在分子量为135.6 kD和121.5 kD;95.6 kD,48.4 kD,39.2 kD和34.9 kD;55.1 kD的蛋白带处存在交叉反应,而这些分子量的蛋白带与其他两属的抗血清均不发生反应.