茶树咖啡碱合成酶CRISPR/Cas9基因组编辑载体的构建

CRISPR/Cas9技术是一门新兴的基因组定点编辑技术,具有操作简单、高效的优点,可轻松实现对目标基因的敲除、替换和定点突变等操作。该技术刚诞生,就受到了全球生命科学领域研究者的关注,不到3年的时间就已经成功应用于多种动、植物当中。然而CRISPR/Cas9技术在茶树中的应用面临载体构建问题,本文以茶树咖啡碱合成酶为例,联合采用常规PCR、Overlapping PCR和Golden Gate Cloning技术,构建了包含茶树咖啡碱合成酶双靶点的CRISPR/Cas9基因编辑载体,为CRISPR/Cas9介导的基因组编辑技术在茶树中的应用奠定了坚实基础。     

小麦面粉中赭曲霉毒素A的CdSe/ZnS荧光量子点免疫探针检测

构建一种基于荧光量子点免疫探针的小麦面粉中赭曲霉毒素A(OTA)的检测方法。以EDC为活化剂将CdSe/ZnS半导体荧光量子点与OTA单克隆抗体偶联,制备OTA检测探针并评价其检测性能。结果表明:制备的检测探针在3d保存期内具有良好的稳定性。当OTA质量浓度为0.1~0.9μg/L时,检测探针的荧光强度与OTA质量浓度呈线性关系,其相关系数为0.996 3,检出限为0.017 7μg/L。与国标法对照的结果说明,此方法具有可行性。小麦面粉中的加标回收试验结果显示,OTA的回收率为96.0%~102.7%,相对标准偏差不超过4.1%,说明准确性良好。可见,OTA的CdSe/ZnS荧光量子点免疫探针检测方法操作简单、准确度高、结果稳定,有望应用于小麦面粉中OTA的检测。     

超高效液相色谱-质谱/质谱法同时测定蔬菜中7种磺胺的残留量

为建立超高效液相色谱-质谱/质谱法(UPLC-MS/MS)同时测定蔬菜中7种磺胺残留量的方法。样品用1%乙酸乙腈提取，以C18色谱柱分离待测物，采用多反应监测(MRM)离子扫描模式，外标法进行定量，线性良好，相关系数均大于0.999。结果显示：7种磺胺的检出限为0.000 4～0.000 6 mg/kg，样品添加回收率为67.5%～94.1%，相对标准偏差均少于5.3%(n＝6)。该方法简单快捷、定量准确，可满足多种蔬菜中7种磺胺的残留检测要求。     

绵羊卵泡发育过程中关键信号通路研究进展

卵泡从原始卵泡发育为成熟卵泡，直至排卵、黄体发育等过程都受到精密的调控，产生大量的优势卵泡是绵羊产多羔及实现快速扩繁的关键因素。研究发现，相关信号通路和转录因子通过影响绵羊卵泡中卵母细胞、颗粒细胞的生长，进而调控卵泡的发育成熟，对这些信号通路进行深入了解，有助于探索卵泡发育的调控机制，早日实现绵羊高效繁育。Notch是卵泡发育过程中发挥重要作用的高度保守信号通路，PI3K/AKT/mTOR信号通路各成员都是广泛存在于细胞内的信号转导分子，在卵泡发育早期发挥了主要作用，还有间隙连接（gap junction，GJ）和跨带突触（transzonal projections，TZPs）等物理连接方式，在细胞间的交流通讯起到重要作用。作者详细介绍了Notch信号通路、PI3K/AKT/mTOR信号通路、间隙连接及跨带突触的结构功能在绵羊卵泡发育中的调控作用，为进一步探明绵羊卵泡发育的调控机制提供参考。     

基于工作过程CAD/CAM工程技术应用课程教学改革实践

针对CAD/CAM工程技术应用的课程体系、理论课程教学内容、教学方法和实验课程教学进行了改革,成功地建立了CAD/CAM工程技术应用课程体系,取得了良好的教学效果。     

大豆胞囊线虫主效抗病基因Rhg4(GmSHMT)的CAPS/dCAPS标记开发和利用

大豆胞囊线虫(soybean cyst nematode,SCN)严重危害世界大豆生产,Rhg4(resistance to Heterodera glycines 4)是控制大豆SCN抗性的2个主效位点之一。本研究针对Rhg4(Gm SHMT)上的2个单核苷酸多态性(single nucleotide polymorphisms,SNP)位点开发了快速、经济、简便易行的CAPS(Rhg4-389)和d CAPS标记(Rhg4-1165),并用开发的2个标记鉴定了以大豆胞囊线虫应用核心种质为主的193份代表性抗感种质。结果表明,Rhg4-389和Rhg4-1165位点间存在显著连锁不平衡(P=0.0001,r2=0.87),可形成4种单倍型。Rhg4-389-G/Rhg4-1165-T和Rhg4-389-C/Rhg4-1165-A为优势单倍型,稀有单倍型Rhg4-389-G/Rhg4-1165-A和Rhg4-389-C/Rhg4-1165-T是在中国抗源中新发现的单倍型。结合193份种质对SCN 3号小种抗性鉴定分析发现,Rhg4-389-G和Rhg4-1165-T主要存在于抗病种质,它们形成的单倍型对抗病种质鉴定效率可达94.1%。本研究开发了可用于辅助大豆SCN抗性鉴定且方便育种家利用的CAPS/d CAPS标记,且用其摸清了应用核心种质等重要抗源在Rhg4位点的"本底",为育种家有效利用这些优异抗源提供了重要信息。     

Comparison of dietary essential amino acid requirements determined from group‐housed versus individually‐housed juvenile bluegill,Lepomis macrochirus

Two 60‐day experiments were conducted sequentially to determine (i) lysine requirement of juvenile bluegill, Lepomis macrochirus based on the dose–response method, (ii) requirements for other essential amino acids (EAAs) using whole‐body amino acid profile and (iii) whether differences in growth rates of group‐housed versus individually‐housed bluegills lead to different lysine requirement levels because of the presence and absence, respectively, of social hierarchies. Seven, semi‐purified, experimental diets (isonitrogenous, isocaloric) were prepared to contain graded levels of digestible lysine (10–31 g kg⁻¹). Experiment‐1 involved group‐housed bluegills (approximately 27 g, n = 10 fish/chamber, 4 chambers/diet) whereas experiment‐2 involved individually‐housed bluegills (approximately 30 g, n = 1 fish/chamber, 14 chambers/diet). Fish were fed twice daily to apparent satiation. Bluegill growth responses in both experiments generally improved (P < 0.05, anova ) with increasing dietary lysine levels from 10 to 16 g kg⁻¹, and then levelled off with further increase in lysine level (P > 0.05). Optimal dietary lysine level (digestible basis) was estimated to be 15 g kg⁻¹ based on broken‐line regression analyses of relative growth rate and feed conversion ratio with no differences being observed between the two rearing methods. Determined dietary requirement levels for other EAAs ranged from 2.4 g kg⁻¹ (tryptophan) to 15.3 g kg⁻¹ (leucine).     

Single and Interactive Effects of Temperature and Light Quality on Four Canola Cultivars

Combined effects of temperature and light quality on plants have received little attention. We investigated the single and interactive effects of temperature and light quality on growth and physiological characteristics of four canola (Brassica napus) cultivars – Clearfield 46A76 (cv₁), Clearfield 45H72 (cv₂), Roundup Ready 45H24 (cv₃) and Roundup Ready 45H21 (cv₄). Plants were grown under lower (24°/20 °C) and higher (30°/26 °C) temperature regimes at low red/far‐red (R/FR), normal R/FR and high R/FR light ratios in environment‐controlled growth chambers (16 h light/8 h dark). Higher temperature reduced stem height and diameter; leaf number and area; dry matter of all plant parts; and specific leaf weight, but increased leaf area ratio; and chlorophyll (Chl) fluorescence (Y). Low R/FR increased stem height; Y; and ethylene, but decreased stem diameter; F_v/F_m; Chl a; Chl b; and carotenoids. Among cultivars, plants from cv₄ were tallest with thickest stems and greatest dry matter. None of the main factors affected gas exchange. Higher temperature at high R/FR caused cv₃ to be shortest, whereas lower temperature at low R/FR caused cv₄ to be tallest. We conclude that heat and other stress factors will adversely affect sensitive crops, but tolerant genotypes should perform well under future climate.     

An experimental study of charcoal degradation in a boreal forest

Degradation rates of pyrogenic carbon (PyC) under natural environmental conditions are largely unknown. Here we present results from a field experiment monitoring the change in mass, C- and N concentrations of a variety of charcoal types in a Norwegian boreal forest over a period of 20 months. The charcoal types represent different feedstock tree species, production temperature regimes, and placements in the forest, i.e. above ground, in the humus layer or in contact with the mineral subsoil. The types of charcoal had different initial C concentrations mainly depending on their production temperature. Nevertheless, all types of charcoal at all placements in the forest showed an initial drop in their C concentrations, which subsequently rose back to reach near initial values in part of the charcoal types. In part of the charcoal types, N concentrations decreased throughout the experiment, exhibiting considerable variation among feedstock species, production temperature regime, and placements in the forest. C/N ratios rose especially in charcoal made from wood of Scots pine (Pinus sylvestris L.), and charcoal that had been stored in contact with the mineral subsoil showed the most rapid mass gain. Our results confirm the important influence of production temperature and feedstock type on the degradation of charcoal, but they also show that microbial activity and environmental conditions play significant roles in charcoal degradation and thus for the fate of pyrogenic carbon under natural conditions.