粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。     

华南农业大学农事训练课程的理论与实践——以养蚕与缫丝课为例

以养蚕与缫丝课程模块为例,对其课程内容安排、教学特点、考核方法进行分析,并对其所面临的问题展开探讨,旨在为进一步提高相关实践教学效果提供借鉴。     

基于PSR模型的黄河陕西段鱼类增殖放流生态安全评价

基于压力、状态、响应模型(PSR)和层次分析法(AHP),确定17项指标通过数据的标准化处理,指标权重赋值、权重一致性检验、评价等级确定以及评价模型的构建,用生态安全评价黄河陕西段鱼类增殖放流效果,分析生态安全所面临的主要问题。结果表明:黄河陕西段2013年增殖放流生态安全度(ESI)评价等级为Ⅱ级,为良好状态;2014年评价等级为Ⅲ级,处于一般状态;2015年评价等级为Ⅳ级,处于较差状态,属于临界不安全状态以下水平。生态安全形势呈现出逐年下降局势。不安全状态受到影响较大的前3个指标是:黄河径流量变化影响、重要生境保持率和公众资源环境保护意识的影响,3个指标下降值占到下降ESI值的64.93%。其次还受到污水排放达标率、鱼类增殖放流量、政策和管理水平、鱼类生物多样性指数、保护区建设、水质综合污染指数、群落结构等诸多因素影响。研究显示,现阶段增殖放流对黄河生态安全有一定影响,还存在一定提升空间。     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

贝母属植物叶绿体基因组候选SNP位点应用分析

贝母属植物是中国中药材的宝库,由于形态上难以区分,在民间应用、临床应用和中药市场中还存在混淆应用、以次充好的现象,急需开发基于新型分子标记位点的精准检测方法。本研究应用多重序列比对、SNP筛选等生物信息学分析手段,对贝母属15种植物28条叶绿体基因组序列进行分析。结果发现,贝母属叶绿体基因组DNA同源性达97.22%,通过分析共发现SNP位点5879个,其中川贝母类鉴别候选位点71个,川贝母鉴别候选位点4个,瓦布贝母鉴别候选位点37个,太白贝母鉴别候选位点147个,浙贝母鉴别候选位点91个,湖北贝母鉴别候选位点271个,平贝母鉴别候选位点1393个,伊贝母鉴别候选位点89个。本研究还对SNP位点在精准鉴定、精确定量应用方面进行了讨论,可为川贝母中药资源鉴定提供技术支撑。     

Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes

Wheat leaf rust,caused by Puccinia triticina(Pt),is an important foliar disease that has an important influence on wheat yield.The most economic,safe and effective way to control the disease is growing resistant cultivars.In the present study,a total of 46 wheat landraces and 34 wheat lines with known Lr(leaf rust resistance)genes were inoculated with 16Pt pathotypes for postulating seedling resistance gene(s)in the greenhouse.These cultivars and five wheat differential lines with adult plant resistance(APR)genes(Lr12,Lr22b,Lr34,Lr35 and Lr37)were also evaluated for identification of slow rusting resistance in the field trials in Baoding,Hebei Province of China in the 2014–2015 and 2015–2016 cropping seasons.Furthermore,10 functional molecular markers closely linked to 10 known Lr genes were used to detect all the wheat genotypes.Results showed that most of the landraces were susceptible to most of the Pt pathotypes at seedling stage.Nonetheless,Lr1 was detected only in Hongtangliangmai.The field experimental test of the two environments showed that 38 landraces showed slow rusting resistance.Seven cultivars possessed Lr34 but none of the landraces contained Lr37 and Lr46.Lr genes namely,Lr9,Lr19,Lr24,Lr28,Lr29,Lr47,Lr51 and Lr53 were effective at the whole plant stage.Lr18,Lr36 and Lr45 had lost resistance to part of pathotypes at the seedling stage but showed high resistance at the adult plant stage.Lr34 as a slowing rusting gene showed good resistance in the field.Four race-specific APR genes Lr12,Lr13,Lr35 and Lr37 conferred good resistance in the field experiments.Seven race-specific genes,Lr2b,Lr2c,Lr11,Lr16,Lr26,Lr33 and LrB had lost resistance.The 38 landraces showed slow rusting resistance to wheat leaf rust can be used as resistance resources for wheat resistance breeding in China.     

基于主成分分析和Copula函数的灌溉水利用系数影响因素研究

以新疆生产建设兵团第十二师中型灌区为研究对象，采用首尾法测算2018年和2019年4个样点灌区的灌溉水利用系数，在采用主成分分析法对指标体系降维处理的基础上，用Copula函数构建PCA-Copula评价分析方法，对灌溉水利用系数各影响因素的影响程度和影响规律进行计算分析。结果表明：两年测算数据表明，4个样点灌区灌溉水利用系数都在0.63以上，且最大值达到0.668，分别高于同期全疆、全国平均水平5.05%、10.15%；主成分分析得出，渠道衬砌率（0.944）、滴灌灌溉面积比（0.746）、作物需水量（0.635）、实际灌溉面积（0.734）等都具有超过60%的正贡献率，而葡萄种植比（-0.586）和灌区毛灌溉用水量（-0.645）等具有超过58%的负贡献率。利用PCA-Copula分析评估方法得出，作物种植比例和节水灌溉工程状况对十二师中型灌区灌溉水有效利用系数影响显著（P<0.05），其中葡萄净灌溉定额和灌区毛灌溉用水量对灌溉水有效利用系数的影响极其显著（P<0.01），相关系数分别为0.875、0.742，同时利用Spearman等级相关系数检验法和线型回归来检验PCA-Copula评价法与熵值法的密切程度，检验结果其相关系数分别为 0.87（P＜0.05）和0. 52（P＜0.001），表明PCA-Copula评价方法适用于研究灌溉水利用系数影响因素。     

代森锰锌液相色谱法成功晋升为CIPAC方法的历程与启示

由利民化学有限责任公司自主研发的代森锰锌液相色谱分析方法,历时4年,终于在2019年德国布伦瑞克召开的第63届国际农药分析协作委员会(CIPAC)年会上顺利通过答辩,成功晋升为CIPAC方法。本文对此过程进行了总结梳理,以为我国农药标准方法走向国际提供借鉴。