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。     

基于多反应监测质谱技术的水稻叶片蛋白绝对定量方法

【目的】建立水稻叶片蛋白的多反应监测(MRM)质谱绝对定量方法。【方法】水稻叶片蛋白经含1.0%十二烷基硫酸钠(SDS)的磷酸盐(PBS)缓冲液提取,丙酮沉淀除杂纯化、胰蛋白酶消化,酶解液经液相色谱分离,MRM质谱监测,外标法定量。【结果】向提取缓冲液中加入1.0%SDS可增强水稻叶片中16种靶蛋白和总蛋白质的提取效果;不同有机试剂处理,总蛋白质沉淀量存在显著差异(P<0.05),沉淀能力从强到弱依次为乙腈>丙酮>异丙醇>甲醇>乙醇;对于16种目标蛋白,总体以丙酮沉淀效果最好,其次是异丙醇和乙腈,甲醇和乙醇效果较差。该方法线性范围均达到3个数量级,定量限为0.1~2.5 nmol/L,灌浆期16种水稻叶片蛋白质含量为6.0~2818.1μg/g,相对标准偏差均小于14%。【结论】SDS可显著提高水稻叶片蛋白提取效果,采用丙酮或乙腈可获得较好的蛋白沉淀效果,但不同蛋白质略有差异。结合MRM质谱监测技术可实现水稻叶片蛋白的绝对定量,方法线性范围宽、敏度高、重复性好。     

杂交稻低播量精量播种育秧及机插取秧特性

【目的】明确杂交稻70 g以下低播量精量穴播和条播对育秧效果及机插特性的影响,突破生产中杂交稻机插的技术瓶颈。【方法】以中浙优1号和甬优1540两个杂交稻品种为材料,利用机插标准9寸盘,设置机械穴播和条播两个精量播种方式,并以机械流水线撒播为对照,穴播规格为16(纵向)×34(横向)穴,条播为纵向16条,以穴播5粒、3粒及2粒的播种量进行播种试验。考查了低播量下不同播种方式对秧苗生长影响及配套取秧效果【结果】1)低播量下不同播种方式对秧苗成苗率的影响不大。2)降低播种量提高秧苗生长一致性,且穴播和条播秧苗生长一致性好于撒播。3)与撒播相比,精量穴播和条播能够在低播量下提高秧苗根系盘结力和成毯性,中浙优1号和甬优1540根系盘结力比撒播平均高75.4%和81.0%,播量每穴3粒时即能有效成毯,穴播和条播差异不大。4)精量穴播和条播能够显著降低低播量下机插漏秧率,中浙优1号和甬优1540机插漏秧率平均分别比撒播低76.3%和74.6%,穴播和条播下,两个品种每穴播量3粒的漏秧率均在1%以下,与撒播相比降幅在10个百分点以上。5)精量穴播和条播机插取秧苗数均匀度比撒播要好,两个品种预期取秧2~5苗(5粒)、1~3苗(3粒)和1~2苗(2粒)比例均达80%以上,中浙优1号和甬优1540机插苗数均匀度平均比撒播高121.2%和67.0%,其中,穴播机插取秧苗数均匀度及预期取秧苗数比例高于条播。【结论】精量穴播和条播可以解决目前杂交稻机插用种量大、漏秧率高和机插取秧苗数均匀度差的问题。     

鸡Prnp基因原核表达载体的构建及其在大肠埃希菌中的表达

试验旨在构建鸡Prnp基因原核表达载体,并在大肠埃希菌中进行表达,为制备鸡朊蛋白单克隆抗体提供材料。根据GenBank已报道的鸡Prnp基因组序列和pET-28a质粒多克隆位点设计引物,以健康的鸡全血基因组DNA为材料,采用PCR的方法扩增鸡的Prnp基因,将目的基因片段与pET-28a载体连接,构建重组原核表达载体。重组菌转化到E. coli BL21(DE3)感受态细胞中,并用异丙基-β-D-硫代半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)进行诱导表达,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定表达的重组蛋白。结果表明,重组蛋白在诱导剂的终浓度为0.08 mmol·L^-1,16 ℃,220 r·min^-1诱导7 h,蛋白表达量最高。综上所述,本研究成功构建了pET-28a-ChPrnp重组表达菌株,为Prnp朊蛋白的结构、生理功能和致病机制研究提供方法。     

四端法土壤电导率传感器恒流源设计与试验

土壤信息对于指导农业生产有着极为重要的作用，土壤电导率反映了土壤含水量、盐分、粘粒含量和类型等土壤信息，准确获取土壤电导率对于实现农业精细化生产意义重大。在各种土壤电导率测量方法中，四端法因其成本低、精度高、测量快速和操作简便而大量应用于实际测量；恒流源是四端法测量仪器的重要组成部分，其性能直接决定着测量仪器的精度以及测量范围。本文对比了3种恒流源对测量仪器测量性能的影响，发现采用Howland恒流源的四端法测量仪器高电导率测量能力强而低电导率测量范围较小，测量精度最高；采用改进型Howland恒流源的四端法测量仪器低电导率测量范围有所扩大，测量精度良好；采用基于差动放大器的恒流源的电导率仪低电导率测量范围最大，高电导率测量范围较优，测量精度良好。     

提取方法对库尔勒香梨果实表皮蜡质提取效果的影响

以库尔勒香梨为试材，三氯甲烷和正己烷为提取剂，采用常温法、加热法和常温加热组合法提取香梨果实表皮蜡质，探讨香梨果皮蜡质的提取方法，为研究香梨果皮蜡质结构及成分提供基础。结果表明：以三氯甲烷为提取剂的效果明显优于正己烷，采取常温结合加热法提取表皮蜡质的含量最高，常温法提取的蜡质含量最低。扫描电镜观察结果表明：采用常温加热结合法，以三氯甲烷作溶剂可充分提取香梨表皮蜡质，而正己烷提取果实表皮呈不明显网状结构，仍有晶体残留；采用加热法，以三氯甲烷提取香梨果实表皮蜡质后，表皮蜡质有小部分残留，而正己烷提取后残留晶体较三氯甲烷提取后多，有不规则的小粒状晶体残存；用常温法则两种溶剂均不能将香梨表皮蜡质提取干净。综合提取蜡质含量和扫描电镜分析的结果得出，以三氯甲烷为溶剂、采用常温加热结合法是有效且易行的提取库尔勒香梨表皮蜡质的方法。     

蔬菜幼苗叶面积快速测定方法筛选与优化

为得到一种快速且准确测定蔬菜幼苗叶面积的方法,以番茄幼苗为试材,以方格法为对照,研究扫描分析法、叶面积仪法、打孔称重法测定叶面积的准确性、精确性与测定速度,然后采用黄瓜、辣椒叶片及纸片的面积对测定结果进行验证,最后基于不同扫描分辨率对筛选出的方法进行优化。结果表明,相比叶面积仪法和打孔称重法,扫描分析法测定结果更接近真实值,且精度更高,用时更短。扫描分析法不同分辨率下测定结果无显著差异,且扫描分辨率越低所需测定时间越短。扫描分辨率50 dpi下的扫描分析法可作为一种快速准确测定蔬菜幼苗叶面积的方法。     

Age estimation of different tree species using a special kind of an electrically recording resistance drill

The determination of tree age is an important issue for urban green planning, forestry and dendrology; finding non-destructive and quasi-non-destructive methods for this purpose is of great theoretical and practical importance. The resistance drilling method is quasi-non-destructive because the average diameter of an opening that remains after drilling does not exceed 3 mm. Do electrically recording resistance drills allow precise assessment of tree age? The aim of this study is to assess the accuracy of determining the number of tree rings based on an examination of this special kind of drilling resistance profiles for three tree species, the pine Pinus sylvestris L., the oak Quercus robur L., and the birch Betula pendula ROTH. In 2015 and 2016, 15 pine trees, 15 oak trees, and 15 birch trees were randomly selected. For each studied tree, a measurement was conducted using the electrically recording resistance drill IML-Resi E400 with a flat-tipped 1.5/3 mm steel needle (research sample), and an increment core was taken (reference sample). The drill used was not a real Resistograph^®. The analysis of the E400-profiles underestimated the number of tree rings; the mean bias error (MBE) values were –6.5, –2.5, and –6.0 years for pine, oak, and birch, respectively. The proportion of investigated trees with less than five years difference between the research and reference samples varied from 38.4 (birch) to 66.7 (oak) percent. The accuracy of tree age determination was lowest for birch and highest for oak. The binomial generalised linear model (GLM) revealed that the most accurate tree age assessments were obtained from tree rings wider than 2 mm. The measurements clearly showed that the electrically recording resistance drill IML-Resi E400 enables a quick, although approximate, tree age assessment. Future research should concentrate on electronically regulating and recording drills, providing a higher spatial and signal resolution, and a stronger correlation to wood density.     

菠萝蜜种子淀粉制备香草兰微胶囊的工艺研究

以菠萝蜜种子淀粉为壁材,采用饱和水溶液法制备香草兰精油微胶囊,以包埋产率为指标,采用响应面分析法对微胶囊的包埋条件进行优化探讨。结果表明:5个单因素中,影响最显著的因素为壁芯材比例、包埋温度和包埋时间。响应面优化得到香草兰精油微胶囊的最佳工艺条件为壁芯材比例为7.5∶1;包埋时间72 min;包埋温度54℃,此条件下的包埋产率为(95.46±0.2)%,包埋率为(76.35±0.6)%,载油量为(27.73±0.3)%。试验证明,此条件结果与模型预测值相吻合,此工艺条件可为菠萝蜜种子淀粉包埋香草兰精油微胶囊工艺提供理论依据。     

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.