霜霉威盐酸盐防治黄瓜霜霉病试验

小区试验每公顷用霜霉威盐酸盐,有效成分600克、800克、1000克,发病初期叶面喷雾,对黄瓜霜霉病的防治效果为69.9%~82.3%,显著优于三乙膦酸铝,每公顷有效成分2250克。大面积示范每公顷用霜霉威盐酸盐有效成分800克,防效达79.6%,增收黄瓜44940千克。     

Comparative Proteomic Analysis Shows an Elevation of Mdhl Associated with Hepatotoxicity Induced by Copper Nanoparticle in Rats

Copper nanoparticle is a new material widely used in biological medicine, animal husbandry and industrial areas, but its potential toxicity to human health and environment remains unclear. In order to study the hepatotoxic mechanisms of nanoparticles copper, two-dimensional gel electrophoresis （2-DE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI/TOF MS） of proteomics technology were used to isolate and identify the differentially expressed proteins from liver, which associated with hepatotoxicity induced by copper nanoparticle in rats. In this study, we have screened 15 kinds of proteins related with hepatotoxicity, of which spot8212 was identified as Malate dehydrogenase （Mdhl）. The mRNA expression trend of Mdhl was consistent with the result of 2-DE by RT-PCR validation. Bioinformatics analysis showed that Mdhl was stable and no signal peptides, subcellular location was in endoplasmic reticulum; it contained many functional sites such as malate dehydrogenase activity signal sites 155LTRLDHNRAKSQI167; a helixes and random coils were the two main elements. Homologous analysis demonstrated high homologous of Mdhl in rats with mouse and human, and the phylogenetic tree of Mdhl was constructed. The result indicated that copper nanoparticle could regulate up the Mdhl protein expression so as to compensate the energy deficit. Energy metabolic disturbance may be a pathway for copper nanoparticle particles to exert the hepatotoxic effects in rats.     

土壤和黄瓜中霜霉威盐酸盐残留量的测定

为了建立测定土壤和黄瓜种霜霉威盐酸盐残留量的液相色谱串联质谱分析方法,采用乙腈和水提取土壤和黄瓜样品,盐析后取上清液,液相色谱串联质谱法测定。经过3个水平的添加回收试验,所建方法的平均回收率在91.7%~98.7%,相对标准偏差1.0%~3.3%,n=5,目标物最低检出浓度0.01 mg·kg-1。所建方法的准确度、灵敏度和精密度均满足农药残留分析的要求。     

基质辅助激光解吸电离飞行时间质谱检测蛋白质分子量的优化研究

【目的】研究采用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）技术检测蛋白质分子量的优化条件。【方法】选择3种不同的基质检测牛血清白蛋白（BSA）的分子量,在选定最佳基质化合物的基础上,通过改变样品蛋白质的浓度,以获得高质量的质谱峰图,确定蛋白质样品的最小和最优检测量。【结果】1 μg蛋白质样品与10 mg/mL含0.1%乙酸（TFA）和50%乙腈（ACN）的芥子酸（SA）基质水溶液混合检测分子量,可产生最高质量的MALDI-TOF-MS峰图,同时蛋白质样品最小检测量应不少于500 ng。【结论】MALDI-TOF-MS技术操作简单、分析快速、灵敏度高,是一种检测蛋白质分子量的可靠方法;研究结果确定了获得最优化质谱峰的最佳组合,这是从高度复杂的蛋白质样品测定准确分子量的必备条件。     

霜霉威杀菌剂产品登记动态概况

霜霉威杀菌剂，法国安万特作物科学公司1997年在我国正式登记商品普力克72．2％水剂，防治黄瓜猝倒病、疫病用量（有效成分、下同），3．6-5．4克／平方米苗床浇灌，防治黄瓜霜霉病用量649．8-1083克／公顷，甜椒疫病、烟草黑胫病用量775．5-1164克／公顷。比利时农化公司1998年在我国临时登记商品扑霉特原药、扑霉特72．2％水剂，     

霜霉威AS防治马铃薯晚疫病效果试验

利用72.2%霜霉威AS800倍液和25%甲霜灵可湿性粉剂两种药剂进行防治马铃薯晚疫病的效果试验,试验设3个处理,每个处理重复4次,随机区组排列。72.2%霜霉威AS按75 g/hm2施药后,防治效果明显,茎叶正常死亡,产量是对照的1.6倍,且对人和作物都很安全,是防治马铃薯晚疫病的有效杀菌剂,可在生产中应用。     

Mitochondrial Proteomic Analysis of Cytoplasmic Male Sterility Line and Its Maintainer in Wheat （Triticum aestivum L.）

To investigate the CMS mechanism of wheat on proteomic level and find the crucial proteins which related to fertility,mitochondria was isolated from young spike of wheat by differential centrifugation and Percoll density-gradient methods.Determined by marker enzyme assays and chlorophyll content,the mainly contaminants in the spike mitochondrial fraction were caused by peroxisomes,plastids and chloroplasts after the first discontinuous Percoll density gradient centrifugation.In order to improve the purity of spike mitochondria,a second 28% Percoll self-forming density gradient centrifugation was further carried out,the result showed that the contaminants were decreased to negligible amount,meanwhile the integrity of mitochondria （88%） was improved to 90%.The spike mitochondria proteins extracted from uninucleate stage of （S）-1376A and （A）-1376B were separated by two-dimensional electrophoresis （2-DE）,and the silver stained gels were analyzed by PDQuest 2-DE software,about 326 protein spots could be visualized on the 2-DE maps,and also revealed a similar pattern between the male sterile line and its maintainer line,except 11 spots were differentially expressed.A total of five differentially expressed proteins were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）,three of them were identified as manganese superoxide dismutase and T5E216 following NCBInr database by the Mascot software.These results may contribute to further understanding of the mechanisms of CMS in wheat.     

霜霉威盐酸盐水剂含量的气相色谱分析

建立以正十五烷为内标物,用气相色谱法测定农药杀菌剂霜霉威盐酸盐水剂的检测方法。样品经衍生化处理后,通过OV-101毛细管柱分离,氢火焰离子化检测器,对霜霉威进行气相色谱分离和测定,加标回收率99.4%~102.7%,相对标准偏差2.0%。     

Immunoproteomic Analysis of Bordetella bronchiseptica Outer Membrane Proteins and Identification of New Immunogenic Proteins

Bordetella bronchiseptica is a Gram-negative pathogen that causes acute and chronic respiratory infection in a variety of animals. To identify useful antigen candidates for diagnosis and subunit vaccine of B. bronchiseptica, immunoproteomic analysis was adopted to analyse outer membrane proteins of it. The outer membrane proteins extracted from B. bronchiseptica were separated by two-dimensional gel electrophoresis and analyzed by Western blotting for their reactivity with the convalescent serum against two strains. Immunogenic proteins were identified by matrix-assisted laser desorption/ionization time of flight-mass spectrometry（MALDI-TOF-MS）, a total of 14 proteins are common immunoreactive proteins, of which 1 was known antigen and 13 were novel immunogenic proteins for B. bronchiseptica. Putative lipoprotein gene was cloned and recombinantly expressed. The recombinant protein induced high titer antibody, but showed low protective indices against challenges with HB（B. bronchiseptica strain isolated from a infected rabbit）. The mortality of mice was 80% compared to 100% of positive controls. The identification of these novel antigenic proteins is an important resource for further development of a new diagnostic test and vaccine for B. bronchiseptica.     

72％霜霉清防治黄瓜霜霉病药效试验

田间试验表明 ,72 %霜霉清可湿性粉剂稀释 60 0～ 80 0倍可较好地控制黄瓜霜霉病的危害 ,明显优于 1 0 %霜脲氰可湿性粉剂 70 0倍液和 70 %代森锰锌可湿性粉剂 80 0倍液的防效     

霜霉威胁迫下黄瓜CsWRKY30表达及功能分析

【目的】从前期转录组测序结果中筛选获得一个在霜霉威(propamocarb)胁迫条件下差异上调表达的基因CsWRKY30,对其进行克隆并分析其在霜霉威胁迫下的功能,了解黄瓜低霜霉威残留的分子机制。【方法】通过PCR技术扩增CsWRKY30全长,利用NCBI和PlantCARE在线工具分别进行该基因编码蛋白的保守结构域分析和启动子序列分析;利用实时荧光定量PCR分析CsWRKY30在霜霉威胁迫及其他胁迫条件下的相关表达模式;通过与GFP蛋白融合对CsWRKY30蛋白进行亚细胞定位;通过花序侵染法将CsWRKY30构建的植物过表达载体转化到哥伦比亚野生型拟南芥中,对获得的纯合转基因株系在霜霉威胁迫条件下的功能进行鉴定。【结果】CsWRKY30的CDS序列全长为1 014 bp,其编码的337个氨基酸中包含1个由60个氨基酸组成的WRKY结构域。CsWRKY30表达模式分析结果显示,黄瓜遭受霜霉威胁迫时,CsWRKY30在低霜霉威残留品系D0351中表达量显著上调,而在高霜霉威残留品系D9320中表达量并没有发生改变;在霜霉威胁迫的0.5-9 h间,该基因在D0351中的表达量一直明显高于对照,然而在24 h以后,该基因的表达不再显著上调。组织特异性表达分析表明,CsWRKY30主要在黄瓜果实中表达。蛋白亚细胞定位结果表明,CsWRKY30定位于细胞核。对CsWRKY30转基因拟南芥进行霜霉威胁迫发现,在未处理条件下,CsWRKY30 转基因拟南芥与野生型拟南芥表型上无明显差异;在2 mmol?L^-1霜霉威处理条件下,CsWRKY30转基因拟南芥萌发率及主根长均明显高于野生型拟南芥。在其他逆境作用下,CsWRKY30对霜霉威和多主棒孢霉菌条件积极响应,对干旱和高盐没有作用,同时受到脱落酸（ABA）信号诱导。【结论】黄瓜CsWRKY30在霜霉威胁迫条件下发挥重要作用,过量表达CsWRKY30可显著提高转基因拟南芥对霜霉威胁迫的抵抗能力。     

Study on Relationship Between Cucumber Germplasm and Propamocarb Residue Using Subjective Rating Technique

Propamocarb(PM)residue in cucumber(Cucumis sativus L.)receives little attention.As is well known to all,high PM residue of cucumber could lead to increase in the violation rates of maximum residue limits and ultimately cause harm to human health.Knowledge of PM residue could help cucumber breeders in developing cultivars with low PM residue and improving cucumber quality.In this study,32 representative cucumber accessions(26 breeding lines and six cultivars)from different regions of China were evaluated for their PM residue in fruit and leaf to provide meaning to the subjective rating,which was highly correlated with PM residual content of fruit(r=0.97)and leaf(r=0.94).In addition,PM residual content of North China ecotype was the highest and Pickling ecotype was the lowest in fruit and leaf of cucumber.The leaf had significantly higher(P<0.01)PM residual content than the fruit,and poor correlation between leaf and fruit was represented.This study verified PM residual relationship between fruit and leaf,and laid the foundation for further identification of germplasm resources and breeding of new varieties for low PM residue of cucumber.     

枯萎病菌胁迫下黄瓜叶片蛋白质组分析

以黄瓜感枯萎病品系1900和抗枯萎病转基因品系1905为试材,采用灌根法接种枯萎病菌分生孢子悬浮液,48h后提取叶片蛋白质,采用差异蛋白质双向电泳技术研究枯萎病胁迫下黄瓜叶片蛋白质组的变化.结果表明:黄瓜幼苗在枯萎病菌胁迫下,感病和抗病品系的双向电泳图谱存在一定差异.通过质谱分析,共鉴定出3-磷酸甘油醛脱氢酶、景天庚酮糖-1,7-二磷酸酶、木葡聚糖内转糖苷酶、聚光复合体叶绿素a/b结合蛋白、假拟蛋白和5类表达差异的蛋白点.这些植物代谢中产生的能量和一些小分子可能激发了黄瓜抗枯萎病基因的表达.     

几种杀菌剂防治黄瓜霜霉病田间试验效果分析

试验表明,72.2%普力克AS、64%杀毒钒WP、72%克露WP、47%加瑞农WP和70%安泰生WP均为防治黄瓜霜霉病较好的药剂品种.其中,以72.2%普力克AS800倍液的药效表现最好,三次药后10 d的防效达到97.89%,显著优于同剂量的其它药剂;72%克露WP800倍液的药效为94.08%,表现次之;47%加瑞农WP800倍液和70%安泰生WP800倍液的药效分别为89.4%和87.88%,二者的药效无差异.     

黄瓜根系蛋白质组分析的双向电泳技术条件优化

双向电泳(2-DE)是蛋白质组学研究的重要支撑技术之一。为建立适于黄瓜根系蛋白质组分析的双向电泳技术,对黄瓜根系蛋白质样品提取方法、裂解缓冲液配方及SDS-PAGE分离胶浓度等参数进行研究。结果表明:采用TCA/丙酮法提取黄瓜根系中的蛋白质,裂解缓冲液为8mol·L-1尿素、2mol·L-1硫脲、2%IPG Buffer、4%CHAPS、1%TBP、65mmol·L-1DTT、2mmol·L-1EDTA、0.001%溴酚蓝和1%鸡尾酒,SDS-PAGE分离胶浓度为11%时,可获得分离效果较好的双向电泳图谱。该技术条件为适合黄瓜根系蛋白分离的较优双向电泳体系。     

化学药剂胁迫下6株生防菌在黄瓜植株的定殖能力分析

[目的]探究化学药剂胁迫对生防菌在黄瓜植株上定殖能力的影响.[方法]设置一个温室试验和两个田间试验,分别在无药剂和无病原菌、无药剂和有病原菌、有药剂和有病原菌的黄瓜植株上处理生防菌.[结果]供试的6株生防菌在黄瓜叶片和根围上均具有良好的定殖效果.在无药剂和无病原菌的温室条件下,各生防菌定殖量均呈现降-增-稳定的趋势,菌...     

不同诱导剂诱导黄瓜愈伤组织凋亡的研究

以黄瓜愈伤组织为材料,比较二甲基亚砜(dimethyl sulfoxide,DMSO)、VK3和CaCl2相同浓度诱导细胞凋亡的差别。将愈伤组织悬浮培养并分离单细胞后进行活性检测和荧光检测,并对其DNA进行琼脂糖凝胶电泳检测,结果表明,3种诱导剂都能诱导黄瓜愈伤组织凋亡,诱导能力VK3>DMSO>CaCl2。     

PP_(333)诱导黄瓜子叶节差异蛋白表达的分析

【目的】了解和寻找PP333处理所导致的差异蛋白表达,为深入研究PP333的作用规律和机理获取有价值的信息。【方法】应用双向凝胶电泳技术研究PP333处理后黄瓜离体子叶节培养物差异蛋白的表达,结合质谱分析和蛋白质数据库检索,确定所检出差异蛋白质的性质和功能。【结果】双向电泳检测到黄瓜离体子叶节培养物蛋白点有1000个左右,质谱(MALDI-TOF-TOF/MS)分析鉴定出5个有信息价值的差异表达蛋白点,分别为14-3-3蛋白(R2)、氨基肽酶(R7)、甲硫氨酸合酶(R11)、60s酸性核糖体蛋白(R20)和PEPC(R21)。这些蛋白质与代谢过程、蛋白质翻译及信号转导等关系密切。【结论】PP333处理导致多种蛋白质表达发生改变,并通过多种蛋白质的相互协调作用调控植物体的生长发育。