鸡白痢沙门氏菌与鸡伤寒沙门氏菌基因组消减文库的构建

为了解鸡白痢沙门氏菌与鸡伤寒沙门氏菌的基因组差别,筛选鸡白痢沙门氏菌特有的序列,利用SSH对鸡白痢沙门氏菌C79-13（实验方）与鸡伤寒沙门氏菌Sg9（驱动方）基因组进行了比较。选择四碱基内切酶Rsa I将C79-13与Sg9基因组DNA酶切,酶切的C79-13基因组DNA分成两份,分别与接头1和接头2R连接,然后进行两轮杂交和两轮PCR ,得到消减混合物, 将其与PCR?2.1克隆载体连接,转化感受态E.coli DH5α建立消减文库。结果共获得400个阳性克隆;筛选240个克隆制备质粒进行PCR检测,均扩增出100～2000 bp大小的片段。差异DNA消减文库的构建为进一步筛选、克隆鸡白痢沙门氏菌特异的未知新基因、建立分子鉴别新体系奠定了基础。     

Determination of multiresidues in rapeseed, rapeseed oil, and rapeseed meal by acetonitrile extraction, low-temperature cleanup, and detection by liquid chromatography with tandem mass spectrometry

A multiresidue method for determining pesticides in rapeseed, rapeseed oil, and rapeseed meal by use of liquid chromatography-tandem mass spectrometry is developed. Samples were extracted with acetonitrile or acidified acetonitrile and cleaned up by a 12 h freezing step. The recovery data were obtained by spiking blank samples at three concentration levels. The recoveries of 27 selected pesticides in rapeseed, rapeseed oil, and rapeseed meal were in the range of 70-118%, at the concentration level of 10 μg kg(-1), with intraday and interday precisions of lower than 22 and 27%, respectively. Linearity was studied between 2 and 500 μg L(-1) with determination coefficients (R(2)) of higher than 0.98 for all compounds in the three matrices. The limits of quantitation (LOQs) of pesticides in rapeseed, rapeseed oil, and rapeseed meal ranged from 0.3 to 18 μg kg(-1). The n-octanol-water partition coefficient showed more influence than water solubility in extracting pesticides by acetonitrile from matrices of high fat content. This method was successfully applied for routine analysis in commercial products.     

铁皮石斛人工繁育技术

以铁皮石斛商品瓶苗和盆栽苗茎段为试材,添加不同浓度6-BA、NAA,诱导茎段腋芽、原球茎和不定根,建立再生体系,快速繁殖铁皮石斛组培苗,以期为秦岭淮河一线以北地区人工繁殖铁皮石斛提供参考依据。结果表明:外植体宜采用70%酒精浸泡20 s,0.1%升汞浸泡8 min消毒;MS+6-BA 3.0~5.0 mg·L^-1+NAA 0.1~1.0 mg·L^-1适合腋芽分化和增殖,分化率92%~100%;1/2MS+6-BA 2.0 mg·L^-1+NAA 0.5 mg·L^-1+土豆泥15%适合原球茎诱导增殖,增殖倍数7.7;1/2MS+NAA 0.3 mg·L^-1生根率为100%;以松针土移栽,成活率83%。     

航空静电喷雾与非静电喷雾沉积效果的比较

利用直升机R44和固定翼飞机Y5B挂载自主研制的航空静电喷雾系统设备进行机场和野外航空静电喷雾与非静电喷雾的比较试验.结果表明:静电喷雾存地面和空中靶标表面、树冠中上部叶片正面的沉积效果明显优于非静电喷雾:雾粒细小、分布均匀、沉积密度提高1倍以上,最高达1.75倍;直升机静电喷雾在叶片背面的沉积率为非静电喷雾的2.15倍,是常规航空喷雾的3.31倍.Y5B静电喷雾在叶片背面的沉积率为非静电喷雾的2.38倍,是常规航空喷雾的3.44倍.     

转基因抗虫保铃棉(Bollgard~ひ)检测方法研究

运用常规 PCR方法和实时荧光 PCR方法对转基因抗虫保铃棉外源基因 Ca MV 35 S启动子、NOS终止子、标记基因NPT II和目的基因 Cry IA(c)进行了检测。建立了快速、准确的抗虫保铃棉籽转基因成分的 PCR检测方法。     

响应面法优化绿色木霉产孢子固体发酵培养基

为探讨绿色木霉固体发酵产孢子的最佳发酵培养基。运用响应面法设计试验,筛选出主要影响因子碳酸钙、硫酸铵及氯化锰,以孢子量为响应值建立二次回归方程,并借助Design Expert 8.0.6软件进行分析数据并确定优化条件。结果表明,在温度28℃,接种量15%,料水比1:0.70的培养条件下发酵7天,孢子量为8.505×109CFU/g。通过优化,最终确定最佳发酵配方为:麸皮(过40目筛)50%,稻壳粉(过20目筛)30%,玉米粉(过80目筛)20%,硫酸铵1.08%,黄豆饼粉2.00%,玉米浆干粉2.00%,碳酸钙0.55%,氯化锰0.37%,pH 6.0～7.0。该配方所用原料廉价易得,发酵水平高,为大规模工业化生产提供保障。     

Immunofluorescence Plaque Assay for African Swine Fever Virus

Suitably diluted cell culture adapted African swine fever virus preparations were inoculated on VERO cell monolayers and grown on coverslips. Gum tragacanth was used as an overlay. After three days incubation at 37°C the infected cultures were fixed with acetone and stained with fluorescent antibody conjugate. Fluorescing plaques consisted of 20-30 infected cells.

Three statistical criteria for a quantitatively reliable assay were met: the Poisson distribution for plaque counts, linearity of the relationship between the concentration of virus and the plaque count and reproducibility of replicate titrations. The method is suitable for counts up to at least 70 plaques per 5 cm² coverslip and computed titers are reproducible within 0.16 log units with a total of 300 plaques enumerated.

     

Quantitative dynamics on stimulating regeneration and sowing seedlings of Larix gmelinii in Daxing''''an Mountains

Introduction4LarixgmeliniiisoneofmajortimberspeciesinthenortheastDaxing'anMountainsinChina.Itplaysanimportantroleinprovidinglumber,maintainingthenaturalenvironmentinthisregionandadjustingecologicalequilibrium.Alargenumberofburnedareasandcut-overareasneedtoregenerateeveryyear.Larixgmeliniihasgreatnaturalregenerationcapabilityandenoughseedsources,andalmostmeetsabumperharvestyeareverythreeyears(Xuetal.1992;Shan1990;Xuetal.1993).Someburnedandloggedareaswouldberapidlyrestoredthroughafewofassista…     

闽南毛竹林合理经营密度的研究

在闽南毛竹分布区的毛竹林分中建立标准地２４７块，利用柯尔莫哥洛夫的正态分布检验法。结合半峰宽理论，分析计算了闽南毛竹林的合理经营密度为１２００株￣１６５０朱／ｈｍ＾２，为闽南毛竹林持续经营提供了理论依据。     

实生板栗高接换冠技术的研究

采用多种嫁接方法在改造大龄实生板栗上进行不同时间和嫁接时对树冠的不同处理的试验，结果表明，应用挖骨皮接、插皮接、切腹接的最佳时期是在树体萌芽前１０ｄ至萌芽期（即３月中旬左右），段木芽接法宜在３月上旬前进行为好；１树１桩嫁接成活率较１树多桩低，当１树少于４桩嫁接时，适当留１～２个辅养枝可提高成活率，且对接芽生长影响不大．