正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

猪带绦虫六钩蚴cDNA文库的构建与筛选

在体外将猪带绦虫虫卵孵化为有活力的六钩蚴。采用商品化试剂盒抽提六钩蚴总 RNA、m RNA,反转录为双链c DNA,再加 Eco R Adapter。在去除小片段后 ,dsc DNA与 λgt11噬菌体 DNA连接 ,经蛋白包装 ,构建了猪带绦虫六钩蚴λgt11c DNA文库。该文库效价为 3.5× 10 9pfu/ m L ,重组 DNA片段大小为 0 .5～ 3.2 kb,平均为 1.6 kb,蓝白斑比 1∶ 9。用抗体探针对文库进行免疫学筛选 ,在消除非特异性反应的基础上筛选约 10 6 重组子 ,共得到 118株强阳性克隆 ;应用 PCR鉴定上述部分阳性克隆 ,均扩增到 0 .5 kb以上的片段。结果显示 ,构建的文库合格 ,含六钩蚴所有抗原基因 ,可用于六钩蚴 c DNA克隆的筛选 ;用免疫学与 PCR联合筛选 c DNA文库 ,可消除假阳性     

Rapid Screening for Aluminum Tolerance in Cereals by Use of the Chlorophyll Fluorescence Test

The effects of aluminum on the photosynthetic apparatus were examined in cereals grown in nutrient solutions (pH 4.5) at two Al levels (0 and 148 μM). The chlorophyll fluorescence kinetics results confirmed that the soft wheat ‘BHG’ cultivar has the potential for growth on acid soils while triticale cultivars ‘Niovi’ and ‘Dada’ appeared to be relatively tolerant. The percentage decrease in F_v/F_m of the less tolerant cultivars after Al-treatment indicated a decrease in the efficiency of the primary photochemistry of PS II, while the decrease in the ratio F_V/F_osuggested that exposure of the cultivars ‘Dio’ and ‘Appulo E’ to aluminum caused injury to the thylakoid structure. The percentage fluctuations of the ratio F_v/F_mwere shown to correlate very closely with the assessment of injury as evaluated by the relative top fresh weight. Measurements of chlorophyll fluorescence in vivo could be used to monitor injury caused by “Al-stress”, and thus they may serve as a rapid screening test for Al tolerance.     

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 10⁸ colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×10³ and 2.3×10⁴ cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R ²=0.74).     

关于编制我国果树种质资源评价系统若干问题的商榷

资源评价是果树种质资源工作的中心环节。编制果树种资源评价系统是发展和提高我国果树种质资源工作水平的一项重要基础工作。“七五”期间,“果树种质资源主要性状鉴定评价”专题组编制１８种果树的《果树种质资源描述符》,于１９９０年在农业出版社出版。本文就其中其中若干问题提出商榷意见。     

平阴玫瑰花粉亚显微形态及品种分类研究

 对平阴玫瑰24个品种的花粉亚显微形态进行了扫描电镜观察研究, 并进行了主成分分析和聚类分析。结果表明, 在花粉亚显微形态特征方面, 各品种之间既有共性, 也存在明显差别, 特别是外壁纹饰的不同可作为玫瑰品种鉴别的重要依据。聚类分析将平阴玫瑰花粉分为两大类, 即玫瑰系和蔷薇系, 与传统形态分类结论基本一致, 但也存在一定的差异。     

草菇栽培原料发酵剂菌株的筛选及应用初探

从广西军区食用菌生产基地、广西大学微生物厂、广西大学牧场等地采集的样品中分离得到多株菌株。再从耐热、耐碱、产酶能力及营养物综合利用等方面考查各菌株，最终筛选得到理想的放线菌1株、细菌3株组成发酵剂菌株，并用于栽培原料发酵后栽培草菇。实验结果显示：加入该菌剂的料堆27h后温度可升至65℃，此时对照55℃，环境40℃，草菇生物学效率比对照提高了4.76％至7．56％。     

菊花18个品种的RAPD分析

 采用RAPD 技术分析了18 个菊花品种DNA 的多态性。从80 个10 碱基随机引物中筛选出多态性频率高的3 个引物。扩增的多态性片段在600 bp～1300 bp 之间。检测出两个品种特有的分子标记, ‘大红托桂’有OPD15 (1200 bp) ,‘玉翎管’缺失OPA17 (1100 bp) 。瓣型一致的品种间基因型相似系数较高。     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

对锰害敏感性不同的两个苹果品种枝条中锰的积累与分布

 以对苹果粗皮病抗性不同的两个苹果品种富士(感病) 和乔纳金(抗病) 为材料,通过田间采样、室内盆栽模拟,结合组织化学、显微鉴定和X- 射线微区分析,研究了枝条内锰的累积、分布与粗皮病抗性的关系。结果表明,锰在韧皮部的含量高于木质部,感病品种含量高于抗病品种;粗皮病病斑部位存在氧化锰沉淀,韧皮部锰的局部累积和分布与病斑的分布相一致;锰在感病品种的枝条横截面上存在局部累积现象,而在抗病品种上分布较均匀。