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     

二十三个柚品种特性评价

对1965年定植的23个柚(Citrus grandis Osbeck)品种进行了20多年的性状比较。筛选出矮化类型6个,早熟和晚熟类型各3个,丰产类型6个,品质优良类型11个。发现仅有部分柚品种富含维生素C;果汁中维生素C含量与酸含量呈明显负相关。证实长寿沙田柚中的冬瓜圈和古老钱两个选系比广西沙田柚更丰产优质;垫江白柚质优,但产量低。简述了各参试品种的利用前途。     

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。     

板栗优良新品种‘黄棚’

 ‘黄棚’是20世纪90年代初从海拔300 m以上的山地栗园选出的实生变异优株。经多年高接鉴定和丰产栽培试验表明, 该品种早实, 丰产, 抗旱, 耐瘠薄, 品质优良, 为中早熟、适应范围广的炒食型板栗新品种。     

黑穗醋栗新品种‘甜蜜’

 ‘甜蜜’是从波兰引进的黑穗醋栗试材中筛选出来的晚熟品种, 生长势强, 果穗长, 单果质量1.47 g, 可溶性固形物14% , 维生素C 1.6 g/kg FM。果实出汁率70%。风味酸甜, 鲜食口味佳。丰产性好。在长春地区7月中下旬成熟。     

梨新品种‘花盖王’

 ‘花盖王’是从‘花盖梨’中选出的梨四倍体芽变新品种。该品种除保持‘花盖梨’的优质、丰产、抗寒等优良性状外，最突出特点是果实大，单果质量是‘花盖梨’的2倍多，最大为250 g。     

早熟大果鲜食枣品种‘七月鲜''

‘七月鲜’是从合阳县农家品种中选出的早熟优良品种，比中熟品种‘梨枣’早熟20d以上；果大，整齐，平均单果质量29．8g，可溶性固形物含量28．9％，可食率97．8％，味甜，肉质细，极宜鲜食，不易裂果，早果性强，抗缩果病，节间短，树体矮小，适宜矮化密植和设施栽培。