Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

孔雀河流域绿洲生态支持系统调控模式研究

生态环境用水量的减少是孔雀河流域近 50年来现代绿洲环境退化的主要原因 ,维持生态与社会经济的合理用水比例是绿洲系统持续发展的前提。以此为基础 ,本文采用水资源承载力的方法探讨了绿洲生态支持子系统和社会经济子系统优化调控模式 ,最后就配置方案及发展模式的可行性进行了分析     

城市绿地系统对经济发展提升作用的机制——与大连市实证研究

城市绿地系统具有生态、经济、社会等多重属性 ,在城市复合系统中占有特殊的位置 ,对城市经济发展亦具有直接、间接的提升作用 :( 1 )改善城市环境质量 ,创造环境优势 ,促进城市地价及布局其间的各种经济成分增值 ;集聚外来资金和高新产业发展 ,带动整个城市产业结构优化升级。( 2 )绿地建设作为城市经济产业的有机组成部分 ,形成环境经济产业链 ,拉动其相关经济产业的发展 ,促进经济增长。 ( 3 )提高城市知名度 ,促进城市旅游业发展 ,集聚高素质人才 ,促进城市经济发展 ,本文以北方海滨城市大连为例 ,在定性分析绿地经济提升作用机制的基础上 ,建立计量经济模型 ,定量分析城市绿地系统的经济提升作用     

ITS—RFLP分析进行昆虫病原线虫斯氏属和异小杆属的分类鉴定

本研究通过PCR扩增和六种限制性内切酶（AluⅠ，HinfⅠ，MboⅠ，RsaⅠ，HaeⅢ和PvuⅡ）酶切，对国内害虫防治上常用的几种昆虫病原线虫，包括斯氏属S.car-pocapsae,S.feltiae和S.glaseri以及异小杆属H.bacteriophora,H.zealandica,H.indicus和H.megidis等8个品系rDNA-ITS进行分析。建立起可以区分各线虫种的标准RFLP图谱。该方法快速简便，稳定可靠，需要的样品量少。可以用于新鲜的，或冻存的样品，甚至分析单条的线虫，不仅可进行昆虫病原线虫的快速分类鉴定。而且进一步可以应用于线虫田间释放的辅助监测。实际田间感染率的测定和线虫毒力的比较。     

氮磷钾混合肥对锥栗坚果产量和质量的影响

以 5年生锥栗为试树 ,进行土壤穴施氮磷钾混合肥试验 ,研究不同氮磷钾肥配比对锥栗坚果产量和质量的影响。结果表明 ,不同组合处理之间 ,除叶片的蛋白质含量和单株果枝数有显著差异外 ,叶绿素含量、单枝结棚数、单株结棚数、栗棚重、坚果重、出实率、单株棚产量和坚果产量的差异均不大。以穴施 0 .2 0kg尿素、0 .1 0kg钙镁磷和 0 .1 5kg氯化钾的混合肥较经济而有效     

Effects of external counterpulsation on renin-angiotensin system and hemodynamics in dogs with myocardial ischemia

AIM:To examine the effect of external counterpulsation(ECP) on renin-angiotensin system(RAS) and hemodynamics in dogs with myocardial ischemia and their relationship. METHODS:Acute myocardial ischemia was induced by occluding the left anterior descending of the dogs. The local renin activity, angiotensin Ⅱ(AngⅡ) level, angiotension-converting enzyme(ACE) activity were tested by biochemical methods and hemodynamics was recorded by a 8-channel physiological recorder.RESULTS:Ischemia could actiivate renin,ACE and AngⅡ in cardiovasculature and ECP reduced them except for renin in ischemic myocardium. Ischemia also could activate RAS in lung and kidney, which play an important role on circulating RAS, and ECP reduced them. Furthermore, ECP could improve hemodynamics and there existed a close relationship between local AngⅡlevel and hemodynamics. CONCLUSION:ECP can reduce local RAS and improve hemodynamics in dogs with myocardial ischemia, which might be one of mechanisms underlying the protective effect of ECP on ischemic myocardium.     

两系稻制种穗期病害发生较重的原因及防治措施

调查分析稻粒黑粉病、穗颈稻瘟等两系杂交稻制种穗期主要病害的发生原因,提出了清除菌源、轮换制种、健身栽培、对口药剂防治等防治措施。     

刺梨及其近缘种PCR实验体系的建立与优化

以刺梨及其近缘种月季为试材,进行了RAPD-PCR实验参数的确立和优化试验。结果表明,刺梨及月季25μL反应体系的最优组成为2.5μL10×反应缓冲液,2mmol/LMg2+,0.2mmol/LdNTP,1.6mg/L模板DNA,0.4μmol/L随机引物和1.2UTaqDNA多聚酶。经PCR扩增验证,此反应体系亦适宜于刺梨的部分近缘种,可有效用于RAPD分析;通过将退火温度提高至50℃或采用“Touchdown”扩增程序,并在50μL反应体系中适当增加特异引物对浓度(1.0μmol/L),模板DNA(4.0mg/L)和TaqDNA多聚酶(3.0U/管)的使用量,建立起适合于刺梨特异DNA片段检测及回收的特异PCR扩增实验体系,为刺梨的分子克隆奠定了技术基础。     

黑杨萎蔫枝叶对棉铃虫生殖行为的影响

研究了黑杨(Populus nigra)萎蔫枝叶及其气味对棉铃虫成虫的隐藏部位、产卵部位、交配率及产卵量等的影响，并对黑杨萎蔫枝把在田间诱集到的棉铃虫雌蛾进行了解剖观察。结果表明，在田间，黑杨萎蔫枝把诱集到的棉铃虫93．3％已经交配。在气候箱中，放置黑杨萎蔫枝叶的环境中配对饲养的棉铃虫，经过1个暗期和2个暗期后，其交配率与对照环境中的均无显著差异。在光期开始时，有51．5％的成虫隐藏在饲养笼内所设置的黑杨叶子圆片下，对照纱布圆片下仅有16．7％。叶子圆片所在区域的日平均落卵量显著大于对照区。黑杨萎蔫枝叶及其气味对棉铃虫的产卵前期、产卵持续期、成虫寿命和产下50％的卵所用的时间无显著影响，但饲养笼内有黑杨枝把或黑杨叶片气味存在时，棉铃虫的产卵量比模拟枝把和对照显著提高。     

十字花科蔬菜-小菜蛾-寄生性天敌三营养级系统研究:不同条件下小菜蛾幼虫被寄生率调查

本文根据在深圳、广州、东莞三地对不同蔬菜、不同农事措施、杀虫剂不同施用水平菜地小菜蛾自然种群幼虫被寄生率调查结果，分析十字花科蔬菜－小菜蛾－菜蛾绒茧蜂三种群系统中，蔬菜品种、农事操作、杀虫剂对菜蛾绒茧蜂寄生行为的影响。结果说明：不同蔬菜品种上，小菜蛾幼虫被寄生率有不同程度的差异，以包心菜菜地寄生率最高，超过３０％。十字花科蔬菜与其他非十字花科蔬菜的间作对绒茧蜂的寄生率也存在一定的影响作用。深圳、东莞、广州三个地区，以广州地区小菜蛾幼虫被寄生率较高。化学杀虫剂降低绒茧蜂的寄生率。小菜蛾幼虫的体色与被寄生率也有一定的关系，以体色与背景色一致的个体被寄生率较低