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 .     

云南醉鱼草属(Buddleja L.) 观赏植物资源的调查研究

 在对云南分布的醉鱼草属观赏资源系统调查和资料整理的基础上, 研究了该属植物在云南的地理分布规律及25 种(包括变种) 醉鱼草的观赏性状, 包括花、叶、香气等观赏特征及其开花期, 指出开发利用面对的问题和应采取的措施。     

云南特有野生果树资源及其分布特点

 云南有各类野生果树资源约500 余种, 其中云南特有的164种, 分属35科54属, 多为热带、亚热带果树和浆果类果树, 形成滇南—滇东南、滇西—滇西北和滇中—滇东北3个特有野生果树集中分布中心。这些果树资源用途广泛, 优异性状明显, 有待深入研究和综合开发利用。     

Detection of Colletotrichum coccodes and Helminthosporium solani in soils by bioassay

The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.     

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。     

水杨酸和乙烯在烟草抗黑胫病中的作用

5mmol/L水杨酸或1g/L乙烯外源处理可诱导烟草对黑胫病的抗性，推迟植株死亡。不能积累水杨酸的转nahG基因植株和不表现乙烯应答反应的转etr1-1基因植株对黑胫病的抗性显著弱于野生型植株。组成性积累高水平水杨酸的转entC和pmsB基因植株对黑胫病的抗性明显强于野生型植株。这些结果说明水杨酸和乙烯增强烟草对黑胫病的抗性，减缓黑胫病的扩展。0．5mmol/L KCN连续浇灌植株4天或浸溃叶片2，4h均不能恢复转nahG基因植株丧失的对黑胫病的抗病性。此结果进一步支持关于KCN处理不能恢复转nahG基因植株丧失的对真菌病害抗病性的假说。     

温度对稻茎毛眼水蝇实验种群增长的影响

试验结果表明:在17～31℃恒温范围内,稻茎毛眼水蝇的发育速率随温度上升呈逻辑斯蒂曲线趋势。幼虫、蛹存活率、成虫产卵量、种群增长指数随温度的上升呈抛物线趋势。卵孵化率在20～31℃范围内无显著差异,均在92％以上。在17℃和35℃时,孵化率下降至86.67％和76.36％。在35℃时,幼虫不能存活,成虫不产卵。     

茉莉花茶加工技术与吸香机理研究现状

为了进一步综合地认识茉莉花茶窨制的工艺本质、完善茉莉花茶窨制技术,本文综述和分析了茉莉花茶加工技术及吸香机理的研究现状,并对后续研究进行了展望。     

土壤肥料学云南省省级精品课程教学改革与实践

为适应高等农业教育改革和推进素质教育的要求,针对云南农业大学涉农专业的基本特点,结合土壤肥料学云南省省级精品课程的建设和云南省土壤与肥料方面的基本情况,以培养学生的创新精神与实践能力为根本目的,在教学内容、教学条件、教学方法等方面进行了该课程教学改革与实践,收到显著成效.     

安化黑毛茶加工现状与机械化发展趋势

本文结合安化黑毛茶初制加工现状及发展黑茶机械化生产的必要性,指出黑茶渥堆机的设计重点及难点,以期达到渥堆过程的量化控制,实现机械化生产,使黑茶生产的环境条件及加工流程符合现代化茶叶加工的要求。