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Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR 总被引:4,自引:1,他引:4
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 . 相似文献
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为延长牡丹切花的观赏时间和提高观赏品质,在对水性丙烯酸树脂和茶树精油喷涂处理浓度和喷涂时间单因素试验的基础上进行Box-Behnken中心组合试验设计,并通过响应面回归分析优化牡丹切花喷涂处理条件。结果表明,牡丹切花喷涂处理的最佳条件为水性丙烯酸82.95 g ? L-1、茶树精油0.65 mL ? L-1,持续喷涂10.28 min。按此条件进行验证,测得牡丹切花常温贮藏5 d后的花瓣总色差值为2.15,对照组色差为6.64;该处理能增加切花的观赏时间,降低失水率和最大花径皱缩率,明显改善花瓣的物理特性,并在一定程度上降低花瓣腐烂指数。 相似文献
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氟吡菌酰胺不同施药方式对水稻拟禾本科根结线虫的防治效果 总被引:1,自引:0,他引:1
为评价41.7%氟吡菌酰胺悬浮剂防治水稻拟禾本科根结线虫Meloidogyne graminicola的应用潜力,将氟吡菌酰胺与吡虫啉种衣剂混合后采用种子包衣法及喷洒法研究其对水稻拟禾本科根结线虫的田间防效,并测定了不同施药方法对水稻生长和产量的影响。结果表明,氟吡菌酰胺4.2、8.3、12.5 g(a.i.)/kg种子与吡虫啉18.0 g(a.i.)/kg种子混合包衣处理,播种后35 d其根结抑制率和防效分别为41.0%~51.8%和47.4%~58.6%,土壤中2龄幼虫减退率为38.6%~40.4%,显著高于单施吡虫啉18.0 g(a.i.)/kg种子处理。水稻播种后连续3次以氟吡菌酰胺250.2、375.3、500.4 g(a.i.)/hm2进行土壤喷洒,最后1次施药后7 d,其根结抑制率和防效分别为81.0%~89.9%和65.9%~74.3%,土壤中2龄幼虫减退率为65.4%~73.4%,均显著高于对照药剂克百威1 800.0 g(a.i.)/hm2处理。氟吡菌酰胺各处理对水稻苗期生长均有较好的保护作用,能显著提高千粒重和有效穗数,产量比空白对照增加50.0%~61.2%,保产效果显著。水稻播种后35 d采用氟吡菌酰胺对稻田进行1次喷洒,对水稻具有一定的保护作用,保产效果不明显。表明采用氟吡菌酰胺与吡虫啉混合包衣种子处理及在水稻苗期进行喷洒处理对水稻拟禾本科根结线虫防效显著,具有显著的保产效果。 相似文献
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壳聚糖涂膜对大蒜保鲜效果的研究 总被引:1,自引:0,他引:1
采用壳聚糖作为涂膜保鲜材料,在常温条件下研究不同浓度壳聚糖的单膜和复合膜对大蒜贮藏期间主要理化特性的变化.结果表明,1.50%壳聚糖单膜和壳聚糖-玉米淀粉复合膜均可有效抑制大蒜的呼吸强度,保持大蒜的水分含量、硬度、可溶性同形物含量和总糖含量,以壳聚糖-玉米淀粉复合膜的保鲜效果最为理想. 相似文献
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Sherry Rachel Jacob MB Arunkumar Madhuban Gopal Chitra Srivastava SN Sinha 《Pest management science》2009,65(7):817-822
BACKGROUND: An efficient delivery system for seed‐protectant chemicals is needed in light of several disadvantages of conventional seed treatment methods. This study evaluates the efficacy of film‐coat application in maintaining the persistence and potency of imidacloprid on Lycopersicon esculentum (L.) Mill. seeds after simultaneous storage under ambient and regulated environment in paper and aluminium packages. RESULTS: High‐performance liquid chromatography (HPLC) revealed 0.135 mg kg?1 of herbage material to be the threshold value beyond which absolute control was obtained, and with film coating the latter was achieved even with half‐dosage seed treatment, irrespective of the storage condition. The technique provided early protection to the crop and also nullified the deleterious effects of ambient storage on the persistence and potency of the pesticide. CONCLUSION: Film coating enabled superior pesticide dosage as well as higher biological efficacy to be achieved. Hence, in addition to being an ecofriendly alternative, the technique would be a more economically viable option for storage of treated seeds. Copyright © 2009 Society of Chemical Industry 相似文献
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玉米茎腐病是世界玉米产区普遍发生的一种土传病害,也是我国玉米产区的主要病害.为了筛选合适的玉米茎腐病病原菌的拮抗菌,并研究其种衣剂对玉米生长的作用,采用平板对峙试验对10株疑似拮抗菌进行筛选,并测定铁载体和吲哚乙酸(IAA)两种生理指标,筛选对玉米种子有促生潜力的菌株,将筛选到的菌株作为微生物种衣剂的活性成分,研究不同... 相似文献