共查询到19条相似文献,搜索用时 54 毫秒
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松材线虫是进境木质包装中的重要检疫对象之一。本研究以进境木质包装中截获的12种伞滑刃属线虫,包括4种不同来源的松材线虫、3种不同来源的拟松材线虫、豆伞滑刃线虫、大尖尾伞滑刃线虫、阿苏里伞滑刃线虫、伯氏伞滑刃线虫及拟小刺伞滑刃线虫为材料,进行实时荧光PCR检测。结果显示,仅松材线虫可观察到明显的荧光强度变化,而其他线虫的荧光强度没有变化。通过研磨样品的方法,提取单条松材线虫的DNA,进行实时荧光PCR试验,检测成功率为100%,且不论是雄虫、雌虫或幼虫均能检测出来。该研究对于我国口岸进境木质包装中松材线虫的检测具有一定的实际意义。 相似文献
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两种分子技术检测松木中松材线虫的效果评价 总被引:1,自引:0,他引:1
为更好地解决当前松材线虫Bursaphelenchus xylophilus检测工作中频现的假阴性问题,通过不同分子技术检测效果的比较评价,摸索出了一套可提高松材线虫检测准确性的技术体系。针对早期感病松木的低线虫量,建立了线虫的最大量分离方法和基于rDNA ITS序列的分子检测体系,该体系可高效检测出单条松材线虫,准确率达93.75%。以来自浙江省不同疫区的96份松木样品为检测材料,比较了SCAR标记和ITS序列2种分子检测技术的阳性率。结果显示,通过首次PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率分别为52.08%和55.21%,SCAR标记的检测阳性率为30.21%;通过第二次或巢式PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率提高到了97.92%和100.00%,SCAR标记的检测阳性率提高到了59.38%,表明基于ITS序列的检测阳性率明显高于SCAR标记,且通过第二次或巢式PCR方法可进一步提高检测灵敏度,降低假阴性率。因此,通过线虫的最大量分离并基于rDNA ITS序列的分子检测可明显提高检测准确性,更适用于松材线虫的常规检测。 相似文献
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口岸截获的日本松材线虫和中国南京松材线虫研究(摘要) 总被引:1,自引:0,他引:1
口岸截获的日本松材线虫和中国南京松材线虫研究(摘要)焦国尧沈培垠徐培方安榆林李红梅付鹏(南京动植物检疫局210009)(南京农业大学210094)数十年来,由松材线虫(Bursaphelenchusxylophilus)引起的松树萎蔫病在日本普遍发... 相似文献
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应用TaqMan探针实时荧光定量PCR技术早期检测稻瘟病 总被引:2,自引:1,他引:2
稻瘟病是一种严重危害水稻生产的真菌病害,早期监测和防治是关键。温室接种大田主栽品种合系39和感病水稻蒙古稻,定时观察发病症状。提取水稻病样总DNA,根据稻瘟菌28S rDNA基因序列,设计特异引物和TaqMan探针,进行荧光定量PCR检测。结果表明稻瘟菌在蒙古稻和大田主栽品种合系39上症状最初出现时间为接种后72 h,蒙古稻典型症状出现时间为接种后168 h,在合系39上出现典型症状的时间为接种后190 h以后。利用TaqMan探针荧光定量PCR技术,在接种12 h的蒙古稻和合系39上均能检测到稻瘟菌DNA,接种48 h,菌量拷贝数达到最高峰,接种72 h,菌量拷贝数开始下降。不同品种中病原菌拷贝数存在差别,在接种12 h的蒙古稻中稻瘟菌含量为7.2×103个拷贝数,在合系39中为4.9×103个拷贝数。本研究结果表明,应用实时荧光定量PCR技术可以在接种后12 h症状未显现之前检测到稻瘟菌,为稻瘟病流行监测和早期防治提供了科学依据。 相似文献
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Bursaphelenchus xylophilus is the causative agent of pine wilt disease. These nematodes disperse in the cortical and wood resin canals of host tree branches in the early stage of infection. To determine whether the dispersal can be regarded as diffusion, we inoculated B. xylophilus into living or boiled Pinus densiflora branch sections and investigated the spatial distribution 24 hr later. Assuming that some nematodes disperse randomly in the axial direction of branch sections and the rest remain at the inoculation site, the coefficient of diffusion (D) by dispersers was estimated to be 23.7–40.7 and 109.4 cm2/day for the living and boiled branches, respectively. The proportion of nondispersers was estimated to be 0.46–0.79 and 0.52 for the living and boiled branches, respectively. When the nematodes were inoculated at the centre of upright living branch sections, they dispersed up and down at similar D values of 7.4 and 7.9 cm2/day, respectively. 相似文献
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松材线虫组DNA条形码筛选 总被引:1,自引:0,他引:1
本文基于Gen Bank中登录的14种松材线虫组线虫的28S、18S和ITS部分基因DNA序列,利用遗传距离法及分子生物学方法评价其各自作为松材线虫组DNA条形码序列的适用性。结果显示,28S(拟松材线虫不分亚种)、28S(拟松材线虫分亚种)、18S和ITS的基因序列种内成对遗传距离平均值分别为0.0071、0.0030、0.0007、0.0043,种间成对遗传距离平均值分别为0.0476、0.0454、0.0052、0.1556,其中28S、18S基因序列种内及种间距离存在一定程度的重叠,ITS具有一定程度的Barcoding gap。3个基因序列构建的NJ进化树显示,28S、ITS构建的系统进化树具有较高的节点支持率,可以有效将14个松材线虫组线虫分离成独立分支,而且28S可以有效鉴别出拟松材线虫的2个亚种;基于18S基因构建的进化树不能对吉拉尼伞滑刃线虫、日本冷杉伞滑刃线虫、拟松材线虫进行有效区分。因此,28S、ITS区具有一定的遗传距离间隔以及相对较高的物种识别率,可以作为松材线虫组线虫候选条形码基因。 相似文献
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热处理杀灭木质包装中松材线虫的技术研究 总被引:6,自引:2,他引:6
近年来,出入境检验检疫部门多次从经热处理的木质包装中截获松材线虫.本实验利用电热恒温箱和热处理窑对携带松材线虫的进口木质包装进行热处理试验,结果表明:在实验室和工厂化热处理试验中,木材中心温度达到56℃时处理4h不能完全杀灭木质包装中的松材线虫;木材中心温度达到60℃时在实验室试验中需要3.5h方能完全杀灭松材线虫,而在工厂化热处理试验中,木材中心温度达到60℃处理2h,尚不能完全杀灭松材线虫;实验室和工厂化试验表明,仅当木材中心温度达到65℃,保持0.5h以上方能100%杀灭松材线虫.本文探讨了实验结果与目前IPPC推荐的国际标准(即木材中心温度56℃时处理30min以上)不一致的原因. 相似文献
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N. Kim H. W. Jeon M. Mannaa S.-I. Jeong J. Kim J. Kim C. Lee A. R. Park J.-C. Kim Y.-S. Seo 《Plant pathology》2019,68(3):434-444
Pine wilt disease (PWD) is the most destructive disease threatening pine worldwide. The disease is mainly caused by the pine wood nematode, Bursaphelenchus xylophilus, which is vectored by pine sawyer longhorn-beetles, Monochamus spp. This study aimed to select resistance-inducing pine endophytic bacteria for management of PWD. To set up a defence-related genes expression pattern for screening, four chemical inducers (salicylic acid, γ-aminobutyric acid (GABA), β-aminobutyric acid and α-aminobutyric acid) were tested in vitro on pine calli and in vivo on pine seedlings. Treatment with GABA had the greatest reduction in PWD severity on pine seedlings. The pattern of defence-related gene expression in calli treated with GABA was used to select potential resistance-inducing bacterial strains. In addition, 92 bacterial strains were isolated from pine tree needles and stems and were tested for expression of defence-related genes in pine calli in vitro. Among the tested strains, 13 showed a similar pattern to GABA treatment in at least four tested defence-related genes and were selected for the seedling assay. From the seedling assay, three bacterial strains (16YSM-E48, 16YSM-P180 and 16YSM-P39) showed significant reduction in PWD severity compared to the untreated control. Moreover, among the selected strains, cell-free culture supernatant of strain 16YSM-P180 significantly reduced PWD severity in inoculated pine seedlings. The selected strains were identified based on the 16S rRNA sequence as Pseudomonas putida 16YSM-E48, Curtobacterium pusillum 16YSM-P180 and Stenotrophomonas rhizophila 16YSM-P39. These selected strains are suggested as potential alternatives for management of PWD by induction of systemic resistance. 相似文献