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Direct amplification by PCR of DNA from ungerminated teliospores of Tilletia species tiltiltia未萌发端孢DNA的PCR直接扩增
Pub Date : 1999-03-01 DOI: 10.1080/07060661.1999.10600076
J. Mcdonald, E. Wong, G. Kristjansson, G. P. White
Abstract A simple technique of conducting polymerase chain reaction (PCR) on single ungerminated teliospores of Telleria species was developed. Teliospores were manually cracked under a stereo microscope prior to adding to the PCR reaction mixture. Amplification product was obtained using primers for either a portion of the nuclear ribosomal intergenic spacer region or a portion of the mitochondria) DNA. Collections of teliospores from Tilletia indica, Tilletia barclayana, Tilletia controversa, Tilletia tritici, Tilletia laevis and an unidentified Tilletia sp, from Lolium varied in the proportion of spores from which amplification product could be detected, with the success rate ranging from 100 to 10%. This technique avoids the difficulty and time delay in having to germinate teliospores prior to extracting DNA from a mycelial matte and thus will be of great value in the application of PCR methods for regulatory testing and phylogenetic studies of Tilletia species.
摘要建立了一种对Telleria (Telleria)未萌发的单孢子进行聚合酶链反应(PCR)的简易方法。在加入PCR反应混合物之前,在立体显微镜下人工裂解远孢子。利用引物对核糖体基因间间隔区或线粒体DNA的一部分进行扩增。来自黑麦草的印度Tilletia、barclayana Tilletia、controversa Tilletia、tritictia tritici、laevis Tilletia和一种未知Tilletia sp .采集的远孢子扩增产物的孢子比例不同,成功率为100 ~ 10%。该技术避免了在提取菌丝体DNA之前萌发端孢子的困难和时间延迟,因此将在PCR方法应用于tiltiltia物种的调节测试和系统发育研究中具有重要价值。
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引用次数: 12
Impact of animal manures on verticillium wilt, potato scab, and soil microbial populations 动物粪便对黄萎病、马铃薯痂和土壤微生物种群的影响
Pub Date : 1999-03-01 DOI: 10.1080/07060661.1999.10600089
K. Conn, G. Lazarovits
Abstract This study examined the effect of chicken, liquid swine, and solid cattle manures on verticillium wilt and potato scab incidence at two commercial farms near Alliston, Ontario. The manures (chicken, 66 t/ha; swine, 55 hLJha; cattle, 100 t/ha) were spread in the spring of 1996 and incorporated with a Rototiller to a depth of 15 cm. Survival of Verticillium dahliae microsclerotia (MS) was determined by burying nylon mesh bags containing MS in the plots. After 4 weeks, the MS were plated on agaz medium. Soil samples collected after incorporation of manures were brought back to the laboratory and used for concurrent laboratory experiments. Ten-gram amounts of soil were placed into test tubes, bags of MS buried in the soil,and survival measured under controlled conditions. Potatoes were planted in the field 1 month after amendment. Potatoes were planted in 1997 and 1998 without any further treatment. verticillium wilt incidence was determined by isolation of V.dahliae from petioles. Scab incidence was...
摘要本研究在安大略省阿利斯顿附近的两个商业农场研究了鸡粪、猪粪液和牛粪固体对黄萎病和马铃薯痂病发病率的影响。鸡粪66吨/公顷;猪,55 hLJha;1996年春,采用旋耕机播种,耕牛100吨/公顷),播种深度为15厘米。采用含MS的尼龙网袋埋置法测定了大丽花黄萎病(Verticillium dahliae microsclerotia, MS)的存活率。4周后,MS在agaz培养基上镀。混合肥料后收集的土壤样品带回实验室并用于同步实验室实验。将10克的土壤放入试管中,将MS袋埋在土壤中,并在受控条件下测量存活率。改良后1个月在地里种上土豆。马铃薯在1997年和1998年种植,没有任何进一步的处理。通过从叶柄中分离大丽花黄萎病菌来确定黄萎病的发病率。结痂发生率…
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引用次数: 136
CHARACTERIZATION OF CHANGES IN POPULATIONS OF PHYTOPHTHORA INFESTANS IN CANADA USING MATING TYPE AND METALAXYL SENSITIVITY MARKERS 利用交偶型和甲螨酯敏感性标记对加拿大疫霉菌种群变化进行表征
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500392
R. Peters, H. W. Platt, R. Hall
Isolates of Phytophthora infestans were obtained from individual fields of potato and tomato and from potato storages across Canada in 1994 (142 samples yielding 555 isolates), 1995 (185 samples yielding 914 isolates), and 1996 (300 samples yielding 1013 isolates). Characterization of these isolates according to mating type and sensitivity to metalaxyl revealed the changing nature of P. infestans populations in Canada. In 1994, isolates of the traditional Al, metalaxyl-sensitive (MS) phenotype were common, but by 1996, they were no longer recovered from any tissue samples. Isolates of the A2 mating type, which were predominantly insensitive to metalaxyl (MI), were prevalent in 1996, except in British Columbia (B.C.). In B.C., Al isolates highly insensitive to metalaxyl predominated. Isolates of the Al, MI phenotype were also found in a sample of tomatoes from Ontario in 1996. New populations composed of the A2 mating type were predominantly insensitive to metalaxyl, but they showed an intermediate respons...
在1994年(142份样品,分离出555株)、1995年(185份样品,分离出914株)和1996年(300份样品,分离出1013株),从加拿大各地的马铃薯和番茄单田和马铃薯储存库中分离出了疫霉菌株。根据这些菌株的交配类型和对甲螨灵的敏感性进行鉴定,揭示了加拿大病原菌种群的变化性质。1994年,传统的Al,甲酰基敏感(MS)表型的分离株很常见,但到1996年,它们不再从任何组织样本中恢复。除不列颠哥伦比亚省外,1996年普遍存在对甲螨灵(MI)不敏感的A2交配型菌株。在bc省,Al菌株对甲螨灵高度不敏感。1996年在安大略省的番茄样品中也发现了Al, MI表型的分离株。由A2交配型组成的新群体对甲螨灵不敏感,但表现出中间反应。
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引用次数: 43
Mapping antigenic epitopes of potato virus A using monoclonal antibodies and overlapping synthetic peptides 利用单克隆抗体和重叠合成肽定位马铃薯病毒A抗原表位
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500387
N. Cerovska, M. Filigarová, T. Moravec, Z. Šubr
Six mouse monoclonal antibodies (MAbs) against potato virus A (PVA) were examined on their reactivity with PVA and its denatured capsid protein (PVA-CP) bound to nitrocellulose membrane. Five MAbs reacted with native PVA, three of them also with PVA-CP. MAb 534 gave no reaction in dot-blot tests. In Western blot analysis only MAbs 151, 290, and 328 reacted with PVA-CP. Competition binding test data confirm mutual spatial proximity of epitopes corresponding to these MAbs. Pepscan (SPOTs tests) with overlapping octapeptides representing the sequence of the first 60 amino acids from the N terminus of PVA-CP showed that the epitopes detected by MAb 151, 328, and 634 are located in this region. MAb 534 was deduced to react with discontinuous CP epitope. Results of analogic peptide synthesis for the MAb 151 epitope indicate that two lysine residues are essential for binding of this MAb to its epitope. Our polyclonal antibodies against PVA reacted with six different regions in this part of the CP.
研究了6种抗马铃薯病毒A (PVA)的小鼠单克隆抗体(mab)与PVA及其与硝化纤维素膜结合的变性衣壳蛋白(PVA- cp)的反应性。5个单抗与天然PVA反应,其中3个也与PVA- cp反应。MAb 534在点印迹试验中无反应。在Western blot分析中,只有mab 151,290和328与PVA-CP反应。竞争结合试验数据证实了这些单克隆抗体对应的表位在空间上相互接近。用重叠的八肽代表PVA-CP N端前60个氨基酸序列的Pepscan (spot试验)表明,MAb检测到的表位位于该区域。推断MAb 534与不连续CP表位反应。单抗151表位的类似肽合成结果表明,两个赖氨酸残基是单抗与表位结合所必需的。我们的抗PVA多克隆抗体与这部分CP的6个不同区域发生反应。
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引用次数: 2
Standardization of toxin nomenclature in the Pyrenophora tritici-repentis/wheat interaction 小麦与小麦互作白热菌毒素命名的标准化
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500415
L. Ciuffetti, L. Francl, G. M. Ballance, W. Bockus, L. Lamari, S. Meinhardt, J. Rasmussen
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引用次数: 87
Molecular aspects of host-pathogen interactions in tan spot of wheat 小麦褐斑病寄主与病原菌相互作用的分子研究
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500416
G. M. Ballance, L. Lamar
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引用次数: 2
Tillage and summerfallow effects on leaf spot diseases of wheat in the semiarid Canadian Prairies 加拿大半干旱草原耕作和夏耕对小麦叶斑病的影响
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500406
M. Fernandez, B. McConkey, R. Zentner
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引用次数: 18
Mendelian inheritance of resistance to tan spot [Pyrenophora tritici-repentis] in selected genotypes of durum wheat (Triticum turgidum) 硬粒小麦(Triticum turgidum)部分基因型抗褐斑病的孟德尔遗传
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500412
F. Gamba, L. Lamari
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引用次数: 72
Differential accumulation of proteins in resistant and susceptible sugar pine (Pinus lambertiana) seedlings inoculated with the white pine blister rust fungus (Cronartium ribicola) 白松水疱锈病菌对糖松(Pinus lambertiana)抗性和易感蛋白积累的影响
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500398
A. Ekramoddoullah, Y. Tan
Foliar and stem proteins from 21 resistant and 21 susceptible sugar pine seedlings inoculated with Cronartium ribicola were resolved on SDS-PAGE and two-dimensional gel electrophoresis. The protein profiles of pooled and individual samples were analyzed. SDS-PAGE analysis showed the presence of a 19-kDa protein band in infected susceptible foliage. Partial N-terminal amino acid sequence analysis revealed that this band was a mixture of histone and a Cro r I protein. The Cro r I protein was fungus- derived, while the origin of histone was not clear. Utilizing fungal-specific monoclonal antibodies, we determined that about 90% of susceptible seedlings had fungal antigens, while only one resistant seedling showed the presence of fungal antigens. This illustrated the potential use of these monoclonal antibodies as probes in screening seedlings for susceptibility. In addition, significantly greater amounts of seasonally regulated Pin I I protein was found in the foliage of resistant versus susceptible seedling...
采用SDS-PAGE和双向凝胶电泳技术,对21株抗性和21株易感糖松苗的叶片和茎部蛋白进行了分离。分析了混合样品和单个样品的蛋白质谱。SDS-PAGE分析显示,受感染的易感叶片中存在一条19 kda的蛋白带。部分n端氨基酸序列分析表明,该条带是组蛋白和Cro r I蛋白的混合物。Cro r I蛋白来源于真菌,而组蛋白的来源尚不清楚。利用真菌特异性单克隆抗体,我们确定约90%的易感幼苗具有真菌抗原,而只有一株抗性幼苗显示真菌抗原的存在。这说明了这些单克隆抗体作为筛选幼苗易感性的探针的潜在用途。此外,抗性幼苗叶片中季节性调节的Pin I I蛋白含量显著高于易感幼苗。
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引用次数: 12
INHERITANCE OF RACE-SPECIFIC NECROTIC AND CHLOROTIC REACTIONS INDUCED BY PYRENOPHORA TRITICI-REPENTIS IN HEXAPLOID WHEATS 小麦六倍体核裂菌诱导的种族特异性坏死和褪绿反应的遗传
Pub Date : 1998-12-01 DOI: 10.1080/07060669809500411
F. Gamba, L. Lamari, A. Brûlé-Babel
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引用次数: 86
期刊
Canadian Journal of Plant Pathology-revue Canadienne De Phytopathologie
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