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Analysis of Grapevine rupestris stem pitting-associated virus in Slovakia Reveals Differences in Intra-Host Population Diversity and Naturally Occurring Recombination Events 斯洛伐克葡萄树茎蚀相关病毒的分析揭示了宿主内种群多样性和自然发生重组事件的差异
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.OA.07.2016.0158
M. Glasa, L. Predajňa, K. Šoltys, N. Sihelská, A. Nagyová, T. Wetzel, S. Sabanadzovic
Grapevine rupestris stem pitting-associated virus (GRSPaV) is a worldwide-distributed pathogen in grapevines with a high genetic variability. Our study revealed differences in the complexity of GRSPaV population in a single host. A single-variant GRSPaV infection was detected from the SK30 grapevine plant. On the contrary, SK704 grapevine was infected by three different GRSPaV variants. Variant-specific RT-PCR detection protocols have been developed in this work to study distribution of the three different variants in the same plant during the season. This study showed their randomized distribution in the infected SK704 grapevine plant. Comparative analysis of fulllength genome sequences of four Slovak GRSPaV isolates determined in this work and 14 database sequences showed that population of the virus cluster into four major phylogenetic lineages. Moreover, our analyses suggest that genetic recombination along with point mutations could play a significant role in shaping evolutionary history of GRSPaV and contributed to its extant genetic diversification.
葡萄茎蚀相关病毒(GRSPaV)是一种分布在世界各地的葡萄病原体,具有很高的遗传变异性。我们的研究揭示了单一宿主中GRSPaV种群复杂性的差异。从葡萄植株SK30中检测到单变异GRSPaV感染。相反,SK704葡萄被三种不同的GRSPaV变体感染。本研究开发了变异特异性RT-PCR检测方案,以研究三种不同变异在同一植物中的季节分布。本研究显示了它们在受感染的葡萄植株SK704中的随机分布。对4个斯洛伐克GRSPaV分离株的全基因组序列与14个数据库序列的比较分析表明,该病毒群可分为4个主要的系统发育谱系。此外,我们的分析表明,基因重组和点突变可能在塑造GRSPaV的进化史中发挥重要作用,并有助于其现存的遗传多样化。
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引用次数: 20
Development of Carrot Medium Suitable for Conidia Production of Venturia nashicola 胡萝卜分生孢子适宜培养基的研制
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.NT.07.2016.0159
E. Choi, Gyoung-Hee Kim, Young Sun Lee, J. Jung, Janghoon Song, Y. Koh
The causal fungus of pear scab, Venturia nashicola, grows slowly and rarely produces conidia on artificial media in the laboratory, but it produced conidia on the Cheongah medium containing Cheongah powder. V. nashicola grew too slow to produce conidia until 15 days after cultivation but produced conidia with 4 × 104 conidia/plate 30 days after cultivation on the Cheongah medium containing 1% Cheongah powder. V. nashicola showed a peak production of conidia with 4.5 × 105 conidia/plate 60 days after cultivation on the carrot medium containing 2% carrot powder, one of the constituents of Cheongah powder. The carrot medium is considered to be the best medium to obtain conidia of V. nashicola in the laboratory until now. This is the first report on the development of a suitable medium for conidia production of V. nashicola, as far as we know.
梨痂病病原菌Venturia nashicola在实验室人工培养基上生长缓慢,很少产生分生孢子,但在含有清菌粉的清菌培养基上产生分生孢子。在含有1%清菌粉的清菌培养基上培养30 d后可产生4 × 104个/板的分生孢子。在含有2%胡萝卜粉(旗袍粉的一种成分)的胡萝卜培养基上培养60 d后,紫紫病菌的分生孢子产量达到峰值,为4.5 × 105个/板。迄今为止,在实验室中,胡萝卜培养基被认为是获得纳什科菌分生孢子的最佳培养基。据我们所知,这是第一次报道了一种适宜的分生孢子培养基的开发。
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引用次数: 6
Enhancement of Seed Dehiscence by Seed Treatment with Talaromyces flavus GG01 and GG04 in Ginseng (Panax ginseng) 黄Talaromyces flavus GG01和GG04种子处理对人参种子开裂的促进作用
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.OA.06.2016.0146
Min-Jeong Kim, C. Shim, Yong-ki Kim, Sung-Jun Hong, Jong-ho Park, Eun-Jung Han, Seok-Cheol Kim
Seed dehiscence of ginseng (Panax ginseng C. A. Mayer) is affected by moisture, temperature, storage conditions and microbes. Several microbes were isolated from completely dehisced seed coat of ginseng cultivars, Chunpoong and Younpoong at Gumsan, Korea. We investigated the potential of five Talaromyces flavus isolates from the dehiscence of ginseng seed in four traditional stratification facilities. The isolates showed antagonistic activities against fungal plant pathogens, such as Cylindrocarpon destructans, Fusarium oxysporum, Rhizoctonia solani, Sclerotinia nivalis, Botrytis cinerea, and Phytophthora capsici. The dehiscence ratios of ginseng seed increased more than 33% by treatment of T. flavus GG01, GG02, GG04, GG12, and GG23 in comparison to control (28%). Among the treatments, the reformulating treatment of T. flavus isolates GG01 and GG04 showed the highest of stratification ratio of ginseng seed. After 16 weeks, the reformulating treatment of T. flavus isolates GG01 and GG04 significantly enhanced dehiscence of ginseng seed by about 81% compared to the untreated control. The candidate’s treatment of T. flavus GG01 and GG04 showed the highest decreasing rate of 93% in seed coat hardness for 112 days in dehiscence period. The results suggested that the pre-inoculation of T. flavus GG01 and GG04 found to be very effective applications in improving dehiscence and germination of ginseng seed.
人参(Panax ginseng C. A. Mayer)的种子开裂受水分、温度、储存条件和微生物的影响。从韩国锦山春丰和永丰两个人参品种完全开裂的种皮中分离出几种微生物。研究了在四种传统分层设施中从人参种子开裂中分离得到的5株黄Talaromyces flavus菌株的潜力。分离物对植物真菌病原菌有拮抗作用,如破坏圆柱碳菌、尖孢镰刀菌、番茄根丝核菌、nival菌核菌、灰霉病菌和辣椒疫霉。与对照(28%)相比,黄曲霉GG01、GG02、GG04、GG12和GG23处理人参种子的开裂率提高了33%以上。其中,黄曲霉分离株GG01和GG04重新配方处理的人参种子分层率最高。16周后,黄曲霉分离株GG01和GG04与未处理的对照相比,显著提高了人参种子的开裂率约81%。黄曲霉候选处理GG01和GG04在开裂期112 d内种皮硬度下降率最高,达93%。结果表明,预接种黄曲霉GG01和GG04对改善人参种子的开裂和萌发具有很好的效果。
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引用次数: 12
Lucerne transient streak virus; a Recently Detected Virus Infecting Alfafa (Medicago sativa) in Central Saudi Arabia 卢塞恩瞬态条纹病毒;a最近在沙特阿拉伯中部发现感染苜蓿(Medicago sativa)的病毒
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.OA.06.2016.0143
A. Raza, I. Al-Shahwan, O. Abdalla, M. Al-Saleh, M. Amer
A survey was conducted to determine the status of Lucerne transient streak virus (LTSV) in three high-yielding alfalfa regions in central Saudi Arabia (Riyadh, Qassim, and Hail) during 2014. Three hundred and eight symptomatic alfalfa, and seven Sonchus oleraceus samples were collected. DAS-ELISA indicated that 59 of these samples were positive to LTSV. Two isolates of LTSV from each region were selected for molecular studies. RT-PCR confirmed the presence of LTSV in the selected samples using a specific primer pair. Percentage identity and homology tree comparisons revealed that all Saudi isolates were more closely related to each other but also closely related to the Canadian isolate-JQ782213 (97.1–97.6%) and the New Zealand isolate-U31286 (95.8–97.1%). Comparing Saudi isolates of LTSV with ten other sobemoviruses based on the coat protein gene sequences confirmed the distant relationship between them. Eleven out of fourteen plant species used in host range study were positive to LTSV. This is the first time to document that Trifolium alexandrinum, Nicotiana occidentalis, Chenopodium glaucum, and Lathyrus sativus are new host plant species for LTSV and that N. occidentalis being a good propagative host for it.
为确定2014年期间沙特阿拉伯中部三个苜蓿高产区(利雅得、卡西姆和哈伊尔)卢塞恩瞬时条纹病毒(LTSV)的状况,开展了一项调查。采集到有症状的紫花苜蓿308份,水蛭7份。DAS-ELISA检测结果显示,59份样品LTSV阳性。从每个地区选取两株LTSV进行分子研究。RT-PCR使用特定的引物对确认了所选样品中LTSV的存在。结果表明,沙特分离株与加拿大分离株jq782213(97.1-97.6%)和新西兰分离株u31286(99.8 - 97.1%)亲缘关系较近。通过将沙特分离的LTSV与其他10种sobemov的外壳蛋白基因序列进行比较,证实了它们之间的远缘关系。在寄主范围研究中,14种植物中有11种对LTSV呈阳性。本研究首次证实亚历山大Trifolium alexandrinum、西烟叶(Nicotiana occidentalis)、Chenopodium glaucum和Lathyrus sativus是LTSV的新寄主植物,而西烟叶是LTSV的良好繁殖寄主。
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引用次数: 4
Erratum: The Effects of High Temperature on Infection by Potato virus Y, Potato virus A, and Potato leafroll virus 勘误:高温对马铃薯Y病毒、马铃薯A病毒和马铃薯卷叶病毒感染的影响
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.ER.12.2015.0259
B. Chung, T. Canto, F. Tenllado, K. Choi, J. Joa, J. Ahn, C. Kim, Ki Seck Do
[This corrects the article on p. 321 in vol. 32, PMID: 27493607.].
[这更正了第32卷第321页的文章,PMID: 27493607]。
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引用次数: 1
Selection of Small Synthetic Antimicrobial Peptides Inhibiting Xanthomonas citri subsp. citri Causing Citrus Canker 抑制柑橘黄单胞菌的合成抗菌肽的筛选。导致柑橘溃疡病
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.NT.09.2015.0188
Jeahyuk Choi, E. Park, Seweon Lee, J. Hyun, K. Baek
Citrus canker disease decreases the fruit quality and yield significantly, furthermore, emerging of streptomycin-resistant pathogens threatens the citrus industry seriously because of a lack of proper control agents. Small synthetic antimicrobial peptides (AMPs) could be a promising alternative. Fourteen hexapeptides were selected by using positional scanning of synthetic peptide combinatorial libraries. Each hexapeptide showed different antimicrobial spectrum against Bacillus, Pseudomonas, Xanthomonas, and Candida species. Intriguingly, BHC10 showed bactericidal activity exclusively on Xanthomonas citri subsp. citri (Xcc), while BHC7 was none-active exclusively against two Pseudomonas spp. at concentration of 100 μg/ml suggesting potential selectivity constrained in hexapeptide frame. Three hexapeptides, BHC02, 06 and 11, showed bactericidal activities against various Xcc strains at concentration of 10 μg/ml. When they were co-infiltrated with pathogens into citrus leaves the disease progress was suppressed significantly. Further study would be needed to confirm the actual disease control capacity of the selected hexapeptides.
柑橘溃疡病严重降低果实品质和产量,而且由于缺乏适当的防治剂,耐链霉素病原体的出现严重威胁着柑橘产业。小合成抗菌肽(AMPs)可能是一个有前途的替代品。利用合成肽组合文库的定位扫描筛选出14条六肽。每种六肽对芽孢杆菌、假单胞菌、黄单胞菌和念珠菌具有不同的抗菌谱。有趣的是,BHC10仅对柑橘黄单胞菌具有抑菌活性。而BHC7在浓度为100 μg/ml时对两种假单胞菌均无特异性活性,提示其潜在的选择性受六肽框架的限制。在浓度为10 μg/ml时,BHC02、06和11 3个六肽对多种Xcc菌株均有抑菌活性。当它们与病原菌共渗进入柑橘叶片时,病害进展明显受到抑制。需要进一步的研究来确认所选六肽的实际疾病控制能力。
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引用次数: 9
Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione 杀真菌剂异丙二酮对互花霉形态发生的影响
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.NT.06.2016.0145
Eunji Kim, H. Lee, Young Ho Kim
Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4–5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.
抗杀菌剂的互交黑穗病阻碍了作物田间互交黑穗病的实际防治。本研究旨在探讨异丙二酮对异丙二酮的细胞学抗性机制。用异丙二酮和不加异丙二酮在马铃薯葡萄糖琼脂(PDA)上培养仙人掌褐斑菌,用光镜和电镜观察了不同生长特征的真菌培养物,分别为不生长、初始生长和完全生长。在不使用杀菌剂的PDA上,菌丝从孵育后1天开始生长,并继续处于充分生长状态(对照-生长,Con-G),而在有异丙二酮的PDA上,前3个DAI没有真菌生长(iprodione-no growth, Ipr-N),但一旦在4-5 DAI开始初始生长(iprodione-initial growth, Ipr-I),菌落就会不断生长扩大,达到充分生长状态(iprodione-growth, Ipr-G)。提示ipr - 1可能是抵抗杀真菌毒性的形态发生变化的转折点。cong形成多细胞分生孢子,有细胞壁和隔,细胞质完整致密。在Ipr-N中,真菌的孢子形成主要是未发育的单细胞分生孢子,细胞质降解和坏死。而在ipr - 1中,产孢过程中发生了明显的细胞变化,形成了具有双层(增厚)细胞壁的多细胞分生孢子,并在分生细胞质中积累了增殖的脂质体,这可能抑制了杀菌剂进入分生细胞,降低了杀菌剂相关的毒性,并可能分别被用作能量和营养来源。使真菌进一步生长形成成熟菌落,如Ipr-G,形成多细胞分生孢子,具有细胞壁和完整的细胞质,具有脂质体,如conn - g。
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引用次数: 11
Construction of a System for the Strawberry Nursery Production towards Elimination of Latent Infection of Anthracnose Fungi by a Combination of PCR and Microtube Hybridization 利用PCR和微管杂交技术构建消除炭疽病真菌潜伏感染的草莓苗圃生产体系
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.NT.05.2016.0132
K. Furuta, S. Nagashima, T. Inukai, C. Masuta
One of the major problems in strawberry production is difficulty in diagnosis of anthracnose caused by Colletotrichum acutatum or Glomerella cingulata in latent infection stage. We here developed a diagnostic tool for the latent infection consisting of initial culturing of fungi, DNA extraction, synthesis of PCR-amplified probes and microtube hybridization (MTH) using a macroarray. The initial culturing step is convenient to lure the fungi out of the plant tissues, and to extract PCR-inhibitor-free DNA directly from fungal hyphae. For specific detection of the fungi, PCR primers were designed to amplify the fungal MAT1-2 gene. The subsequent MTH step using the PCR products as probes can replace the laborious electrophoresis step providing us sequence information and high-throughput screening. Using this method, we have conducted a survey for a few thousands nursery plants every year for three consecutive years, and finally succeeded in eliminating latent infection in the third year of challenge.
草莓生产中存在的主要问题之一是潜伏感染期炭疽菌或带状肾小球引起的炭疽病诊断困难。我们在此开发了一种潜伏性感染的诊断工具,包括真菌的初始培养、DNA提取、pcr扩增探针的合成和使用宏阵列的微管杂交(MTH)。最初的培养步骤便于将真菌诱导出植物组织,并直接从真菌菌丝中提取不含pcr抑制剂的DNA。为了对真菌进行特异性检测,设计PCR引物扩增真菌MAT1-2基因。随后的MTH步骤使用PCR产物作为探针,可以取代费力的电泳步骤,为我们提供序列信息和高通量筛选。利用这种方法,我们连续三年每年对几千株苗木进行调查,最终在攻毒的第三年成功地消灭了潜伏侵染。
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引用次数: 5
Effects of Soil Textures on Infectivity of Root-Knot Nematodes on Carrot 土壤质地对胡萝卜根结线虫侵染力的影响
Pub Date : 2017-02-01 DOI: 10.5423/PPJ.OA.07.2016.0155
Eunji Kim, Y. Seo, Yong Su Kim, Yong-Keun Park, Young Ho Kim
This study was conducted to examine infectivity (penetration and gall and egg-mass formations) of the root-knot nematodes, Meloidogyne incognita and M. hapla, on carrots grown in soil conditions of 5 different soil textures consisting of bed-soil (b) and sand (s) mixtures (b-s mixtures) at the ratios of 10:0, 7:3, 5:5, 3:7, and 0:10. For M. incognita, the nematode penetration rates in b-s of 0:10 (100% sand) were significantly higher than in the other b-s mixtures, more greatly at 2 and 5 days after inoculation than at 10 DAI, while no significant differences in the penetration rates were mostly shown for M. hapla at the above DAI. However, for both nematodes, gall and egg-mass formations were remarkably increased in the b-s mixture of 0:10, compared to the other b-s mixtures, which is coincided with the general aspects of severe nematode infestations in sandy soils. This suggests the increased gall and egg-mass formations of M. incognita should be derived from the increased penetration rates in the sandy soil conditions, which provide a sufficient aeration due to coarse soil nature for the nematodes, leading to their mobility increased for the enhanced root penetration. For M. hapla, it is suggested that the sandy soil conditions affect positively on the healthy plant growth with little accumulation of the inhibitory materials and sufficient aeration, enhancing the nematode growth and feeding activities. All of these aspects provide information reliable for the development screening techniques efficient for the evaluation of the nematode resistance in the breeding programs.
研究了根结线虫(Meloidogyne incognita)和根结线虫(M. hapla)在5种不同土壤质地(床土(b)和沙(s)混合物(b-s混合物)配比为10:0、7:3、5:5、3:7和0:10)的土壤条件下对胡萝卜的侵染性(渗透、瘿和卵团形成)。在0∶10(100%沙子)的b-s混合物中,黑纹田鼠的线虫渗透率显著高于其他b-s混合物,接种后2天和5天的线虫渗透率显著高于10 DAI,而在上述DAI中,黑纹田鼠的线虫渗透率无显著差异。然而,对于这两种线虫,与其他b-s混合物相比,0∶10的b-s混合物显著增加了瘿和卵团的形成,这与沙质土壤中严重线虫侵染的一般方面相吻合。这表明,在沙质土壤条件下,粗质土壤为线虫提供了充足的通气性,从而增加了线虫的移动性,从而增强了根系的渗透能力,从而增加了线虫的胆和卵团形成。结果表明,沙质土壤条件下抑制物质积累较少,通气性充足,有利于线虫的生长和取食活性。这些方面都为线虫抗性评价筛选技术的开发提供了可靠的信息。
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引用次数: 17
Simultaneous Detection of Three Bacterial Seed-Borne Diseases in Rice Using Multiplex Polymerase Chain Reaction 多重聚合酶链反应同时检测水稻三种细菌性种传病害
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.NT.05.2016.0118
I. Kang, Mi-Hyung Kang, T. Noh, H. Shim, D. Shin, Su-Jin Heu
Burkholderia glumae (bacterial grain rot), Xanthomonas oryzae pv. oryzae (bacterial leaf blight), and Acidovorax avenae subsp. avenae (bacterial brown stripe) are major seedborne pathogens of rice. Based on the 16S and 23S rDNA sequences for A. avenae subsp. avenae and B. glumae, and transposase A gene sequence for X. oryzae pv. oryzae, three sets of primers had been designed to produce 402 bp for B. glumae, 490 bp for X. oryzae, and 290 bp for A. avenae subsp. avenae with the 63°C as an optimum annealing temperature. Samples collected from naturally infected fields were detected with two bacteria, B. glumae and A. avenae subsp. avenae but X. oryzae pv. oryzae was not detected. This assay can be used to identify pathogens directly from infected seeds, and will be an effective tool for the identification of the three pathogens in rice plants.
葡萄伯克霍尔德菌(细菌性谷粒腐病),米黄单胞菌。稻瘟病菌(细菌性叶枯萎病)和青霉病亚种。褐条病菌是水稻主要的种传病原菌。基于A. avenae亚种的16S和23S rDNA序列。稻瘟病弧菌的转座酶A基因序列。共设计了3套引物,分别对B. glumae、X. oryzae和A. avenae分别产生402 bp、490 bp和290 bp的特异序列。以63℃为最佳退火温度。从自然感染地采集的样品中检测到2种细菌,即葡萄芽孢杆菌和绿芽孢杆菌亚种。但X. oryzae pv。未检测到Oryzae。该方法可用于直接从侵染种子中鉴定病原菌,并将成为鉴定水稻三种病原菌的有效工具。
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引用次数: 8
期刊
The Plant Pathology Journal
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