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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
Digital Infrared Thermal Imaging of Crape Myrtle Leaves Infested with Sooty Mold 黑霉侵染紫薇叶片的数字红外热成像研究
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.NT.04.2016.0112
Jiyeon Kim, Si-gyun Kweon, Junhyung Park, Harim Lee, Ki Woo Kim
The spatial patterns for temperature distribution on crape myrtle leaves infested with sooty mold were investigated using a digital infrared thermal imaging camera. The mean temperatures of the control and sooty regions were 26.98°C and 28.44°C, respectively. In the thermal images, the sooty regions appeared as distinct spots, indicating that the temperatures in these areas were higher than those in the control regions on the same leaves. This suggests that the sooty regions became warmer than their control regions on the adaxial leaf surface. Neither epidermal penetration nor cell wall dissolution by the fungus was observed on the adaxial leaf surface. It is likely that the high temperature of black leaves have an increased cooling load. To our knowledge, this is the first report on elevated temperatures in sooty regions, and the results show spatial heterogeneity in temperature distribution across the leaf surface.
利用数字红外热像仪研究了紫薇黑霉侵染后叶片温度的空间分布规律。对照区和黑烟区平均温度分别为26.98°C和28.44°C。在热像图上,黑烟区呈明显的斑点状,表明这些区域的温度高于同一叶片上的对照区域。这表明在叶片正面,黑烟区比对照区变暖。在叶片正面未观察到真菌对表皮的渗透和细胞壁的溶解。这很可能是黑叶的高温增加了冷负荷。据我们所知,这是第一个关于烟灰地区温度升高的报告,结果显示了叶片表面温度分布的空间异质性。
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引用次数: 6
Altering Conidial Dispersal of Alternaria solani by Modifying Microclimate in Tomato Crop Canopy 改变番茄作物冠层小气候对茄斑孢孢子扩散的影响
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.OA.06.2015.0101
P. Jambhulkar, N. Jambhulkar, M. Meghwal, G. S. Ameta
Early blight of tomato caused by Alternaria solani, is responsible for severe yield losses in tomato. The conidia survive on soil surface and old dry lower leaves of the plant and spread when suitable climatic conditions are available. Macroclimatic study reveals that highest inoculum concentration of Alternaria spores appeared in May 2012 to 2013 and lowest concentration during January 2012 to 2013. High night temperature positively correlated and significantly (P < 0.01) involved in conidial spore dispersal and low relative humidity (RH) displayed significant (P < 0.05) but negative correlation with conidial dispersal. The objective of the study was to modify microclimatic conditions of tomato crop canopy which may hamper conidial dispersal and reduce disease severity. We evaluated effect of marigold intercropping and plastic mulching singly and in consortia on A. solani conidial density, tomato leaf damage and microclimatic parameters as compar to tomato alone (T). Tomato-marigold intercropping–plastic mulching treatment (T + M + P) showed 35–39% reduction in disease intensity as compared to tomato alone. When intercropped with tomato, marigold served as barrier to conidial movement and plastic mulching prevented evapotranspiration and reduced the canopy RH that resulted in less germination of A. solani spores. Marigold intercropping and plastic mulching served successfully as physical barrier against conidial dissemination to diminish significantly the tomato foliar damage produced by A. solani.
番茄早疫病是番茄产量损失最严重的病害之一。分生孢子在土壤表面和植物下部干燥的老叶片上存活,并在适宜的气候条件下扩散。宏观气候研究表明,2012年5月至2013年5月接种量最高,2012年1月接种量最低。高夜间温度与孢子扩散呈极显著正相关(P < 0.01),低相对湿度与孢子扩散呈极显著负相关(P < 0.05)。研究的目的是改变番茄作物冠层中可能阻碍分生孢子扩散和降低病害严重程度的小气候条件。研究了金盏花间作与地膜覆盖对茄蚜孢子密度、番茄叶片损伤和小气候参数的影响,结果表明,与单独覆盖番茄相比,间作与地膜覆盖对茄蚜病强的影响降低了35 ~ 39%。当与番茄间作时,万寿菊对分生孢子的运动起屏障作用,地膜覆盖抑制了蒸散作用,降低了冠层相对湿度,导致茄灰孢子萌发率降低。万寿菊间作和地膜覆盖作为物理屏障,有效地抑制了分生孢子的传播,显著降低了茄蚜对番茄叶片的危害。
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引用次数: 14
D-PSA-K: A Model for Estimating the Accumulated Potential Damage on Kiwifruit Canes Caused by Bacterial Canker during the Growing and Overwintering Seasons D-PSA-K:猕猴桃生长期和越冬期细菌溃疡病对猕猴桃植株累积潜在危害的估计模型
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.OA.03.2016.0068
Ki Seok Do, B. Chung, J. Joa
We developed a model, termed D-PSA-K, to estimate the accumulated potential damage on kiwifruit canes caused by bacterial canker during the growing and overwintering seasons. The model consisted of three parts including estimation of the amount of necrotic lesion in a non-frozen environment, the rate of necrosis increase in a freezing environment during the overwintering season, and the amount of necrotic lesion on kiwifruit canes caused by bacterial canker during the overwintering and growing seasons. We evaluated the model’s accuracy by comparing the observed maximum disease incidence on kiwifruit canes against the damage estimated using weather and disease data collected at Wando during 1994–1997 and at Seogwipo during 2014–2015. For the Hayward cultivar, D-PSA-K estimated the accumulated damage as approximately nine times the observed maximum disease incidence. For the Hort16A cultivar, the accumulated damage estimated by D-PSA-K was high when the observed disease incidence was high. D-PSA-K could assist kiwifruit growers in selecting optimal sites for kiwifruit cultivation and establishing improved production plans by predicting the loss in kiwifruit production due to bacterial canker, using past weather or future climate change data.
我们开发了一个称为D-PSA-K的模型来估计猕猴桃生长和越冬季节细菌溃疡病对猕猴桃甘蔗造成的累积潜在损害。该模型由三部分组成,分别是对非冷冻环境下猕猴桃坏死灶数量的估计、越冬季节冷冻环境下猕猴桃坏死灶增加速度的估计以及越冬和生长季节猕猴桃菌类溃疡病引起的猕猴桃甘蔗坏死灶数量的估计。我们通过比较观察到的猕猴桃甘蔗最大疾病发病率与使用1994-1997年在莞岛和2014-2015年在西归浦收集的天气和疾病数据估计的损害来评估该模型的准确性。对于海沃德品种,D-PSA-K估计累积损害约为观察到的最大疾病发生率的9倍。对于Hort16A品种,D-PSA-K估算的累积伤害值在观察到病害发生率高时较高。D-PSA-K可以利用过去的天气或未来的气候变化数据,预测由于细菌溃疡病导致的猕猴桃产量损失,从而帮助猕猴桃种植者选择最佳的猕猴桃种植地点,并制定改进的生产计划。
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引用次数: 2
Overexpression of a Pathogenesis-Related Protein 10 Enhances Biotic and Abiotic Stress Tolerance in Rice 致病相关蛋白10的过表达增强了水稻的生物和非生物胁迫耐受性
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.OA.06.2016.0141
Jingni Wu, Sang Gon Kim, K. Kang, Ju-Gon Kim, Sangryeol Park, Ravi Gupta, Yong Hwan Kim, Yiming Wang, S. Kim
Pathogenesis-related proteins play multiple roles in plant development and biotic and abiotic stress tolerance. Here, we characterize a rice defense related gene named “jasmonic acid inducible pathogenesis-related class 10” (JIOsPR10) to gain an insight into its functional properties. Semi-quantitative RT-PCR analysis showed up-regulation of JIOsPR10 under salt and drought stress conditions. Constitutive over-expression JIOsPR10 in rice promoted shoot and root development in transgenic plants, however, their productivity was unaltered. Further experiments exhibited that the transgenic plants showed reduced susceptibility to rice blast fungus, and enhanced salt and drought stress tolerance as compared to the wild type. A comparative proteomic profiling of wild type and transgenic plants showed that overexpression of JIOsPR10 led to the differential modulation of several proteins mainly related with oxidative stresses, carbohydrate metabolism, and plant defense. Taken together, our findings suggest that JIOsPR10 plays important roles in biotic and abiotic stresses tolerance probably by activation of stress related proteins.
致病相关蛋白在植物发育、生物和非生物胁迫耐受中发挥着多种作用。在这里,我们对水稻防御相关基因“茉莉酸诱导致病相关类10”(JIOsPR10)进行了表征,以深入了解其功能特性。半定量RT-PCR分析显示,在盐胁迫和干旱胁迫条件下,JIOsPR10基因表达上调。JIOsPR10在水稻中的组成型过表达促进了转基因植株的茎和根发育,但对其产量没有影响。进一步的实验表明,与野生型相比,转基因植株对稻瘟病菌的敏感性降低,对盐和干旱胁迫的耐受性增强。野生型和转基因植物的比较蛋白质组学分析表明,JIOsPR10的过表达导致了几种主要与氧化胁迫、碳水化合物代谢和植物防御相关的蛋白质的差异调节。综上所述,我们的研究结果表明,JIOsPR10可能通过激活应激相关蛋白在生物和非生物胁迫耐受中发挥重要作用。
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引用次数: 69
Management of Powdery Mildew in Squash by Plant and Alga Extract Biopesticides 植物和藻类提取物生物农药防治南瓜白粉病的研究
Pub Date : 2016-12-01 DOI: 10.5423/PPJ.OA.05.2016.0131
Shouan Zhang, Z. Mersha, G. Vallad, Cheng-Hua Huang
Although many fungicides are registered for use to control powdery mildew on cucurbits, management of resistance to fungicides in pathogen populations still remains a major challenge. Two biopesticides Regalia SC and HMO 736 were evaluated in the greenhouse and field for their efficacy against powdery mildew in squash. In greenhouses, Regalia SC alone significantly (P < 0.05) reduced powdery mildew compared to the nontreated control, and was as effective as the chemical standard Procure 480SC (triflumizole). In alternation with Procure 480SC, Regalia SC demonstrated greater or equivalent effects on reducing the disease. HMO 736 alone showed varying levels of disease control, but alternating with Procure 480SC significantly improved control efficacy. In addition, application of Regalia SC or HMO 736 each in alternation with Procure 480SC significantly increased the chlorophyll content in leaves and the total fresh weight of squash plants, when compared with the water control, Regalia SC and HMO 736 alone. In field trials, application of Regalia SC and HMO 736 each alone significantly reduced disease severity in one of two field trials during the early stage of disease development, but not during later stages when disease pressure became high. Both Regalia SC and HMO 736 each applied in alternation with Procure 480SC significantly improved the control efficacy compared to Procure 480SC alone. Results from this study demonstrated that an integrated management program can be developed for powdery mildew in squash by integrating the biopesticides Regalia SC, HMO 736 with the chemical fungicide Procure 480SC.
尽管许多杀菌剂已登记用于控制葫芦上的白粉病,但病原菌种群对杀菌剂的抗性管理仍然是一个主要挑战。对两种生物农药Regalia SC和HMO 736在温室和田间对南瓜白粉病的防治效果进行了评价。在温室中,与未经处理的对照相比,Regalia SC单独使用显著(P < 0.05)降低了白粉病的发病率,其效果与化学标准品(三氟唑)相同。与采购480SC交替,Regalia SC在减少疾病方面表现出更大或同等的效果。单独使用HMO 736显示不同程度的疾病控制,但与采购480SC交替使用可显著提高控制效果。此外,与单独施用Regalia SC和HMO 736相比,交替施用Regalia SC或HMO 736显著提高了南瓜植株叶片叶绿素含量和总鲜重。在田间试验中,在两项田间试验中的一项中,单独施用Regalia SC和HMO 736分别显著降低了疾病发展早期的疾病严重程度,但在疾病压力变得高的后期阶段则没有效果。Regalia SC和HMO 736分别与采购480SC交替使用,与单独采购480SC相比,显著提高了控制效果。本研究结果表明,将生物农药Regalia SC、HMO 736与化学杀菌剂Procure 480SC结合使用,可以开发出一套综合治理南瓜白粉病的方案。
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引用次数: 19
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The Plant Pathology Journal
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