西安市樱桃病毒病调查及检测研究

对樱桃病毒病的发生情况进行了田间调查,运用鉴别寄主反应型、病原传播特性、负染法电镜观察、琼脂双扩散法测定血清学反应等方法进行检测,查明多数为李属坏死环斑病毒单独感染,其余为李属坏死环斑病毒和苹果退绿叶斑病毒或苹果茎沟病毒混合感染,总感染率为93.94%,并对其感染的症状进行描述。     

微茎尖培养脱除樱桃2种病毒试验

樱桃树受到病毒侵害,造成树势衰弱,果实畸形,观赏和经济价值大大降低,其中危害严重的病毒有苹果褪绿叶斑病毒(ACLSV)和李属坏死环斑病毒(PNRSV)2种.本文采用微茎尖组织培养法脱除苹果褪绿叶斑病毒和李属坏死环斑病毒,并用RT-PCR技术检测脱毒效果.     

李属坏死环斑病毒(PNRSV)新疆巴旦木分离物外壳蛋白基因(CP)片断的克隆与序列分析

【目的】为了查明新疆巴旦木果树病原病毒的种类,为病毒的分子检测提供基础。【方法】采用双抗体夹心酶联免疫吸附法(DAS-ELISA),从巴旦木果树叶片中检测到李属坏死环斑病毒(PNRSV)。以阳性样品的总RNA为模板进行PNRSV外壳蛋白基因(CP)RT-PCR扩增,对预期430 bp大小的4个扩增产物进行克隆、测序和序列分析。【结果】结果表明,PNRSV新疆巴旦木分离物CP基因的核苷酸和氨基酸序列与世界各地已报道的分离物具有较高的同源性,分别为80.0%～97.2%和80.1%～94.1%,其中与阿根廷分离物AY217的同源性最高;而与同属03亚组的苹果花叶病毒(ApMV)同源性较低,仅为52.9%～55.4%和45.6%～48.6%。新疆PNRSV各分离物之间CP基因高度同源。序列比对与系统发生树的分析显示,PNRSV新疆巴旦木分离物的株系属于GroupⅠ组。【结论】确定了PNRSV为侵染新疆巴旦木果树的病原病毒。这是PNRSV在新疆发现的首次报道。     

3个苹果新品种病毒病的发生状况及ASSVd序列分析

【目的】近年来，苹果病毒病的发生日趋严重，对产业的健康发展造成了严重影响，但是其在苹果新品种瑞阳、瑞雪、瑞香红上的发病状况尚不明确。【方法】在2021—2022年采用RT-PCR方法对白水地区198份新品种样品进行病毒病检测，并对感染苹果锈果类病毒（apple scar skin viroid,ASSVd）植株的果实症状进行了调查和基因克隆、测序。【结果】检测结果显示白水地区新品种潜隐性病毒的感病率显著高于非潜隐性病毒，其中苹果褪绿叶斑病毒（apple chlorotic leaf spot virus,ACLSV）的检出率最高，达到了91.72%，其次是苹果茎沟病毒（apple stem grooving virus,ASGV）和苹果茎痘病毒（apple stem pitting virus,ASPV），检出率分别达到了77.16%、37.13%，苹果坏死花叶病毒（apple necrotic mosaic virus,ApNMV）的检出率为30.31%，苹果锈果类病毒和苹果凹果类病毒（apple dimple fruit viroid, ADFVd）的检出率接近10%；通过对新品种...     

我国发现苹果花叶新病毒

正前不久,中国农科院植物保护研究所研究团队与中国农业大学以及日本果树病毒专家合作,利用高通量测序技术从苹果花叶样品中鉴定出一种新的病毒,命名为苹果坏死花叶病毒(Apple necrotic mosaic virus,ApNMV),并证实这种病毒跟我国苹果花叶病高度相关,颠覆了人们对苹果花叶病病原的常规认知。     

两步多重RT-PCR快速检测苹果潜隐性病毒

【目的】为建立检测3种苹果潜隐性病毒更为快速准确的方法,【方法】以携带苹果茎沟病毒(ASGV)、苹果褪绿叶斑病毒(ACLSV)和苹果茎痘病毒(ASPV)的成龄苹果树韧皮部为试材,根据基因库中苹果茎痘病毒(Apple stempitting virus,ASPV)的外壳蛋白基因序列,设计合成了2对特异性引物,分别与合成的苹果茎沟病毒(ASGV)引物和苹果褪绿叶斑病毒(ACLSV)的引物组合配伍,筛选出最佳的引物对组合。对cDNA的合成所用引物和总RNA模板进行遴选和量化,对PCR过程中cDNA的用量、引物对的终浓度、退火温度等主要影响因素进行优化。【结果】结果表明,反转录引物为Oligo(dT)18、总RNA 0.1～3.0μL时,可以得到较好的cDNA;ASPV-FR与ASGV-Pch、ACLSV-Pch引物组合优于ASPV-Pch,并且筛选出4组最佳的终浓度处理组合;cDNA用量为2.0~4.0μL、退火温度为50.0~52.9℃、循环数为35时,扩增效果较好。采用优化的多重RT-PCR体系对采自陕西和山东两省的样品进行检测,并用3种病毒的单重RT-PCR体系进行验证。【结论】结果呈现高度一致性,充分印证了该多重RT-PCR体系的准确性,适用于大量样品的快速检测。     

新疆无花果花叶病病毒鉴定与发生调查

【目的】无花果花叶病是危害无花果的主要病害之一,在新疆地区发生普遍,但导致该病害发生的病原种类尚不明确,因此进行了新疆无花果花叶病的病原鉴定与发生情况调查,为进一步明确不同无花果病毒的潜在危害以及病害防控提供依据。【方法】对新疆8个地区21个果园及路边的盆栽风景无花果进行病毒病的调查,通过高通量测序对感病无花果样品进行病毒初步鉴定,利用PCR或RT-PCR法,对采集自新疆8个地区的、不同花叶病症状及无症状的135份叶片样品进行病毒检测。【结果】高通量测序分析表明,感病样品中存在5种无花果病毒,分别为无花果花叶病毒(Fig mosaic virus,FMV)、无花果杆状病毒1(Fig badnavirus 1,FBV-1)、无花果杆状病毒2(Fig badnavirus 2,FBV-2)、无花果叶斑相关病毒4(Fig leaf mottle-associated virus 4,FLMaV-4)和无花果斑点相关病毒(Fig fleck-associated virus,FFkaV)。PCR及RT-PCR检测结果表明,135份无花果样品中存在6种无花果病毒,其检出率分别为FMV(100%)、FBV-1(100%)、FBV-2(99.3%)、FLMaV-4(74.8%)、FFkaV(31.9%)、无花果叶斑相关病毒1(Fig leaf mottle-associated virus 1,FLMaV-1)(19.3%)。新疆无花果病毒普遍存在复合侵染现象,感染4种以上病毒的样品有127份,占94%。2种、3种、4种和5种病毒复合侵染检出率分别为0.7%、5.2%、62.2%和31.9%。FMV、FBV-1、FBV-2在不同地区的发病率没有明显差异,并且症状为环斑、褪绿斑驳、褶皱及无症状样品中都能检测出FMV、FBV-1、FBV-2,这3种病毒可能是侵染新疆无花果的主要病毒。【结论】鉴定到侵染新疆无花果花叶病样的病毒性病原有6种,病毒检出率较前人报道的具有明显提升,并且普遍存在2种以上病毒复合侵染的现象,其中FMV、FBV-1、FBV-2检出率最高。     

3种苹果潜隐病毒多重RT-PCR检测体系的建立

 研究建立了能同时检测苹果茎沟病毒(Apple stem grooving virus, ASGV) 、苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus, ACLSV) 和苹果茎痘病毒(Apple stem pitting virus, ASPV) 的多重RT-PCR方法。以复合感染3种病毒的苹果‘望山红’组培苗为试材, 对影响多重PCR的反应条件进行了一系列的调整和优化。多重RT-PCR体系灵敏性测验显示, 最低能从RNA总量187.5 ng的样品中检测3种病毒的存在。多重RT-PCR产物的序列与报道的病毒序列有较高的同源性, 12个田间苹果样品的多重RT-PCR检测结果与单一PCR的结果一致, 初步证明了多重RT-PCR检测的准确性。     

通辽市塞外红苹果中3种苹果潜隐病毒的检测及变异分析

【目的】明确通辽地区塞外红苹果（Malus pumila ‘Saiwaihong’）中苹果褪绿叶斑病毒（ACLSV）、苹果茎痘病毒（ASPV）和苹果茎沟病毒（ASGV）3种苹果潜隐病毒的普遍率和遗传多样性。【方法】从通辽市开鲁县8个乡镇随机采集了96份塞外红苹果枝条，采用RT-PCR法对样品进行3种苹果潜隐病毒检测。对检测为阳性的样品进行病毒基因克隆、测序，并进行序列一致性分析和系统发育分析。【结果】检测结果表明，被检测的塞外红苹果中3种苹果潜隐病毒均有不同程度的发生，并且存在2种或3种病毒的复合侵染。基于CP基因对病毒一致性和变异情况进行分析，结果表明，来源于塞外红苹果的29个ACLSV分离物的核苷酸序列一致性为78.1%～99.9%，与其他15个已知的ACLSV分离物的一致性为69.1%～93.6%；来源于塞外红苹果的12个ASPV分离物的核苷酸序列一致性为83.6%～99.7%，与其他15个已知的ASPV分离物的一致性为75.4%～91.5%；来源于塞外红苹果的21个ASGV分离物的核苷酸序列一致性为96.4%～100.0%，与其他已知的40个ASGV分离物的一致性为89.9%～...     

基于宏病毒组测序技术的苹果病毒病鉴定与分析

采用宏病毒组测序技术（Virome Sequencing Technology）对河南地区染病苹果树进行病毒检测与分析,从建库测序的苹果叶片中共检测出13种病毒,其中有5种苹果病毒：苹果茎沟病毒（apple stem grooving virus,ASGV）、苹果茎痘病毒（apple stem pitting virus,ASPV）、苹果褪绿叶斑病毒（apple chlorotic leafspotvirus,ACLSV）、苹果坏死花叶病毒（applenecroticmosaicvirus,Ap NMV）和苹果绿皱缩相关病毒（apple green crinkle associated virus,AGCa V）。对病毒检测结果中的5种苹果病毒进行RT-PCR验证,扩增后均获得目的条带,与宏病毒组测序检测结果一致,表明宏病毒组测序检测结果精准可靠。根据测序结果设计特异性引物对ASPV全长进行扩增,得到1条长度为9 246 nt的ASPV-HN分离物基因组,与NCBI数据库中已报道的19个ASPV分离物的基因组核苷酸序列一致性为75.48%～79.85%;宏病毒组测序拼接所得的两条长度均...     

Antifungal effects of iron sulfate on grapevine fungal pathogens

The present study aimed to determine the most efficient experimental conditions of iron sulfate use leading to optimal inhibition in the development of fungal pathogens. Assays have been focused on fungal species inducing severe grapevine diseases. FeSO₄ directly inhibited the in vitro mycelial growth of Botrytis cinerea, Eutypa lata, Phaeomoniella chlamydospora, Phaeoacremonium aleophilum, Diplodia seriata, and Neofusicoccum parvum with variable efficiency in the range of 0.5–10 mM. The development was always completely inhibited at 20 mM. This inhibitory effect was greatly increased at acidic pH values. The anionic moiety of the molecule was of importance since bromide, chloride and sulfate were highly active, whereas acetate and oxalate showed a small effect. Electron microscope observations on E. lata and B. cinerea showed that a treatment with FeSO₄ induced dramatic changes in the hyphal organization leading to cell death. No toxicity was observed on grapevine leaves following repeated FeSO₄ sprays in the antifungal concentration range. Therefore, FeSO₄ may be proposed to effectively replace the long-term pollutant use of CuSO₄ as an antifungal agent, with the additional advantage of iron being an important plant micronutrient.     

戊唑醇与代森锰锌对苹果3种主要病菌的联合毒力

为了弄清戊唑醇与代森锰锌混配对苹果轮纹病菌等3种苹果主要病菌的抑制作用,找出2种药剂混配的最佳配比,为田间使用提供依据,采用共毒系数法测定了戊唑醇与代森锰锌不同配比对苹果轮纹菌、苹果炭疽菌和苹果腐烂菌的联合毒力。结果表明,戊唑醇∶代森锰锌以1∶4比例混配对苹果轮纹病菌和苹果炭疽病菌的抑制效果最好,共毒系数分别为351.42和405.82;戊唑醇∶代森锰锌以4∶1比例混配对苹果腐烂病菌的抑制效果最好,共毒系数为169.49,增效作用显著。     

Modelling the spread of tree pests and pathogens in urban forests

Understanding the potential dynamics of tree pests and pathogens is a vital component for creating resilient urban treescapes. Epidemiologically relevant features include differences in environmental stress and tree management between street and garden trees, and variation in the potential for human-mediated spread due to intensity of human activity, traffic flow and buildings. We extend a standard spatially explicit raster-based model for pest and pathogen spread by dividing the urban tree population into roadside trees and park/garden trees. We also distinguish between naturally-driven radial spread of pests and pathogens and human-mediated linear spread along roads. The model behaviour is explored using landscape data for tree locations in an exemplar UK town. Two main sources of landscape data were available: commercially collated aerial data, which have high coverage but no information on species; and, an urban tree inventory, with low, non-random, coverage but with some species data. The data were insufficient to impute a species-specific host landscape accurately; however, by combining the two data sources, and applying either random or Matérn cluster point process driven selection of a subset of all trees, we create two sets of potential host landscapes. We find that combining the two mechanisms of dispersal has a non-additive effect, with the enhanced linear dispersal enabling new foci of infection to be established more rapidly than with radial dispersal alone; and clustering of trees by species slows down the expansion of epidemics when compared with random distribution of tree species within known host locations.     

KC提取液对几种植物病原菌的抑制作用的研究

采用生长速率法和孢子囊萌发法测试了KC提取液对马铃薯晚疫病菌、赤霉病菌、山药炭疽病菌、梨黑斑病菌菌丝生长和孢子囊萌发的抑制作用。结果表明,KC提取液对马铃薯晚疫病菌菌丝生长有明显的抑制作用,同时对马铃薯晚疫病菌孢子囊萌发也有一定抑制作用。对赤霉病菌、山药炭疽病菌、梨黑斑病菌菌丝生长没有抑制作用。     

柑橘炭疽病菌的分离、鉴定及在果实上的潜伏侵染特性

【目的】明确柑橘炭疽病原菌的种类及其潜伏侵染特性,为柑橘病害防控提供理论依据。【方法】对采自湖北省3个地区的炭疽病样品进行病原菌的分离、形态学特征观察和致病性测定并采用r DNA-ITS序列分析法对代表菌株T4进行分子鉴定。此外,通过组织分离法,研究炭疽病菌在柑橘花器和发育期果实上的潜伏情况。【结果】获得的19株致病菌均为胶孢炭疽病菌(Colletotrichum gloeosporioides),但菌株间的致病力存在明显差异;T4菌株的测序结果与NCBI已登录的胶孢炭疽菌序列(FJ459917.1,FJ459916.1)比对发现,一致性达100%。柑橘炭疽病菌在花器和果实上的潜伏侵染呈一定的规律性:花器官中,花瓣的带菌率最高;果实上,果蒂和脐部带菌率最高,其次为果柄部位。【结论】研究区域的柑橘花和果实均携带胶孢炭疽病菌,该病菌具有潜伏侵染的特性。生产上要重视花期的病害防控,果实采后贮藏时需重视对果蒂部、脐部及果柄部位的消毒处理。     

21种杀菌剂对6种蔬菜病原菌的离体抑制效果

为减少农药使用量,选用高效且广谱的杀菌剂,采用菌丝生长速率法和最低抑制浓度法测定21种杀菌剂对6种蔬菜病原菌的抑制效果.结果表明,当质量浓度为10μg·mL-1时,苯醚甲环唑·吡唑醚菌酯和申嗪霉素对立枯丝核菌、尖孢镰刀菌苦瓜专化型和瓜果腐霉的抑制率超过50％;吡唑萘菌胺和乙蒜素·代森锰锌对立枯丝核菌的抑制率大于95％....     

Spectral quality affects disease development of three pathogens on hydroponically grown plants

Plants were grown under light-emitting diode (LED) arrays with various spectra to determine the effects of light quality on the development of diseases caused by tomato mosaic virus (ToMV) on pepper (Capsicum annuum L.), powdery mildew [Sphaerotheca fuliginea (Schlectend:Fr.) Pollaci] on cucumber (Cucumis sativus L.), and bacterial wilt (Pseudomonas solanacearum Smith) on tomato (Lycopersicon esculentum Mill.). One LED (660) array supplied 99% red light at 660 nm (25 nm bandwidth at half-peak height) and 1% far-red light between 700 to 800 nm. A second LED (660/735) array supplied 83% red light at 660 nm and 17% far-red light at 735 nm (25 nm bandwidth at half-peak height). A third LED (660/BF) array supplied 98% red light at 660 nm, 1% blue light (BF) between 350 to 550 nm, and 1% far-red light between 700 to 800 nm. Control plants were grown under broad-spectrum metal halide (MH) lamps. Plants were grown at a mean photon flux (300 to 800 nm) of 330 micromoles m-2 s-1 under a 12-h day/night photoperiod. Spectral quality affected each pathosystem differently. In the ToMV/pepper pathosystem, disease symptoms developed slower and were less severe in plants grown under light sources that contained blue and UV-A wavelengths (MH and 660/BF treatments) compared to plants grown under light sources that lacked blue and UV-A wavelengths (660 and 660/735 LED arrays). In contrast, the number of colonies per leaf was highest and the mean colony diameters of S. fuliginea on cucumber plants were largest on leaves grown under the MH lamp (highest amount of blue and UV-A light) and least on leaves grown under the 660 LED array (no blue or UV-A light). The addition of far-red irradiation to the primary light source in the 660/735 LED array increased the colony counts per leaf in the S. fuliginea/cucumber pathosystem compared to the red-only (660) LED array. In the P. solanacearum/tomato pathosystem, disease symptoms were less severe in plants grown under the 660 LED array, but the effects of spectral quality on disease development when other wavelengths were included in the light source (MH-, 660/BF-, and 660/735-grown plants) were equivocal. These results demonstrate that spectral quality may be useful as a component of an integrated pest management program for future space-based controlled ecological life support systems.     

Integrating soil solarization with grafting on resistant rootstocks for management of soil-borne pathogens of eggplant

Summary

Soil solarization and grafting onto a resistant tomato rootstock (‘Brigeor F1‘), and the integrated use of both methods were evaluated in four field trials carried out in Cyprus, for management of soil-borne pathogens and pests of eggplant. Soil solarization was highly effective against Verticillium wilt (V. dahliae) but only partially effective against corky root rot (Pyrenochaeta lycopersici) and root-knot nematodes (Meloidogyne spp.); it also controlled adequately most annual weeds. By contrast, grafting provided complete protection from corky root rot and root-knot, but only partial protection from Verticillium wilt. Complete resistance to nematodes, however, occurred only in trials with greenhouse-grown (winter) crops. In open-field (summer) crops, about 50% of the grafted plants sustained slight nematode infection, apparently because of breakdown of resistance at high soil temperatures. Solarization, or grafting gave significant yield increases over that of the untreated check. However, a combination of both was much more effective than either alone. Together, they provided complete protection from all three diseases. Most importantly, they had an additive effect on yield, resulting in a significant increase over that obtained by either method alone. The average yield (kg plant-1) obtained in two greenhouse trials was 9002E5 for the control, 16.1 for grafting alone, 14.1 for solarization alone and 20.2 for the combination of the two methods. The integrated use of solarization and grafting appears to be a sustainable alternative to methyl bromide fumigation.     

独活提取物抑制柑橘病原真菌活性成分的分离与鉴定

【目的】为了探明独活抑制柑橘病原真菌的活性成分。【方法】采用生长速率法、溶剂萃取、硅胶柱层析、气相色谱-质谱联用(GC-MS)等方法研究独活乙醇提取物对3种柑橘贮藏病害病原真菌的抑菌作用,并分离和鉴定其抑菌活性成分。【结果】独活乙醇提取物中含有高效抑制柑橘炭疽菌的活性物质。GC-MS鉴定B2组分中含有4种香豆素类化合物—异佛手柑内酯、牛防风素、茴芹内酯、异茴芹内酯。异佛手柑内酯、茴芹内酯、异茴芹内酯3种化合物对柑橘炭疽菌的EC50在45 mg.L-1左右,其中茴芹内酯的抑制活性最好,其EC50为41.521 6 mg.L-1。显微观察发现,茴芹内酯处理导致柑橘炭疽菌菌丝体分支增多、畸形,部分隔膜消失等变化。【结论】异佛手柑内酯、茴芹内酯、异茴芹内酯为独活抑制柑橘病原真菌的主要活性物质。     

云南泸西苹果病虫害调查与病原及害虫鉴定

云南红河州泸西县为云南重要的苹果新产区,调查鉴定泸西县苹果病虫害的发生情况,对泸西县的苹果病虫害防控,苹果产业发展具有重要意义。本研究系统调查和鉴定了云南红河州泸西县向阳乡习峨村、三塘乡、向阳乡沙马村等3个苹果产区的病虫害。通过室内病原显微观察,对真菌病原进行形态鉴定,应用RT-PCR检测技术与测序鉴定病毒,害虫形态鉴定。最终结果表明:云南泸西苹果病虫害共8种,其中真菌病害3种,有斑点落叶病、枝干轮纹病、褐斑病;病毒病害2种,有苹果茎沟病毒病、苹果花叶病毒病;虫害3种,蚜虫、蓟马和金纹细蛾3种虫害。其中斑点落叶病和褐斑病为主要病害,病毒病害其次,虫害发生率较小。其次通过调查与鉴定,否定了危险性病害苹果炭疽叶枯病在云南泸西的存在。