Synergistic Disease in Pepper Caused by the Mixed Infection of Cucumber mosaic virus and Pepper mottle virus

ABSTRACT The occurrence of more than one virus species in a single plant is not uncommon in cultivated and native plant species. A mixed virus infection may lead to greater disease severity than individual viral components and this is sometimes referred to as a synergistic disease. Although, in some cases, synergism has been demonstrated for various plant growth parameters such as plant height, weight, and yield, proof of synergy typically has not been demonstrated for symptom severity when the mixed virus infection was not lethal. We demonstrated synergy in bell pepper plants co-infected with Cucumber mosaic virus (CMV) and Pepper mottle virus (PepMoV) relative to each virus alone for stem height (two of three trials) and aboveground fresh weight (one of three trials) using factorial analysis and Abbott's equation for synergy. This approach allowed affirmation of the type of response (i.e., synergistic rather than antagonistic) and statistical proof of synergy. A detailed evaluation of symptom severity for each viral treatment revealed three phases associated with host plant developmental stages. A numerical symptom severity rating scale was developed and used in each of two equations to demonstrate statistical proof for synergy based on symptom severity for co-infected plants. Virus accumulation in noninoculated leaves was determined by enzyme-linked immunosorbent assay. In singly infected plants, CMV titers declined in mildly symptomatic leaves representing later stages of plant development, but titers increased in similar leaves of co-infected plants. In contrast, PepMoV titers did not differ in singly or co-infected plants.     

矿物油和维生素B1对马铃薯病毒病的防效研究

多数马铃薯病毒可以借助蚜虫传播, 并通过块茎世代积累, 导致马铃薯种性退化, 严重影响块茎的产量和品质?为了筛选新型?环保的马铃薯病毒病防治药剂, 本研究通过3个季节的田间试验, 对矿物油?维生素B1和杀虫剂吡虫啉在防治马铃薯病毒病中的效果进行了评价?结果表明, 通过马铃薯出苗后间隔10 d连续3次喷施, 矿物油能够控制马铃薯卷叶病(potato leaf-roll virus, PLRV)的发生, 对马铃薯M病毒(potato virus M, PVM)和马铃薯S病毒(potato virus S, PVS)的平均防效也分别达到66.72%和70.40%, 但对马铃薯Y病毒(potato virus Y, PVY)和马铃薯A病毒(potato virus A, PVA)在不同的年份和季节的防效不稳定, 平均防效为27.34%和65.02%?维生素B1对PLRV?PVM和PVS的防效也比较明显, 分别达83.36%?83.33%和73.32%, 而对PVY同样防效不稳定, 对PVA防效不明显?杀虫剂吡虫啉对PLRV?PVS和PVM的防效也不稳定, 且对PVY和PVA的防效均不显著?本研究中马铃薯X病毒(potato virus X, PVX)发生频率极低, 未进行病毒病的防效比较?综上所述, 矿物油和维生素B1对马铃薯主要病毒病的综合防效较吡虫啉好, 同时它们的增产效果更明显, 产投比高于化学药剂, 值得推广?     

The effects of <Emphasis Type="Italic">Gibberella zeae</Emphasis>, <Emphasis Type="Italic">Barley Yellow Dwarf Virus</Emphasis>, and co-infection on <Emphasis Type="Italic">Rhopalosiphum padi</Emphasis> olfactory preference and performance

Insect-borne viruses promote several changes in plant phenotype, which can modify plant-vector interactions in favor of virus survival and dissemination. Although co-infections commonly occur in the field, little is known about their effects on interactions with the vector. The ecological interactions between Barley Yellow Dwarf Virus (BYDV) and its aphid vector, Rhopalosiphum padi, have been investigated extensively, but the vector’s behavior in more complex scenarios has yet to be examined. We assessed olfactory response and performance of R. padi to wheat singly and doubly infected by the pathogenic fungus Giberella zeae and BYDV. Non-viruliferous aphids preferred odors of BYDV-infected wheat over healthy wheat, as previously reported in the literature, and they were still preferentially attracted to BYDV-infected plant during co-infection. However, around 35% more non-viruliferous aphids chose healthy wheat over G. zeae-infected wheat. Viruliferous aphids did not show any preference to the treatments. BYDV-infected wheat was a superior host than healthy wheat for the aphids whose population increased in 25%. We observed a synergistic effect of the co-infected wheat, which was the best host for aphids, and promoted an elevation of 42% on population growth. Our results indicate that co-infection might be beneficial for virus spread as does not interfere with aphid olfactory preference and provides greater colony growth than in singly infected plants.     

Arabidopsis thaliana,an experimental host for tomato yellow leaf curl disease‐associated begomoviruses by agroinoculation and whitefly transmission

Tomato yellow leaf curl disease is one of the most devastating viral diseases affecting tomato crops worldwide. This disease is caused by several begomoviruses (genus Begomovirus, family Geminiviridae), such as Tomato yellow leaf curl virus (TYLCV), that are transmitted in nature by the whitefly vector Bemisia tabaci. An efficient control of this vector‐transmitted disease requires a thorough knowledge of the plant–virus–vector triple interaction. The possibility of using Arabidopsis thaliana as an experimental host would provide the opportunity to use a wide variety of genetic resources and tools to understand interactions that are not feasible in agronomically important hosts. In this study, it is demonstrated that isolates of two strains (Israel, IL and Mild, Mld) of TYLCV can replicate and systemically infect A. thaliana ecotype Columbia plants either by Agrobacterium tumefaciens‐mediated inoculation or through the natural vector Bemisia tabaci. The virus can also be acquired from A. thaliana‐infected plants by B. tabaci and transmitted to either A. thaliana or tomato plants. Therefore, A. thaliana is a suitable host for TYLCV–insect vector–plant host interaction studies. Interestingly, an isolate of the Spain (ES) strain of a related begomovirus, Tomato yellow leaf curl Sardinia virus (TYLCSV‐ES), is unable to infect this ecotype of A. thaliana efficiently. Using infectious chimeric viral clones between TYLCV‐Mld and TYLCSV‐ES, candidate viral factors involved in an efficient infection of A. thaliana were identified.     

Control of air-borne field spread of tulip breaking virus,lily symptomless virus and lily virus X in lilies by mineral oils,synthetic pyrethroids,and a nematicide in the Netherlands

The inhibitory effect on the spread of viruses in lilies viz., tulip breaking virus (TBV; nonpersistently aphid-borne, potyvirus,) lily symptomless virus (LSV; non-persistently alphidborne, carlavirus), and lily virus X (LVX; potexvirus of unknown etiology), was studied of brands of mineral oil (Luxan oil H and Duphar-7E oil) and synthetic pyrethroid insecticides (l-cyhalothrin and deltamethrin), and a nematicide (aldicarb) in crops in which virus-infected plants were present as virus sources. The spread of TBV and LSV were controlled by sprays of mineral oil and insecticide, while that of LSV was also limited by the soil-applied nematicide. The spread of LVX was reduced by the insecticides and, not effectively by the mineral-oil spraying, by which data the mode of transmission may be presumed to be by an insect in the persistent or semi-persistent manner.Mixtures of mineral oil and pyrethroid were more effective in the reduction of spread of TBV and LSV than either components tested alone. The mineral oil was the most effective component in the mixtures in which pyrethroid added a slight extra effect. The addition of pyrethroid did not mask either the lower efficacy of the oil brand Duphar-7E oil, or the diminished inhibitory effect of low dosages of oil. The normal rate of mineral oil gave similar control to that of a mixture of mineral oil at half rate plus the pyrethroid at full dosage. Low rates of oil, or even synthetic pyrethroids alone may be used on cultivars which suffer of the loss of bulb weight by the use of normal or decreased rates of oil. Weekly sprays were more effective than fortnightly sprays. The rate of control by the weekly sprays ranged between 90 and 95% for Luxan oil H at half dosage plus the full rate of pyrethroid. Weekly sprayed synthetic pyrethroids alone onto the virus sources and the plants to be infected gave 60–70% control. The weight ratios tended to be slightly reduced if the half dosage of the efficient Luxan oil H was used. Factors which affect the control of the air-borne field spread of viruses by mineral oils and synthetic pyrethroid insecticides in lilies are discussed.     

Control of potato virus Y (PVY) in seed potatoes by oil spraying,straw mulching and intercropping

Potato virus Y (PVY) is the potato virus with the highest economic impact on seed potato production. Insecticides are efficient in controlling aphids, which are the vectors of this virus, but rarely limit virus spread in the field. Straw mulching and mineral oil spraying are known as alternatives to insecticides to reduce PVY incidence, but important year‐to‐year variation in efficacy has been observed with both of these techniques. Preliminary studies revealed the efficacy of intercropping in controlling PVY spread, but more data are needed to validate this observation. A four‐year field trial was conducted in Switzerland to assess the potential synergistic effect of combining mineral oil spraying with straw mulching to increase the protection of seed potato crops against PVY spread. Furthermore, the efficacy of intercropping with oat and hairy vetch was examined as a novel way to control in‐field PVY spread. The present work demonstrates that the modes of action of mineral oil and straw mulching are complementary and reduce the year‐to‐year variation observed with oil and straw when used alone as PVY control agents. The results also demonstrate the efficacy of intercropping for the control of PVY, and the mode of action of this novel control method is discussed. Overall, this work shows that it is possible to increase the protection of potato fields against PVY spread by combining control strategies with different modes of action that complement each other, such as mulching, oil spraying and intercropping.     

The genus <Emphasis Type="Italic">Luteovirus</Emphasis> from infection to disease

Luteoviruses are economically important plant viruses. Specifically, barley yellow dwarf virus is epiphytotic to almost all small-grain cereal growing areas. The disease cycle is complex. This luteovirus has evolved several intelligent mechanisms to communicate with both plant and phloem-feeding insect-vector aphid. Environmental cues influence disease severity, aphid infestation and viral load. Within an aphid, virus circulates persistently in a non-propagative manner and is transmitted selectively to the host plants. Selection of viruses within aphids has a role in virus isolate prevalence over a specific area. In the host-plant system, the virus has to release its single sense-strand RNA genome (approx. 5.6 to 6 kb), translate and subsequently replicate its genome using its own replicase and host machinery. This review summarizes our current understanding of disease epidemiology and reviews the current literature encompassing viral infectivity, economic impact and control measures.     

一种蚜虫持久性传病的长豇豆黄化型病毒

 1983年6月,在南京郊区的长豇豆上采到1株表现植株矮缩症状的C-7病毒分离物。接种试验证明,它不能摩擦接种传病,但可以由豆蚜(Aphis craccivora)、棉蚜(A.gossypii)和桃蚜(Myzus persicae)以持久性方式传病。寄主范围测定的结果表明:分离物可以侵染长豇豆、豇豆、蚕豆、大豆、菜豆、豌豆、赤豆、利马豆、苜蓿、红三叶、地三叶、绛三叶、葫芦巴,紫云英和苕子等15种豆科植物和曼陀罗1种茄科植物。这些植物大都出现植株矮化,叶片扭曲,卷缩或僵缩,不能开花结实等症状。豆蚜的传病性状中,获毒饲育的最短传病时间为3小时,接毒饲育最短传病时间为10分钟,循回期是24小时左右。但是,传病率最高的获毒饲育时间是2~3天,接毒饲育时间在1天以上。接种1头蚜虫就具有传病能力,5头蚜虫能达到100%的传病率。蚜虫可以终身传毒,蜕皮不影响其传毒力,但传毒有间歇性。根据它的基本性状,病毒C-7分离物是一种豆科植物的黄化型病毒,可能是属于大麦黄矮病毒组(Luteovirus Group)的成员。     

Evidence of <Emphasis Type="Italic">olive mild mosaic virus</Emphasis> transmission by <Emphasis Type="Italic">Olpidium brassicae</Emphasis>

Transmission of three strains of OMMV by an Olpidium sp. was evaluated and compared. The three strains were 1) an OMMV wild type (WT) recovered from olive trees, 2) an OMMV variant (L11) obtained after 15 serial passages of single local lesions induced in Chenopodium murale plants, and 3) a construct OMMV/OMMVL11 in which the coat protein (CP) gene replaced that of the wild type. A single-sporangial culture derived from Chinese cabbage (Brassica pekinensis) used as a bait plant grown in soil of an olive orchard, was identified as Olpidium brassicae based on the size and sequence of the generated amplicon in PCR specific tests. Each of the three virus strains was soil transmitted to cabbage roots in the absence of the fungus at similar rates of 30 to 40%. Separate plant inoculation by O. brassicae zoospores incubated with each viral strain resulted in enhanced transmission of OMMV, reaching 86% of infection whereas that of the other two strains remained practically unaffected at ca. 34%. Binding assays showed that the amount of virus bound to zoospores, estimated spectrophotometrically, was 7% in the case of OMMV, and practically nil in the case of the other two viral strains. Substitution of the coat protein (CP) gene of OMMV by that of the OMMV L11 strain, drastically reduced viral transmissibility in the presence of zoospores to the level of that observed in their absence. Our data shows that OMMV soil transmission is greatly enhanced by O. brassicae zoospores and that the viral CP plays a significant role in this process, most likely by facilitating virus binding and later entrance into the host plant roots.     

内质网应激因子NAC089突变体的创建及其对病毒侵染胁迫的响应

 有研究表明烟草花叶病毒(Tobacco mosaic virus, TMV)或黄瓜花叶病毒(Cucumber mosaic virus, CMV)侵染烟草能够激活转录因子NbNAC089,从ER膜移至细胞核。为进一步阐释内质网应激因子NbNAC089对病毒侵染胁迫的响应机制,利用CRISPR/Cas9技术构建敲除载体,经烟草遗传转染获得NbNAC089基因突变植株。植株接种病毒后采用qRT-PCR检测病毒CP基因和寄主UPR基因的表达。结果表明:CRISPR/Cas9系统定点敲除NbNAC089基因后,目的基因靶位点序列有碱基的置换与缺失。正常生长条件下,转基因植株与野生型无差异。植株接种TMV-GFP后24~96 h,突变体中UPR基因(BiP、bZIP28、bZIP60)的表达量显著高于野生型;接种TMV-GFP后2~6 d突变体中病毒的积累量和扩展速度显著高于野生型。表明NbNAC089为UPR的抑制因子,对病毒增殖具有负调控作用。     

Crop border and mineral oil sprays used in combination as physical control methods of the aphid‐transmitted potato virus Y in potato

BACKGROUND: The objectives of this work were to determine if the control of potato virus Y (PVY, genus Potyvirus, family Potyviridae) in seed potato could be improved by combining border crops and mineral oil sprays, and if the border crop acts as a barrier or a virus sink. RESULTS: Field tests over 3 years confirmed that mineral oils alone are an effective barrier to PVY, and showed that borders alone act as a PVY sink. Combining the familiar mineral oil and the more recent crop border methods was almost twice as effective in reducing PVY incidence as either one used alone. The combination provided consistently high PVY control compared with the variable and often lower level of control by either method alone. The contribution of the oil to PVY reduction was similar whether it was applied to the border, the center seed plot, or both. Oil application to the border alone should not affect efficacy and would help keep control costs down. CONCLUSION: Combining border and oil provided the best reduction in PVY incidence 3 years out of 3, providing producers with a tool to reduce year‐to‐year variation in the effectiveness of crop borders or oil sprays used separately. © Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri‐Food Canada. Published by John Wiley and Sons, Ltd.     

Presence of Grapevine leafroll-associated virus 3 in primary salivary glands of the mealybug vector Planococcus citri suggests a circulative transmission mechanism

Grapevine leafroll-associated virus 3 (GLRaV-3) is a mealybug-transmissible ampelovirus. Though the transmission mechanism has been described as semipersistent on the basis of temporal parameters, definitive proof of this mechanism has never been provided. In the present study, we carried out preliminary assays to establish the location of the virus in its vector, Planococcus citri. After dissecting the insects, GLRaV-3 was detected by means of IC-RT-PCR in the salivary glands, intestine and Malpighian tubes, but not in the sucking apparatus. Immunogold labelling of the capsid protein revealed the presence of the virus in some cells of the primary salivary glands, but not in the alimentary channel of the stylet, or in the accessory salivary glands. The strong labelling of the electron-dense secretion vesicles in some cells of the primary salivary glands, together with the non-detection of the virus in the sucking apparatus suggests that the transmission mechanism may be different from that previously described. We propose a circulative transmission mechanism based on a specific transportation route for the viral particles from the midgut or hindgut to the salivary glands. As the transmission mechanism is generally a common feature of a viral genus, the existence of a circulative transmission mechanism for other mealybug-transmitted ampeloviruses is expected. Organ by organ analysis of GLRaV-1, another ampelovirus not transmissible by P. citri, showed the absence of the virus in the salivary glands, thus providing further, though indirect, evidence in favour of circulative transmission for this virus genus.     

Reservoirs of plant virus disease: Occurrence of wheat dwarf virus and barley/cereal yellow dwarf viruses in Sweden

Non-crop plants such as grasses and volunteer plants are an inseparable part of the flora of crop fields and can influence virus incidence in crop plants. The presence of grasses as virus reservoirs can lead to a higher probability of virus incidence in crop plants. However, the role of reservoirs as an inoculum source in agricultural fields has not been well studied for many viral diseases of crops. Grasses have been found to constitute potential reservoirs for cereal-infecting viruses in different parts of the world. This study revealed that cereal-infecting viruses such as wheat dwarf virus (WDV), barley yellow dwarf viruses (BYDVs), and cereal yellow dwarf virus-RPV (CYDV-RPV) can be found among ryegrass growing in or around winter wheat fields. Phylogenetic analysis showed that a WDV isolate from ryegrass was a typical WDV-E isolate that infects wheat. Similarly, a ryegrass isolate of barley yellow dwarf virus-PAV (BYDV-PAV) grouped in a clade together with other BYDV-PAV isolates. Inoculation experiments under greenhouse conditions confirmed that annual ryegrass of various genotypes can be infected with WDV to a very low titre. Moreover, leafhoppers were able to acquire WDV from infected ryegrass plants, despite the low titre, and transmit the virus to wheat, resulting in symptoms. Information from the grass reservoir may contribute to improving strategies for controlling plant virus outbreaks in the field. Knowledge of the likely levels of virus in potential reservoir plants can be used to inform decisions on insect vector control strategies and may help to prevent virus disease outbreaks in the future.     

Movement of aphid-transmitted Sugarcane yellow leaf virus (ScYLV) within and between sugarcane plants

Sugarcane yellow leaf virus (ScYLV) is distributed worldwide and has been shown to be the cause of the disease sugarcane yellow leaf syndrome (YLS). This study was an investigation of the transmission and spread of ScYLV in Hawaii. Several aphids are known to transmit the virus, but investigation of infestation and transmission efficiency showed Melanaphis sacchari to be the only vector important for field spread of the disease. The initial multiplication of ScYLV in a virus-free plant occurred exclusively in very young sink tissues. When a single leaf was inoculated on a plant, that leaf and all older leaves remained virus-free, based on tissue-blot immunoassay, whereas meristems and all subsequently formed new leaves became infected. Therefore, only after those leaves which had already developed before inoculation had been shed, did the complete plant contain ScYLV. Spread of the viral infection to neighbouring plants in the plantation fields via aphids was relatively slow and in the range of a few metres per year. No indication of long-distance transfer could be seen. This indicates that it may be possible to produce and use virus-free seed cane for planting of high-yielding but YLS-susceptible cultivars.     

Effect of temperature, vector life stage, and plant access period on transmission of banana bunchy top virus to banana

The study of the transmission biology of insect-borne plant viruses is important to develop disease control practices. We characterized the transmission of a nanovirus, Banana bunchy top virus (BBTV), by its aphid vector Pentalonia nigronervosa Coquerel (Hemiptera, Aphididae) with respect to temperature, vector life stage, and plant access time. Adult aphids transmitted BBTV more efficiently than third instar nymphs at all temperatures tested. Adult aphids transmitted the virus more efficiently at 25 and 30 degrees C than at 20 degrees C, but temperature had no impact on transmission efficiency by nymphs. By decoupling the relationship between temperature and aphid BBTV acquisition or inoculation, we determined that temperature affected inoculation events more strongly than acquisition. Longer plant access periods increased viral acquisition and inoculation efficiencies in a range of 60 min to 24 h. Both BBTV acquisition and inoculation efficiencies peaked after 18 h of plant access period. We also show that BBTV transmission by P. nigronervosa requires a latent period. Our results demonstrate that vector transmission of BBTV is affected by temperature, vector life stage, and plant access period.     

An infectious cDNA clone of a radish-infecting <Emphasis Type="Italic">Turnip mosaic virus</Emphasis> strain

Turnip Mosaic Virus (TuMV) is an economically important potyvirus for which hundreds of hosts have been reported, thus making it a rather exceptional case in the genus. Several viral infectious clones have been generated over the years, which have been useful in deciphering the viral elements involved in the interactions of this virus with the host plant, such as different forms of resistance, gene silencing suppression, host range or host developmental alterations. However, all infectious clones obtained so far correspond to viral isolates within the same phylogenetic cluster, a circumstance biasing our understanding of the peculiarities of this potyvirus. In particular, members of one viral cluster of radish-infecting isolates have been especially reluctant to be copied into infectious clones. This paper reports the construction of an infectious clone of the TuMV isolate JPN 1, belonging to this cluster. The infectious clone maintains all the distinctive biological properties previously described for this viral isolate. The availability of this infectious clone opens the door to many additional studies on the virus, which should allow a deeper understanding of the differential responses to different strains of TuMV in several different hosts.     

Drought as a modulator of plant–virus–vector interactions: Effects on symptom expression,plant immunity and vector behaviour

Plant–virus interactions are affected by environmental conditions that determine plant vulnerability to pathogens and the population dynamics of insect vectors. We hypothesize that drought enhances horizontal transmission by dampening the basal immunity of plants, which triggers symptom expression and vector manipulation. The potato yellow vein virus (PYVV) causes potato yellow vein disease (PYVD), a re-emerging epidemic of potato crops in South America, and is transmitted horizontally by the greenhouse whitefly (GWF), Trialeurodes vaporariorum (Hemiptera: Aleyrodidae), or vertically through infected seed tubers. We investigated the role of drought and temperature as modulators of PYVD symptom expression, plant immune response, and vector survival, development and host preference. We found that drought induced symptom expression, suppressed the salicylic acid pathway and increased PYVV replication. GWF survival was reduced on PYVV-infected hosts and development was slowest when they fed on plants with PYVD symptoms, which also triggered adults’ attraction to PYVV-infected plants. However, adults previously fed on infected plants showed the opposite effect, being more attracted to PYVV-free plants. We propose a theoretical model that explains the role of drought in modulating potato–PYVV–GWF interactions and provides new insights into plant–virus–vector coevolution.