Studies on poly(acrylic acid) induced resistance to viral and non-viral plant pathogens

Studies on polymer size, concentration and mode of application, either as foliar spray or soil drench, in relation to the induction of resistance to tobacco mosaic virus (TMV) in Nicotiana tabacum cv xanthi-nc by poly(acrylic acid) (PA) are reported. PA also induced resistance to TMV in N. glutinosa, to pelargonium leaf curl virus in Datura stramonium, to cucumber mosaic virus in Vigna sinensis and to tobacco ring-spot virus in N. tabacum cv White Burley. No TMV was detected in PA-treated tomato cv Virocross 11 days after inoculation; but the susceptible cultivar Craigella became infected. PA treatment had no effect on TMV replication in White Burley tobacco but resistance was induced to Peronospora tabacina, a fungal pathogen of N. tabacum cv xanthi-nc. The potential of PA-induced resistance as a control measure for viruses and fungi is discussed.     

Systemic infection of some N-gene-carrying Nicotiana species and cultivars after inoculation with tobacco mosaic virus

Since July 1974Nicotiana tabacum ‘Samsun NN’ plants, inoculated with the common strain of tobacco mosaic virus (TMV), have occasionally been found to develop necrosis on non-inoculated upper leaves 2–7 days after the local necrotic lesions had appeared on the lower leaves. All these plants had been kept in a growth chamber at 17–20°C. Other tobacco species and cultivars carrying theN gene, such asN. glutinosa andN. tabacum ‘Xanthi-nc’, showed the same phenomenon. Substantial amounts of TMV could be recovered from leaves with systemic symptoms. The systemic necrosis somewhat resembled that caused by tobacco rattle virus (TRV). A number of possible causes, such as high concentration of the inoculum, contamination with another strain of TMV or with TRV, change in the genetic composition of the host plants and certain growing conditions (soil, water, pesticides) were investigated. None of these factors could be held fully responsible for the abnormal systemic reaction, although there was evidence that the soil could sometimes play an important rôle.     

应用A-蛋白诱捕修饰和羊抗兔血清诱捕双修饰法研究马铃薯X病毒

A-蛋白和羊抗兔血清应用于免疫电镜,检测马铃薯X病毒(PVX)粗汁液,灵敏度比诱捕修饰法高;而且加羊抗兔血清后,病毒粒体比诱捕修饰法明显加粗,在电镜放大2500倍时,就能清晰地观察到。用诱捕双修饰法检测PVX,证实在寄主植物的叶、茎和根中均存在,以叶内含量最高。接种在普通烟上第三天,接种叶就能检测到PVX,接种15天后,寄主体内病毒能达到较高浓度,而且高浓度一直可保持2个月以上;PVX在不同寄主中的含量不同,以普通烟中病毒浓度最高,心叶烟、番茄和黄花烟中次之;昆诺阿藜、千日红和大椒中含毒量极抵。用诱捕双修饰法检测马铃薯薯块休眠芽中的病毒获得成功。     

Exogenous application of melatonin improves plant resistance to virus infection

In this study, melatonin (MEL)-mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti-virus infection activity to 37.4% in virus-infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT-PCR and virus titres measured by dot-ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT-PCR, in virus-infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H₂O₂) in the virus-infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL-induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT-PCR and virus titres analysed by dot-ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL-mediated plant resistance to TMV is proposed. MEL-mediated plant resistance to viruses provides a new avenue to control plant viral diseases.     

Tobacco streak virus in sunflower (Helianthus annuus)

Tobacco streak virus (TSV) was isolated from a plant of sunflower (Helianthus annuus) showing severe necrosis and chlorosis in the leaves. The virus was identified as TSV by serology and, to some extent, by host range. The type of symptoms varied with the host plant in which the virus had been propagated in successive transfers. Test plants inoculated with the virus propagated inNicotiana rustica produced symptoms which very much differed from those brought about by the virus from eitherN. clevelandii orChenopodium amaranticolor.The significance of the host-mediated variation in symptoms is discussed.Samenvatting Tabaksstrepenvirus (TSV) werd geïsoleerd uit zonnebloem (Helianthus annuus) die sterke necrose en chlorose van de bladeren vertoonde. De identiteit van het virus werd vastgesteld op grond van serologische reacties en, tot op zekere hoogte, de symptomatologie. Het type symptoom op de toetsplanten bleek echter sterk afhankelijk te zijn van de plant waarvan het inoculum afkomstig was. Was het virus verschillende malen achtereen vermeerderd inNicotiana rustica dan waren de symptomen op de toetsplanten zeer verschillend van die, welke werden veroorzaakt door virus vermeerderd inN. clevelandii ofChenopodium amaranticolor. De betekenis van deze door de waardplant bewerkstelligde variatie in symptomen wordt besproken.     

侵染扶桑的烟草花叶病毒分离物鉴定

从表现叶斑驳症状的扶桑病株上获得一病毒分离物,电镜下可见约300 nm×18 nm的杆状粒子,其与烟草花叶病毒抗血清呈明显的阳性反应,dsRNA约为6.4 kbp。根据烟草花叶病毒(tobacco.mosaic virus,TMV)的RNA序列设计引物,进行RT-PCR检测,扩增出约800 bp的预期特异片段。将PCR产物连接pMD18-T载体,转化大肠杆菌DH5α,得到了含有目的片段的重组子。序列分析表明,与周雪平等报道的序列(GenBank AJ011933.1)同源性达99％。通过生物学、病毒粒子观察、血清学以及分子生物学实验结果,确定该病毒分离物为TMV。     

The identification of two new strains of bean common mosaic virus

Two sap-transmissible virus isolates from bean (Phaseolus vulgaris L.) were identified as bean common mosaic virus (BCMV) on the basis of particle size and morphology, serology, non-persistent aphid transmission, very limited host range, and symptoms and seed transmission in bean. In bean varietal reaction both isolates differed from each other and all six Dutch BCMV strains described before. From literature data it may be concluded that they also differ from thirteen other strains described elsewhere. The isolate from Peruvian seed may be related to strains reported from Costa Rica and Peru, but these have been described incompletely. The two isolates obtained at Wageningen are therefore described as new strains and designated BCMV-NL7 and BCMV-NL8. The latter seems unusual in its extremely high dilution end point, in its serological affinity to both BCMV and BYMV, and in not being infectious toChenopodium amaranticolor andC. quinoa. Tetragonia expansa proved to be a new local lesion host of BCMV. There is an urgent need for international standardization of strains of BCMV.     

Determination of the infection pressure of potato virus YN

In 1976 consecutive series of plants ofNicotiana tabacum ‘White Burley’ replaced weekly, were used as bait plants to determine the infection pressure of potato virus Y^N (PVY^N) in a crop of ware potatoes in the centre of the Netherlands. The first PVY^N-infected tobacco plants were found mid May. The course of infection of the tobacco plants was not correlated with the flight ofMyzus persicae, which started towards the end of June. Aphid species other thanMyzus persicae presumably are responsible for the infection observed early.Rhopalosiphum padi andAcyrthosiphon pisum flew much earlier thanMyzus persicae and are vectors of PVY^N.     

Influence of an m-type thioredoxin in maize on potyviral infection

Expression of many host genes can be altered during virus infection. In a previous study of sugarcane mosaic virus (SCMV) infection in maize (Zea mays), we observed that expression of ZmTrm2, a gene encoding thioredoxin m, was up-regulated at about 10 days post-inoculation (dpi). In this present study we determined that ZmTrm2 silencing in maize by virus-induced gene silencing significantly enhanced systemic SCMV infection. In contrast transient over-expression of ZmTrm2 in maize protoplasts inhibited accumulation of SCMV viral RNA. Furthermore, we found that in inoculated Nicotiana tabacum leaves transient expression of ZmTrm2 inhibited accumulation of the RNA of tobacco vein-banding mosaic virus (TVBMV), a potyvirus infecting dicotyledonous plants. Interestingly in ZmTrm2 transiently expressed N. tabacum leaves, we detected by semi-quantitative RT-PCR a reduced level of the mRNA of class I beta-1, 3-glucanase (GluI), a protein known to have a role in cell wall callose deposition and viral movement. Our data indicate that the maize ZmTrm2 plays an inhibitory role during infection of plants by SCMV and TVBMV.     

进境荷兰郁金香种苗中南芥菜花叶病毒的鉴定

为了防止南芥菜花叶病毒(Arabis mosaic virus,ArMV)传入我国,采用血清学、分子生物学、电镜观察及生物学接种等方法对从荷兰进口的郁金香种苗进行了ArMV检测。结果表明:ArMV血清学反应为强阳性;RT-PCR反应扩增出370bp的特异性目标条带;RT-PCR产物与已报道的3个ArMV部分外壳蛋白基因的核苷酸序列同源性为91.00%～93.24%,氨基酸序列同源性为99%～100%;免疫电镜观察到叶片病汁液中含有直径约30nm的球状病毒粒体;该病毒在黄花烟、白肋烟、矮牵牛和昆诺藜等鉴别寄主上引起坏死和褪绿等典型症状,经DAS-ELISA检测,这些接种寄主均呈ArMV血清学阳性反应。故将该病毒鉴定为南芥菜花叶病毒。     

Effect of two satellite RNAs on Nicotiana glutinosa infected with Cucumber mosaic virus (CMV)

Two types of satellite RNAs (XJs1 and XJs2) were found to be associated with the two Cucumber mosaic virus (CMV) beet isolates XJ1 and XJ2 consisting of 384 nucleotides and 336 nucleotides, respectively. Our work showed that XJs1 and XJs2 could attenuate symptoms on Nicotiana glutinosa induced by CMV and the effect of XJs1 on symptoms was stronger than XJs2. Co-inoculation of CMV with XJs1 produced lighter oxidative stress than that with XJs2. Oxygen uptake rates through the alternative pathway in leaves infected with CMV and XJs2 were always higher than that with CMV and XJs1 and lower than infection with CMV alone. The non-photochemical fluorescence quenching (qN) increased more in N. glutinosa infected with CMV and XJs2 than that with CMV and XJs1. The relationship among helper virus, satRNAs and the host plants is discussed. Two closely-related satRNAs from the same plant has not been reported so far. The effects of these two satRNAs on virus-infected N. glutinosa were significantly different. The difference may be caused by deletion and mutation of the satRNAs.     

海带多糖对烟草花叶病毒的抑制作用及其对烟草酶活性的影响

为明确海带多糖抗病毒剂对烟草抗烟草花叶病毒（Tobacco mosaic virus,TMV）病的诱导抗性及其对TMV感染后的保护作用,采用间接酶联免疫吸附法（ELISA）研究0.5 mg/mL海带多糖水剂、0.5 mg/mL香菇多糖水剂、6.25 mg/L氨基寡糖素水剂和0.1 mg/mL盐酸吗啉胍.铜可湿性粉剂对TMV侵染的预防保护作用和对TMV感染植株的治疗作用,并检测施用海带多糖后烟草体内过氧化物酶（POD）、苯丙氨酸解氨酶（PAL）及超氧化物歧化酶（SOD）活性的变化和海带多糖对感染TMV后烟草叶绿素含量的影响。接种前喷施0.5 mg/mL海带多糖后,再接种TMV病毒,其抑制率可达42.42%,与喷施6.25 mg/L氨基寡糖素抑制率44.96%的效果相当,预防保护作用较好;但两种药剂处理感病植株的治疗效果较差,其抑制率仅分别为38.93%和40.13%。海带多糖能够系统地诱导烟草体内POD、PAL及SOD活性,从而控制了感染TMV后烟草叶绿素含量下降。说明海带多糖可诱导植物产生抗病性。     

油菜花叶病毒15号和菸花叶病毒间干扰作用的研究

 TMV与YMV₁₅在系统感染寄主——菸(Nicotiana tabacum)上的相互干扰作用是很微弱的。同时接种二病毒时,在感染的早期(接种后2-3天) YMV₁₅占优势,在此时间以后TMV的浓度就超过YMV₁₅。二病毒以相同的侵染力同时接种心叶菸(Nicotiana glutinosa)时,则大部分的局部斑点是由YMV₁₅形成的。上述结果表明,YMV₁₅竞争侵染点的能力较TMV为大。试验并证明完整的YMV₁₅及其蛋白都能干扰TMV侵染菜豆(Phaseoluss vulgaris),因而这种干扰似决定于病毒蛋白质部分对于侵染点附着的特异性。     

Accessions of Australian Nicotiana species suitable as indicator hosts in the diagnosis of plant virus diseases

When screening the genusNicotiana for sensitive and differential hosts for a group of mechanically transmissible plant viruses with narrow host ranges, development of systemic symptoms was alsmost exclusively observed in species of three closely related sections of the subgenusPetunioides. These species wereN. miersii (sectionAcuminatae),N. bigelovii andN. clevelandii (Bigelovianae) andN. benthamiana, N. cavicola, N. ingulba, N. occidentalis, N. rosulata andN. rotundifolia (Suaveolentes). Except forN. benthamiana andN. clevelandii, which are already known for their large virus ranges, they are new experimental hosts that appeared very useful for detection of viruses and for differentiation of viruses that closely resemble each other in host range. Accessions of the same species often varied largely in local and systemic viral response. EspeciallyN. benthamiana-9,N. miersii-33 andN. occidentalis-37B (code numbers given by Tobacco Research Laboratory, Oxford, N.C., USA) are recommended for routine inoculation tests. The sensitiveSuaveolentes species mentioned are native to the arid parts of Australia. Collections of these species deserve attention in studies on virus diseases of unknown etiology where experimental hosts are lacking.Samenvatting In het genusNicotiana werden vatbare, gevoelige en differentiële toetsplanten opgespoord door toetsing van vooral de collectie van het Tobacco Research Laboratory (Oxford, N.C., VS) met een aantal virussen waarvoor weinig of geen bruikbare toetsplanten bekend waren. Systemische reacties werden bijna uitsluitend in soorten van drie nauw verwante secties van het subgenusPetunioides waargenomen. Deze soorten warenN. miersii (sectieAcuminatae),N. bigelovii enN. clevelandii (Bigelovianae) enN. benthamiana, N. cavicola, N. ingulba, N. occidentalis, N. rosulata enN. rotundifolia (Suaveolentes). Behalve voor virusvermeerdering bleken deze soorten ook zeer geschikt te zijn voor differentiatie van virussen of virusstammen met een bijna gelijke waardplantenreeks.Collectienummers van één en dezelfde soort konden sterk in lokale en systemische reactie verschillen. VooralN. benthamiana-9,N. miersii-33 enN. occidentalis-37B kunnen voor routine-inoculaties worden aanbevolen. Met uitzondering vanN. benthamiana enN. clevelandii zijn de genoemde soorten of genotypen nieuwe experimentele waardplanten voor de virologie. De soorten die van de sectieSuaveolentes worden genoemd, zijn inheems in de woestijnachtige gebieden van Australië. Verzamelingen daarvan verdienen nadere aandacht bij de diagnostiek van virusziekten waarvoor nog geen experimentele waarplanten voorhanden zijn.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

Effect of 6-azauracil on infection with tobacco mosaic virus

6-Azauracil caused a marked reduction in the number and size of local lesions on excisedNicotiana glutinosa leaves or leaf discs inoculated with tobacco mosaic virus or its nucleic acid. The amount of infectivity recovered from tobacco (N. tabacum White Burley) leaf discs floated on the pyrimidine analogue solution and subsequently ground and assayed 24, 48, 72, and 96 h after inoculation with intact virus was reduced in the 24, 48, and 72 h series but reached the same level as that of the water-treated control discs in the 96 h series.By contrast, the amount of infectivity in stripped epidermal tissue ofN. glutinosa leaves inoculated with nucleic acid was not reduced in strips floated on the analogue for 24, 50, and 70 h as compared with water-floated controls. The analogue had no effect on infectivity of the virus in vitro and did not act as an inhibitor of infection.Part of this paper was presented at the International Symposium on Plant Pathology, held in New Delhi from December 27, 1966 till January 1, 1967. A summary will be published in the symposium volume entitled Plant disease problems.     

Investigation ofHippeastrum mosaic virus inHippeastrum hybridum

An attempt was made to identify a mosaic disease inHippeastrum hybridum. Infectious virus material could be demonstrated in roots, leaves, stem, perianth, stamen and pistil. Inclusion bodies were found in the epidermis of leaves, stem, spatha leaves and in the perianth. Virus concentration in a young stage was high but decreased by aging of the perianth. Efforts to transmit the virus by aphids failed. However, the virus was transmitted by seed in a few cases. Plants of 30 species reacted negatively upon inoculation with the virus.Hippeastrum hybridum, Gomphrena globosa, Chlorophytum spec. andLycopersicum esculentum could be infected experimentally. On account of the host range and presence of inclusion bodies the mosaic symptoms inHippeastrum are not caused by tomato spotted wilt virus or Cucumber mosaic virus. Results suggest that the virus under investigation is theHippeastrum mosaic virus. Dr.M. K. Corbett, Wageningen, succeeded in purifying the virus by density gradient centrifugation. Whe preparation contained flexuous rod particles. Plants ofDatura stramonium, Nicotiana glutinosa andN. tabacum Samsun could be infected. Within two weeks after inoculation with purified virus solution these plants showed systemic symptoms.

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Samenvatting In een kwekerij te Hoorn werden planten vanHippeastrum hybridum aangetroffen, die mozaïekverschijnselen vertoonden. De oorzaak hiervan werd nagegaan. Infectieus virusmateriaal kon worden aangetoond in wortels, bladeren, bloemstengel, bloemdekbladen, meeldraden en stijl. Celinsluitsels kwamen voor in de epidermis van bladeren en stengel en in de bloemschede. De aanwezigheid van insluitsels in het bloemdek was afhankelijk van de ouderdom van de bloem. Er schijnt een omgekeerd evenredige relatie te bestaan tussen de virusconcentratie en het aantal insluitsels in bloemen.Pogingen om het virus over te brengen door bladluizen mislukten. In enkele gevallen had zaadoverdracht plaats.Dertig plantesoorten reageerden negatief op een inoculatie met virushoudend sap.Hippeastrum hybridum, Gomphrena globosa, Chlorophytum spec. enLycopersicum esculentum konden wel worden geïnfecteerd. Gezien de waardplantenreeks en het voorkomen van celinsluitsels kunnen de mozaïeksymptomen inHippeastrum niet toegeschreven worden aan Tomato spotted wilt-virus of het komkommer-mozaïek-virus, maar is het waarschijnlijk dat zij worden veroorzaakt door hetHippeastrum-mozaïek-virus, reeds beschreven doorBrierley (1948),Johnson (1951) enProcenko & Procenko (1964).Dr.M. K. Corbett, Wageningen, slaagde erin het virus te zuiveren door middel van density gradient-centrifugering.Datura stramonium, N. glutinosa enN. tabacum Samsun werden systemisch ziek binnen twee weken na inoculatie met de gezuiverde virus-oplossing.

     

Occurrence of viruses in field-grown pepper crops and some of their reservoir weed hosts in Samsun, Turkey

Pepper production is affected by several viral diseases in Samsun, Turkey. To determine the identity of these viruses, a total of 313 samples from field-grown peppers were collected during surveys in 1998 and 1999, and tested by enzyme linked immunosorbent assay (ELISA). Six viruses,Alfalfa mosaic virus (AMV),Cucumber mosaic virus (CMV),Potato virus Y (PVY),Tomato mosaic virus (ToMV),Tobacco mosaic virus (TMV) andTomato spotted wilt virus (TSWV) were detected in the samples. Of 313 plants tested, 42 were doubly infected, and TMV+PVY (15.4%) was the most common double infection. This is also the first report of AMV in pepper fields in Turkey. The effect of some weed species that may act as reservoir of these viruses was also investigated in the region and of 24 weed species belonging to 15 families tested, 16 were found to be infected with at least one virus.Amaranthus retroflexus (Redroot pigweed) appeared to be a common host of CMV, PVY, ToMV, TMV and TSWV, whereasHibiscus trionum (Venice mallow) was recorded as a new weed host of PVY and TSWV. The majority of weed species found to be virus infected were very common in the pepper growing areas of the region. This indicates that pepper fields contaminated with these weeds are under risk of viral infections. http://www.phytoparasitica.org posting July 21, 2005.